123 results on '"Gerard Wagemaker"'
Search Results
2. Lentiviral Hematopoietic Stem Cell Gene Therapy Corrects Murine Pompe Disease
- Author
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Merel Stok, Helen de Boer, Marshall W. Huston, Edwin H. Jacobs, Onno Roovers, Trudi P. Visser, Holger Jahr, Dirk J. Duncker, Elza D. van Deel, Arnold J.J. Reuser, Niek P. van Til, and Gerard Wagemaker
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murine Pompe disease ,acid α-glucosidase ,lentiviral vector ,hematopoietic stem cell transplantation ,skeletal muscle ,central nervous system ,Genetics ,QH426-470 ,Cytology ,QH573-671 - Abstract
Pompe disease is an autosomal recessive lysosomal storage disorder characterized by progressive muscle weakness. The disease is caused by mutations in the acid α-glucosidase (GAA) gene. Despite the currently available enzyme replacement therapy (ERT), roughly half of the infants with Pompe disease die before the age of 3 years. Limitations of ERT are immune responses to the recombinant enzyme, incomplete correction of the disease phenotype, lifelong administration, and inability of the enzyme to cross the blood-brain barrier. We previously reported normalization of glycogen in heart tissue and partial correction of the skeletal muscle phenotype by ex vivo hematopoietic stem cell gene therapy. In the present study, using a codon-optimized GAA (GAAco), the enzyme levels resulted in close to normalization of glycogen in heart, muscles, and brain, and in complete normalization of motor function. A large proportion of microglia in the brain was shown to be GAA positive. All astrocytes contained the enzyme, which is in line with mannose-6-phosphate receptor expression and the key role in glycogen storage and glucose metabolism. The lentiviral vector insertion site analysis confirmed no preference for integration near proto-oncogenes. This correction of murine Pompe disease warrants further development toward a cure of the human condition.
- Published
- 2020
- Full Text
- View/download PDF
3. Preclinical Efficacy and Safety Evaluation of Hematopoietic Stem Cell Gene Therapy in a Mouse Model of MNGIE
- Author
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Rana Yadak, Raquel Cabrera-Pérez, Javier Torres-Torronteras, Marianna Bugiani, Joost C. Haeck, Marshall W. Huston, Elly Bogaerts, Steffi Goffart, Edwin H. Jacobs, Merel Stok, Lorena Leonardelli, Luca Biasco, Robert M. Verdijk, Monique R. Bernsen, George Ruijter, Ramon Martí, Gerard Wagemaker, Niek P. van Til, and Irenaeus F.M. de Coo
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MNGIE ,thymidine phosphorylase ,hematopoietic stem cells ,lentiviral vectors ,gene therapy ,Genetics ,QH426-470 ,Cytology ,QH573-671 - Abstract
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is an autosomal recessive disorder caused by thymidine phosphorylase (TP) deficiency resulting in systemic accumulation of thymidine (d-Thd) and deoxyuridine (d-Urd) and characterized by early-onset neurological and gastrointestinal symptoms. Long-term effective and safe treatment is not available. Allogeneic bone marrow transplantation may improve clinical manifestations but carries disease and transplant-related risks. In this study, lentiviral vector-based hematopoietic stem cell gene therapy (HSCGT) was performed in Tymp−/−Upp1−/− mice with the human phosphoglycerate kinase (PGK) promoter driving TYMP. Supranormal blood TP activity reduced intestinal nucleoside levels significantly at low vector copy number (median, 1.3; range, 0.2–3.6). Furthermore, we covered two major issues not addressed before. First, we demonstrate aberrant morphology of brain astrocytes in areas of spongy degeneration, which was reversed by HSCGT. Second, long-term follow-up and vector integration site analysis were performed to assess safety of the therapeutic LV vectors in depth. This report confirms and supplements previous work on the efficacy of HSCGT in reducing the toxic metabolites in Tymp−/−Upp1−/− mice, using a clinically applicable gene transfer vector and a highly efficient gene transfer method, and importantly demonstrates phenotypic correction with a favorable risk profile, warranting further development toward clinical implementation.
- Published
- 2018
- Full Text
- View/download PDF
4. In vivo expansion of co-transplanted T cells impacts on tumor re-initiating activity of human acute myeloid leukemia in NSG mice.
- Author
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Malte von Bonin, Martin Wermke, Kadriye Nehir Cosgun, Christian Thiede, Martin Bornhauser, Gerard Wagemaker, and Claudia Waskow
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Medicine ,Science - Abstract
Human cells from acute myeloid leukemia (AML) patients are frequently transplanted into immune-compromised mouse strains to provide an in vivo environment for studies on the biology of the disease. Since frequencies of leukemia re-initiating cells are low and a unique cell surface phenotype that includes all tumor re-initiating activity remains unknown, the underlying mechanisms leading to limitations in the xenotransplantation assay need to be understood and overcome to obtain robust engraftment of AML-containing samples. We report here that in the NSG xenotransplantation assay, the large majority of mononucleated cells from patients with AML fail to establish a reproducible myeloid engraftment despite high donor chimerism. Instead, donor-derived cells mainly consist of polyclonal disease-unrelated expanded co-transplanted human T lymphocytes that induce xenogeneic graft versus host disease and mask the engraftment of human AML in mice. Engraftment of mainly myeloid cell types can be enforced by the prevention of T cell expansion through the depletion of lymphocytes from the graft prior transplantation.
- Published
- 2013
- Full Text
- View/download PDF
5. In memory of Professor Boris Afanasyev (August 28, 1947 – March 16, 2020)
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Boris Fehse, Sergey F. Bagnenko, Rüdiger Hehlmann, Alexei B. Chukhlovin, Ludmila S. Zubarovskaya, Inna V. Markova, Gerard Wagemaker, Alexander D. Kulagin, Axel R. Zander, and Ivan S. Moiseev
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Transplantation ,Molecular Medicine - Published
- 2020
6. SUL-109 Protects Hematopoietic Stem Cells from Apoptosis Induced by Short-Term Hypothermic Preservation and Maintains Their Engraftment Potential
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Özgür Özyüncü, F. Duygu Uçkan-Çetinkaya, Aynura Mammadova, Fatima Aerts-Kaya, Gerard Wagemaker, Trudi P. Visser, and Burcu Pervin
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CD34 ,Cold storage ,Antigens, CD34 ,Apoptosis ,Hypothermia ,Andrology ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Animals ,Humans ,Medicine ,Viability assay ,Chromans ,Transplantation ,business.industry ,Hematopoietic Stem Cell Transplantation ,Hematology ,Fetal Blood ,Hematopoietic Stem Cells ,Haematopoiesis ,medicine.anatomical_structure ,Cell culture ,030220 oncology & carcinogenesis ,Bone marrow ,Stem cell ,business ,030215 immunology - Abstract
The newly developed 6-hydroxychromanol derivate SUL-109 was shown to provide protection during hypothermic storage of several cell lines, but has not been evaluated in hematopoietic stem cells (HSCs). Hypothermic preservation of HSCs would be preferred over short-term cryopreservation to prevent cell loss during freezing/thawing and would be particularly useful for short-term storage, such as during conditioning of patients or transport of HSC transplants. Here we cultured human CD34+ umbilical cord blood (UCB) cells and lineage-depleted (Lin–) Balb/c bone marrow (BM) cells for up to 7 days in serum-free HSC expansion medium with hematopoietic growth factors. SUL-109-containing cultures were stored at 4°C for 3 to 14 days. The UCB cells were tested for viability, cell cycle, and reactive oxygen species (ROS). DMSO-cryopreserved Lin– BM cells or Lin– BM cells maintained for 14 days at 4°C were transplanted into RAG2−/− Balb/c mice and engraftment was followed for 6 months. The addition of SUL-109 during the hypothermic storage of expanded CD34+ UCB cells provided a significant improvement in cell survival of the immature CD34+/CD38- fraction after 7 days of hypothermic storage through scavenging of hypothermia-induced ROS and was able to preserve the multilineage capacity of human CD34+ UCB cells for up to 14 days of cold storage. In addition, SUL-109 protected murine BM Lin– cells from 14 days of hypothermic preservation and maintained their engraftment potential after transplantation in immune-deficient RAG2−/− mice. Our data indicate that SUL-109 is a promising novel chemical for use as a protective agent during cold storage of human and murine HSCs to prevent hypothermia-induced apoptosis and promote cell viability.
- Published
- 2020
7. Partial Hemopoietic Chimerism Described by Means of a Mathematical Model of the Erythroid Pathway
- Author
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Jenne J. Wielenga, Gerard Wagemaker, and A. van Rotterdam
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Haematopoiesis ,Biology ,Cell biology - Published
- 2020
8. Lentiviral Hematopoietic Stem Cell Gene Therapy Corrects Murine Pompe Disease
- Author
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Marshall W. Huston, Edwin H. Jacobs, Gerard Wagemaker, Onno Roovers, Elza D. van Deel, Helen de Boer, Arnold J.J. Reuser, Dirk J. Duncker, Niek P. van Til, Holger Jahr, Trudi P. Visser, Merel Stok, Pediatrics, Hematology, Clinical Genetics, Molecular Genetics, Orthopedics and Sports Medicine, and Cardiology
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0301 basic medicine ,murine Pompe disease ,lcsh:QH426-470 ,Receptor expression ,medicine.medical_treatment ,Genetic enhancement ,Hematopoietic stem cell transplantation ,Article ,Viral vector ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Genetics ,medicine ,skeletal muscle ,lcsh:QH573-671 ,Molecular Biology ,Glycogen ,lcsh:Cytology ,business.industry ,lentiviral vector ,Hematopoietic stem cell ,Skeletal muscle ,Enzyme replacement therapy ,central nervous system ,lcsh:Genetics ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,030220 oncology & carcinogenesis ,hematopoietic stem cell transplantation ,acid α-glucosidase ,Cancer research ,Molecular Medicine ,business - Abstract
Pompe disease is an autosomal recessive lysosomal storage disorder characterized by progressive muscle weakness. The disease is caused by mutations in the acid α-glucosidase (GAA) gene. Despite the currently available enzyme replacement therapy (ERT), roughly half of the infants with Pompe disease die before the age of 3 years. Limitations of ERT are immune responses to the recombinant enzyme, incomplete correction of the disease phenotype, lifelong administration, and inability of the enzyme to cross the blood-brain barrier. We previously reported normalization of glycogen in heart tissue and partial correction of the skeletal muscle phenotype by ex vivo hematopoietic stem cell gene therapy. In the present study, using a codon-optimized GAA (GAAco), the enzyme levels resulted in close to normalization of glycogen in heart, muscles, and brain, and in complete normalization of motor function. A large proportion of microglia in the brain was shown to be GAA positive. All astrocytes contained the enzyme, which is in line with mannose-6-phosphate receptor expression and the key role in glycogen storage and glucose metabolism. The lentiviral vector insertion site analysis confirmed no preference for integration near proto-oncogenes. This correction of murine Pompe disease warrants further development toward a cure of the human condition., Graphical Abstract, This publication reports that stem cell gene therapy using a codon-optimized gene encoding acid α-glucosidase (GAA) cures the mouse model of Pompe disease, a lysosomal storage disorder.
- Published
- 2020
9. Preclinical Efficacy and Safety Evaluation of Hematopoietic Stem Cell Gene Therapy in a Mouse Model of MNGIE
- Author
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Ramon Martí, Niek P. van Til, Joost C. Haeck, Lorena Leonardelli, Luca Biasco, Elly Bogaerts, Irenaeus F.M. de Coo, Merel Stok, Raquel Cabrera-Pérez, Gerard Wagemaker, Steffi Goffart, Marshall W. Huston, Marianna Bugiani, George J G Ruijter, Edwin H. Jacobs, Javier Torres-Torronteras, Monique R. Bernsen, Rana Yadak, Robert M. Verdijk, Pathology, Amsterdam Neuroscience - Cellular & Molecular Mechanisms, Neurology, Radiology & Nuclear Medicine, and Clinical Genetics
- Subjects
0301 basic medicine ,lcsh:QH426-470 ,Genetic enhancement ,lentiviral vectors ,thymidine phosphorylase ,Article ,Viral vector ,03 medical and health sciences ,chemistry.chemical_compound ,Gene therapy ,Genetics ,Medicine ,Vector (molecular biology) ,lcsh:QH573-671 ,Thymidine phosphorylase ,Molecular Biology ,Phosphoglycerate kinase ,lcsh:Cytology ,business.industry ,Hematopoietic stem cell ,Lentiviral vectors ,gene therapy ,humanities ,Deoxyuridine ,hematopoietic stem cells ,3. Good health ,lcsh:Genetics ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,MNGIE ,Cancer research ,Molecular Medicine ,business ,Thymidine ,Hematopoietic stem cells - Abstract
Altres ajuts: The authors acknowledge the financial support for this study by Join4energy, Ride4Kids, the Sophia Foundation (SSW0645), Stichting NeMo, in the context of funding provided by the European Commission's 5th, 6th, and 7th Framework Programs(contracts QLK3-CT-2001-00427-INHERINET, LSHB-CT-2004-005242-CONSERT, LSHB-CT-2006-19038 Magselectofection, and grant agreements 222878-PERSIST and 261387 CELL-PID), and by the Netherlands Health Research and Development Organization ZonMw (Translational Gene Therapy program projects 43100016 and 43400010). We thank Dr. Michio Hirano (Department of Neurology, Columbia University Medical Center, New York, USA) for providing the murine model, Louis Boon (Epirus Biopharmaceuticals, Utrecht, the Netherlands) for kindly providing anti-B220 antibody, Prof. Peter A.E. Sillevis Smitt (Department of Neurology, Erasmus MC, Rotterdam, the Netherlands), Pier.G. Mastroberardino and Chiara Milanese (Department of Molecular Genetics, Erasmus MC), Kees Schoonderwoerd (Department of Clinical Genetics, Erasmus MC), and Jeroen de Vrij (Department of Neurosurgery, Erasmus MC) for valuable discussions, Lidia Hussaarts (Department of Clinical Genetics, Erasmus MC) for technical support, King Lam (Department of Pathology, Erasmus MC) for pathology evaluation, and F. Dionisio and A. Aiuti from HSR-TIGET, Milan, for the support to the integration site analysis. Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is an autosomal recessive disorder caused by thymidine phosphorylase (TP) deficiency resulting in systemic accumulation of thymidine (d-Thd) and deoxyuridine (d-Urd) and characterized by early-onset neurological and gastrointestinal symptoms. Long-term effective and safe treatment is not available. Allogeneic bone marrow transplantation may improve clinical manifestations but carries disease and transplant-related risks. In this study, lentiviral vector-based hematopoietic stem cell gene therapy (HSCGT) was performed in Tymp −/− Upp1 −/− mice with the human phosphoglycerate kinase (PGK) promoter driving TYMP. Supranormal blood TP activity reduced intestinal nucleoside levels significantly at low vector copy number (median, 1.3; range, 0.2-3.6). Furthermore, we covered two major issues not addressed before. First, we demonstrate aberrant morphology of brain astrocytes in areas of spongy degeneration, which was reversed by HSCGT. Second, long-term follow-up and vector integration site analysis were performed to assess safety of the therapeutic LV vectors in depth. This report confirms and supplements previous work on the efficacy of HSCGT in reducing the toxic metabolites in Tymp −/− Upp1 −/− mice, using a clinically applicable gene transfer vector and a highly efficient gene transfer method, and importantly demonstrates phenotypic correction with a favorable risk profile, warranting further development toward clinical implementation.
- Published
- 2018
10. Lentiviral Hematopoietic Stem Cell Gene Therapy in Inherited Immune and Lysoso- mal Enzyme Deficiencies
- Author
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Gerard Wagemaker
- Subjects
Transplantation ,business.industry ,Transgene ,Genetic enhancement ,Hematopoietic stem cell ,medicine.disease ,Clinical trial ,Metachromatic leukodystrophy ,medicine.anatomical_structure ,Immune system ,Immunology ,medicine ,Molecular Medicine ,Adrenoleukodystrophy ,business ,Gene - Abstract
Rare diseases affect millions of people worldwide. Many of those are inherited disorders resulting in chronic disability and requiring cost-intensive care. Hematopoietic stem cell gene therapy has been developed over more than 20 years. At the state of the art, gene therapy is within reach for diseases in which (i) the genetic defect is identified, (ii) the diagnosis is made sufficiently early for a meaningful therapeutic intervention, (iii) a specific animal model is available for efficacy and safety evaluation. Appropriate therapeutic transgenes should also comply with certain biological criteria. Third-generation lentiviral vectors have been made self-inactivating (SIN) by deletion of enhancer regions from the LTR sequences thus reducing the risk of influencing nearby genes, resulting in favorable safety profiles. At the present time, lentiviral hematopoietic stem cell gene therapy has entered the stage of initial clinical implementation for immune deficiencies and lysosomal storage disorders. We discuss initial clinical trials using these vectors for selected metabolic storage disorders, which include adrenoleukodystrophy, metachromatic leukodystrophy, Hurler (MPS I), Pompe (GSD II), and Fabry diseases. This brief review summarizes the development and current clinical implementation of these approaches.
- Published
- 2016
11. Efficacy Of Lentivirus-Mediated Gene Therapy In An Omenn Syndrome Recombination-Activating Gene 2 Mouse Model Is Not Hindered By Inflammation And Immune Dysregulation
- Author
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Anna Villa, Paola Carrera, Marita Bosticardo, Elena Fontana, Maria Carmina Castiello, Sara Penna, Monica Zanussi, Lucia Sergi Sergi, Elena Draghici, Luigi Poliani, Nicolò Sacchetti, Gerard Wagemaker, Valentina Capo, Barbara Cassani, Luigi D. Notarangelo, Niek P. van Til, Kerry Dobbs, Rosita Rigoni, Paolo Uva, İç Hastalıkları, Hematology, Capo, V, Castiello, M, Fontana, E, Penna, S, Bosticardo, M, Draghici, E, Poliani, L, Sergi Sergi, L, Rigoni, R, Cassani, B, Zanussi, M, Carrera, P, Uva, P, Dobbs, K, Sacchetti, N, Notarangelo, L, van Til, N, Wagemaker, G, and Villa, A
- Subjects
Male ,0301 basic medicine ,Rag gene ,Allergy ,T-Lymphocytes ,Genetic enhancement ,Immunology ,Autoimmunity ,Mice, Transgenic ,Article ,Gene therapy ,Lentiviral vector ,Omenn syndrome ,Rag genes ,Immunology and Allergy ,03 medical and health sciences ,RAG2 ,medicine ,Animals ,Lymphocyte Count ,Immunodeficiency ,Inflammation ,B-Lymphocytes ,Severe combined immunodeficiency ,business.industry ,Lentivirus ,Genetic Therapy ,medicine.disease ,Autoimmune regulator ,3. Good health ,DNA-Binding Proteins ,Mice, Inbred C57BL ,Transplantation ,Disease Models, Animal ,030104 developmental biology ,Primary immunodeficiency ,Female ,Severe Combined Immunodeficiency ,business - Abstract
Background Omenn syndrome (OS) is a rare severe combined immunodeficiency associated with autoimmunity and caused by defects in lymphoid-specific V(D)J recombination. Most patients carry hypomorphic mutations in recombination-activating gene ( RAG ) 1 or 2. Hematopoietic stem cell transplantation is the standard treatment; however, gene therapy (GT) might represent a valid alternative, especially for patients lacking a matched donor. Objective We sought to determine the efficacy of lentiviral vector (LV)–mediated GT in the murine model of OS (Rag2 R229Q/R229Q ) in correcting immunodeficiency and autoimmunity. Methods Lineage-negative cells from mice with OS were transduced with an LV encoding the human RAG2 gene and injected into irradiated recipients with OS. Control mice underwent transplantation with wild-type or OS-untransduced lineage-negative cells. Immunophenotyping, T-dependent and T-independent antigen challenge, immune spectratyping, autoantibody detection, and detailed tissue immunohistochemical analyses were performed. Results LV-mediated GT allowed immunologic reconstitution, although it was suboptimal compared with that seen in wild-type bone marrow (BM)−transplanted OS mice in peripheral blood and hematopoietic organs, such as the BM, thymus, and spleen. We observed in vivo variability in the efficacy of GT correlating with the levels of transduction achieved. Immunoglobulin levels and T-cell repertoire normalized, and gene-corrected mice responded properly to challenges in vivo . Autoimmune manifestations, such as skin infiltration and autoantibodies, dramatically improved in GT mice with a vector copy number/genome higher than 1 in the BM and 2 in the thymus. Conclusions Our data show that LV-mediated GT for patients with OS significantly ameliorates the immunodeficiency, even in an inflammatory environment.
