46 results on '"Parenza M"'
Search Results
2. HOXB7: a key factor for tumor-associated angiogenic switch
- Author
-
Care\', A., Felicetti, F., Meccia, F., Bottero, L., Parenza, M., Stoppacciaro, Antonella, Peschle, C., and Colombo, M. P.
- Subjects
Vascular Endothelial Growth Factor A ,Transplantation, Heterologous ,Mice, Nude ,Breast Neoplasms ,Endothelial Growth Factors ,Adenocarcinoma ,Gene Expression Regulation, Enzymologic ,Mice ,Transduction, Genetic ,Angiopoietin-1 ,Tumor Cells, Cultured ,Animals ,Humans ,Protein Isoforms ,Receptors, Growth Factor ,Glucuronidase ,Homeodomain Proteins ,Lymphokines ,Membrane Glycoproteins ,Neovascularization, Pathologic ,Reverse Transcriptase Polymerase Chain Reaction ,Vascular Endothelial Growth Factors ,Coculture Techniques ,Up-Regulation ,Gene Expression Regulation, Neoplastic ,Matrix Metalloproteinase 9 ,Matrix Metalloproteinase 2 ,Endothelium, Vascular ,Neoplasm Transplantation - Abstract
We had demonstrated previously a functional bridge between altered homebox (HOX) gene expression and tumor progression through HOXB7 transactivation of basic fibroblast growth factor. Here, we have studied whether HOXB7, in addition to basic fibroblast growth factor, may induce other genes directly or indirectly related to neoangiogenesis and tumor invasion. Parental, beta-galactosidase-transduced, and HOXB7-transduced SkBr3 cell lines were examined for the expression of several growth factors and growth factor receptors involved in the proliferative and angiogenic processes. Vascular endothelial growth factor, melanoma growth-stimulatory activity/growth-related oncogenene alpha, interleukin-8, and angiopoietin-2 were up-regulated by HOXB7 transduction. The exception was angiopoietin-1 expression that was abrogated. Additional analyses included the expression levels of enzymes such as matrix metalloprotease (MMP)-2 and MMP-9 and heparanase, capable of proteolytic degradation of extracellular matrix and basement membranes. Results showed an induction of only MMP-9. The functional implication of such a finding was tested using an in vitro coculture assay in a three-dimensional matrix. A delay of differentiation with persistent nests of proliferating cells was found in endothelial cells cocultured with HOXB7-transduced SkBr3 cells. Tumorigenicity of these cells has been evaluated in vivo. Xenograft into athymic nude mice showed that SkBr3/HOXB7 cells developed tumors in mice, either irradiated or not, whereas parental SkBr3 cells did not show any tumor take unless mice were sublethally irradiated. Comparison of tumor nodules for vascularization by CD-31 and CD-34 immunostaining revealed an increased number of blood vessels in tumors expressing HOXB7. Together, the results indicate HOXB7 as a key factor up-regulating a variety of proangiogenic stimuli. Thus, HOXB7 gene or protein is a target to aim at to inhibit tumor-associated neoangiogenesis, considering the number and the redundancy of proangiogenic molecules that should be targeted one by one to theoretically achieve the same effect.
- Published
- 2001
3. Granulocyte colony-stimulating factor (G-CSF) gene transduction in murine adenocarcinoma drives neutrophil-mediated tumor inhibition in vivo. Neutrophils discriminate between G-CSF-producing and G-CSF-nonproducing tumor cells
- Author
-
Mario Paolo Colombo, Lombardi, L., Stoppacciaro, A., Melani, C., Parenza, M., Bottazzi, B., and Parmiani, G.
- Subjects
Immunity, Cellular ,Mice, Inbred BALB C ,Neutrophils ,Immunology ,Adenocarcinoma ,Chemotaxis, Leukocyte ,Mice ,Microscopy, Electron ,Transduction, Genetic ,Colonic Neoplasms ,Granulocyte Colony-Stimulating Factor ,Cell Adhesion ,Tumor Cells, Cultured ,Immunology and Allergy ,Animals - Abstract
We have previously demonstrated that the murine colon adenocarcinoma C-26 cell line transduced with the human gene for the granulocyte CSF (G-CSF) loses tumorigenic activity through a mechanism that involved massive targeting of neutrophils at the site of tumor injection. The suppression of tumorigenicity by G-CSF was limited to the G-CSF-producing cells and was not transferred to nonproducing C-26 cells in a mixed tumor transplantation assay. We present direct evidence that neutrophils are involved in this phenomenon. We firstly examined, by electron microscopy (EM), the morphology of tumor infiltrates obtained 2, 5, and 10 days after s.c. injection of a mixture of G-CSF-producing and -nonproducing C-26 cells into syngeneic BALB/c mice. The EM analysis showed at 5, but not at 2 or 10 days, the presence of neutrophils in intimate contact with tumor cells. We then investigated whether neutrophils discriminate between G-CSF-producing and -nonproducing C-26 cells. To this aim, C-26 cells were transduced, via retroviral vector, with the Escherichia coli LacZ gene and mixed tumor transplantation assays were performed by injecting a mixture of G-CSF-producing beta-gal- and G-CSF-nonproducing beta-gal+ C-26 cells at different ratios. Histologic and EM analysis of the tumors growing at the site of injection were carried out. Five days after injection, treatment with x-gal revealed, at the histochemical level, the presence of neutrophils around G-CSF producing beta-gal- cells; cell-cell contacts and fusion of cell membranes were detected by EM only between neutrophils and G-CSF-producing cells. In vitro experiments, performed in Boyden chambers, confirmed that the G-CSF produced by C-26 cells was a chemoattractant for neutrophils. In addition, a colorimetric, cytostatic assay revealed that neutrophils were able to inhibit the growth of G-CSF-producing but not of G-CSF-nonproducing C-26 cells. Thus the tumor take after injection of G-CSF-producing C-26 cells seems to be controlled in situ through two major mechanisms namely neutrophil chemotaxis and neutrophil-mediated tumor inhibition. The results indicate that neutrophils can discriminate between G-CSF-producing and -nonproducing tumor cells and that neutrophils infiltrate the tumor mixture as long as G-CSF-producing cells are present.