- Published
- 2018
12. Assessing the carcinogenic potential of low-dose exposures to chemical mixtures in the environment: the challenge ahead
- Author
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Dustin G. Brown, Tove Hultman, Judith Weisz, H. Kim Lyerly, Paola A. Marignani, Ann-Karin Olsen, Rabindra Roy, Kim Moorwood, Masoud H. Manjili, Monica Vaccari, Jesse Roman, Hasiah Ab Hamid, Kalan R. Prudhomme, Periyadan K. Krishnakumar, Chenfang Dong, Tiziana Guarnieri, Leandro S. D'Abronzo, Gloria M. Calaf, Amelia K Charles, Emanuela Corsini, Yunus A. Luqmani, Graeme Williams, Louis Vermeulen, Pankaj Vadgama, Sarah N Bay, Véronique Maguer-Satta, Sabine A. S. Langie, Christian C. Naus, Le Jian, Gladys N. Nangami, Lorenzo Memeo, Stephanie C. Casey, Thomas Sanderson, Takemi Otsuki, Nichola Cruickshanks, William H. Bisson, Sudjit Luanpitpong, Jonathan Whitfield, Ahmed Lasfar, Yon Rojanasakul, A. Ivana Scovassi, Shelley A. Harris, Ferdinando Chiaradonna, Richard Ponce-Cusi, Gregory T. Wolf, Valérian Dormoy, Roslida Abd Hamid, Hyun Ho Park, Matilde E. Lleonart, William K. Decker, Maria Romano, Leroy Lowe, Fabio Marongiu, Jan Vondráček, Chiara Mondello, Luc Leyns, Josiah Ochieng, Pratima Nangia-Makker, Edward A. Ratovitski, Zhiwei Hu, Jayadev Raju, Hemad Yasaei, Rafaela Andrade-Vieira, Jordan Woodrick, Hideko Sone, Harini Krishnan, W. Kimryn Rathmell, Andrew Collins, Luoping Zhang, Barry J. Barclay, Amaya Azqueta, Laura Soucek, Marc A. Williams, David O. Carpenter, Roberta Palorini, Rita Nahta, Juan Fernando Martinez-Leal, Firouz Darroudi, Rita Dornetshuber-Fleiss, James E. Klaunig, Elizabeth P. Ryan, Qiang Shawn Cheng, Arthur Berg, Andrew Ward, Gudrun Koppen, Tao Chen, Petr Heneberg, Michael Gilbertson, Amedeo Amedei, Sakina E. Eltom, Ezio Laconi, Joseph Christopher, Hiroshi Kondoh, Neetu Singh, Danielle J Carlin, Marion Chapellier, Michalis V. Karamouzis, Rekha Mehta, Tae-Jin Lee, Annamaria Colacci, Venkata S. Sabbisetti, Mark Wade, Micheline Kirsch-Volders, Patricia Ostrosky-Wegman, Isabelle R. Miousse, Patricia A. Thompson, Philippa D. Darbre, Frederik J. van Schooten, Sofia Pavanello, Igor Koturbash, Binhua P. Zhou, Ranjeet Kumar Sinha, Anna C. Salzberg, Mahara Valverde, Fahd Al-Mulla, Julia Kravchenko, Nicole Kleinstreuer, Carolyn J. Baglole, Menghang Xia, Samira A. Brooks, Amancio Carnero, Gunnar Brunborg, Sandra S. Wise, Daniel C. Koch, John Pierce Wise, Rabeah Al-Temaimi, Laetitia Gonzalez, Lisa J. McCawley, R. Brooks Robey, Gary S. Goldberg, Thierry Massfelder, Linda S M Gulliver, Olugbemiga Ogunkua, Emilio Rojas, Eun-Yi Moon, Lin Li, Silvana Papagerakis, Nik van Larebeke, Adela Lopez de Cerain Salsamendi, Staffan Eriksson, Simona Romano, Dean W. Felsher, Paramita M. Ghosh, Karine A. Cohen-Solal, Paul Dent, Jun Sun, Carmen Blanco-Aparicio, Riccardo Di Fiore, Chia-Wen Hsu, Mahin Khatami, Kannan Badri Narayanan, Francis Martin, Colleen S. Curran, Dale W. Laird, William H. Goodson, Abdul Manaf Ali, Valerie Odero-Marah, Michael J. Gonzalez, Renza Vento, Liang Tzung Lin, Clement G. Yedjou, Hosni Salem, Hsue-Yin Hsu, Zhenbang Chen, Nuzhat Ahmed, Gerard Wagemaker, Sandra Ryeom, Stefano Forte, Debasish Roy, Nancy B. Kuemmerle, Robert C. Castellino, Po Sing Leung, Wilhelm Engström, National Institute of Environmental Health Sciences (US), Research Council of Norway, Ministerio de Economía y Competitividad (España), Instituto de Salud Carlos III, Red Temática de Investigación Cooperativa en Cáncer (España), European Commission, Junta de Andalucía, Ministerio de Educación y Ciencia (España), Ministero dell'Istruzione, dell'Università e della Ricerca, University of Oslo, Regione Emilia Romagna, National Institutes of Health (US), Consejo Nacional de Ciencia y Tecnología (México), Associazione Italiana per la Ricerca sul Cancro, National Research Foundation of Korea, Ministry of Education, Science and Technology (South Korea), Fondo Nacional de Desarrollo Científico y Tecnológico (Chile), Ministry of Education, Culture, Sports, Science and Technology (Japan), Japan Science and Technology Agency, Ministry of Science and Technology (Taiwan), Arkansas Biosciences Institute, Czech Science Foundation, Fundación Fero, Swim Across America, American Cancer Society, Research Foundation - Flanders, Austrian Science Fund, Institut National de la Santé et de la Recherche Médicale (France), Natural Sciences and Engineering Research Council of Canada, Farmacologie en Toxicologie, RS: NUTRIM - R4 - Gene-environment interaction, Goodson, William H, Lowe, Leroy, Carpenter, David O, Gilbertson, Michael, Manaf Ali, Abdul, Lopez de Cerain Salsamendi, Adela, Lasfar, Ahmed, Carnero, Amancio, Azqueta, Amaya, Amedei, Amedeo, Charles, Amelia K, Collins, Andrew R, Ward, Andrew, Salzberg, Anna C, Colacci, Annamaria, Olsen, Ann Karin, Berg, Arthur, Barclay, Barry J, Zhou, Binhua P, Blanco Aparicio, Carmen, Baglole, Carolyn J, Dong, Chenfang, Mondello, Chiara, Hsu, Chia Wen, Naus, Christian C, Yedjou, Clement, Curran, Colleen S, Laird, Dale W, Koch, Daniel C, Carlin, Danielle J, Felsher, Dean W, Roy, Debasish, Brown, Dustin G, Ratovitski, Edward, Ryan, Elizabeth P, Corsini, Emanuela, Rojas, Emilio, Moon, Eun Yi, Laconi, Ezio, Marongiu, Fabio, Al Mulla, Fahd, Chiaradonna, Ferdinando, Darroudi, Firouz, Martin, Francis L, Van Schooten, Frederik J, Goldberg, Gary S, Wagemaker, Gerard, Nangami, Gladys N, Calaf, Gloria M, Williams, Graeme, Wolf, Gregory T, Koppen, Gudrun, Brunborg, Gunnar, Lyerly, H. Kim, Krishnan, Harini, Ab Hamid, Hasiah, Yasaei, Hemad, Sone, Hideko, Kondoh, Hiroshi, Salem, Hosni K, Hsu, Hsue Yin, Park, Hyun Ho, Koturbash, Igor, Miousse, Isabelle R, Scovassi, A. Ivana, Klaunig, James E, Vondráček, Jan, Raju, Jayadev, Roman, Jesse, Wise, John Pierce, Whitfield, Jonathan R, Woodrick, Jordan, Christopher, Joseph A, Ochieng, Josiah, Martinez Leal, Juan Fernando, Weisz, Judith, Kravchenko, Julia, Sun, Jun, Prudhomme, Kalan R, Narayanan, Kannan Badri, Cohen Solal, Karine A, Moorwood, Kim, Gonzalez, Laetitia, Soucek, Laura, Jian, Le, D'Abronzo, Leandro S, Lin, Liang Tzung, Li, Lin, Gulliver, Linda, Mccawley, Lisa J, Memeo, Lorenzo, Vermeulen, Loui, Leyns, Luc, Zhang, Luoping, Valverde, Mahara, Khatami, Mahin, Romano, MARIA FIAMMETTA, Chapellier, Marion, Williams, Marc A, Wade, Mark, Manjili, Masoud H, Lleonart, Matilde E, Xia, Menghang, Gonzalez, Michael J, Karamouzis, Michalis V, Kirsch Volders, Micheline, Vaccari, Monica, Kuemmerle, Nancy B, Singh, Neetu, Cruickshanks, Nichola, Kleinstreuer, Nicole, van Larebeke, Nik, Ahmed, Nuzhat, Ogunkua, Olugbemiga, Krishnakumar, P. K, Vadgama, Pankaj, Marignani, Paola A, Ghosh, Paramita M, Ostrosky Wegman, Patricia, Thompson, Patricia A, Dent, Paul, Heneberg, Petr, Darbre, Philippa, Sing Leung, Po, Nangia Makker, Pratima, Cheng, Qiang Shawn, Robey, R. Brook, Al Temaimi, Rabeah, Roy, Rabindra, Andrade Vieira, Rafaela, Sinha, Ranjeet K, Mehta, Rekha, Vento, Renza, Di Fiore, Riccardo, Ponce Cusi, Richard, Dornetshuber Fleiss, Rita, Nahta, Rita, Castellino, Robert C, Palorini, Roberta, Abd Hamid, Roslida, Langie, Sabine A. S, Eltom, Sakina E, Brooks, Samira A, Ryeom, Sandra, Wise, Sandra S, Bay, Sarah N, Harris, Shelley A, Papagerakis, Silvana, Romano, Simona, Pavanello, Sofia, Eriksson, Staffan, Forte, Stefano, Casey, Stephanie C, Luanpitpong, Sudjit, Lee, Tae Jin, Otsuki, Takemi, Chen, Tao, Massfelder, Thierry, Sanderson, Thoma, Guarnieri, Tiziana, Hultman, Tove, Dormoy, Valérian, Odero Marah, Valerie, Sabbisetti, Venkata, Maguer Satta, Veronique, Rathmell, W. Kimryn, Engström, Wilhelm, Decker, William K, Bisson, William H, Rojanasakul, Yon, Luqmani, Yunu, Chen, Zhenbang, Hu, Zhiwei, Goodson, W., Lowe, L., Carpenter, D., Gilbertson, M., Ali, A., de Cerain Salsamendi, A., Lasfar, A., Carnero, A., Azqueta, A., Amedei, A., Charles, A., Collins, A., Ward, A., Salzberg, A., Colacci, A., Olsen, A., Berg, A., Barclay, B., Zhou, B., Blanco-Aparicio, C., Baglole, C., Dong, C., Mondello, C., Hsu, C., Naus, C., Yedjou, C., Curran, C., Laird, D., Koch, D., Carlin, D., Felsher, D., Roy, D., Brown, D., Ratovitski, E., Ryan, E., Corsini, E., Rojas, E., Moon, E., Laconi, E., Marongiu, F., Al-Mulla, F., Chiaradonna, F., Darroudi, F., Martin, F., Van Schooten, F., Goldberg, G., Wagemaker, G., Nangami, G., Calaf, G., Williams, G., Wolf, G., Koppen, G., Brunborg, G., Kim Lyerly, H., Krishnan, H., Hamid, H., Yasaei, H., Sone, H., Kondoh, H., Salem, H., Hsu, H., Park, H., Koturbash, I., Miousse, I., Ivana Scovassi, A., Klaunig, J., Vondráček, J., Raju, J., Roman, J., Wise, J., Whitfield, J., Woodrick, J., Christopher, J., Ochieng, J., Martinez-Leal, J., Weisz, J., Kravchenko, J., Sun, J., Prudhomme, K., Narayanan, K., Cohen-Solal, K., Moorwood, K., Gonzalez, L., Soucek, L., Jian, L., D'Abronzo, L., Lin, L., Li, L., Gulliver, L., Mccawley, L., Memeo, L., Vermeulen, L., Leyns, L., Zhang, L., Valverde, M., Khatami, M., Romano, M., Chapellier, M., Williams, M., Wade, M., Manjili, M., Lleonart, M., Xia, M., Gonzalez, M., Karamouzis, M., Kirsch-Volders, M., Vaccari, M., Kuemmerle, N., Singh, N., Cruickshanks, N., Kleinstreuer, N., Van Larebeke, N., Ahmed, N., Ogunkua, O., Krishnakumar, P., Vadgama, P., Marignani, P., Ghosh, P., Ostrosky-Wegman, P., Thompson, P., Dent, P., Heneberg, P., Darbre, P., Leung, P., Nangia-Makker, P., Cheng, Q., Brooks Robey, R., Al-Temaimi, R., Roy, R., Andrade-Vieira, R., Sinha, R., Mehta, R., Vento, R., Di Fiore, R., Ponce-Cusi, R., Dornetshuber-Fleiss, R., Nahta, R., Castellino, R., Palorini, R., Hamid, R., Langie, S., Eltom, S., Brooks, S., Ryeom, S., Wise, S., Bay, S., Harris, S., Papagerakis, S., Romano, S., Pavanello, S., Eriksson, S., Forte, S., Casey, S., Luanpitpong, S., Lee, T., Otsuki, T., Chen, T., Massfelder, T., Sanderson, T., Guarnieri, T., Hultman, T., Dormoy, V., Odero-Marah, V., Sabbisetti, V., Maguer-Satta, V., Kimryn Rathmell, W., Engström, W., Decker, W., Bisson, W., Rojanasakul, Y., Luqmani, Y., Chen, Z., Hu, Z., Goodson, W.H., Carpenter, D.O., Ali, A.M., de Cerain Salsamendi, A.L., Charles, A.K., Collins, A.R., Salzberg, A.C., Olsen, A.-K., Barclay, B.J., Zhou, B.P., Baglole, C.J., Hsu, C.-W., Naus, C.C., Curran, C.S., Laird, D.W., Koch, D.C., Carlin, D.J., Felsher, D.W., Brown, D.G., Ryan, E.P., Moon, E.-Y., Martin, F.L., Van Schooten, F.J., Goldberg, G.S., Calaf, G.M., Wolf, G.T., Hamid, H.A., Salem, H.K., Hsu, H.-Y., Park, H.H., Miousse, I.R., Klaunig, J.E., Vondracek, J., Wise, J.P., Whitfield, J.R., Christopher, J.A., Martinez-Leal, J.F., Prudhomme, K.R., Narayanan, K.B., Cohen-Solal, K.A., D'Abronzo, L.S., Lin, L.-T., Mccawley, L.J., Romano, M.F., Williams, M.A., Manjili, M.H., Gonzalez, M.J., Karamouzis, M.V., Kuemmerle, N.B., Krishnakumar, P.K., Marignani, P.A., Ghosh, P.M., Leung, P.S., Cheng, Q.S., Sinha, R.K., Castellino, R.C., Hamid, R.A., Langie, S.A.S., Brooks, S.A., Wise, S.S., Bay, S.N., Harris, S.A., Casey, S.C., Lee, T.-J., Engstrom, W., Decker, W.K., Bisson, W.H., sans affiliation, Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA), Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), We gratefully acknowledge the support of the National Institute of Health-National Institute of Environmental Health Sciences (NIEHS) conference grant travel support (R13ES023276), Glenn Rice, Office of Research and Development, United States Environmental Protection Agency, Cincinnati, OH, USA also deserves thanks for his thoughtful feedback and inputs on the manuscript, William H.Goodson III was supported by the California Breast Cancer Research Program, Clarence Heller Foundation and California Pacific Medical Center Foundation, Abdul M.Ali would like to acknowledge the financial support of the University of Sultan Zainal Abidin, Malaysia, Ahmed Lasfar was supported by an award from the Rutgers Cancer Institute of New Jersey, Ann-Karin Olsen and Gunnar Brunborg were supported by the Research Council of Norway (RCN) through its Centres of Excellence funding scheme (223268/F50), Amancio Carnero’s lab was supported by grants from the Spanish Ministry of Economy and Competitivity, ISCIII (Fis: PI12/00137, RTICC: RD12/0036/0028) co-funded by FEDER from Regional Development European Funds (European Union), Consejeria de Ciencia e Innovacion (CTS-1848) and Consejeria de Salud of the Junta de Andalucia (PI-0306-2012), Matilde E. Lleonart was supported by a trienal project grant PI12/01104 and by project CP03/00101 for personal support. Amaya Azqueta would like to thank the Ministerio de Educacion y Ciencia (‘Juande la Cierva’ programme, 2009) of the Spanish Government for personal support, Amedeo Amedei was supported by the Italian Ministry of University and Research (2009FZZ4XM_002), and the University of Florence (ex60%2012), Andrew R.Collins was supported by the University of Oslo, Annamaria Colacci was supported by the Emilia-Romagna Region - Project ‘Supersite’ in Italy, Carolyn Baglole was supported by a salary award from the Fonds de recherche du Quebec-Sante (FRQ-S), Chiara Mondello’s laboratory is supported by Fondazione Cariplo in Milan, Italy (grant n. 2011-0370), Christian C.Naus holds a Canada Research Chair, Clement Yedjou was supported by a grant from the National Institutes of Health (NIH-NIMHD grant no. G12MD007581), Daniel C.Koch is supported by the Burroughs Wellcome Fund Postdoctoral Enrichment Award and the Tumor Biology Training grant: NIH T32CA09151, Dean W. Felsher would like to acknowledge the support of United States Department of Health and Human Services, NIH grants (R01 CA170378 PQ22, R01 CA184384, U54 CA149145, U54 CA151459, P50 CA114747 and R21 CA169964), Emilio Rojas would like to thank CONACyT support 152473, Ezio Laconi was supported by AIRC (Italian Association for Cancer Research, grant no. IG 14640) and by the Sardinian Regional Government (RAS), Eun-Yi Moon was supported by grants from the Public Problem-Solving Program (NRF-015M3C8A6A06014500) and Nuclear R&D Program (#2013M2B2A9A03051296 and 2010-0018545) through the National Research Foundation of Korea (NRF) and funded by the Ministry of Education, Science and Technology (MEST) in Korea, Fahd Al-Mulla was supported by the Kuwait Foundation for the Advancement of Sciences (2011-1302-06), Ferdinando Chiaradonna is supported by SysBioNet, a grant for the Italian Roadmap of European Strategy Forum on Research Infrastructures (ESFRI) and by AIRC (Associazione Italiana Ricerca sul Cancro, IG 15364), Francis L.Martin acknowledges funding from Rosemere Cancer Foundation, he also thanks Lancashire Teaching Hospitals NHS trust and the patients who have facilitated the studies he has undertaken over the course of the last 10 years, Gary S.Goldberg would like to acknowledge the support of the New Jersey Health Foundation, Gloria M.Calaf was supported by Fondo Nacional de Ciencia y Tecnología (FONDECYT), Ministerio de Educación de Chile (MINEDUC), Universidad de Tarapacá (UTA), Gudrun Koppen was supported by the Flemish Institute for Technological Research (VITO), Belgium, Hemad Yasaei was supported from a triennial project grant (Strategic Award) from the National Centre for the Replacement, Refinement and Reduction (NC3Rs) of animals in research (NC.K500045.1 and G0800697), Hiroshi Kondoh was supported in part by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, Japan Science and Technology Agency and by JST, CREST, Hsue-Yin Hsu was supported by the Ministry of Science and Technology of Taiwan (NSC93-2314-B-320-006 and NSC94-2314-B-320-002), Hyun Ho Park was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) of the Ministry of Education, Science and Technology (2012R1A2A2A01010870) and a grant from the Korea Healthcare Technology R&D project, Ministry of Health and Welfare, Republic of Korea (HI13C1449), Igor Koturbash is supported by the UAMS/NIH Clinical and Translational Science Award (UL1TR000039 and KL2TR000063) and the Arkansas Biosciences Institute, the major research component of the Arkansas Tobacco Settlement Proceeds Act of 2000, Jan Vondráček acknowledges funding from the Czech Science Foundation (13-07711S), Jesse Roman thanks the NIH for their support (CA116812), John Pierce Wise Sr. and Sandra S.Wise were supported by National Institute of Environmental Health Sciences (ES016893 to J.P.W.) and the Maine Center for Toxicology and Environmental Health, Jonathan Whitfield acknowledges support from the FERO Foundation in Barcelona, Spain, Joseph Christopher is funded by Cancer Research UK and the International Journal of Experimental Pathology, Julia Kravchenko is supported by a philanthropic donation by Fred and Alice Stanback, Jun Sun is supported by a Swim Across America Cancer Research Award, Karine A.Cohen-Solal is supported by a research scholar grant from the American Cancer Society (116683-RSG-09-087-01-TBE), Laetitia Gonzalez received a postdoctoral fellowship from the Fund for Scientific Research–Flanders (FWO-Vlaanderen) and support by an InterUniversity Attraction Pole grant (IAP-P7-07), Laura Soucek is supported by grant #CP10/00656 from the Miguel Servet Research Contract Program and acknowledges support from the FERO Foundation in Barcelona, Spain, Liang-Tzung Lin was supported by funding from the Taipei Medical University (TMU101-AE3-Y19), Linda Gulliver is supported by a Genesis Oncology Trust (NZ) Professional Development Grant, and the Faculty of Medicine, University of Otago, Dunedin, New Zealand, Louis Vermeulen is supported by a Fellowship of the Dutch Cancer Society (KWF, UVA2011-4969) and a grant from the AICR (14–1164), Mahara Valverde would like to thank CONACyT support 153781, Masoud H. Manjili was supported by the office of the Assistant Secretary of Defense for Health Affairs (USA) through the Breast Cancer Research Program under Award No. W81XWH-14-1-0087 Neetu Singh was supported by grant #SR/FT/LS-063/2008 from the Department of Science and Technology, Government of India, Nicole Kleinstreuer is supported by NIEHS contracts (N01-ES 35504 and HHSN27320140003C), P.K. Krishnakumar is supported by the Funding (No. T.K. 11-0629) of King Abdulaziz City for Science and Technology, Riyadh, Saudi Arabia, Paola A.Marignani is supported by the Dalhousie Medical Research Foundation, The Beatrice Hunter Cancer Institute and CIHR and the Nova Scotia Lung Association, Paul Dent is the holder of the Universal Inc.Chair in Signal Transduction Research and is supported with funds from PHS grants from the NIH (R01-CA141704, R01-CA150214, R01-DK52825 and R01-CA61774), Petr Heneberg was supported by the Charles University in Prague projects UNCE 204015 and PRVOUK P31/2012, and by the Czech Science Foundation projects P301/12/1686 and 15-03834Y, Po Sing Leung was supported by the Health and Medical Research Fund of Food and Health Bureau, Hong Kong Special Administrative Region, Ref. No: 10110021, Qiang Cheng was supported in part by grant NSF IIS-1218712, R. Brooks Robey is supported by the United States Department of Veterans Affairs, Rabindra Roy was supported by United States Public Health Service Grants (RO1 CA92306, RO1 CA92306-S1 and RO1 CA113447), Rafaela Andrade-Vieira is supported by the Beatrice Hunter Cancer Research Institute and the Nova Scotia Health Research Foundation, Renza Vento was partially funded by European Regional Development Fund, European Territorial Cooperation 2007–13 (CCI 2007 CB 163 PO 037, OP Italia-Malta 2007–13) and grants from the Italian Ministry of Education, University and Research (MIUR) ex-60%, 2007, Riccardo Di Fiore was a recipient of fellowship granted by European Regional Development Fund, European Territorial Cooperation 2007–2013 (CCI 2007 CB 163 PO 037, OP Italia-Malta 2007–2013), Rita Dornetshuber-Fleiss was supported by the Austrian Science Fund (FWF, project number T 451-B18) and the Johanna Mahlke, geb.-Obermann-Stiftung, Roberta Palorini is supported by a SysBioNet fellowship, Roslida Abd Hamid is supported by the Ministry of Education, Malaysia-Exploratory Research Grant Scheme-Project no: ERGS/1-2013/5527165, Sabine A.S.Langie is the beneficiary of a postdoctoral grant from the AXA Research Fund and the Cefic-LRI Innovative Science Award 2013, Sakina Eltom is supported by NIH grant SC1CA153326, Samira A.Brooks was supported by National Research Service Award (T32 ES007126) from the National Institute of Environmental Health Sciences and the HHMI Translational Medicine Fellowship, Sandra Ryeom was supported by The Garrett B. Smith Foundation and the TedDriven Foundation, Thierry Massfelder was supported by the Institut National de la Santé et de la Recherche Médicale INSERM and Université de Strasbourg, Thomas Sanderson is supported by the Canadian Institutes of Health Research (CIHR, MOP-115019), the Natural Sciences and Engineering Council of Canada (NSERC, 313313) and the California Breast Cancer Research Program (CBCRP, 17UB-8703), Tiziana Guarnieri is supported by a grant from Fundamental Oriented Research (RFO) to the Alma Mater Studiorum University of Bologna, Bologna, Italy and thanks the Fondazione Cassa di Risparmio di Bologna and the Fondazione Banca del Monte di Bologna e Ravenna for supporting the Center for Applied Biomedical Research, S.Orsola-Malpighi University Hospital, Bologna, Italy, W.Kimryn Rathmell is supported by the V Foundation for Cancer Research and the American Cancer Society, William K.Decker was supported in part by grant RP110545 from the Cancer Prevention Research Institute of Texas, William H.Bisson was supported with funding from the NIH P30 ES000210, Yon Rojanasakul was supported with NIH grant R01-ES022968, Zhenbang Chen is supported by NIH grants (MD004038, CA163069 and MD007593), Zhiwei Hu is grateful for the grant support from an institutional start-up fund from The Ohio State University College of Medicine and The OSU James Comprehensive Cancer Center (OSUCCC) and a Seed Award from the OSUCCC Translational Therapeutics Program., Sans affiliation, Courcelles, Michel, Goodson, W, Lowe, L, Carpenter, D, Gilbertson, M, Ali, A, de Cerain Salsamendi, A, Lasfar, A, Carnero, A, Azqueta, A, Amedei, A, Charles, A, Collins, A, Ward, A, Salzberg, A, Colacci, A, Olsen, A, Berg, A, Barclay, B, Zhou, B, Blanco Aparicio, C, Baglole, C, Dong, C, Mondello, C, Hsu, C, Naus, C, Yedjou, C, Curran, C, Laird, D, Koch, D, Carlin, D, Felsher, D, Roy, D, Brown, D, Ratovitski, E, Ryan, E, Corsini, E, Rojas, E, Moon, E, Laconi, E, Marongiu, F, Al Mulla, F, Chiaradonna, F, Darroudi, F, Martin, F, Van Schooten, F, Goldberg, G, Wagemaker, G, Nangami, G, Calaf, G, Williams, G, Wolf, G, Koppen, G, Brunborg, G, Kim Lyerly, H, Krishnan, H, Hamid, H, Yasaei, H, Sone, H, Kondoh, H, Salem, H, Hsu, H, Park, H, Koturbash, I, Miousse, I, Ivana Scovassi, A, Klaunig, J, Vondráček, J, Raju, J, Roman, J, Wise, J, Whitfield, J, Woodrick, J, Christopher, J, Ochieng, J, Martinez Leal, J, Weisz, J, Kravchenko, J, Sun, J, Prudhomme, K, Narayanan, K, Cohen Solal, K, Moorwood, K, Gonzalez, L, Soucek, L, Jian, L, D'Abronzo, L, Lin, L, Li, L, Gulliver, L, Mccawley, L, Memeo, L, Vermeulen, L, Leyns, L, Zhang, L, Valverde, M, Khatami, M, Romano, M, Chapellier, M, Williams, M, Wade, M, Manjili, M, Lleonart, M, Xia, M, Gonzalez, M, Karamouzis, M, Kirsch Volders, M, Vaccari, M, Kuemmerle, N, Singh, N, Cruickshanks, N, Kleinstreuer, N, Van Larebeke, N, Ahmed, N, Ogunkua, O, Krishnakumar, P, Vadgama, P, Marignani, P, Ghosh, P, Ostrosky Wegman, P, Thompson, P, Dent, P, Heneberg, P, Darbre, P, Leung, P, Nangia Makker, P, Cheng, Q, Brooks Robey, R, Al Temaimi, R, Roy, R, Andrade Vieira, R, Sinha, R, Mehta, R, Vento, R, Di Fiore, R, Ponce Cusi, R, Dornetshuber Fleiss, R, Nahta, R, Castellino, R, Palorini, R, Hamid, R, Langie, S, Eltom, S, Brooks, S, Ryeom, S, Wise, S, Bay, S, Harris, S, Papagerakis, S, Romano, S, Pavanello, S, Eriksson, S, Forte, S, Casey, S, Luanpitpong, S, Lee, T, Otsuki, T, Chen, T, Massfelder, T, Sanderson, T, Guarnieri, T, Hultman, T, Dormoy, V, Odero Marah, V, Sabbisetti, V, Maguer Satta, V, Kimryn Rathmell, W, Engström, W, Decker, W, Bisson, W, Rojanasakul, Y, Luqmani, Y, Chen, Z, and Hu, Z
- Subjects
Cancer Research ,Carcinogenesis ,[SDV]Life Sciences [q-bio] ,METHOXYCHLOR-INDUCED ALTERATIONS ,Review ,Pharmacology ,MESH: Carcinogens, Environmental ,Carcinogenic synergies ,Chemical mixtures ,Neoplasms ,MESH: Animals ,MESH: Neoplasms ,Carcinogenesi ,Risk assessment ,Cancer ,ACTIVATED PROTEIN-KINASES ,Medicine (all) ,Low dose ,1. No poverty ,Cumulative effects ,BREAST-CANCER CELLS ,General Medicine ,Environmental exposure ,MESH: Carcinogenesis ,BIO/10 - BIOCHIMICA ,EPITHELIAL-MESENCHYMAL TRANSITION ,3. Good health ,[SDV] Life Sciences [q-bio] ,Environmental Carcinogenesis ,ESTROGEN-RECEPTOR-ALPHA ,Human ,MESH: Environmental Exposure ,ENDOCRINE-DISRUPTING CHEMICALS ,TARGETING TISSUE FACTOR ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Biology ,Prototypical chemical disruptors ,Exposure ,[SDV.CAN] Life Sciences [q-bio]/Cancer ,Environmental health ,medicine ,[SDV.EE.SANT] Life Sciences [q-bio]/Ecology, environment/Health ,Carcinogen ,Environmental carcinogenesis ,[SDV.EE.SANT]Life Sciences [q-bio]/Ecology, environment/Health ,MESH: Humans ,Animal ,POLYBROMINATED DIPHENYL ETHERS ,Environmental Exposure ,medicine.disease ,MESH: Hazardous Substances ,Carcinogens, Environmental ,MIGRATION INHIBITORY FACTOR ,VASCULAR ENDOTHELIAL-CELLS ,Hazardous Substance ,Neoplasm - Abstract
Goodson, William H. et al., © The Author 2015. Lifestyle factors are responsible for a considerable portion of cancer incidence worldwide, but credible estimates from the World Health Organization and the International Agency for Research on Cancer (IARC) suggest that the fraction of cancers attributable to toxic environmental exposures is between 7% and 19%. To explore the hypothesis that low-dose exposures to mixtures of chemicals in the environment may be combining to contribute to environmental carcinogenesis, we reviewed 11 hallmark phenotypes of cancer, multiple priority target sites for disruption in each area and prototypical chemical disruptors for all targets, this included dose-response characterizations, evidence of low-dose effects and cross-hallmark effects for all targets and chemicals. In total, 85 examples of chemicals were reviewed for actions on key pathways/ mechanisms related to carcinogenesis. Only 15% (13/85) were found to have evidence of a dose-response threshold, whereas 59% (50/85) exerted low-dose effects. No dose-response information was found for the remaining 26% (22/85). Our analysis suggests that the cumulative effects of individual (non-carcinogenic) chemicals acting on different pathways, and a variety of related systems, organs, tissues and cells could plausibly conspire to produce carcinogenic synergies. Additional basic research on carcinogenesis and research focused on low-dose effects of chemical mixtures needs to be rigorously pursued before the merits of this hypothesis can be further advanced. However, the structure of the World Health Organization International Programme on Chemical Safety 'Mode of Action' framework should be revisited as it has inherent weaknesses that are not fully aligned with our current understanding of cancer biology., We gratefully acknowledge the support of the National Institute of Health-National Institute of Environmental Health Sciences (NIEHS) conference grant travel support (R13ES023276); Glenn Rice, Office of Research and Development, United States Environmental Protection Agency, Cincinnati, OH, USA also deserves thanks for his thoughtful feedback and inputs on the manuscript; William H.Goodson III was supported by the California Breast Cancer Research Program, Clarence Heller Foundation and California Pacific Medical Center Foundation; Abdul M.Ali would like to acknowledge the financial support of the University of Sultan Zainal Abidin, Malaysia; Ahmed Lasfar was supported by an award from the Rutgers Cancer Institute of New Jersey; Ann-Karin Olsen and Gunnar Brunborg were supported by the Research Council of Norway (RCN) through its Centres of Excellence funding scheme (223268/F50), Amancio Carnero’s lab was supported by grants from the Spanish Ministry of Economy and Competitivity, ISCIII (Fis: PI12/00137, RTICC: RD12/0036/0028) co-funded by FEDER from Regional Development European Funds (European Union), Consejeria de Ciencia e Innovacion (CTS-1848) and Consejeria de Salud of the Junta de Andalucia (PI-0306-2012); Matilde E. Lleonart was supported by a trienal project grant PI12/01104 and by project CP03/00101 for personal support. Amaya Azqueta would like to thank the Ministerio de Educacion y Ciencia (‘Juande la Cierva’ programme, 2009) of the Spanish Government for personal support; Amedeo Amedei was supported by the Italian Ministry of University and Research (2009FZZ4XM_002), and the University of Florence (ex60%2012); Andrew R.Collins was supported by the University of Oslo; Annamaria Colacci was supported by the Emilia-Romagna Region - Project ‘Supersite’ in Italy; Carolyn Baglole was supported by a salary award from the Fonds de recherche du Quebec-Sante (FRQ-S); Chiara Mondello’s laboratory is supported by Fondazione Cariplo in Milan, Italy (grant n. 2011-0370); Christian C.Naus holds a Canada Research Chair; Clement Yedjou was supported by a grant from the National Institutes of Health (NIH-NIMHD grant no. G12MD007581); Daniel C.Koch is supported by the Burroughs Wellcome Fund Postdoctoral Enrichment Award and the Tumor Biology Training grant: NIH T32CA09151; Dean W. Felsher would like to acknowledge the support of United States Department of Health and Human Services, NIH grants (R01 CA170378 PQ22, R01 CA184384, U54 CA149145, U54 CA151459, P50 CA114747 and R21 CA169964); Emilio Rojas would like to thank CONACyT support 152473, Ezio Laconi was supported by AIRC (Italian Association for Cancer Research, grant no. IG 14640) and by the Sardinian Regional Government (RAS); Eun-Yi Moon was supported by grants from the Public Problem-Solving Program (NRF-015M3C8A6A06014500) and Nuclear R&D Program (#2013M2B2A9A03051296 and 2010-0018545) through the National Research Foundation of Korea (NRF) and funded by the Ministry of Education, Science and Technology (MEST) in Korea; Fahd Al-Mulla was supported by the Kuwait Foundation for the Advancement of Sciences (2011-1302-06); Ferdinando Chiaradonna is supported by SysBioNet, a grant for the Italian Roadmap of European Strategy Forum on Research Infrastructures (ESFRI) and by AIRC (Associazione Italiana Ricerca sul Cancro; IG 15364); Francis L.Martin acknowledges funding from Rosemere Cancer Foundation; he also thanks Lancashire Teaching Hospitals NHS trust and the patients who have facilitated the studies he has undertaken over the course of the last 10 years; Gary S.Goldberg would like to acknowledge the support of the New Jersey Health Foundation; Gloria M.Calaf was supported by Fondo Nacional de Ciencia y Tecnología (FONDECYT), Ministerio de Educación de Chile (MINEDUC), Universidad de Tarapacá (UTA); Gudrun Koppen was supported by the Flemish Institute for Technological Research (VITO), Belgium; Hemad Yasaei was supported from a triennial project grant (Strategic Award) from the National Centre for the Replacement, Refinement and Reduction (NC3Rs) of animals in research (NC.K500045.1 and G0800697); Hiroshi Kondoh was supported in part by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, Japan Science and Technology Agency and by JST, CREST; Hsue-Yin Hsu was supported by the Ministry of Science and Technology of Taiwan (NSC93-2314-B-320-006 and NSC94-2314-B-320-002); Hyun Ho Park was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) of the Ministry of Education, Science and Technology (2012R1A2A2A01010870) and a grant from the Korea Healthcare Technology R&D project, Ministry of Health and Welfare, Republic of Korea (HI13C1449); Igor Koturbash is supported by the UAMS/NIH Clinical and Translational Science Award (UL1TR000039 and KL2TR000063) and the Arkansas Biosciences Institute, the major research component of the Arkansas Tobacco Settlement Proceeds Act of 2000; Jan Vondráček acknowledges funding from the Czech Science Foundation (13-07711S); Jesse Roman thanks the NIH for their support (CA116812), John Pierce Wise Sr. and Sandra S.Wise were supported by National Institute of Environmental Health Sciences (ES016893 to J.P.W.) and the Maine Center for Toxicology and Environmental Health; Jonathan Whitfield acknowledges support from the FERO Foundation in Barcelona, Spain; Joseph Christopher is funded by Cancer Research UK and the International Journal of Experimental Pathology; Julia Kravchenko is supported by a philanthropic donation by Fred and Alice Stanback; Jun Sun is supported by a Swim Across America Cancer Research Award; Karine A.Cohen-Solal is supported by a research scholar grant from the American Cancer Society (116683-RSG-09-087-01-TBE); Laetitia Gonzalez received a postdoctoral fellowship from the Fund for Scientific Research–Flanders (FWO-Vlaanderen) and support by an InterUniversity Attraction Pole grant (IAP-P7-07); Laura Soucek is supported by grant #CP10/00656 from the Miguel Servet Research Contract Program and acknowledges support from the FERO Foundation in Barcelona, Spain; Liang-Tzung Lin was supported by funding from the Taipei Medical University (TMU101-AE3-Y19); Linda Gulliver is supported by a Genesis Oncology Trust (NZ) Professional Development Grant, and the Faculty of Medicine, University of Otago, Dunedin, New Zealand; Louis Vermeulen is supported by a Fellowship of the Dutch Cancer Society (KWF, UVA2011-4969) and a grant from the AICR (14–1164); Mahara Valverde would like to thank CONACyT support 153781; Masoud H. Manjili was supported by the office of the Assistant Secretary of Defense for Health Affairs (USA) through the Breast Cancer Research Program under Award No. W81XWH-14-1-0087 Neetu Singh was supported by grant #SR/FT/LS-063/2008 from the Department of Science and Technology, Government of India; Nicole Kleinstreuer is supported by NIEHS contracts (N01-ES 35504 and HHSN27320140003C); P.K. Krishnakumar is supported by the Funding (No. T.K. 11-0629) of King Abdulaziz City for Science and Technology, Riyadh, Saudi Arabia; Paola A.Marignani is supported by the Dalhousie Medical Research Foundation, The Beatrice Hunter Cancer Institute and CIHR and the Nova Scotia Lung Association; Paul Dent is the holder of the Universal Inc.Chair in Signal Transduction Research and is supported with funds from PHS grants from the NIH (R01-CA141704, R01-CA150214, R01-DK52825 and R01-CA61774); Petr Heneberg was supported by the Charles University in Prague projects UNCE 204015 and PRVOUK P31/2012, and by the Czech Science Foundation projects P301/12/1686 and 15-03834Y; Po Sing Leung was supported by the Health and Medical Research Fund of Food and Health Bureau, Hong Kong Special Administrative Region, Ref. No: 10110021; Qiang Cheng was supported in part by grant NSF IIS-1218712; R. Brooks Robey is supported by the United States Department of Veterans Affairs; Rabindra Roy was supported by United States Public Health Service Grants (RO1 CA92306, RO1 CA92306-S1 and RO1 CA113447); Rafaela Andrade-Vieira is supported by the Beatrice Hunter Cancer Research Institute and the Nova Scotia Health Research Foundation, Renza Vento was partially funded by European Regional Development Fund, European Territorial Cooperation 2007–13 (CCI 2007 CB 163 PO 037, OP Italia-Malta 2007–13) and grants from the Italian Ministry of Education, University and Research (MIUR) ex-60%, 2007; Riccardo Di Fiore was a recipient of fellowship granted by European Regional Development Fund, European Territorial Cooperation 2007–2013 (CCI 2007 CB 163 PO 037, OP Italia-Malta 2007–2013); Rita Dornetshuber-Fleiss was supported by the Austrian Science Fund (FWF, project number T 451-B18) and the Johanna Mahlke, geb.