- Published
- 1992
4. 2013 POSTER Bone marrow-derived TNF-a promotes tumour growth in a spontaneous model of mammary carcinogenesis
- Author
-
Chiodoni, C., primary, Sangaletti, S., additional, Parenza, M., additional, Ratti, C., additional, and Colombo, M.P., additional
- Published
- 2007
- Full Text
- View/download PDF
5. Regression of an established tumor genetically modified to release granulocyte colony-stimulating factor requires granulocyte-T cell cooperation and T cell-produced interferon gamma.
- Author
-
Stoppacciaro, A, primary, Melani, C, additional, Parenza, M, additional, Mastracchio, A, additional, Bassi, C, additional, Baroni, C, additional, Parmiani, G, additional, and Colombo, M P, additional
- Published
- 1993
- Full Text
- View/download PDF
6. Granulocyte colony-stimulating factor (G-CSF) gene transduction in murine adenocarcinoma drives neutrophil-mediated tumor inhibition in vivo. Neutrophils discriminate between G-CSF-producing and G-CSF-nonproducing tumor cells.
- Author
-
Colombo, M P, primary, Lombardi, L, additional, Stoppacciaro, A, additional, Melani, C, additional, Parenza, M, additional, Bottazzi, B, additional, and Parmiani, G, additional
- Published
- 1992
- Full Text
- View/download PDF
7. Granulocyte colony-stimulating factor gene transfer suppresses tumorigenicity of a murine adenocarcinoma in vivo.
- Author
-
Colombo, M P, primary, Ferrari, G, additional, Stoppacciaro, A, additional, Parenza, M, additional, Rodolfo, M, additional, Mavilio, F, additional, and Parmiani, G, additional
- Published
- 1991
- Full Text
- View/download PDF
8. Alloreactivity and Tumor Antigens: Generation of Syngeneic Antilymphoma Killer Lymphocytes by Alloimmunization of Mice With Normal Cells<xref ref-type='fn' rid='fn2'>2</xref><xref ref-type='fn' rid='fn3'>3</xref>
- Author
-
Sensi Ml, Parenza M, and Giorgio Parmiani
- Subjects
Cancer Research ,Lymphokine-activated killer cell ,biology ,Chemistry ,Lymphocyte ,T lymphocyte ,Major histocompatibility complex ,Immune system ,medicine.anatomical_structure ,Oncology ,Antigen ,Immunology ,medicine ,biology.protein ,Cytotoxic T cell ,Cytotoxicity - Abstract
The possibility of obtaining a syngeneic antitumor effect by immunization with normal allogeneic cells was investigated by tests of the lytic activity of peritoneal exudate cells (PEC) of BALB/c mice immunized with lymphoid cells of either a single strain or a pool of six different allogeneic strains on the syngeneic Moloney virus-induced lymphoma YC8 target and on one of its clones designated YC8-D1. Significant cytotoxicity on both targets but not on two other BALB/c lymphomas was obtained with PEC of BALB/c mice singly immunized to the non-H-2-incompatible but H-2-compatible DBA/2 or B10.D2 lymphoid cells. The lack of lysis of YC8 cells by PEC of BALB/c mice immune to B10.A (H-2k,d) suggests that the in vitro killing was restricted by Kd-IEd region products of the major histocompatibility complex. Pool immunization was effective in generating antitumor cytotoxic lymphocytes only when DBA/2 lymphoid cells were included in the pool. The pattern of reactivity of effectors elicited in (BALB/c x DBA/2)F1 and in (BALB/c x B10.D2)F1 mice by immunization with DBA/2 and B10.D2 cells showed that at least two sets of antigens are recognized on YC8 targets, one shared by DBA/2 and B10.D2 tissues and the other expressed by DBA/2 cells only. Cold target blocking experiments indicated that the same effectors recognized non-H-2 antigens of DBA/2 and the cross-reacting YC8 determinants. The antitumor effect was mediated by T-cells, since it was abrogated by treatment of effectors with anti-Thy 1.2 serum plus complement. These data indicate that determinants defined by cytotoxic T-lymphocytes are expressed on the BALB/c lymphoma YC8 and cross-react with non-H-2 antigens of DBA/2 and B10.D2 strains.
- Published
- 1983
9. Alloreactivity and tumor antigens: generation of syngeneic antilymphoma killer lymphocytes by alloimmunization of mice with normal cells
- Author
-
Marialuisa Sensi, Parenza, M., and Parmiani, G.
- Subjects
Cytotoxicity, Immunologic ,Killer Cells, Natural ,Mice ,Mice, Inbred BALB C ,Lymphoma ,Antigens, Neoplasm ,Mice, Inbred DBA ,H-2 Antigens ,Animals ,Ascitic Fluid ,Neoplasms, Experimental ,Cross Reactions ,Antilymphocyte Serum - Abstract
The possibility of obtaining a syngeneic antitumor effect by immunization with normal allogeneic cells was investigated by tests of the lytic activity of peritoneal exudate cells (PEC) of BALB/c mice immunized with lymphoid cells of either a single strain or a pool of six different allogeneic strains on the syngeneic Moloney virus-induced lymphoma YC8 target and on one of its clones designated YC8-D1. Significant cytotoxicity on both targets but not on two other BALB/c lymphomas was obtained with PEC of BALB/c mice singly immunized to the non-H-2-incompatible but H-2-compatible DBA/2 or B10.D2 lymphoid cells. The lack of lysis of YC8 cells by PEC of BALB/c mice immune to B10.A (H-2k,d) suggests that the in vitro killing was restricted by Kd-IEd region products of the major histocompatibility complex. Pool immunization was effective in generating antitumor cytotoxic lymphocytes only when DBA/2 lymphoid cells were included in the pool. The pattern of reactivity of effectors elicited in (BALB/c x DBA/2)F1 and in (BALB/c x B10.D2)F1 mice by immunization with DBA/2 and B10.D2 cells showed that at least two sets of antigens are recognized on YC8 targets, one shared by DBA/2 and B10.D2 tissues and the other expressed by DBA/2 cells only. Cold target blocking experiments indicated that the same effectors recognized non-H-2 antigens of DBA/2 and the cross-reacting YC8 determinants. The antitumor effect was mediated by T-cells, since it was abrogated by treatment of effectors with anti-Thy 1.2 serum plus complement. These data indicate that determinants defined by cytotoxic T-lymphocytes are expressed on the BALB/c lymphoma YC8 and cross-react with non-H-2 antigens of DBA/2 and B10.D2 strains.
- Published
- 1983
10. Interleukin 12 potentiates the curative effect of a vaccine based on interleukin 2-transduced tumor cells
- Author
-
Vagliani, M., Rodolfo, M., Federica Cavallo, Parenza, M., Melani, C., Parmiani, G., Forni, G., and Colombo, M. P.
- Subjects
Mice, Inbred BALB C ,Vaccines ,DNA, Complementary ,T-Lymphocytes ,Granulocyte-Macrophage Colony-Stimulating Factor ,Drug Synergism ,Adenocarcinoma ,Combined Modality Therapy ,Immunotherapy, Adoptive ,Interleukin-12 ,Killer Cells, Natural ,Mice ,Lymphocytes, Tumor-Infiltrating ,Chemotherapy, Adjuvant ,Transduction, Genetic ,Colonic Neoplasms ,Animals ,Humans ,Interleukin-2 ,Cell Division - Abstract
The purpose of these studies was to determine whether systemic administration of recombinant interleukin 12 (rIL-12) is able to potentiate an initial, but insufficient T-cell antitumor response. Mice challenged with carcinoma cells engineered to release interleukin 2 (IL-2) and displaying such a response received single or multiple i.p. injections of rIL-12. This combination of systemic rIL-12 and local IL-2 increased the percentage of mice that rejected two different IL-2 gene-transduced tumors. In another set of experiments more closely resembling a clinical situation, IL-2 gene-transduced tumors were used as vaccines in an attempt to cure mice bearing wild-type parental tumors. The combination of these vaccines with systemic rIL-12 cured mice more effectively than rIL-12 and IL-2 gene-transduced tumor vaccines alone.
11. Interleukin 12 gene therapy of MHC-negative murine melanoma metastases
- Author
-
Nanni, P., Rossi, I., Giovanni, C., Landuzzi, L., Nicoletti, G., antonella stoppacciaro, Parenza, M., Colombo, M. P., and Lollini, P. -L
- Subjects
Cytotoxicity, Immunologic ,Male ,Lung Neoplasms ,Neovascularization, Pathologic ,H-2 Antigens ,Melanoma, Experimental ,Mice, Nude ,Genetic Therapy ,Cancer Vaccines ,Interleukin-12 ,Major Histocompatibility Complex ,Mice, Inbred C57BL ,Immunocompromised Host ,Mice ,Transduction, Genetic ,Animals ,Neoplasm Metastasis - Abstract
Immunological gene therapy of cancer relies heavily on the activation of T cells, but tumors with defects in MHC gene expression are not recognized by MHC-restricted T cells. To investigate the potential of cytokine genes for the therapy of MHC-negative tumors, we transduced B78H1, a class I-negative murine melanoma clone, with a polycistronic vector carrying murine interleukin (IL)-12 genes. The clones studied produced 400-25,000 pg/ml IL-12; their in vitro growth properties were similar to those of parental cells. A complete inhibition of growth was observed in vivo both after s.c. and i.v. administration of all IL-12 clones. IL-12-transduced cells were also used as a therapeutic vaccine in mice bearing micrometastases by nontransduced parental cells. A significant (80-90%) reduction in the number of lung nodules was obtained. Immunohistochemical analysis and studies in immunocompromised hosts showed that T cells and natural killer cells had a significant role in the elimination of IL-12-releasing cells. In situ hybridization with cytokine probes detected a strong increase in the proportion of leukocytes positive for IFN-gamma, tumor necrosis factor alpha, IL-1beta, and IFN-inducible protein 10 at the site of rejection of IL-12-engineered tumor cells. However, it was clear that the loss of in vivo growth was also due to T-cell- and natural killer cell-independent factors, possibly related to the antiangiogenic properties of IL-12. In conclusion, tumor therapy based on IL-12 gene transduction was effective on a MHC-negative metastatic tumor, suggesting a possible application to MHC-defective human neoplasms.
12. Regression of an established tumor genetically modified to release granulocyte colony-stimulating factor requires granulocyte-T cell cooperation and T cell-produced interferon γ
- Author
-
Stoppacciaro, A., Melani, C., Parenza, M., Mastracchio, A., Bassi, C., Baroni, C., Parmiam, G., and Mario Paolo Colombo
13. HOXB7: A key factor for tumor-associated angiogenic switch
- Author
-
Alessandra Carè, Felicetti, F., Meccia, E., Bottero, L., Parenza, M., Stoppacciaro, A., Peschle, C., and Colombo, M. P.
14. Imatinib spares cKit expressing prostate neuroendocrine tumors, whereas kills seminal vesicle epithelial-stromal tumors targeting PDGFR-β
- Author
-
Jachetti E, Alice Rigoni, Bongiovanni L, Arioli I, Botti L, Parenza M, Cancila V, Chiodoni C, Festinese F, Bellone M, Tardanico R, Tripodo C, and Mp, Colombo
15. Amount of interleukin 12 available at the tumor site is critical for tumor regression
- Author
-
Colombo, M. P., Vagliani, M., Spreafico, F., Parenza, M., Claudia Chiodoni, Melani, C., and Stoppacciaro, A.