-Obermann-Stiftung; Roberta Palorini is supported by a SysBioNet fellowship; Roslida Abd Hamid is supported by the Ministry of Education, Malaysia-Exploratory Research Grant Scheme-Project no: ERGS/1-2013/5527165; Sabine A.S.Langie is the beneficiary of a postdoctoral grant from the AXA Research Fund and the Cefic-LRI Innovative Science Award 2013; Sakina Eltom is supported by NIH grant SC1CA153326; Samira A.Brooks was supported by National Research Service Award (T32 ES007126) from the National Institute of Environmental Health Sciences and the HHMI Translational Medicine Fellowship; Sandra Ryeom was supported by The Garrett B. Smith Foundation and the TedDriven Foundation; Thierry Massfelder was supported by the Institut National de la Santé et de la Recherche Médicale INSERM and Université de Strasbourg; Thomas Sanderson is supported by the Canadian Institutes of Health Research (CIHR; MOP-115019), the Natural Sciences and Engineering Council of Canada (NSERC; 313313) and the California Breast Cancer Research Program (CBCRP; 17UB-8703); Tiziana Guarnieri is supported by a grant from Fundamental Oriented Research (RFO) to the Alma Mater Studiorum University of Bologna, Bologna, Italy and thanks the Fondazione Cassa di Risparmio di Bologna and the Fondazione Banca del Monte di Bologna e Ravenna for supporting the Center for Applied Biomedical Research, S.Orsola-Malpighi University Hospital, Bologna, Italy; W.Kimryn Rathmell is supported by the V Foundation for Cancer Research and the American Cancer Society; William K.Decker was supported in part by grant RP110545 from the Cancer Prevention Research Institute of Texas; William H.Bisson was supported with funding from the NIH P30 ES000210; Yon Rojanasakul was supported with NIH grant R01-ES022968; Zhenbang Chen is supported by NIH grants (MD004038, CA163069 and MD007593); Zhiwei Hu is grateful for the grant support from an institutional start-up fund from The Ohio State University College of Medicine and The OSU James Comprehensive Cancer Center (OSUCCC) and a Seed Award from the OSUCCC Translational Therapeutics Program.
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- 2015
13. Enhancement of mouse hematopoietic stem/progenitor cell function via transient gene delivery using integration-deficient lentiviral vectors
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Maria E, Alonso-Ferrero, Niek P, van Til, Kerol, Bartolovic, Márcia F, Mata, Gerard, Wagemaker, Dale, Moulding, David A, Williams, Christine, Kinnon, Simon N, Waddington, Michael D, Milsom, and Steven J, Howe
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Homeodomain Proteins ,Recombinant Fusion Proteins ,Genetic Vectors ,Graft Survival ,Lentivirus ,Hematopoietic Stem Cell Transplantation ,Hematopoietic Stem Cells ,Article ,Hematopoiesis ,Mice, Inbred C57BL ,Mice ,Angiopoietin-like Proteins ,Gene Expression Regulation ,Genes, Reporter ,Transduction, Genetic ,Radiation Chimera ,Animals ,Humans ,Cell Lineage ,Transgenes ,K562 Cells ,Angiopoietin-Like Protein 3 ,Transcription Factors - Abstract
Highlights • Integration-deficient vectors (IdLVs) express genes transiently in dividing stem cells. • Hematopoietic stem/progenitor cells (HSPCs) can be programmed using IdLVs. • HOXB4 or Angptl3 expression from IdLVs improves engraftment of transplanted HSPCs. • Short-term gene delivery avoids the side effects associated with constitutive expression., Integration-deficient lentiviruses (IdLVs) deliver genes effectively to tissues but are lost rapidly from dividing cells. This property can be harnessed to express transgenes transiently to manipulate cell biology. Here, we demonstrate the utility of short-term gene expression to improve functional potency of hematopoietic stem and progenitor cells (HSPCs) during transplantation by delivering HOXB4 and Angptl3 using IdLVs to enhance the engraftment of HSPCs. Constitutive overexpression of either of these genes is likely to be undesirable, but the transient nature of IdLVs reduces this risk and those associated with unsolicited gene expression in daughter cells. Transient expression led to increased multilineage hematopoietic engraftment in in vivo competitive repopulation assays without the side effects reported in constitutive overexpression models. Adult stem cell fate has not been programmed previously using IdLVs, but we demonstrate that these transient gene expression tools can produce clinically relevant alterations or be applied to investigate basic biology.
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- 2017
14. Pretransplant Mobilization with Granulocyte Colony-Stimulating Factor Improves B-Cell Reconstitution by Lentiviral Vector Gene Therapy in SCID-X1 Mice
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Adriaan R.A. Riegman, Rana Yadak, Marshall W. Huston, Helen de Boer, Niek P. van Til, Yvette van Helsdingen, Gerard Wagemaker, İç Hastalıkları, Neurology, Clinical Genetics, and Hematology
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Male ,Transplantation Conditioning ,Genetic enhancement ,medicine.medical_treatment ,T-Lymphocytes ,Genetic Vectors ,Hematopoietic stem cell transplantation ,Research & Experimental Medicine ,X-Linked Combined Immunodeficiency Diseases ,Lymphocyte Depletion ,Bone Marrow Stem Cell Transplantation ,Mice ,Transduction, Genetic ,Granulocyte Colony-Stimulating Factor ,Genetics ,medicine ,Animals ,Molecular Biology ,Hematopoietic Stem Cell Mobilization ,Research Articles ,Genetics & Heredity ,Mice, Knockout ,Severe combined immunodeficiency ,B-Lymphocytes ,business.industry ,Lentivirus ,Hematopoietic Stem Cell Transplantation ,Hematopoietic stem cell ,Genetic Therapy ,medicine.disease ,Hematopoietic Stem Cells ,Granulocyte colony-stimulating factor ,Disease Models, Animal ,medicine.anatomical_structure ,Biotechnology & Applied Microbiology ,Immunology ,Molecular Medicine ,Female ,Bone marrow ,business ,Interleukin Receptor Common gamma Subunit - Abstract
Hematopoietic stem cell (HSC) gene therapy is a demonstrated effective treatment for X-linked severe combined immunodeficiency (SCID-X1), but B-cell reconstitution and function has been deficient in many of the gene therapy treated patients. Cytoreductive preconditioning is known to improve HSC engraftment, but in general it is not considered for SCID-X1 since the poor health of most of these patients at diagnosis and the risk of toxicity preclude the conditioning used in standard bone marrow stem cell transplantation. We hypothesized that mobilization of HSC by granulocyte colony-stimulating factor (G-CSF) should create temporary space in bone marrow niches to improve engraftment and thereby B-cell reconstitution. In the present pilot study supplementing our earlier preclinical evaluation (Huston et al., 2011), Il2rg(-/-) mice pretreated with G-CSF were transplanted with wild-type lineage negative (Lin(-)) cells or Il2rg(-/-) Lin(-) cells transduced with therapeutic IL2RG lentiviral vectors. Mice were monitored for reconstitution of lymphocyte populations, level of donor cell chimerism, and antibody responses as compared to 2 Gy total body irradiation (TBI), previously found effective in promoting B-cell reconstitution. The results demonstrate that G-CSF promotes B-cell reconstitution similar to low-dose TBI and provides proof of principle for an alternative approach to improve efficacy of gene therapy in SCID patients without adverse effects associated with cytoreductive conditioning.
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- 2014
15. Lentiviral Stem Cell Gene Therapy for Pompe Disease
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Qiushi, Liang, Merel, Stok, Yvette, van Helsdingen, Guus, van der Velden, Ed, Jacobs, Dirk, Duncker, Arnold, Reuser, Ans, van der Ploeg, Arnold, Vulto, Niek P, van Til, and Gerard, Wagemaker
- Published
- 2016
16. 5-Androstene-3β,17β-diol Promotes Recovery of Immature Hematopoietic Cells Following Myelosuppressive Radiation and Synergizes With Thrombopoietin
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Fatima S.F. Aerts-Kaya, Dwight R. Stickney, Gerard Wagemaker, James M. Frincke, Shazia Arshad, Trudi P. Visser, Chris L. Reading, and Hematology
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Androstenediol ,Cancer Research ,Bone Marrow Cells ,Spleen ,Granulocyte ,Colony-Forming Units Assay ,Blood cell ,Mice ,Bone Marrow ,Granulocyte Colony-Stimulating Factor ,Animals ,Medicine ,Radiology, Nuclear Medicine and imaging ,Progenitor cell ,Thrombopoietin ,Mice, Inbred BALB C ,Blood Cells ,Radiation ,business.industry ,Drug Synergism ,Total body irradiation ,Hematopoietic Stem Cells ,Haematopoiesis ,medicine.anatomical_structure ,Oncology ,Radiology Nuclear Medicine and imaging ,Immunology ,Cancer research ,Drug Therapy, Combination ,Dose Fractionation, Radiation ,Bone marrow ,business ,Whole-Body Irradiation - Abstract
Purpose 5-Androstene-3β,17β-diol (5-AED) stimulates recovery of hematopoiesis after exposure to radiation. To elucidate its cellular targets, the effects of 5-AED alone and in combination with (pegylated) granulocyte colony-stimulating factor and thrombopoietin (TPO) on immature hematopoietic progenitor cells were evaluated following total body irradiation. Methods and Materials BALB/c mice were exposed to radiation delivered as a single or as a fractionated dose, and recovery of bone marrow progenitors and peripheral blood parameters was assessed. Results BALB/c mice treated with 5-AED displayed accelerated multilineage blood cell recovery and elevated bone marrow (BM) cellularity and numbers of progenitor cells. The spleen colony-forming unit (CFU-S) assay, representing the life-saving short-term repopulating cells in BM of irradiated donor mice revealed that combined treatment with 5-AED plus TPO resulted in a 20.1-fold increase in CFU-S relative to that of placebo controls, and a 3.7 and 3.1-fold increase in comparison to 5-AED and TPO, whereas no effect was seen of Peg-G-CSF with or without 5-AED. Contrary to TPO, 5-AED also stimulated reconstitution of the more immature marrow repopulating (MRA) cells. Conclusions 5-AED potently counteracts the hematopoietic effects of radiation-induced myelosuppression and promotes multilineage reconstitution by stimulating immature bone marrow cells in a pattern distinct from, but synergistic with TPO.
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- 2012
17. Mobilization of hepatic mesenchymal stem cells from human liver grafts
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Jaap Kwekkeboom, Fatima S. F. Aerts Kaya, Hugo W. Tilanus, Monique M A Verstegen, Suomi M. G. Fouraschen, Andrew P. Stubbs, Wilfred F. J. van IJcken, Geert Kazemier, Qiuwei Pan, Harry L.A. Janssen, Mario Pescatori, Herold J. Metselaar, Antoine van der Sloot, Jeroen de Jonge, Gerard Wagemaker, Luc J. W. van der Laan, Ron Smits, Gastroenterology & Hepatology, Surgery, Hematology, Pathology, and Cell biology
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Pathology ,medicine.medical_specialty ,T cell ,medicine.medical_treatment ,Clinical uses of mesenchymal stem cells ,Mice, SCID ,Biology ,Liver transplantation ,Mesenchymal Stem Cell Transplantation ,Mice ,Liver disease ,SDG 3 - Good Health and Well-being ,Mice, Inbred NOD ,Osteogenesis ,Adipocytes ,medicine ,Animals ,Humans ,Hematopoietic Stem Cell Mobilization ,Cell Proliferation ,Stem cell transplantation for articular cartilage repair ,Transplantation ,Hepatology ,Gene Expression Profiling ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Flow Cytometry ,medicine.disease ,Liver Transplantation ,Perfusion ,medicine.anatomical_structure ,Liver ,Hepatocytes ,Surgery ,Stem cell - Abstract
Extensive studies have demonstrated the potential applications of bone marrow-derived mesenchymal stem cells (BM-MSCs) as regenerative or immunosuppressive treatments in the setting of organ transplantation. The aims of the present study were to explore the presence and mobilization of mesenchymal stem cells (MSCs) in adult human liver grafts and to compare their functional capacities to those of BM-MSCs. The culturing of liver graft preservation fluids (perfusates) or end-stage liver disease tissues resulted in the expansion of MSCs. Liver-derived mesenchymal stem cells (L-MSCs) were equivalent to BM-MSCs in adipogenic and osteogenic differentiation and in wingless-type-stimulated proliferative responses. Moreover, the genome-wide gene expression was very similar, with a 2-fold or greater difference found in only 82 of the 32,321 genes (0.25%). L-MSC differentiation into a hepatocyte lineage was demonstrated in immunodeficient mice and in vitro by the ability to support a hepatitis C virus infection. Furthermore, a subset of engrafted MSCs survived over the long term in vivo and maintained stem cell characteristics. Like BM-MSCs, L-MSCs were found to be immunosuppressive; this was shown by significant inhibition of T cell proliferation. In conclusion, the adult human liver contains an MSC population with a regenerative and immunoregulatory capacity that can potentially contribute to tissue repair and immunomodulation after liver transplantation. Liver Transpl 17:596-609, 2011. (C) 2011 AASLD.
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- 2011
18. Magselectofection: an integrated method of nanomagnetic separation and genetic modification of target cells
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Niek P. van Til, Marshall W. Huston, Gerard Wagemaker, J. Henk de Jong, Ian C.D. Johnston, Arzu Cengizeroglu, Yolanda Sanchez-Antequera, Zygmunt Pojda, Martina Anton, Christian Plank, Olga Mykhaylyk, and Hematology
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Immunology ,Cell ,Population ,Genetic Vectors ,02 engineering and technology ,Cell Separation ,Gene delivery ,Biology ,Transfection ,Biochemistry ,Jurkat cells ,03 medical and health sciences ,Transduction (genetics) ,Jurkat Cells ,Magnetics ,Mice ,medicine ,Animals ,Antigens, Ly ,Humans ,education ,030304 developmental biology ,0303 health sciences ,education.field_of_study ,Mesenchymal stem cell ,Gene Transfer Techniques ,Hematopoietic stem cell ,Membrane Proteins ,Mesenchymal Stem Cells ,Cell Biology ,Hematology ,021001 nanoscience & nanotechnology ,Hematopoietic Stem Cells ,Molecular biology ,medicine.anatomical_structure ,Nanoparticles ,0210 nano-technology ,K562 Cells ,Interleukin Receptor Common gamma Subunit - Abstract
Research applications and cell therapies involving genetically modified cells require reliable, standardized, and cost-effective methods for cell manipulation. We report a novel nanomagnetic method for integrated cell separation and gene delivery. Gene vectors associated with magnetic nanoparticles are used to transfect/transduce target cells while being passaged and separated through a high gradient magnetic field cell separation column. The integrated method yields excellent target cell purity and recovery. Nonviral and lentiviral magselectofection is efficient and highly specific for the target cell population as demonstrated with a K562/Jurkat T-cell mixture. Both mouse and human enriched hematopoietic stem cell pools were effectively transduced by lentiviral magselectofection, which did not affect the hematopoietic progenitor cell number determined by in vitro colony assays. Highly effective reconstitution of T and B lymphocytes was achieved by magselectofected murine wild-type lineage-negative Sca-1+ cells transplanted into Il2rg−/− mice, stably expressing GFP in erythroid, myeloid, T-, and B-cell lineages. Furthermore, nonviral, lentiviral, and adenoviral magselectofection yielded high transfection/transduction efficiency in human umbilical cord mesenchymal stem cells and was fully compatible with their differentiation potential. Upscaling to a clinically approved automated cell separation device was feasible. Hence, once optimized, validated, and approved, the method may greatly facilitate the generation of genetically engineered cells for cell therapies.