- Subjects
CD4-Positive T-Lymphocytes ,Mice ,Mice, Inbred BALB C ,Base Sequence ,Colonic Neoplasms ,Molecular Sequence Data ,Animals ,Adenocarcinoma ,Immunologic Memory ,Interleukin-12 ,Recombinant Proteins ,DNA Primers - Abstract
The C26 colon carcinoma is resistant to systemic recombinant interleukin 12 (rIL-12) therapy. Transduction of C26 with genes encoding the two subunits of murine IL-12 to release 30-80 pg/ml resulted in delayed tumor onset after injection of 5 x 10(4) cells into syngeneic BALB/c mice and in 40% tumor regression after injection into CD4-depleted mice. Here, we analyzed the activity of rIL-12 (1 microgram/day) against C26 grown into CD4-depleted mice. Like in mice given injections of interleukin 12 (IL-12) gene-transduced C26 cells, depletion of CD4+ cells led to tumor regression in 6 of 14 mice, and immumocytochemical characterization of tumor-infiltrating leukocytes showed abundant infiltration by CD8+ T cells and asialoGM1+ natural killer cells, which were scanty in tumors from nondepleted mice. On the basis of the percentage of tumor regression and leukocyte infiltration we can conclude that, in the C26 system, systemic rIL-12 (1 pmicrogramg/day) produces the same results as 30-80 pg/ml IL-12 released at the tumor site. A new polycistronic retroviral vector was then used to increase the amount of IL-12 produced by C26-transduced cells. C26 cells releasing 5 ng/ml IL-12, nearly 100 times more than the above-mentioned transduced cells, were tumorigenic in less than 50% of the mice given injections of 5 x 10(4) cells. In mice given injections of 5 x 10(5) cells, an initial tumor take of 100% followed by a complete tumor regression. Tumor regression was associated with infiltration of CD8+ and asialoGM1+ cells, and mice that remained tumor free were immune to a rechallenge of nontransduced C26 cells. The results indicate that the amount of IL-12 made available at the tumor site may determine both the type and number of infiltrating leukocytes and the events leading to tumor regression as well as it may overcame host immunosuppression.
16. Interleukin 12 potentiates the curative effect of a vaccine based on interleukin 2-transduced tumor cells
- Author
-
Vagliani, M., Rodolfo, M., Cavallo, F., Parenza, M., Melani, C., Parmiani, G., Forni, G., and Colombo, M.P.
- Subjects
Interleukins -- Health aspects ,Cancer -- Care and treatment ,Vaccines -- Research ,Business ,Health care industry - Abstract
According to the authors' abstract of an article published in Cancer Research, 'The purpose of these studies was to determine whether systemic administration of recombinant interleukin 12 (rIL-12) is able [...]
- Published
- 1996
17. Mesenchymal Transition of High-Grade Breast Carcinomas Depends on Extracellular Matrix Control of Myeloid Suppressor Cell Activity
- Author
-
Rosaria Orlandi, Mario P. Colombo, Claudia Chiodoni, Mariella Parenza, Sabina Sangaletti, Barbara Cappetti, Claudio Tripodo, Nadia Castioni, Paola Portararo, Elda Tagliabue, Alessandra Santangelo, Laura Botti, Alessandro Gulino, Sangaletti, S., Tripodo, C., Santangelo, A., Castioni, N., Portararo, P., Gulino, A., Botti, L., Parenza, M., Cappetti, B., Orlandi, R., Tagliabue, E., Chiodoni, C., and Colombo, M.
- Subjects
0301 basic medicine ,Myeloid ,MDSC ,Gene Expression ,medicine.disease_cause ,T-Lymphocytes, Regulatory ,Polyethylene Glycols ,Extracellular matrix ,Mice ,Breast cancer ,Myeloid Cells ,Osteonectin ,Mast Cells ,lcsh:QH301-705.5 ,Mice, Knockout ,Antigen Presentation ,Mice, Inbred BALB C ,EMT ,epithelial to mesenchymal transition ,COX-2 ,CXCL12 ,ECM ,G-CSF ,GM-CSF ,SPARC ,aminobisphosphonates ,cyclooxygenase-2 ,extracellular matrix ,granulocyte colony-stimulating factor ,granulocyte-macrophage colony-stimulating factor ,myeloid-derived suppressor cells ,Granulocyte macrophage colony-stimulating factor ,medicine.anatomical_structure ,Female ,medicine.drug ,Epithelial-Mesenchymal Transition ,Antineoplastic Agents ,Breast Neoplasms ,Biology ,Settore MED/08 - Anatomia Patologica ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Epithelial–mesenchymal transition ,Mesenchymal stem cell ,Xenograft Model Antitumor Assays ,Isogenic human disease models ,030104 developmental biology ,lcsh:Biology (General) ,Celecoxib ,Doxorubicin ,Immunology ,Cancer research ,Myeloid-derived Suppressor Cell ,aminobisphosphonate ,Neoplasm Grading ,Carcinogenesis - Abstract
SummaryThe extracellular matrix (ECM) contributes to the biological and clinical heterogeneity of breast cancer, and different prognostic groups can be identified according to specific ECM signatures. In high-grade, but not low-grade, tumors, an ECM signature characterized by high SPARC expression (ECM3) identifies tumors with increased epithelial-to-mesenchymal transition (EMT), reduced treatment response, and poor prognosis. To better understand how this ECM3 signature is contributing to tumorigenesis, we expressed SPARC in isogenic cell lines and found that SPARC overexpression in tumor cells reduces their growth rate and induces EMT. SPARC expression also results in the formation of a highly immunosuppressive microenvironment, composed by infiltrating T regulatory cells, mast cells, and myeloid-derived suppressor cells (MDSCs). The ability of SPARC to induce EMT depended on the localization and suppressive function of myeloid cells, and inhibition of the suppressive function MDSCs by administration of aminobisphosphonates could revert EMT, rendering SPARC-overexpressing tumor cells sensitive to Doxil. We conclude that that SPARC is regulating the interplay between MDSCs and the ECM to drive the induction of EMT in tumor cells.