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- 2011
19. Correction of Murine SCID-X1 by Lentiviral Gene Therapy Using a Codon-optimized IL2RG Gene and Minimal Pretransplant Conditioning
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Axel Schambach, Niek P. van Til, Shazia Arshad, Michael P. Blundell, Ali Nowrouzi, Adrian J. Thrasher, Claudia Cattoglio, Fang Zhang, Trudi P. Visser, Christof von Kalle, Gerard Wagemaker, Marshall W. Huston, Monique M.A. Verstegen, Yuedan Li, Fulvio Mavilio, Martijn H. Brugman, Manfred Schmidt, Christopher Baum, and Hematology
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Genetic enhancement ,T-Lymphocytes ,Receptors, Antigen, T-Cell ,Mice, SCID ,Biology ,SCID ,Viral vector ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Immune system ,Antigen ,Receptors ,Drug Discovery ,Genetics ,medicine ,Animals ,Codon ,Molecular Biology ,Gene ,030304 developmental biology ,Pharmacology ,0303 health sciences ,Severe combined immunodeficiency ,Phosphoglycerate kinase ,B-Lymphocytes ,Drug Discovery3003 Pharmaceutical Science ,Lentivirus ,Hematopoietic Stem Cell Transplantation ,Original Articles ,Genetic Therapy ,T-Cell ,medicine.disease ,Virology ,Molecular biology ,3. Good health ,030220 oncology & carcinogenesis ,Antibody Formation ,Molecular Medicine ,Severe Combined Immunodeficiency ,Interleukin Receptor Common gamma Subunit ,Ex vivo - Abstract
Clinical trials have demonstrated the potential of ex vivo hematopoietic stem cell gene therapy to treat X-linked severe combined immunodeficiency (SCID-X1) using retroviral vectors, leading to immune system functionality in the majority of treated patients without pretransplant conditioning. The success was tempered by insertional oncogenesis in a proportion of the patients. To reduce the genotoxicity risk, a self-inactivating (SIN) lentiviral vector (LV) with improved expression of a codon optimized human interleukin-2 receptor gamma gene (IL2RG) cDNA (co gamma c), regulated by its 1.1 kb promoter region (gamma cPr), was compared in efficacy to the viral spleen focus forming virus (SF) and the cellular phosphoglycerate kinase (PGK) promoters. Pretransplant conditioning of IL2rg(-/-) mice resulted in long-term reconstitution of T and B lymphocytes, normalized natural antibody titers, humoral immune responses, ConA/IL-2 stimulated spleen cell proliferation, and polyclonal T-cell receptor gene rearrangements with a clear integration preference of the SF vector for proto-oncogenes, contrary to the PGK and gamma cPr vectors. We conclude that SIN lentiviral gene therapy using co gamma c driven by the gamma cPr or PGK promoter corrects the SCID phenotype, potentially with an improved safety profile, and that low-dose conditioning proved essential for immune competence, allowing for a reduced threshold of cell numbers required. Received 23 February 2011; accepted 31 May 2011; published online 12 July 2011. doi:10.1038/mt.2011.127
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- 2011
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20. Lentiviral gene therapy of murine hematopoietic stem cells ameliorates the Pompe disease phenotype
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Edwin H. Jacobs, Pascal van der Wegen, Fatima S. F. Aerts Kaya, Bob J. Scholte, Merel Stok, Trudi P. Visser, Ans T. van der Ploeg, Bart N. Lambrecht, Dirk J. Duncker, Arnold J. J. Reuser, Gerard Wagemaker, Monique Willart, Elnaz Farahbakhshian, Monique C. de Waard, Niek P. van Til, Marian A. Kroos, Monique M A Verstegen, Hematology, Molecular Genetics, Clinical Genetics, Pulmonary Medicine, Cell biology, Cardiology, and Pediatrics
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Hematopoietic System ,Genetic enhancement ,medicine.medical_treatment ,Genetic Vectors ,Immunology ,Gene Expression ,Hematopoietic stem cell transplantation ,Motor Activity ,Biology ,Chimerism ,Biochemistry ,Mice ,03 medical and health sciences ,0302 clinical medicine ,SDG 3 - Good Health and Well-being ,Transduction, Genetic ,Glycogen storage disease type II ,medicine ,Animals ,Humans ,Respiratory function ,Cells, Cultured ,030304 developmental biology ,Mice, Knockout ,0303 health sciences ,Glycogen Storage Disease Type II ,Lentivirus ,Hematopoietic Stem Cell Transplantation ,Hematopoietic stem cell ,alpha-Glucosidases ,Genetic Therapy ,Cell Biology ,Hematology ,Enzyme replacement therapy ,Hematopoietic Stem Cells ,medicine.disease ,3. Good health ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Acid alpha-glucosidase ,Stem cell ,Glycogen - Abstract
Pompe disease (acid α-glucosidase deficiency) is a lysosomal glycogen storage disorder characterized in its most severe early-onset form by rapidly progressive muscle weakness and mortality within the first year of life due to cardiac and respiratory failure. Enzyme replacement therapy prolongs the life of affected infants and supports the condition of older children and adults but entails lifelong treatment and can be counteracted by immune responses to the recombinant enzyme. We have explored the potential of lentiviral vector–mediated expression of human acid α-glucosidase in hematopoietic stem cells (HSCs) in a Pompe mouse model. After mild conditioning, transplantation of genetically engineered HSCs resulted in stable chimerism of approximately 35% hematopoietic cells that overexpress acid α-glucosidase and in major clearance of glycogen in heart, diaphragm, spleen, and liver. Cardiac remodeling was reversed, and respiratory function, skeletal muscle strength, and motor performance improved. Overexpression of acid α-glucosidase did not affect overall hematopoietic cell function and led to immune tolerance as shown by challenge with the human recombinant protein. On the basis of the prominent and sustained therapeutic efficacy without adverse events in mice we conclude that ex vivo HSC gene therapy is a treatment option worthwhile to pursue.
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- 2010
21. Gammaretrovirus-mediated correction of SCID-X1 is associated with skewed vector integration site distribution in vivo
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Frank J. T. Staal, Fang Zhang, Jinhua Bayford, Manuela Wissler, Adrian J. Thrasher, H. Bobby Gaspar, Rachel Peraj, Hanno Glimm, Kerstin Schwarzwaelder, Martijn H. Brugman, Steven J. Howe, Annette Deichmann, Sonja Schmidt, Joanna Sinclair, Gerard Wagemaker, Ulrich Abel, Douglas King, Kathryn L. Parsley, Manfred Schmidt, Claudia Prinz, Christof von Kalle, Kimberly Gilmour, Christine Kinnon, Karin Pike-Overzet, Dick de Ridder, Hematology, and Immunology
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Severe combined immunodeficiency ,biology ,CD3 ,CD34 ,General Medicine ,biology.organism_classification ,medicine.disease ,Molecular biology ,Phosphorus metabolism ,Transplantation ,Gene expression ,Immunology ,medicine ,biology.protein ,Progenitor cell ,Gammaretrovirus - Abstract
We treated 10 children with X-linked SCID (SCID-X1) using gammaretrovirus-mediated gene transfer. Those with sufficient follow-up were found to have recovered substantial immunity in the absence of any serious adverse events up to 5 years after treatment. To determine the influence of vector integration on lymphoid reconstitution, we compared retroviral integration sites (RISs) from peripheral blood CD3+ T lymphocytes of 5 patients taken between 9 and 30 months after transplantation with transduced CD34+ progenitor cells derived from 1 further patient and 1 healthy donor. Integration occurred preferentially in gene regions on either side of transcription start sites, was clustered, and correlated with the expression level in CD34+ progenitors during transduction. In contrast to those in CD34+ cells, RISs recovered from engrafted CD3+ T cells were significantly overrepresented within or near genes encoding proteins with kinase or transferase activity or involved in phosphorus metabolism. Although gross patterns of gene expression were unchanged in transduced cells, the divergence of RIS target frequency between transduced progenitor cells and post-thymic T lymphocytes indicates that vector integration influences cell survival, engraftment, or proliferation.
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- 2007
22. Ectopic retroviral expression of LMO2, but not IL2R gamma, blocks human T-cell development from CD34+cells: implications for leukemogenesis in gene therapy
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Mjt Reinders, Martijn H. Brugman, Adrian J. Thrasher, Gerard Wagemaker, Miranda R. M. Baert, Denise T. D. de Ridder, J J M van Dongen, Monique M.A. Verstegen, Frank Jakob Theodor Staal, Floor Weerkamp, Steven J. Howe, Karin Pike-Overzet, Immunology, and Hematology
- Subjects
LMO2 ,Cancer Research ,T-Lymphocytes ,Antigens, CD34 ,Biology ,medicine.disease_cause ,LYL1 ,Antigens, CD ,hemic and lymphatic diseases ,Proto-Oncogene Proteins ,Metalloproteins ,medicine ,Humans ,Leukemia-Lymphoma, Adult T-Cell ,Progenitor cell ,Growth Substances ,Adaptor Proteins, Signal Transducing ,Severe combined immunodeficiency ,Leukemia ,Oncogene ,Receptors, Interleukin-2 ,Hematology ,Genetic Therapy ,LIM Domain Proteins ,medicine.disease ,DNA-Binding Proteins ,Mutagenesis, Insertional ,Retroviridae ,Oncology ,Cancer research ,Carcinogenesis ,TAL1 - Abstract
The occurrence of leukemia in a gene therapy trial for SCID-X1 has highlighted insertional mutagenesis as an adverse effect. Although retroviral integration near the T-cell acute lymphoblastic leukemia (T-ALL) oncogene LIM-only protein 2 (LMO2) appears to be a common event, it is unclear why LMO2 was preferentially targeted. We show that of classical T-ALL oncogenes, LMO2 is most highly transcribed in CD34+ progenitor cells. Upon stimulation with growth factors typically used in gene therapy protocols transcription of LMO2, LYL1, TAL1 and TAN1 is most prominent. Therefore, these oncogenes may be susceptible to viral integration. The interleukin-2 receptor gamma chain (IL2Rgamma), which is mutated in SCID-X1, has been proposed as a cooperating oncogene to LMO2. However, we found that overexpressing IL2Rgamma had no effect on T-cell development. In contrast, retroviral overexpression of LMO2 in CD34+ cells caused severe abnormalities in T-cell development, but B-cell and myeloid development remained unaffected. Our data help explain why LMO2 was preferentially targeted over many of the other known T-ALL oncogenes. Furthermore, during T-cell development retrovirus-mediated expression of IL2Rgamma may not be directly oncogenic. Instead, restoration of normal IL7-receptor signaling may allow progression of T-cell development to stages where ectopic LMO2 expression causes aberrant thymocyte growth.
- Published
- 2007
23. An inventory of shedding data from clinical gene therapy trials
- Author
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Gerard Wagemaker, Chris H. Bangma, Marjolein M. K. B. van Mierlo, Leonie C. M. Kaptein, Ellen A. M. Schenk-Braat, Urology, and Hematology
- Subjects
Time Factors ,Genetic enhancement ,Genetic Vectors ,Bioinformatics ,medicine.disease_cause ,Viral vector ,Neoplasms ,Drug Discovery ,Genetics ,Animals ,Humans ,Medicine ,Vector (molecular biology) ,Viral shedding ,Molecular Biology ,Adeno-associated virus ,Genetics (clinical) ,Clinical Trials as Topic ,Information Dissemination ,business.industry ,Genetic Therapy ,Assay sensitivity ,Virus Shedding ,Clinical trial ,Immunology ,Molecular Medicine ,business ,Risk assessment - Abstract
Viruses are the most commonly used vectors for clinical gene therapy. The risk of dissemination of a viral vector into the environment via excreta from the treated patient, a phenomenon called shedding, is a major safety concern for the environment. Despite the significant number of clinical gene therapy trials that have been conducted worldwide, there is currently no overview of actual shedding data available. In this article, an inventory of shedding data obtained from a total of 100 publications on clinical gene therapy trials using retroviral, adenoviral, adeno-associated viral and pox viral vectors is presented. In addition, the experimental set-up for shedding analysis including the assays used and biological materials tested is summarized. The collected data based on the analysis of 1619 patients in total demonstrate that shedding of these vectors occurs in practice, mainly determined by the type of vector and the route of vector administration. Due to the use of non-quantitative assays, the lack of information on assay sensitivity in most publications, and the fact that assay sensitivity is expressed in various ways, general conclusions cannot be made as to the level of vector shedding. The evaluation of the potential impact and consequences of the observations is complicated by the high degree of variety in the experimental design of shedding analysis between trials. This inventory can be supportive to clinical gene therapy investigators for the establishment of an evidence-based risk assessment to be included in a clinical protocol application, as well as to national regulatory authorities for the ongoing development of regulatory guidelines regarding gene therapy.
- Published
- 2007
24. The potential for chemical mixtures from the environment to enable the cancer hallmark of sustained proliferative signalling
- Author
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Kim Moorwood, Amedeo Amedei, Jayadev Raju, Rabeah Al-Temaimi, Monica Vaccari, Andrew Ward, Dustin G. Brown, Roslida Abd Hamid, Pankaj Vadgama, Rabindra Roy, Rekha Mehta, Stefano Forte, Gerard Wagemaker, James E. Klaunig, William H. Bisson, Tove Hultman, Michalis V. Karamouzis, Philippa D. Darbre, A. Ivana Scovassi, Thomas Sanderson, Chiara Mondello, Fahd Al-Mulla, Wilhelm Engström, Annamaria Colacci, Neetu Singh, Hosni Salem, Lorenzo Memeo, Jordan Woodrick, Linda S M Gulliver, Hideko Sone, Elizabeth P. Ryan, Staffan Eriksson, Kök Hücre, Swedish University of Agricultural Sciences (SLU), University of Reading (UOR), University of Otago [Dunedin, Nouvelle-Zélande], National and Kapodistrian University of Athens (NKUA), Indiana University [Bloomington], Indiana University System, Health Canada, University of Bath [Bath], Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), National Institute for Environmental Studies (NIES), Queen Mary University of London (QMUL), Hacettepe University = Hacettepe Üniversitesi, King George's Medical University, Kuwait University, UNIVERSITY OF FIRENZE (UNIVERSITY OF FIRENZE), Università degli Studi di Firenze = University of Florence [Firenze] (UNIFI), Center for Environmental Carcinogenesis and Risk Assessment Environmental Protection and Health Prevention Agency Emilia Romagna Region Viale Filopanti 20/22, National Research Council [Italy] (CNR), Universiti Putra Malaysia, Mediterranean Institute of Oncology, Georgetown University Medical Center, Cairo University, Department of Environmental and Radiological Health Sciences, Colorado State University [Fort Collins] (CSU), Oregon State University (OSU), Fondazione Cariplo (2011-0370 to C.M.), Kuwait Institute for the Advancement of Sciences (2011-1302-06 to F.A.-M.), Grant University Scheme (RUGs) Ministry of Education Malaysia (04-02-12-2099RU to R.A.H.), Italian Ministry of University and Research (2009FZZ4XM_002 to A.A.), the University of Florence (2012 to A.A.), US Public Health Service Grants (RO1 CA92306, RO1 CA92306-S1, RO1 CA113447 to R.R.), Department of Science and Technology, Government of India (SR/FT/LS-063/2008 to N.S.), National Institute for Health Research Grant (II-ES-0511-21005), Nanosilver based catheters (to P.V.), Medical Research Council UK (MR/L007215/1 to A.W.), WEM Consulting (to T.H.), and Getting to know cancer (to the entire team).
- Subjects
Cancer Research ,medicine.drug_class ,[SDV]Life Sciences [q-bio] ,MESH: Neoplasms/chemically induced ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Review ,Biology ,Cell cycle ,MESH: Cell Proliferation/drug effects ,Hazardous Substances ,MESH: Environmental Exposure/adverse effects ,MESH: Neoplasms/etiology ,Neoplasms ,medicine ,Animals ,Humans ,MESH: Animals ,MESH: Carcinogens, Environmental/adverse effects ,MESH: Signal Transduction/drug effects ,Carcinogen ,Cancer ,Cell Proliferation ,MESH: Humans ,Cell growth ,General Medicine ,Environmental exposure ,Environmental Exposure ,medicine.disease ,Estrogen ,Carcinogens, Environmental ,Cell biology ,Signalling ,Kök Hücre ,Signal transduction ,MESH: Hazardous Substances/adverse effects ,Signal Transduction - Abstract
International audience; The aim of this work is to review current knowledge relating the established cancer hallmark, sustained cell proliferation to the existence of chemicals present as low dose mixtures in the environment. Normal cell proliferation is under tight control, i.e. cells respond to a signal to proliferate, and although most cells continue to proliferate into adult life, the multiplication ceases once the stimulatory signal disappears or if the cells are exposed to growth inhibitory signals. Under such circumstances, normal cells remain quiescent until they are stimulated to resume further proliferation. In contrast, tumour cells are unable to halt proliferation, either when subjected to growth inhibitory signals or in the absence of growth stimulatory signals. Environmental chemicals with carcinogenic potential may cause sustained cell proliferation by interfering with some cell proliferation control mechanisms committing cells to an indefinite proliferative span.
- Published
- 2015
25. Lentiviral Stem Cell Gene Therapy for Pompe Disease
- Author
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Merel Stok, Ed H. Jacobs, Ans T. van der Ploeg, Gerard Wagemaker, Niek P. van Til, Dirk J. Duncker, Arnold G. Vulto, Yvette van Helsdingen, Qiushi Liang, Arnold J. J. Reuser, Guus van der Velden, Hematology, Clinical Genetics, Cardiology, and Pharmacy
- Subjects
medicine.medical_specialty ,Glycogen ,business.industry ,Genetic enhancement ,Metabolic disorder ,Disease ,Enzyme replacement therapy ,medicine.disease ,Gastroenterology ,chemistry.chemical_compound ,Immune system ,Neurology ,chemistry ,Internal medicine ,medicine ,Neurology (clinical) ,Stem cell ,business ,Alglucosidase alfa ,medicine.drug - Abstract
Pompe disease is a rare autosomal recessive metabolic disorder caused by defi ciency of lysosomal hydrolase acid α-glucosidase (GAA). GAA degrades glycogen to glucose, and defi ciency results in generalized tissue glycogen accumulation leading to cardiorespiratory failure in the early-onset patients within the fi rst year of life. Enzyme replacement therapy (ERT) by administration of recombinant acid α-glucosidase (alglucosidase alfa, Myozyme®) is currently the only effective treatment, requiring highdose biweekly administration. Although of considerable bene it to many patients, ERT is not curative, requires life-long administration, may result in immune responses to the recombinant enzyme and, partly due to the high doses required for clinical ef icacy, the costs are extremely high. Therefore, a corrective intervention with curative intent represents an unmet medical need.
- Published
- 2015
26. Overcoming promoter competition in packaging cells improves production of self-inactivating retroviral vectors
- Author
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Rainer Loew, D Mueller, Axel Schambach, Christopher Baum, Monique M A Verstegen, Melanie Galla, Gerard Wagemaker, and Jens Bohne
- Subjects
Male ,T-Lymphocytes ,viruses ,Genetic enhancement ,Genetic Vectors ,Antigens, CD34 ,Transfection ,Cell Line ,Viral vector ,Transduction (genetics) ,Transduction, Genetic ,Transcription (biology) ,Genetics ,Animals ,Northern blot ,Promoter Regions, Genetic ,Molecular Biology ,Rous sarcoma virus ,biology ,Terminal Repeat Sequences ,Genetic Therapy ,Flow Cytometry ,biology.organism_classification ,Macaca mulatta ,Molecular biology ,Titer ,Gene Expression Regulation ,Virus Inactivation ,Molecular Medicine ,Gammaretrovirus ,Genetic Engineering ,Plasmids - Abstract
Retroviral vectors with self-inactivating (SIN) long-terminal repeats not only increase the autonomy of the internal promoter but may also reduce the risk of insertional upregulation of neighboring alleles. However, gammaretroviral as opposed to lentiviral packaging systems produce suboptimal SIN vector titers, a major limitation for their clinical use. Northern blot data revealed that low SIN titers were associated with abundant transcription of internal rather than full-length transcripts in transfected packaging cells. When using the promoter of Rous sarcoma virus or a tetracycline-inducible promoter to generate full-length transcripts, we obtained a strong enhancement in titer (up to 4 x 10(7) transducing units per ml of unconcentrated supernatant). Dual fluorescence vectors and Northern blots revealed that promoter competition is a rate-limiting step of SIN vector production. SIN vector stocks pseudotyped with RD114 envelope protein had high transduction efficiency in human and non-human primate cells. This study introduces a new generation of efficient gammaretroviral SIN vectors as a platform for further optimizations of retroviral vector performance.
- Published
- 2006
27. Human thymus contains multipotent progenitors with T/B lymphoid, myeloid, and erythroid lineage potential
- Author
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Martijn H. Brugman, Willem A. Dik, Gerard Wagemaker, Edwin F. E. de Haas, Floor Weerkamp, Miranda R. M. Baert, Trudi P. Visser, Jacques J.M. van Dongen, Christianne J.M. de Groot, Frank J. T. Staal, Immunology, Hematology, and Obstetrics & Gynecology
- Subjects
Myeloid ,Lineage (genetic) ,Cellular differentiation ,T-Lymphocytes ,Immunology ,Antigens, CD34 ,Mice, SCID ,Thymus Gland ,Biology ,Gene Rearrangement, T-Lymphocyte ,Biochemistry ,Antigens, CD1 ,Colony-Forming Units Assay ,Mice ,Mice, Inbred NOD ,hemic and lymphatic diseases ,medicine ,Animals ,Humans ,Cell Lineage ,Lymphopoiesis ,B-Lymphocytes ,integumentary system ,Multipotent Stem Cells ,Hematopoietic Stem Cell Transplantation ,Receptors, Antigen, T-Cell, gamma-delta ,Cell Biology ,Hematology ,Hematopoietic Stem Cells ,Molecular biology ,Transplantation ,Thymocyte ,Haematopoiesis ,medicine.anatomical_structure ,Multipotent Stem Cell - Abstract
It is a longstanding question which bone marrow–derived cell seeds the thymus and to what level this cell is committed to the T-cell lineage. We sought to elucidate this issue by examining gene expression, lineage potential, and self-renewal capacity of the 2 most immature subsets in the human thymus, namely CD34+CD1a– and CD34+CD1a+ thymocytes. DNA microarrays revealed the presence of several myeloid and erythroid transcripts in CD34+CD1a– thymocytes but not in CD34+CD1a+ thymocytes. Lineage potential of both subpopulations was assessed using in vitro colony assays, bone marrow stroma cultures, and in vivo transplantation into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. The CD34+CD1a– subset contained progenitors with lymphoid (both T and B), myeloid, and erythroid lineage potential. Remarkably, development of CD34+CD1a– thymocytes toward the T-cell lineage, as shown by T-cell receptor δ gene rearrangements, could be reversed into a myeloid-cell fate. In contrast, the CD34+CD1a+ cells yielded only T-cell progenitors, demonstrating their irreversible commitment to the T-cell lineage. Both CD34+CD1a– and CD34+CD1a+ thymocytes failed to repopulate NOD/SCID mice. We conclude that the human thymus is seeded by multipotent progenitors with a much broader lineage potential than previously assumed. These cells resemble hematopoietic stem cells but, by analogy with murine thymocytes, apparently lack sufficient self-renewal capacity.
- Published
- 2006
28. Overestimation of hematopoietic stem cell frequencies in human liver grafts
- Author
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Luc J. W. van der Laan, Sean R.R. Hall, Alexander Pedroza-Gonzalez, Gerard Wagemaker, Qiuwei Pan, Hugo W. Tilanus, Jeroen de Jonge, Surgery, Gastroenterology & Hepatology, and Hematology
- Subjects
Pathology ,medicine.medical_specialty ,medicine.anatomical_structure ,Hepatology ,Human liver ,medicine ,Hematopoietic stem cell ,Biology - Published
- 2013
29. Chance or necessity? Insertional mutagenesis in gene therapy and its consequences
- Author
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Gerard Wagemaker, Frank J. T. Staal, Floor Weerkamp, Christof von Kalle, Boris Fehse, Zhixiong Li, M Schmidt, David A. Williams, Stefan Karlsson, Christopher Baum, Immunology, and Hematology
- Subjects
Leukemia, T-Cell ,Transgene ,Genetic enhancement ,Genetic Vectors ,Mutagenesis (molecular biology technique) ,Biology ,Proto-Oncogene Mas ,Risk Assessment ,Leukemogenic ,Insertional mutagenesis ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Proto-Oncogene Proteins ,Metalloproteins ,Proto-Oncogenes ,Drug Discovery ,Genetics ,Animals ,Humans ,Leukemia-Lymphoma, Adult T-Cell ,Molecular Biology ,Gene ,Adaptor Proteins, Signal Transducing ,030304 developmental biology ,Pharmacology ,0303 health sciences ,Hematopoietic Stem Cell Transplantation ,Genetic Therapy ,LIM Domain Proteins ,Up-Regulation ,3. Good health ,DNA-Binding Proteins ,Mutagenesis, Insertional ,Haematopoiesis ,Retroviridae ,030220 oncology & carcinogenesis ,Cancer research ,Molecular Medicine ,Adult stem cell - Abstract
Recently, unusual forms of leukemias have developed as complications following retroviral transfer of potentially therapeutic genes into hematopoietic cells. A crucial component in the pathogenesis of these complications was the upregulation of a cellular proto-oncogene by random insertion of the retroviral gene transfer vector. These findings have great implications for the genetic manipulation of somatic stem cells in medicine. This review discusses the extent to which the random oncogene activation may have required disease-specific stimuli of the transgene and the hematopoietic milieu to become leukemogenic. Based on these considerations, we propose approaches to risk prediction and prevention.
- Published
- 2004
30. New TPO treatment schedules of increased safety and efficacy: pre-clinical validation of a thrombopoiesis simulation model
- Author
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Hila Harpak, Zvia Agur, Anton Ianovski, Moshe Vardi, Gerard Wagemaker, Huub H.D.M. van Vliet, Simone C. C. Hartong, Trudi P. Visser, and Kirill Skomorovski
- Subjects
Chemotherapy ,business.industry ,Immunogenicity ,medicine.medical_treatment ,food and beverages ,Stimulation ,Hematology ,Pharmacology ,Reduced dose ,Immunology ,Medicine ,Platelet ,Thrombopoiesis ,Dosing ,business ,Thrombopoietin - Abstract
Summary. Thrombopoietin (TPO) immunogenicity hampers its development as a therapeutic agent for attenuating thrombocytopenia and improving platelet harvest in donors. This work was aimed at validating, in mouse and in monkey experiments, a thrombopoiesis computer-model prediction that platelet counts, similar to those obtained with accepted TPO dose scheduling, can also be achieved by new and safer schedules of significantly reduced doses. To this end we compared, in a two-arm mouse experiment, platelet increases obtained with a single intraperitoneal dosing of recombinant mouse TPO (17·5 μg/kg), with those obtained by the model-suggested protocol of a significantly reduced dose (2 μg/kg on 4 consecutive days). The two TPO regimens generated similar platelet profiles, peaking at ca. 2700 × 109/l platelets. In rhesus monkeys, treated by rhesus monkey recombinant TPO (5 μg/kg on 4 consecutive days), the suggested protocol yielded effective platelet stimulation with significantly reduced immunogenicity. The model's ability to predict individual monkey responses to several new TPO administration protocols was further validated, proving sufficient robustness in providing good predictions with limited input data. The simulation tool could be used for testing the effects of different therapeutic agents on thrombopoiesis. Human trials are warranted for testing the suggested improved TPO protocol, possibly in conjunction with chemotherapy.
- Published
- 2003
31. Delayed effects of accidental cutaneous radiation exposure: fifteen years of follow-up after the chernobyl accident
- Author
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Gerard Wagemaker, Marianna Steinert, Irina Galstian, Theodor M. Fliedner, Ralf U. Peter, Petra Gottlöber, Melanie Weiss, Natalia Nadejina, Oleg Gergel, Vladimir Bebeshko, David Belyi, and Hematology
- Subjects
Male ,medicine.medical_specialty ,Pathology ,Neoplasms, Radiation-Induced ,Skin Neoplasms ,Time Factors ,Dermatology ,Telangiectases ,Risk Assessment ,Severity of Illness Index ,Skin Diseases ,Russia ,Cohort Studies ,Nuclear Reactors ,Fibrosis ,Severity of illness ,Humans ,Radiodermatitis ,Medicine ,Stage (cooking) ,Skin ,business.industry ,Environmental Exposure ,medicine.disease ,Accidental ,Female ,Skin cancer ,Radioactive Hazard Release ,Ukraine ,business ,Environmental Monitoring ,Follow-Up Studies ,Cohort study - Abstract
Background During the Chernobyl accident in 1986, 237 individuals were identified as having the most severe exposure to ionizing radiation. In the period between 1998 and 2000, 99 long term survivors out of this group were reassessed for radiation-induced cutaneous lesions. Objective To identify sequelae of accidental cutaneous irradiation. Methods Detailed dermatologic examinations, including biopsies of suspicious cutaneous lesions for histopathologic examination and 20 MHz sonography, were performed in all patients. Results Twenty-two of the 99 patients displayed radiation-induced cutaneous lesions. Epidermal atrophy, telangiectases, and pigment alterations were present in all these individuals. Keratotic lesions were found in 14 patients. Cutaneous fibrosis was documented in 8 individuals by the use of 20 MHz sonography, while a radiation ulcer was found in 5. In one patient, two basal cell carcinomas were found. Conclusion The life-long follow-up of irradiated persons is of great importance in order to identify cutaneous neoplasms at an early treatable stage.
- Published
- 2003
32. Thrombopoietin is a major limiting factor for selective outgrowth of human umbilical cord blood cells in non-obese diabetic/severe combined immunodeficient recipient mice
- Author
-
Gerard Wagemaker, Monique M A Verstegen, and A. W. Wognum
- Subjects
Recombinant Human Thrombopoietin ,CD34 ,food and beverages ,hemic and immune systems ,Hematology ,Biology ,Transplantation ,Andrology ,Haematopoiesis ,fluids and secretions ,Immunophenotyping ,medicine.anatomical_structure ,embryonic structures ,Immunology ,medicine ,Bone marrow ,Progenitor cell ,Stem cell - Abstract
Summary. A single dose (0AE3 lg) of recombinant human thrombopoietin (TPO) was injected into sublethal irradiated non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice immediately after transplantation of 1AE5 · 10 5 purified CD34 + umbilical cord blood (UCB) cells. Bone marrow (BM) was analysed for human cells by immunophenotyping and colony culture at d 35. TPO treatment produced a two- to sixfold increase in the frequency and number of human CD45 + cells. The lineage distributions among the human cells were similar irrespective of TPO treatment; however, a prominent increase was observed in CD71 + GpA ‐ cells, reflecting the proliferative stimulus provided by TPO. The frequency of immature CD34 + cells and human granulocyte‐macrophage colony-forming units and erythroid burst-forming units in TPO-treated mice was similar to that of untreated mice, but their absolute numbers had increased proportionally to the increase in human cells. The results demonstrate that human TPO is a major limiting factor for multilineage outgrowth of human UCB cells in NOD/SCID mice and can be conveniently supplemented by single-dose treatment immediately after transplantation. TPO did not affect the survival of mice after transplantation and did not significantly increase the number of immature CD34 + CD38 ‐ cells; secondary transplantation revealed that TPO administration also had no significant effect on longterm repopulation. The findings demonstrate that human TPO is required for proper outgrowth of human haematopoietic stem cells after transplantation. In addition, a single administration of TPO may improve the efficiency and reproducibility of the NOD/SCID mouse assay for human immature transplantable progenitor cells.
- Published
- 2003
33. Co-administration of Flt-3 ligand counteracts the actions of thrombopoietin in myelosuppressed rhesus monkeys
- Author
-
Gerard Wagemaker, Karen J. Neelis, and Simone C. C. Hartong
- Subjects
Chemotherapy ,medicine.medical_specialty ,business.industry ,medicine.medical_treatment ,CD34 ,Hematology ,Total body irradiation ,Placebo ,medicine.anatomical_structure ,Endocrinology ,Internal medicine ,embryonic structures ,Medicine ,Platelet ,Bone marrow ,Progenitor cell ,business ,Thrombopoietin - Abstract
Summary. This placebo-controlled study evaluated the efficacy of Flt-3 ligand (FL) combined with TPO in myelosuppressed rhesus monkeys. The monkeys were subjected to 5 Gy total body irradiation (TBI), resulting in 3 weeks of profound pancytopenia, and received either 5 µg/kg of rhesus TPO i.v. on d 1 (n = 4) and 100 µg/kg/d s.c. human FL (n = 4) or FL alone (n = 4) for 14 consecutive days and were compared with results from a concomitant study involving the administration of TPO alone (n = 4) or placebo (carrier; n = 4). The TPO/FL combination was considerably less effective than TPO alone, with a more profound nadir and a slower recovery to thrombocyte counts > 100 × 109/l, approaching recovery patterns of placebo controls. Leucocyte regeneration was similar in all animals. Monkeys treated with FL alone displayed a regeneration of reticulocytes and thrombocytes in the lower range of those of the placebo controls. Recovery of bone marrow (BM) cellularity was slightly accelerated in the TPO/FL-treated monkeys, but was not reflected by an increase in progenitor cells, in contrast to TPO alone. Monkeys treated with FL alone showed a BM reconstitution similar to placebo-treated controls. FL by itself was not effective as a therapeutic agent in this model for myelosuppression. As FL also suppressed BM CD34+ cell reconstitution, we concluded that FL competed with TPO at the level of immature cell differentiation.
- Published
- 2003
34. Long-Term Effects of Irradiation Before Adulthood on Reproductive Function in the Male Rhesus Monkey1
- Author
-
Roelof Dol, Annemarie van Duijn-Goedhart, Dirk G. de Rooij, Gerard Wagemaker, Paul P.W. van Buul, Henk J. G. van de Kant, Frank H. de Jong, and Johan J. Broerse
- Subjects
medicine.medical_specialty ,Cell Biology ,General Medicine ,Total body irradiation ,Biology ,Sertoli cell ,Epididymis ,Dose–response relationship ,Endocrinology ,medicine.anatomical_structure ,Seminiferous tubule ,Reproductive Medicine ,Internal medicine ,medicine ,Stem cell ,Spermatogenesis ,Testosterone - Abstract
Today, many patients, who are often young, undergo total body irradiation (TBI) followed by bone marrow transplantation. This procedure can have serious consequences for fertility, but the long-term intratesticular effects of this treatment in primates have not yet been studied. Testes and epididymides of rhesus monkeys that received doses of 4-8.5 Gy of TBI at 2-4 yr of age were studied 3-8 yr after irradiation. In all irradiated monkeys, at least some seminiferous tubule cross-sections lacked germ cells, indicating extensive stem cell killing that was not completely repaired by enhanced stem cell renewal, even after many years. Testes totally devoid of germ cells were only found in monkeys receiving doses of 8 Gy or higher and in both monkeys that received two fractions of 6 Gy each. By correlating the percentage of repopulated tubules (repopulation index) with testicular weight, it could be deduced that considerable numbers of proliferating immature Sertoli cells were killed by the irradiation. Because of their finite period of proliferation, Sertoli cell numbers did not recover, and potential adult testis size decreased from approximately 23 to 13 g. Most testes showed some dilated seminiferous tubules, indicating obstructed flow of the tubular fluid at some time after irradiation. Also, in 8 of the 29 irradiated monkeys, aberrant, densely packed Sertoli cells were found. The irradiation did not induce stable chromosomal translocations in spermatogonial stem cells. No apparent changes were seen in the epididymides of the irradiated monkeys, and the size of the epididymis adjusted itself to the size of the testis. In the irradiated monkeys, testosterone and estradiol levels were normal, whereas FSH levels were higher and inhibin levels lower when testicular weight and spermatogenic repopulation were low. It is concluded that irradiation before adulthood has considerable long-term effects on the testis. Potential testis size is reduced, repopulation of the seminiferous epithelium is generally not complete, and aberrant Sertoli cells and dilated tubules are formed. The latter two phenomena may have further consequences at still longer intervals after irradiation.