- Published
- 2016
18. Imatinib spares cKit-expressing prostate neuroendocrine tumors, whereas kills seminal vesicle epithelial-stromal tumors by targeting PDGFR-β
- Author
-
Regina Tardanico, Fabrizio Festinese, Mario P. Colombo, Matteo Bellone, Mariella Parenza, Alice Rigoni, Claudia Chiodoni, Valeria Cancila, Ivano Arioli, Claudio Tripodo, Laura Botti, Lucia Bongiovanni, Elena Jachetti, Jachetti, E., Rigoni, A., Bongiovanni, L., Arioli, I., Botti, L., Parenza, M., Cancila, V., Chiodoni, C., Festinese, F., Bellone, M., Tardanico, R., Tripodo, C., and Colombo, M
- Subjects
0301 basic medicine ,Male ,Cancer Research ,Receptor tyrosine kinase ,Antineoplastic Agent ,Prostate cancer ,Mice ,0302 clinical medicine ,Prostate ,biology ,Seminal Vesicles ,Immunohistochemistry ,Gene Expression Regulation, Neoplastic ,Neuroendocrine Tumors ,Proto-Oncogene Proteins c-kit ,medicine.anatomical_structure ,Oncology ,030220 oncology & carcinogenesis ,Imatinib Mesylate ,Female ,Neuroendocrine Tumor ,medicine.drug ,Tramp ,Human ,Signal Transduction ,PCA3 ,medicine.medical_specialty ,Stromal cell ,Xenograft Model Antitumor Assay ,Protein Kinase Inhibitor ,Antineoplastic Agents ,Mice, Transgenic ,Receptor, Platelet-Derived Growth Factor beta ,03 medical and health sciences ,Internal medicine ,Seminal Vesicle ,medicine ,Animals ,Humans ,Protein Kinase Inhibitors ,Animal ,Prostatic Neoplasms ,Imatinib ,Biomarker ,medicine.disease ,Xenograft Model Antitumor Assays ,Disease Models, Animal ,030104 developmental biology ,Endocrinology ,Imatinib mesylate ,Prostatic Neoplasm ,biology.protein ,Cancer research ,Biomarkers - Abstract
Prostate cancer is a leading cause of cancer-related death in males worldwide. Indeed, advanced and metastatic disease characterized by androgen resistance and often associated with neuroendocrine (NE) differentiation remains incurable. Using the spontaneous prostate cancer TRAMP model, we have shown that mast cells (MCs) support in vivo the growth of prostate adenocarcinoma, whereas their genetic or pharmacologic targeting favors prostate NE cancer arousal. Aiming at simultaneously targeting prostate NE tumor cells and MCs, both expressing the cKit tyrosine kinase receptor, we have tested the therapeutic effect of imatinib in TRAMP mice. Imatinib-treated TRAMP mice experience a partial benefit against prostate adenocarcinoma, because of inhibition of supportive MCs. However, they show an unexpected outgrowth of prostate NE tumors, likely because of defective signaling pathway downstream of cKit receptor. Also unexpected but very effective was the inhibition of epithelial–stromal tumors of the seminal vesicles achieved by imatinib treatment. These tumors normally arise in the seminal vesicles of TRAMP mice, independently of the degree of prostatic glandular lesions, and resemble phyllodes tumors found in human prostate and seminal vesicles, and in breast. In both mice and in patients, these tumors are negative for cKit but express PDGFR-β, another tyrosine kinase receptor specifically inhibited by imatinib. Our results imply a possible detrimental effect of imatinib in prostate cancer patients but suggest a promising therapeutic application of imatinib in the treatment of recurrent or metastatic phyllodes tumors. Mol Cancer Ther; 16(2); 365–75. ©2016 AACR.
- Published
- 2017
19. Trabectedin Overrides Osteosarcoma Differentiative Block and Reprograms the Tumor Immune Environment Enabling Effective Combination with Immune Checkpoint Inhibitors
- Author
-
Cecilia Garofalo, Barbara Cappetti, Alessia Burocchi, Silvia Galvan, Laura Botti, Claudia Chiodoni, Mario P. Colombo, Chiara Ratti, Ilaria Torselli, Lucia Bongiovanni, Maria Cristina Manara, Katia Scotlandi, Sabina Sangaletti, Claudio Tripodo, Valeria Cancila, Mariella Parenza, Cesare Valenti, Ratti, C., Botti, L., Cancila, V., Galvan, S., Torselli, I., Garofalo, C., Manara, M., Bongiovanni, L., Valenti, C., Burocchi, A., Parenza, M., Cappetti, B., Sangaletti, S., Tripodo, C., Scotlandi, K., Colombo, M., and Chiodoni, C.
- Subjects
0301 basic medicine ,Cancer Research ,Lung Neoplasms ,medicine.medical_treatment ,Cellular differentiation ,T-Lymphocytes ,Programmed Cell Death 1 Receptor ,Bone Neoplasms ,Core Binding Factor Alpha 1 Subunit ,Dioxoles ,Biology ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Cell Line, Tumor ,Tetrahydroisoquinolines ,medicine ,Tumor Microenvironment ,Humans ,Trabectedin ,Tumor microenvironment ,Osteosarcoma ,Cancer ,Cell Differentiation ,Immunotherapy ,medicine.disease ,Cellular Reprogramming ,Primary tumor ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,Immunology ,Cancer research ,Osteosarcoma, Trabectedin, tumor mouse models, immune cells, immune checkpoint inhibitors ,Tumor Suppressor Protein p53 ,medicine.drug - Abstract
Purpose: Osteosarcoma, the most common primary bone tumor, is characterized by an aggressive behavior with high tendency to develop lung metastases as well as by multiple genetic aberrations that have hindered the development of targeted therapies. New therapeutic approaches are urgently needed; however, novel combinations with immunotherapies and checkpoint inhibitors require suitable preclinical models with intact immune systems to be properly tested. Experimental Design: We have developed immunocompetent osteosarcoma models that grow orthotopically in the bone and spontaneously metastasize to the lungs, mimicking human osteosarcoma. These models have been used to test the efficacy of trabectedin, a chemotherapeutic drug utilized clinically for sarcomas and ovarian cancer. Results: Trabectedin, as monotherapy, significantly inhibited osteosarcoma primary tumor growth and lung metastases by both targeting neoplastic cells and reprogramming the tumor immune microenvironment. Specifically, trabectedin induced a striking differentiation of tumor cells by favoring the recruitment of Runx2, the master genetic regulator of osteoblastogenesis, on the promoter of genes involved in the physiologic process of terminal osteoblast differentiation. Differentiated neoplastic cells, as expected, showed reduced proliferation rate. Concomitantly, trabectedin enhanced the number of tumor-infiltrating T lymphocytes, with local CD8 T cells, however, likely post-activated or exhausted, as suggested by their high expression of the inhibitory checkpoint molecule PD-1. Accordingly, the combination with a PD-1–blocking antibody significantly increased trabectedin efficacy in controlling osteosarcoma progression. Conclusions: These results demonstrate the therapeutic efficacy of trabectedin in osteosarcoma treatment, unveiling its multiple activities and providing a solid rationale for its combination with immune checkpoint inhibitors. Clin Cancer Res; 23(17); 5149–61. ©2017 AACR.
- Published
- 2016
20. Ultrasound-guided intra-tumor injection of combined immunotherapy cures mice from orthotopic prostate cancer
- Author
-
Mario P. Colombo, Claudia Chiodoni, Ivano Arioli, Claudio Tripodo, Giorgio Mauri, Mariella Parenza, Mauri, G, Chiodoni, C, Parenza, M, Arioli, I, Tripodo, C, and Colombo, MP.
- Subjects
Male ,Cancer Research ,Pathology ,medicine.medical_specialty ,Stromal cell ,medicine.medical_treatment ,Immunology ,Fluorescent Antibody Technique ,Gene delivery ,CD8-Positive T-Lymphocytes ,Injections, Intralesional ,prostate cancer ,immunotherapy ,Adenoviridae ,Immunoenzyme Techniques ,Prostate cancer ,Mice ,Tumor Cells, Cultured ,Immunology and Allergy ,Medicine ,Animals ,Humans ,Cell Proliferation ,Ultrasonography ,Tumor microenvironment ,business.industry ,Antibodies, Monoclonal ,Prostatic Neoplasms ,Immunotherapy ,T lymphocyte ,Genetic Therapy ,medicine.disease ,Combined Modality Therapy ,Interleukin-10 ,Mice, Inbred C57BL ,Oncology ,Oligodeoxyribonucleotides ,Chemokines, CC ,Systemic administration ,business ,CD8 - Abstract
Intra-tumor injection of immunotherapeutic agents is often the most effective, likely because of concomitant modification of tumor microenvironment. We tested an immunotherapeutic regimen consisting of CpG oligonucleotides and of adenovirus-mediated gene delivery of CCL16 chemokine directly into orthotopically implanted prostate tumors by ultrasound-guided injection, followed by systemic administration of an anti-IL-10R antibody. This combination treatment induced rapid stromal rearrangement, characterized by massive leukocyte infiltration and large areas of necrosis, a scenario that eventually led to complete tumor rejection and systemic immunity in 75 % of the treated mice. In vivo T lymphocyte depletion experiments demonstrated that the efficacy of CCL16/CpG/anti-IL-10R combination treatment relies upon CD8 T lymphocytes whereas CD4 T cells are dispensable. The results underlie the feasibility of echo-guided local immunotherapy of tumors located in visceral organs that are not easily accessible.
- Published
- 2013
21. Neutrophil extracellular traps mediate transfer of cytoplasmic neutrophil antigens to myeloid dendritic cells toward ANCA induction and associated autoimmunity
- Author
-
Caterina Vitali, Sabina Sangaletti, Claudio Tripodo, Mario P. Colombo, Carla Guarnotta, Patrizia Casalini, Cristiana Guiducci, Claudia Chiodoni, Barbara Cappetti, Mariella Parenza, Silvia Piconese, Sangaletti, S, Tripodo, C, Chiodoni, C, Guarnotta, C, Cappetti, B, Casalini, P, Piconese, S, Parenza, M, Guiducci, C, Vitali, C, and Colombo, MP
- Subjects
Myeloid ,Neutrophils ,Apoptosis ,Autoimmunity ,medicine.disease_cause ,Autoantigens ,Biochemistry ,Immunoenzyme Techniques ,Mice ,Cytosol ,Myeloid Cells ,Skin ,Mice, Inbred BALB C ,Reverse Transcriptase Polymerase Chain Reaction ,ANCA ,Cell Differentiation ,Hematology ,Flow Cytometry ,Acquired immune system ,Cell biology ,Respiratory burst ,medicine.anatomical_structure ,Female ,Neutrophil extracellular traps ,myeloid dendritic cells ,autoimmunity ,Programmed cell death ,Blotting, Western ,Immunology ,Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis ,Enzyme-Linked Immunosorbent Assay ,Biology ,Real-Time Polymerase Chain Reaction ,Antibodies, Antineutrophil Cytoplasmic ,Antigen ,medicine ,Animals ,Humans ,RNA, Messenger ,cardiovascular diseases ,Cell Proliferation ,Anti-neutrophil cytoplasmic antibody ,Dendritic Cells ,Cell Biology ,myeloid dendritic cell ,Mice, Inbred C57BL ,Immunization ,Neutrophil extracellular trap - Abstract
Antineutrophil cytoplasmic antibodies (ANCAs) target proteins normally retained within neutrophils, indicating that cell death is involved in the autoimmunity process. Still, ANCA pathogenesis remains obscure. ANCAs activate neutrophils inducing their respiratory burst and a peculiar form of cell death, named NETosis, characterized by formation of neutrophil extracellular traps (NETs), decondensed chromatin threads decorated with cytoplasmic proteins endorsed with antimicrobial activity. NETs have been consistently detected in ANCA-associated small-vessel vasculitis, and this association prompted us to test whether the peculiar structure of NET favors neutrophil proteins uploading into myeloid dendritic cells and the induction of ANCAs and associated autoimmunity. Here we show that myeloid DCs uploaded with and activated by NET components induce ANCA and autoimmunity when injected into naive mice. DC uploading and autoimmunity induction are prevented by NET treatment with DNAse, indicating that NET structural integrity is needed to maintain the antigenicity of cytoplasmic proteins. We found NET intermingling with myeloid dendritic cells also positive for neutrophil myeloperoxidase in myeloperoxidase-ANCA-associated microscopic poliangiitis providing a potential correlative picture in human pathology. These data provide the first demonstration that NET structures are highly immunogenic such to trigger adaptive immune response relevant for autoimmunity.
- Published
- 2012
22. SPARC oppositely regulates inflammation and fibrosis in bleomycin-induced lung damage
- Author
-
Ivano Arioli, Patrizia Casalini, Mario P. Colombo, Mariella Parenza, Sabina Sangaletti, Silvia Miotti, Claudio Tripodo, Alessandra Santangelo, Barbara Cappetti, Claudia Chiodoni, Silvia Piconese, Sangaletti, S, Tripodo, C, Cappetti, B, Casalini, P, Chiodoni, C, Piconese, S, Santangelo, A, Parenza, M, Arioli, I, Miotti, S, and Colombo, MP.