- Published
- 2002
35. Human mesenchymal stem cells support B-cell and T-cell differentiation of cord blood CD34+ cells under optimized conditions
- Author
-
Gerard Wagemaker, Duygu Uckan-Cetinkaya, and Fatima Aerts-Kaya
- Subjects
Cancer Research ,Mesenchymal stem cell ,Amniotic stem cells ,Cell Biology ,Hematology ,Biology ,Cord lining ,Cell biology ,Endothelial stem cell ,Cord blood ,Genetics ,Stem cell ,Molecular Biology ,Adult stem cell ,Stem cell transplantation for articular cartilage repair - Published
- 2017
36. Hypothermic storage of hematopoetic stem cells can be used as an alternative to short-term cryopreservation
- Author
-
Gerard Wagemaker, Fatima Aerts-Kaya, Trui Visser, Burcu Pervin, and Duygu Uckan-Cetinkaya
- Subjects
Cancer Research ,030219 obstetrics & reproductive medicine ,0402 animal and dairy science ,04 agricultural and veterinary sciences ,Cell Biology ,Hematology ,Biology ,040201 dairy & animal science ,Cryopreservation ,Term (time) ,Cell biology ,03 medical and health sciences ,0302 clinical medicine ,Genetics ,Stem cell ,Molecular Biology - Published
- 2017
37. Anniversary issue on gene and cell therapy in the Netherlands
- Author
-
Gerard Wagemaker
- Subjects
medicine.medical_specialty ,education.field_of_study ,Severe combined immunodeficiency ,business.industry ,Marrow transplantation ,Genetic enhancement ,Population ,Cell- and Tissue-Based Therapy ,Environmental ethics ,Disease ,Genetic Therapy ,medicine.disease ,Cell therapy ,Family medicine ,Genetics ,medicine ,Molecular Medicine ,Animals ,Humans ,Allogeneic BMT ,business ,education ,Molecular Biology ,Gene ,Netherlands - Abstract
It is a privilege to congratulate Mary Ann Liebert and Jim Wilson and his staff on the 25th anniversary of Human Gene Therapy on behalf of the Netherlands’ Society of Gene and Cell Therapy (NVGCT), and to introduce this special issue directed at gene and cell therapy in The Netherlands on the occasion of the 22nd annual meeting of the European Society of Gene and Cell Therapy in The Hague. The Netherlands has a long scientific tradition, which originates from its 17th-century so-called ‘‘Golden Age’’ after establishment of the Republic of the United Netherlands in 1588. Due to its, in those days, exceptional climate of intellectual and religious tolerance, the republic attracted craftsmen, philosophers, artists, and scientists from all over Europe, in particular, to the renowned Leiden University (established 1575). This included the Russian czar Peter the Great, who modernized his country with many elements of the Dutch model. As a consequence, the Dutch made numerous seminal contributions to the world’s science, technology and engineering, medicine, and agriculture, and still today has a scientific output disproportionate to the small size of the country and its population. This issue of Human Gene Therapy pictures a highly varied landscape of Dutch gene and cell therapy from oncolytic viruses in cancer treatment (Dong et al.) to stem cell gene therapy for rare inherited diseases such as Pompe’s disease (Wagemaker), exon skipping for Duchenne muscular dystrophy (Aartsma-Rus et al.), cellular reprogramming and its therapeutic options (Mikkers et al.), and coping with the regulatory challenges of our time (Aartsma-Rus et al.; Schagen et al.). ‘‘Human Gene Therapy Briefs’’ completes the picture with this year’s achievements in ‘‘natural gene therapy,’’ DNA editing, synthetic gene transfer vectors, T cell engineering for antitumor therapy, and the corporate developments in Prosensa and uniQure. As memorized in the review summarizing the shift from oncogenic to oncolytic viruses (Belcaid et al.), much of the gene and cell therapy efforts, in particular oncolytic viruses and hematopoietic stem cell gene therapy, are branches from the same roots. Van der Eb’s laboratory in Leiden demonstrated efficient transfer of DNA into target cells (Graham and Van der Eb, 1973), and Van Bekkum’s Radiobiological Institute in Rijswijk developed allogeneic bone marrow transplantation (BMT) (summarized in Van Bekkum and De Vries, 1967) and pioneered it in immune deficiency patients in collaboration with the Leiden Children’s Hospital (De Koning et al., 1969), simultaneously with R.A. Good and coworkers at the University of Minnesota (Meuwissen et al., 1969). Both van der Eb and Van Bekkum are honorary members of the NVGCT. It is no coincidence that two of the same four European institutes that subsequently practiced allogeneic BMT for immune deficiencies (Fischer et al., 1986) developed the seminal X-linked severe combined immunodeficiency (SCID) gene therapy trials two decades later. However, at this time, the Leiden Children’s Hospital no longer served as one of the European referral hospitals for inherited immune deficiencies, while the incidence of SCID in The Netherlands is extremely low. The continued collaborative efforts of the Radiobiological Institute and the Leiden Children’s Hospital to apply BMT to diseases such as lysosomal storage diseases resulted in the discovery that microglia descendants of hematopoietic stem cells are capable of passing the blood– brain barrier (Hoogerbrugge et al., 1988), which is nowadays applied successfully in stem cell gene therapy for such disorders. The present achievements in translational gene therapy research would not have been possible without funding by the Netherlands Organization for Health Research and Development ZonMw, in particular by program grants in its Translational Gene Therapy and Adult Stem Cell Therapy Research programs, and in its Priority Medicines Rare Diseases program. Notwithstanding the indispensable national funding, The Netherlands is not a scientific island and much of the present results could also only be achieved in the context of large-scale collaborative projects funded by the Framework Programs of the European Commission, for which I had the privilege of serving as a coordinator for 22 consecutive years, 10 of which were in the field of gene therapy. Summaries of these projects in the 7th Framework Program, in which leading institutes need to collaborate in an open atmosphere of data sharing, have been published in this year’s June issue of Human Gene Therapy Clinical
- Published
- 2014
38. Lentiviral Hematopoietic Stem Cell Gene Therapy in Inherited Metabolic Disorders
- Author
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Gerard Wagemaker
- Subjects
medicine.medical_treatment ,Genetic enhancement ,Treatment outcome ,Genetic Vectors ,Disease ,Hematopoietic stem cell transplantation ,Biology ,Bioinformatics ,Genetic therapy ,Mice ,Immune system ,Genetics ,medicine ,Brief Reviews ,Animals ,Humans ,Molecular Biology ,Lentivirus ,Hematopoietic Stem Cell Transplantation ,Hematopoietic stem cell ,Genetic Therapy ,biology.organism_classification ,Hematopoietic Stem Cells ,Disease Models, Animal ,medicine.anatomical_structure ,Treatment Outcome ,Immunology ,Molecular Medicine ,Metabolism, Inborn Errors - Abstract
After more than 20 years of development, lentiviral hematopoietic stem cell gene therapy has entered the stage of initial clinical implementation for immune deficiencies and storage disorders. This brief review summarizes the development and applications, focusing on the lysosomal enzyme deficiencies, especially Pompe disease.
- Published
- 2014
39. Lentiviral gene transduction of mouse and human hematopoietic stem cells
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Niek P, van Til and Gerard, Wagemaker
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Mice ,HEK293 Cells ,Transduction, Genetic ,Genetic Vectors ,Lentivirus ,Animals ,Humans ,Hematopoietic Stem Cells ,Polymerase Chain Reaction - Abstract
Lentiviral vectors can be used to genetically modify a broad range of cells. Hematopoietic stem cells (HSCs) are particularly suitable for lentiviral gene augmentation, because these cells can be enriched with relative ease from mouse bone marrow and human hematopoietic sources, and in principle require relatively limited cell numbers to completely reconstitute the hematopoietic system in vivo. Furthermore, lentiviral vectors are very efficient if pseudotyped with broad tropism envelope proteins. This chapter focuses on gene modification by the use of self-inactivating third-generation human immunodeficiency virus-derived lentiviral vectors for ex vivo HSC modification for both mouse and human application.
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- 2014
40. Lentiviral Gene Transduction of Mouse and Human Hematopoietic Stem Cells
- Author
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Gerard Wagemaker and Niek P. van Til
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Transduction (genetics) ,Haematopoiesis ,medicine.anatomical_structure ,HEK 293 cells ,Cell ,medicine ,Bone marrow ,Biology ,Stem cell ,Ex vivo ,Tropism ,Cell biology - Abstract
Lentiviral vectors can be used to genetically modify a broad range of cells. Hematopoietic stem cells (HSCs) are particularly suitable for lentiviral gene augmentation, because these cells can be enriched with relative ease from mouse bone marrow and human hematopoietic sources, and in principle require relatively limited cell numbers to completely reconstitute the hematopoietic system in vivo. Furthermore, lentiviral vectors are very efficient if pseudotyped with broad tropism envelope proteins. This chapter focuses on gene modification by the use of self-inactivating third-generation human immunodeficiency virus-derived lentiviral vectors for ex vivo HSC modification for both mouse and human application.
- Published
- 2014
41. The Outcome of Local Radiation Injuries: 14 Years of Follow-up after the Chernobyl Accident
- Author
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Theodor M. Fliedner, Stefani Fh, Vladimir Bebeshko, Marianne Steinert, Petra Gottlöber, Ralf U. Peter, Natalia Nadejina, Melanie Weiss, Gerard Wagemaker, David Belyi, and Hematology
- Subjects
medicine.medical_specialty ,Pediatrics ,Biophysics ,Ionizing radiation ,Cohort Studies ,Radioactive contamination ,medicine ,Humans ,Radiology, Nuclear Medicine and imaging ,Radiation Injuries ,Survival analysis ,Skin ,Cause of death ,Radiation ,business.industry ,Bone marrow failure ,medicine.disease ,Surgery ,Radiation sickness ,Accidental ,Skin cancer ,Radioactive Hazard Release ,Ukraine ,business ,Follow-Up Studies ,Power Plants - Abstract
The Chernobyl nuclear power plant accident on April 26, 1986 was the largest in the history of the peaceful use of nuclear energy. Of the 237 individuals initially suspected to have been significantly exposed to radiation during or in the immediate aftermath of the accident, the diagnosis of acute radiation sickness (ARS) could be confirmed in 134 cases on the basis of clinical symptoms. Of these, 54 patients suffered from cutaneous radiation syndrome (CRS) to varying degrees. Among the 28 patients who died from the immediate consequences of accidental radiation exposure, acute hemopoietic syndrome due to bone marrow failure was the primary cause of death only in a minority. In 16 of these 28 deaths, the primary cause was attributed to CRS. This report describes the characteristic cutaneous sequelae as well as associated clinical symptoms and diseases of 15 survivors of the Chernobyl accident with severe localized exposure who were systematically followed up by our groups between 1991 and 2000. All patients presented with CRS of varying severity, showing xerosis, cutaneous telangiectasias and subungual splinter hemorrhages, hemangiomas and lymphangiomas, epidermal atrophy, disseminated keratoses, extensive dermal and subcutaneous fibrosis with partial ulcerations, and pigmentary changes including radiation lentigo. Surprisingly, no cutaneous malignancies have been detected so far in those areas that received large radiation exposures and that developed keratoses; however, two patients first presented in 1999 with basal cell carcinomas on the nape of the neck and the right lower eyelid, areas that received lower exposures. During the follow-up period, two patients were lost due to death from myelodysplastic syndrome in 1995 and acute myelogenous leukemia in 1998, respectively. Other radiation-induced diseases such as dry eye syndrome (3/15), radiation cataract (5/15), xerostomia (4/15) and increased FSH levels (7/15) indicating impaired fertility were also documented. This study, which analyzes 14 years in the clinical course of a cohort of patients with a unique exposure pattern, corroborates the requirement for long-term, if not life-long, follow-up not only in atomic bomb survivors, but also after predominantly local radiation exposure.
- Published
- 2001
42. Single administration of thrombopoietin to lethally irradiated mice prevents infectious and thrombotic events leading to mortality
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Marie-Hélène Gaugler, Marie Vandamme, Anne Van der Meeren, Gerard Wagemaker, Marc-André Mouthon, Patrick Gourmelon, and Hematology
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Male ,Cancer Research ,Neutrophils ,medicine.medical_treatment ,Drug Evaluation, Preclinical ,Hemorrhage ,Spleen ,Blood cell ,Leukocyte Count ,Mice ,Bone Marrow ,Fibrinolysis ,Genetics ,medicine ,Animals ,RNA, Messenger ,Bone Marrow Diseases ,Molecular Biology ,Thrombopoietin ,Platelet Count ,business.industry ,Immunologic Deficiency Syndromes ,Fibrinogen ,Thrombosis ,Bacterial Infections ,Cell Biology ,Hematology ,Blood Coagulation Disorders ,Total body irradiation ,Platelet Activation ,medicine.disease ,Endotoxemia ,Recombinant Proteins ,Mice, Inbred C57BL ,Radiation Injuries, Experimental ,Haematopoiesis ,medicine.anatomical_structure ,Immunology ,Disease Susceptibility ,Bone marrow ,business ,Biomarkers ,Whole-Body Irradiation - Abstract
Objective A sufficiently high dose of thrombopoietin to overcome initial c-mpl–mediated clearance stimulates hematopoietic reconstitution following myelosuppressive treatment. We studied the efficacy of thrombopoietin on survival after supralethal total body irradiation (9 Gy) of C57BL6/J mice and the occurrence of infectious and thrombotic complications in comparison with a bone marrow graft or prophylactic antibiotic treatment. Methods and Results Administration of 0.3 μg thrombopoietin, 2 hours after irradiation, protected 62% of the mice as opposed to no survival in placebo controls. A graft with a supraoptimal number of syngeneic bone marrow cells (10 6 cells) fully prevented mortality, whereas antibiotic treatment was ineffective. Blood cell recovery was observed in the thrombopoietin-treated mice but not in the placebo or antibiotic-treated group. Bone marrow and spleen cellularity as well as colony-forming unit granulocyte-macrophage and burst-forming unit erythroid were considerably increased in thrombopoietin-treated mice relative to controls. Histologic examination at day 11 revealed numerous petechiae and vascular obstructions within the brain microvasculature of placebo-treated mice, which was correlated with hypercoagulation and hypofibrinolysis. Thrombopoietin treatment prevented coagulation/fibrinolysis disorder and vascular thrombosis. High fibrinogen levels were related to bacterial infections in 67% of placebo-treated mice and predicted mortality, whereas the majority of the thrombopoietin-treated mice did not show high fibrinogen levels and endotoxin was not detectable in plasma. Conclusion We conclude that thrombopoietin administration prevents mortality in mice subjected to 9-Gy total body irradiation both by interfering in the cascade leading to thrombotic complications and by amelioration of neutrophil and platelet recovery and thus protects against infections and hemorrhages.
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- 2001
43. Lack of efficacy of thrombopoietin and granulocyte-macrophage colony-stimulating factor after total body irradiation and autologous bone marrow transplantation in Rhesus monkeys
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Gerard Wagemaker, Karen J. Neelis, Simone C. C. Hartong, Trudy P. Visser, and Hematology
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Male ,Cancer Research ,medicine.medical_specialty ,Cell Separation ,Granulocyte ,Internal medicine ,Genetics ,medicine ,Animals ,Molecular Biology ,Thrombopoietin ,Bone Marrow Transplantation ,business.industry ,Granulocyte-Macrophage Colony-Stimulating Factor ,Cell Biology ,Hematology ,Total body irradiation ,medicine.disease ,Flow Cytometry ,Pancytopenia ,Macaca mulatta ,Lymphocyte Subsets ,Transplantation ,Haematopoiesis ,medicine.anatomical_structure ,Endocrinology ,Granulocyte macrophage colony-stimulating factor ,Drug Therapy, Combination ,Bone marrow ,business ,Whole-Body Irradiation ,medicine.drug - Abstract
Objective If administered in a sufficiently high dose to overcome receptor-mediated clearance and in a well-scheduled manner, thrombopoietin (TPO) prominently stimulates hematopoietic reconstitution following myelosuppressive treatment and potentiates the efficacy of both granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF). However, TPO alone is not effective after bone marrow transplantation. Based on results of GM-CSF and TPO treatment after myelosuppression that resulted in augmented thrombocyte, reticulocyte, and leukocyte regeneration, we evaluated TPO/GM-CSF treatment after lethal irradiation followed by autologous bone marrow transplantation. Materials and Methods Young adult Rhesus monkeys were subjected to 8-Gy total body irradiation (TBI) (x-rays) followed by transplantation of 10 7 /kg unfractionated bone marrow cells. TPO 5 μg/kg was administered intravenously at day 0 to obtain rapidly high levels. Animals then were treated with 5 μg/kg Rhesus TPO and 25 μg/kg GM-CSF given SC on days 1 to 14 after TBI. Results The grafts shortened the profound pancytopenia induced by 8-Gy TBI from 5–6 weeks to 3 weeks. The combination of TPO and GM-CSF did not significantly influence the recovery patterns of thrombocytes (p = 0.39), reticulocytes (p = 0.08), white blood cells (p = 0.08), or bone marrow progenitors compared to TPO alone. Conclusions The present study demonstrates that, after high-dose TBI and transplantation of a limited number of unfractionated bone marrow cells, simultaneous administration of TPO and GM-CSF after TBI is ineffective in preventing pancytopenia. This result contrasts sharply with the prominent stimulation observed in a 5-Gy TBI myelosuppression model, despite a similar level of pancytopenia in the 8-Gy model of the present study. The discordant results of this growth factor combination in these two models may imply codependence of the hematopoietic response to TPO and/or GM-CSF on other factors or cytokines.
- Published
- 2000
44. Stimulation of mouse bone marrow cells with kit ligand FLT3 ligand, and thrombopoietin leads to efficient retrovirus-mediated gene transfer to stem cells, whereas interleukin 3 and interleukin 11 reduce transduction of short- and long-term repopulating ce
- Author
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Marti F.A. Bierhuizen, Trudy P. Visser, Kathelijn Peters, Gerard Wagemaker, Albertus W. Wognum, and Hematology
- Subjects
Blood Platelets ,Male ,Erythrocytes ,Time Factors ,Light ,Genetic Vectors ,Green Fluorescent Proteins ,Stem cell factor ,Bone Marrow Cells ,Cell Separation ,Biology ,Immunophenotyping ,Mice ,alpha-Thalassemia ,Transduction, Genetic ,Genetics ,medicine ,Animals ,Scattering, Radiation ,Progenitor cell ,Molecular Biology ,Interleukin 3 ,Mice, Inbred BALB C ,Stem Cell Factor ,Stem Cells ,Gene Transfer Techniques ,Membrane Proteins ,3T3 Cells ,Flow Cytometry ,Interleukin-11 ,Molecular biology ,Transplantation ,Interleukin 11 ,Haematopoiesis ,Luminescent Proteins ,medicine.anatomical_structure ,Retroviridae ,Thrombopoietin ,Molecular Medicine ,Female ,Interleukin-3 ,Bone marrow ,Stem cell - Abstract
The effects of cytokine stimulation during retroviral transduction on in vivo reconstitution of mouse hematopoietic stem cells was tested in a murine competitive repopulation assay with alpha-thalassemia as a marker to distinguish donor and recipient red blood cells (RBCs) and the enhanced green fluorescent protein (EGFP) as a marker for gene transfer. After transplantation, EGFP was detected in up to 90% of circulating RBCs, platelets, and leukocytes, and in primitive progenitors in bone marrow (BM), spleen, and thymus of individual transplanted mice for observation periods of more than 6 months. Large quantitative differences in reconstitution were observed after transplantation with graded numbers (1000-30, 000) of EGFP(+) cells preconditioned with various combinations of Kit ligand (KL), FLT-3 ligand (FL), thrombopoietin (TPO), interleukin 3 (IL-3), and IL-11. Relative to nonmanipulated BM cells, repopulation of EGFP(+) cells was maintained by KL/FL/TPO stimulation, but approximately 30-fold reduced after KL/FL/TPO/IL-3, or KL/FL/IL-3/IL-11. These differences were not caused by changes in the ability of immature hematopoietic cells to home to the BM, which was only moderately reduced. In conclusion, these quantitative transplantation studies of mice demonstrate the importance of optimal ex vivo cytokine stimulation for gene transfer to stem cells with retention of their in vivo hematopoietic potential, and also emphasize that overall in vitro transduction frequency does not predict gene transfer to repopulating stem cells.