- Subjects
Pathology ,medicine.medical_specialty ,Animals ,Bleomycin ,Bone Marrow Cells ,Chimera ,Collagen ,Down-Regulation ,Fibroblasts ,Leukocytes ,Macrophages ,Mice ,Mice, Inbred BALB C ,Osteonectin ,Pneumonia ,Pulmonary Fibrosis ,Transforming Growth Factor beta ,Tumor Necrosis Factor-alpha ,Inflammation ,Biology ,Pathology and Forensic Medicine ,Fibrosis ,Tumor necrosis factor production ,Pulmonary fibrosis ,medicine ,Inbred BALB C ,Matricellular protein ,Regular Article ,SPARC ,Transforming growth factor beta ,BLEOMYCIN ,medicine.disease ,LUNG DAMAGE ,Cancer research ,biology.protein ,Tumor necrosis factor alpha ,medicine.symptom - Abstract
Fibrosis results from inflammatory tissue damage and impaired regeneration. In the context of bleomycin-induced pulmonary fibrosis, we demonstrated that the matricellular protein termed secreted protein acidic and rich in cysteine (SPARC) distinctly regulates inflammation and collagen deposition, depending on its cellular origin. Reciprocal Sparc(-/-) and wild-type (WT) bone marrow chimeras revealed that SPARC expression in host fibroblasts is required and sufficient to induce collagen fibrosis in a proper inflammatory environment. Accordingly, Sparc(-/-) >WT chimeras showed exacerbated inflammation and fibrosis due to the inability of Sparc(-/-) macrophages to down-regulate tumor necrosis factor production because of impaired responses to tumor growth factor-β. Hence, the use of bone marrow cells expressing a dominant-negative form of tumor growth factor-β receptor type II under the monocyte-specific CD68 promoter, as a decoy, phenocopied Sparc(-/-) donor chimeras. Our results point to an unexpected dual role of SPARC in oppositely influencing the outcome of fibrosis.
- Published
- 2011
23. Mast cell targeting hampers prostate adenocarcinoma development but promotes the occurrence of highly malignant neuroendocrine cancers
- Author
-
Mariella Parenza, Giorgio Mauri, Paola Pittoni, Mario P. Colombo, Alice Rigoni, Sabina Sangaletti, Claudio Tripodo, Silvia Piconese, Pittoni, P., Tripodo, C., Piconese, S., Mauri, G., Parenza, M., Rigoni, A., Sangaletti, S., and Colombo, M.
- Subjects
Male ,Oncology ,Cancer Research ,medicine.medical_specialty ,Epithelial-Mesenchymal Transition ,Mice, Transgenic ,Adenocarcinoma ,Biology ,medicine.disease_cause ,Cell Degranulation ,Mice ,Prostate cancer ,Prostate ,Cell Line, Tumor ,Internal medicine ,Tumor Microenvironment ,medicine ,Mast Cell ,Animals ,Humans ,Mast Cells ,Receptors, Tumor Necrosis Factor, Member 25 ,Tumor microenvironment ,Carcinoma, Neuroendocrine ,Disease Progression ,Matrix Metalloproteinase 9 ,Mice, Inbred C57BL ,Prostatic Neoplasms ,Proto-Oncogene Proteins c-kit ,Animal ,Cancer ,medicine.disease ,humanities ,medicine.anatomical_structure ,Tumor progression ,Prostatic Neoplasm ,Carcinogenesis ,Human ,Tramp - Abstract
Mast cells (MC) are c-Kit–expressing cells, best known for their primary involvement in allergic reactions, but recently reappraised as important players in either cancer promotion or inhibition. Here, we assessed the role of MCs in prostate tumor development. In prostate tumors from both tumor-prone transgenic adenocarcinoma of the mouse prostate (TRAMP) mice and human patients, MCs are specifically enriched and degranulated in areas of well-differentiated (WD) adenocarcinoma but not around poorly differentiated (PD) foci that coexist in the same tumors. We derived novel TRAMP tumor cell lines, representative of WD and PD variants, and through pharmacologic stabilization or genetic ablation of MCs in recipients mice, we showed that MCs promote WD adenocarcinoma growth but are dispensable for PD tumors. WD tumors rely on MCs for matrix metalloprotease 9 (MMP-9) provision, as reconstitution of MC-deficient mice with wild-type but not MMP-9−/− MCs was sufficient to promote their growth. In contrast, PD tumors are MMP-9 self-competent, consistently with epithelial-to-mesenchymal transition. Such a dual source of MMP-9 was confirmed in human tumors, suggesting that MCs could be a good target for early-stage prostate cancer. Interestingly, in testing whether MC targeting could block or delay tumorigenesis in tumor-prone TRAMP mice, we observed a high incidence of early and aggressive tumors, characterized by a neuroendocrine (NE) signature and c-Kit expression. Taken together, these data underscore the contribution of MCs in tumor progression and uncover a new, opposite role of MCs in protecting against the occurrence of aggressive NE variants in prostate cancer. Cancer Res; 71(18); 5987–97. ©2011 AACR.
- Published
- 2011
24. Antibody-mediated blockade of JMJD6 interaction with collagen I exerts antifibrotic and antimetastatic activities.
- Author
-
Miotti S, Gulino A, Ferri R, Parenza M, Chronowska A, Lecis D, Sangaletti S, Tagliabue E, Tripodo C, and Colombo MP
- Subjects
- Animals, Cell Line, Tumor, Cell Nucleus metabolism, Collagen Type I genetics, Enzyme-Linked Immunosorbent Assay, Extracellular Matrix genetics, Extracellular Matrix metabolism, Humans, Immunohistochemistry, Mice, Mice, Inbred BALB C, Mice, Knockout, Osteonectin genetics, Osteonectin metabolism, Peptide Library, Protein Binding, Receptors, Cell Surface genetics, Signal Transduction genetics, Signal Transduction physiology, Xenograft Model Antitumor Assays, Antibodies, Monoclonal metabolism, Collagen Type I metabolism, Receptors, Cell Surface metabolism
- Abstract
JMJD6 is known to localize in the nucleus, exerting histone arginine demethylase and lysyl hydroxylase activities. A novel localization of JMJD6 in the extracellular matrix, resulting from its secretion as a soluble protein, was unveiled by a new anti-JMJD6 mAb called P4E11, which was developed to identify new targets in the stroma. Recombinant JMJD6 binds with collagen type I (Coll-I), and distinct JMJD6 peptides interfere with collagen fibrillogenesis, collagen-fibronectin interaction, and adhesion of human tumor cells to the collagen substrate. P4E11 and collagen binding to JMJD6 are mutually exclusive because the amino acid sequences of JMJD6 necessary for the interaction with Coll-I are part of the conformational epitope recognized by P4E11. In mice injected with mouse 4T1 breast carcinoma cells, treatment with P4E11 reduced fibrosis at the primary tumor and prevented lung metastases. Reduction of fibrosis has also been documented in human breast and ovarian tumors (MDA-MB-231 and IGROV1, respectively) xenotransplanted into immunodeficient mice treated with P4E11. In summary, this study uncovers a new localization and function for JMJD6 that is most likely independent from its canonical enzymatic activities, and demonstrates that JMJD6 can functionally interact with Coll-I. P4E11 mAb, inhibiting JMJD6/Coll-I interaction, represents a new opportunity to target fibrotic and tumor diseases.-Miotti, S., Gulino, A., Ferri, R., Parenza, M., Chronowska, A., Lecis, D., Sangaletti, S., Tagliabue, E., Tripodo, C., Colombo, M. P. Antibody-mediated blockade of JMJD6 interaction with collagen I exerts antifibrotic and antimetastatic activities., (© FASEB.)