- Published
- 2000
45. Efficient detection and selection of immature rhesus monkey and human CD34+ hematopoietic cells expressing the enhanced green fluorescent protein (EGFP)
- Author
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Gerard Wagemaker, Yvonne Westerman, Marti F.A. Bierhuizen, A. W. Wognum, Simone C. C. Hartong, and Trudy P. Visser
- Subjects
Male ,Cancer Research ,Recombinant Fusion Proteins ,Genetic Vectors ,Green Fluorescent Proteins ,CD34 ,Gene Expression ,Antigens, CD34 ,Bone Marrow Cells ,Cell Separation ,Biology ,Transfection ,Cell Line ,Immunophenotyping ,Flow cytometry ,Colony-Forming Units Assay ,Genes, Reporter ,medicine ,Animals ,Humans ,Cytotoxic T cell ,Cell Lineage ,medicine.diagnostic_test ,Membrane Proteins ,Hematology ,Flow Cytometry ,Hematopoietic Stem Cells ,Macaca mulatta ,Molecular biology ,Hematopoietic Stem Cell Mobilization ,Luminescent Proteins ,Haematopoiesis ,Retroviridae ,medicine.anatomical_structure ,Oncology ,Cell culture ,Immunology ,Feasibility Studies ,Bone marrow ,Stem cell ,Biomarkers - Abstract
The feasibility of using the enhanced green fluorescent protein (EGFP) as a selectable reporter molecule of retroviral-mediated gene transfer in immature rhesus monkey and human CD34+ hematopoietic cells was examined. Retroviral transduction with the MFG-EGFP retroviral vector resulted in readily detectable EGFP expression in 27% of human and 11-35% of rhesus monkey bone marrow cells, and in 17-38% of rhesus monkey peripheral blood cells mobilized with FLT3 ligand (FL) and granulocyte colony-stimulating factor (G-CSF). In addition, we used the human CD34+ KG1A cell line as a model to study viability and growth of successfully transduced cells. Cultures of mock- and EGFP-transduced KG1A cells generated equal viable cell numbers for at least 1 month, indicating the absence of a cytotoxic effect of EGFP expression in these cells. FACS selection on the basis of EGFP and CD34 expression resulted in enriched subsets (> or = 87%) of CD34+ EGFP-negative and CD34+ EGFP-positive KG1A, rhesus monkey and human bone marrow cells, demonstrating the potential of obtaining almost pure populations of transduced immature hematopoietic cells. EGFP expression was also readily demonstrated in erythroid and granulocyte/macrophage colonies derived from the CD34+ EGFP-positive rhesus monkey and human bone marrow cells by either inverted fluorescence microscopy or flow cytometry. Using four-color flow cytometry, EGFP expression could also be demonstrated in viable and phenotypically defined immature subpopulations of the CD34+ cells, ie those expressing little or no HLA-DR (rhesus monkey) or CD38 (human) antigens at the cell surface. These results demonstrate that EGFP is a very useful marker to monitor gene transfer efficiency in phenotypically defined immature rhesus monkey and human hematopoietic cell types and to select for these cells by multicolor flow cytometry prior to transplantation.
- Published
- 1999
46. Multilineage outgrowth of both malignant and normal hemopoietic progenitor cells from individual chronic myeloid leukemia patients in immunodeficient mice
- Author
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Gerard Wagemaker, A. W. Wognum, Monique M A Verstegen, Wim Terpstra, Jeroen J. L. M. Cornelissen, and Hematology
- Subjects
Cancer Research ,Myeloid ,Transplantation, Heterologous ,CD34 ,Mice, SCID ,Biology ,Immunocompromised Host ,Mice ,Mice, Inbred NOD ,Leukemia, Myelogenous, Chronic, BCR-ABL Positive ,hemic and lymphatic diseases ,medicine ,Animals ,Humans ,Cytotoxic T cell ,Cell Lineage ,Philadelphia Chromosome ,Progenitor cell ,In Situ Hybridization, Fluorescence ,Tumor Stem Cell Assay ,Graft Survival ,Myeloid leukemia ,Cell Differentiation ,Hematology ,Hematopoietic Stem Cells ,Haematopoiesis ,medicine.anatomical_structure ,Oncology ,Radiation Chimera ,Leukemia, Myeloid, Chronic-Phase ,Immunology ,Neoplastic Stem Cells ,Cancer research ,Bone marrow ,Stem cell ,Blast Crisis ,Neoplasm Transplantation - Abstract
In this study the ability of malignant and normal progenitors in peripheral blood (PB) and bone marrow (BM) of CML patients in chronic phase to proliferate and produce mature progeny after transplantation into hereditary immunodeficient (SCID and NOD/SCID) mice was examined. Engraftment in NOD/SCID mice preconditioned by total body irradiation (TBI) alone was 10-fold higher than in SCID mice preconditioned by macrophage depletion and TBI, demonstrating that NOD/SCID mice are more suitable for engraftment of chronic phase CML cells. Low-density cells at cell doses of 10-30 x 10 6 and purified CD34 + cells at doses of approximately 0.2 x 10 6 engrafted NOD/SCID mice, with levels of 2 to 20% CD45 + cells with production of monocytes, granulocytes, erythroid cells, B-lymphocytes, CD34 + cells and variable frequencies of erythroid and myeloid colony-forming cells. As demonstrated by fluorescent in situ hybridization (FISH) analysis, purified human myeloid, B-lymphoid, erythroid and CD34 + cells from chimeric mouse BM contained Philadelphia-chromosome (Ph)-positive cells and Ph - cells in similar frequencies as primary cells from the CML patients. These results demonstrate that production of mature normal as well as malignant cells of multiple lineages were supported with similar efficiency. In contrast, all human erythroid and myeloid clonogenic cells detected in the mice were Ph - , which can be attributed to less efficient maintenance or more rapid differentiation of immature Ph + cells in the mouse microenvironment. CM blast crisis cells also grew well In NOD/SCID mice, with 80-90% of human cells produced containing the Ph-chromosome. The availability of an in vivo assay that supports outgrowth of normal and malignant stem cells from chronic phase and blast crisis CML patients will facilitate examination of differential effects of growth factors, inhibitory cytokines and cytotoxic drugs on survival of normal and malignant stem cells in vivo and on progression of chronic phase CM towards blast crisis.
- Published
- 1999
47. Highly Efficient Transduction of the Green Fluorescent Protein Gene in Human Umbilical Cord Blood Stem Cells Capable of Cobblestone Formation in Long-Term Cultures and Multilineage Engraftment of Immunodeficient Mice
- Author
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Ana Limón, Jose A. Cancelas, Paula B. van Hennik, Gerard Wagemaker, Rob E. Ploemacher, A. W. Wognum, Marti F.A. Bierhuizen, Monique M A Verstegen, Jordi Barquinero, and Hematology
- Subjects
Cellular differentiation ,Immunology ,Cell Biology ,Hematology ,CD38 ,Biology ,Virology ,Molecular biology ,Biochemistry ,Transplantation ,Haematopoiesis ,medicine.anatomical_structure ,Cell culture ,medicine ,Bone marrow ,Progenitor cell ,Stem cell - Abstract
Purified CD34+ and CD34+CD38− human umbilical cord blood (UCB) cells were transduced with the recombinant variant of Moloney murine leukemia virus (MoMLV) MFG-EGFP or with SF-EGFP, in which EGFP expression is driven by a hybrid promoter of the spleen focus-forming virus (SFFV) and the murine embryonic stem cell virus (MESV). Infectious MFG-EGFP virus was produced by an amphotropic virus producer cell line (GP+envAm12). SF-EGFP was produced in the PG13 cell line pseudotyped for the gibbon ape leukemia virus (GaLV) envelope proteins. Using a 2-day growth factor prestimulation, followed by a 2-day, fibronectin fragment CH-296–supported transduction, CD34+ and CD34+CD38− UCB subsets were efficiently transduced using either vector. The use of the SF-EGFP/PG13 retroviral packaging cell combination consistently resulted in twofold higher levels of EGFP-expressing cells than the MFG-EGFP/Am12 combination. Transplantation of 105 input equivalent transduced CD34+ or 5 × 103input equivalent CD34+CD38− UCB cells in nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice resulted in median engraftment percentages of 8% and 5%, respectively, which showed that the in vivo repopulating ability of the cells had been retained. In addition, mice engrafted after transplantation of transduced CD34+ cells using the MFG-EGFP/Am12 or the SF-EGFP/PG13 combination expressed EGFP with median values of 2% and 23% of human CD45+ cells, respectively, which showed that the NOD/SCID repopulating cells were successfully transduced. EGFP+ cells were found in all human hematopoietic lineages produced in NOD/SCID mice including human progenitors with in vitro clonogenic ability. EGFP-expressing cells were also detected in the human cobblestone area–forming cell (CAFC) assay at 2 to 6 weeks of culture on the murine stromal cell line FBMD-1. During the transduction procedure the absolute numbers of CAFC week 6 increased 5- to 10-fold. The transduction efficiency of this progenitor cell subset was similar to the fraction of EGFP+ human cells in the bone marrow of the NOD/SCID mice transplanted with MFG-EGFP/Am12 or SF-EGFP/PG13 transduced CD34+ cells, ie, 6% and 27%, respectively. The study thus shows that purified CD34+ and highly purified CD34+CD38− UCB cells can be transduced efficiently with preservation of repopulating ability. The SF-EGFP/PG13 vector/packaging cell combination was much more effective in transducing repopulating cells than the MFG-EGFP/Am12 combination.
- Published
- 1998
48. The Efficacy of Recombinant Thrombopoietin in Murine and Nonhuman Primate Models for Radiation‐Induced Myelosuppression and Stem Cell Transplantation
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Albertus W. Wognum, Simone C. C. Hartong, Gerard Wagemaker, Karen J. Neelis, Dan L. Eaton, G. Roger Thomas, Paul J. Fielder, and Hematology
- Subjects
Primates ,medicine.medical_treatment ,CD34 ,Hematopoietic stem cell transplantation ,Biology ,Pharmacology ,Mice ,In vivo ,medicine ,Animals ,Humans ,Thrombopoietin ,Hematopoietic Stem Cell Transplantation ,Immunity ,Cell Biology ,Thrombocytopenia ,Recombinant Proteins ,Transplantation ,Disease Models, Animal ,Haematopoiesis ,medicine.anatomical_structure ,Immunology ,Molecular Medicine ,Bone marrow ,Stem cell ,Developmental Biology - Abstract
Radiation-induced pancytopenia proved to be a suitable model system in mice and rhesus monkeys for studying thrombopoietin (TPO) target cell range and efficacy. TPO was highly effective in rhesus monkeys exposed to the mid-lethal dose of 5 Gy (300 kV x-rays) TBI, a model in which it alleviated thrombocytopenia, promoted red cell reconstitution, accelerated reconstitution of immature CD34+ bone marrow cells, and potentiated the response to growth factors such as GM-CSF and G-CSF. In contrast to the results in the 5 Gy TBI model, TPO was ineffective following transplantation of limited numbers of autologous bone marrow or highly purified stem cells in monkeys conditioned with 8 Gy TBI. In the 5 Gy model, a single dose of TPO augmented by GM-CSF 24 h after TBI was effective in preventing thrombocytopenia. The strong erythropoietic stimulation may result in iron depletion, and TPO treatment should be accompanied by monitoring of iron status. This preclinical evaluation thus identified TPO as a potential major therapeutic agent for counteracting radiation-induced pancytopenia and demonstrated pronounced stimulatory effects on the reconstitution of immature CD34+ hemopoietic cells with multilineage potential. The latter observation explains the potentiation of the hematopoietic responses to G-CSF and GM-CSF when administered concomitantly. It also predicts the effective use of TPO to accelerate reconstitution of immature hematopoietic cells as well as possible synergistic effects in vivo with various other growth factors acting on immature stem cells and their direct lineage-committed progeny. The finding that a single dose of TPO might be sufficient for a clinically significant response emphasizes its potency and is of practical relevance. The heterogeneity of the TPO response encountered in the various models used for evaluation points to multiple mechanisms operating on the TPO response and heterogeneity of its target cells. Mechanistic mouse studies made apparent that the response of multilineage cells shortly after TBI to a single administration of TPO is quantitatively more important for optimal efficacy than the lineage-restricted response obtained at later intervals after TBI and emphasized the importance of a relatively high dose of TPO to overcome initial c-mpl-mediated clearance. Further elucidation of mechanisms determining efficacy might very well result in a further improvement, e.g., following transplantation of limited numbers of stem cells. Adverse effects of TPO administration to myelosuppressed or stem cell transplanted experimental animals were not observed.
- Published
- 1998
49. 279. Efficient and Safe Lentiviral Vector-Mediated Hematopoietic Stem Cell Gene Therapy in MNGIE Mice
- Author
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Rana Yadak, Elly Bogaerts, Jordi Barquinero, George de Ruijter, René F.M. de Coo, Ramon Martí, Javier Torres Torronteras, Cabrera Pérez Raquel, Gerard Wagemaker, Niek P. van Til, Bert Smeets, and Marshall W. Huston
- Subjects
Pharmacology ,Genetic enhancement ,Hematopoietic stem cell ,Biology ,Total body irradiation ,Molecular biology ,Viral vector ,Transplantation ,medicine.anatomical_structure ,Immunology ,Drug Discovery ,medicine ,Genetics ,Molecular Medicine ,Bone marrow ,Stem cell ,Thymidine phosphorylase ,Molecular Biology - Abstract
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) patients are deficient in thymidine phosphorylase(TP) resulting in systemic thymidine(Thd) and deoxyuridine(dUrd) accumulation affecting mtDNA replication and causing mitochondrial dysfunction. Common symptoms are gastrointestinal dysmotility, progressive ophthalmoplegia and leukoencephalopathy. Allogenic hematopoietic stem cell (HSC) transplantation has been shown to reduce disease symptoms, but is not well tolerated due to the inherent toxicity of the procedure. Therefore, syngeneic ex vivo lentiviral vector HSC gene therapy overexpressing the native cDNA or the codon optimized (TPco) sequence driven by the phosphoglycerate kinase (PGK) or spleen focus forming virus (SFFV) promoters in Tp−/-Upp−/- double knockout mice, a model for MNGIE disease, was investigated. At 1 month post transplantation after sublethal total body irradiation, very low TP activity was detected in blood of control wild type mice (0.07±0.03nmoles/h/mg), but enzyme activities in PGK treated mice were at least 90-fold higher (PGK-TP = 150±4 and PGK-TPco = 96±4 nmoles/h/mg), and in SFFV recipient mice 400-fold higher (450±5 nmoles/h/mg). Consequently, a significant reduction of plasma and urine Thd and dUrd levels was observed. Long-term follow up (14 months) showed on average 1.2-fold wild type TP activity levels increase in LV-PGK-TP and LV-PGK-TPco and 36-fold in SFFV-TPco treated mice. This was sufficient for sustained reduction of plasma and urine nucleoside levels, which was achieved at 76.5±8.2% donor chimerism levels with low LV vector copy numbers (1.0±1.1VCN/donor cell). The LV integration profile in bone marrow cells of primary recipients was analyzed; LVs displayed the expected tendency to integrate within highly expressed genes and the integration pattern did not differ from that of other SIN-LV vectors in other disease models (primary immune deficiencies and lysosomal enzyme storage disorders). Overall, stem cell gene therapy provided stable TP expression and long-term biochemical correction in MNGIE mice without genotoxicity or apparent phenotoxicity, which will be further evaluated for somatic and neurological phenotype correction and optimized to develop a clinical protocol to treat MNGIE patients.
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- 2015
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50. A Single Dose of Thrombopoietin Shortly After Myelosuppressive Total Body Irradiation Prevents Pancytopenia in Mice by Promoting Short-Term Multilineage Spleen-Repopulating Cells at the Transient Expense of Bone Marrow–Repopulating Cells
- Author
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Duane C. Bloedow, Paul J. Fielder, Wati Dimjati, Karen J. Neelis, Gerard Wagemaker, Trudi P. Visser, G. Roger Thomas, and Dan L. Eaton
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medicine.medical_specialty ,Hematology ,business.industry ,Immunology ,Cell Biology ,Total body irradiation ,Pharmacology ,Biochemistry ,Transplantation ,Haematopoiesis ,medicine.anatomical_structure ,Pharmacokinetics ,White blood cell ,Internal medicine ,medicine ,Bone marrow ,business ,Thrombopoietin - Abstract
Thrombopoietin (TPO) has been used in preclinical myelosuppression models to evaluate the effect on hematopoietic reconstitution. Here we report the importance of dose and dose scheduling for multilineage reconstitution after myelosuppressive total body irradiation (TBI) in mice. After 6 Gy TBI, a dose of 0.3 μg TPO/mouse (12 μg/kg) intraperitoneally (IP), 0 to 4 hours after TBI, prevented the severe thrombopenia observed in control mice, and in addition stimulated red and white blood cell regeneration. Time course studies showed a gradual decline in efficacy after an optimum within the first hours after TBI, accompanied by a replacement of the multilineage effects by lineage dominant thrombopoietic stimulation. Pharmacokinetic data showed that IP injection resulted in maximum plasma levels 2 hours after administration. On the basis of the data, we inferred that a substantial level of TPO was required at a critical time interval after TBI to induce multilineage stimulation of residual bone marrow cells. A more precise estimate of the effect of dose and dose timing was provided by intravenous administration of TPO, which showed an optimum immediately after TBI and a sharp decline in efficacy between a dose of 0.1 μg/mouse (4 μg/kg; plasma level 60 ng/mL), which was fully effective, and a dose of 0.03 μg/mouse (1.2 μg/kg; plasma level 20 ng/mL), which was largely ineffective. This is consistent with a threshold level of TPO required to overcome initial c-mpl–mediated clearance and to reach sufficient plasma levels for a maximum hematopoietic response. In mice exposed to fractionated TBI (3 × 3 Gy, 24 hours apart), IP administration of 0.3 μg TPO 2 hours after each fraction completely prevented the severe thrombopenia and anemia that occurred in control mice. Using short-term transplantation assays, ie, colony-forming unit–spleen (CFU-S) day 13 (CFU-S-13) and the more immature cells with marrow repopulating ability (MRA), it could be shown that TPO promoted CFU-S-13 and transiently depleted MRA. The initial depletion of MRA in response to TPO was replenished during long-term reconstitution followed for a period of 3 months. Apart from demonstrating again that MRA cells and CFU-S-13 are separate functional entities, the data thus showed that TPO promotes short-term multilineage repopulating cells at the expense of more immature ancestral cells, thereby preventing pancytopenia. The short time interval available after TBI to exert these effects shows that TPO is able to intervene in mechanisms that result in functional depletion of its multilineage target cells shortly after TBI and emphasizes the requirement of dose scheduling of TPO in keeping with these mechanisms to obtain optimal clinical efficacy. © 1998 by The American Society of Hematology.
- Published
- 1998
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