- Published
- 2017
- Full Text
- View/download PDF
25. Trabectedin Overrides Osteosarcoma Differentiative Block and Reprograms the Tumor Immune Environment Enabling Effective Combination with Immune Checkpoint Inhibitors.
- Author
-
Ratti C, Botti L, Cancila V, Galvan S, Torselli I, Garofalo C, Manara MC, Bongiovanni L, Valenti CF, Burocchi A, Parenza M, Cappetti B, Sangaletti S, Tripodo C, Scotlandi K, Colombo MP, and Chiodoni C
- Subjects
- Bone Neoplasms genetics, Bone Neoplasms immunology, Bone Neoplasms pathology, Cell Differentiation drug effects, Cell Line, Tumor, Cellular Reprogramming drug effects, Cellular Reprogramming immunology, Core Binding Factor Alpha 1 Subunit genetics, Dioxoles administration & dosage, Humans, Immunotherapy methods, Lung Neoplasms genetics, Lung Neoplasms pathology, Lung Neoplasms secondary, Osteosarcoma genetics, Osteosarcoma immunology, Osteosarcoma pathology, Programmed Cell Death 1 Receptor antagonists & inhibitors, Programmed Cell Death 1 Receptor immunology, T-Lymphocytes drug effects, T-Lymphocytes immunology, Tetrahydroisoquinolines administration & dosage, Trabectedin, Tumor Microenvironment drug effects, Tumor Microenvironment immunology, Tumor Suppressor Protein p53 genetics, Bone Neoplasms drug therapy, Dioxoles adverse effects, Lung Neoplasms drug therapy, Osteosarcoma drug therapy, Tetrahydroisoquinolines adverse effects
- Abstract
Purpose: Osteosarcoma, the most common primary bone tumor, is characterized by an aggressive behavior with high tendency to develop lung metastases as well as by multiple genetic aberrations that have hindered the development of targeted therapies. New therapeutic approaches are urgently needed; however, novel combinations with immunotherapies and checkpoint inhibitors require suitable preclinical models with intact immune systems to be properly tested. Experimental Design: We have developed immunocompetent osteosarcoma models that grow orthotopically in the bone and spontaneously metastasize to the lungs, mimicking human osteosarcoma. These models have been used to test the efficacy of trabectedin, a chemotherapeutic drug utilized clinically for sarcomas and ovarian cancer. Results: Trabectedin, as monotherapy, significantly inhibited osteosarcoma primary tumor growth and lung metastases by both targeting neoplastic cells and reprogramming the tumor immune microenvironment. Specifically, trabectedin induced a striking differentiation of tumor cells by favoring the recruitment of Runx2, the master genetic regulator of osteoblastogenesis, on the promoter of genes involved in the physiologic process of terminal osteoblast differentiation. Differentiated neoplastic cells, as expected, showed reduced proliferation rate. Concomitantly, trabectedin enhanced the number of tumor-infiltrating T lymphocytes, with local CD8 T cells, however, likely post-activated or exhausted, as suggested by their high expression of the inhibitory checkpoint molecule PD-1. Accordingly, the combination with a PD-1-blocking antibody significantly increased trabectedin efficacy in controlling osteosarcoma progression. Conclusions: These results demonstrate the therapeutic efficacy of trabectedin in osteosarcoma treatment, unveiling its multiple activities and providing a solid rationale for its combination with immune checkpoint inhibitors. Clin Cancer Res; 23(17); 5149-61. ©2017 AACR ., (©2017 American Association for Cancer Research.)
- Published
- 2017
- Full Text
- View/download PDF
26. Imatinib Spares cKit-Expressing Prostate Neuroendocrine Tumors, whereas Kills Seminal Vesicle Epithelial-Stromal Tumors by Targeting PDGFR-β.
- Author
-
Jachetti E, Rigoni A, Bongiovanni L, Arioli I, Botti L, Parenza M, Cancila V, Chiodoni C, Festinese F, Bellone M, Tardanico R, Tripodo C, and Colombo MP
- Subjects
- Animals, Biomarkers, Disease Models, Animal, Female, Humans, Immunohistochemistry, Male, Mice, Mice, Transgenic, Neuroendocrine Tumors drug therapy, Neuroendocrine Tumors pathology, Prostatic Neoplasms drug therapy, Prostatic Neoplasms pathology, Protein Kinase Inhibitors pharmacology, Proto-Oncogene Proteins c-kit metabolism, Receptor, Platelet-Derived Growth Factor beta genetics, Receptor, Platelet-Derived Growth Factor beta metabolism, Seminal Vesicles metabolism, Seminal Vesicles pathology, Signal Transduction drug effects, Xenograft Model Antitumor Assays, Antineoplastic Agents pharmacology, Gene Expression Regulation, Neoplastic drug effects, Imatinib Mesylate pharmacology, Neuroendocrine Tumors genetics, Neuroendocrine Tumors metabolism, Prostatic Neoplasms genetics, Prostatic Neoplasms metabolism, Proto-Oncogene Proteins c-kit genetics, Receptor, Platelet-Derived Growth Factor beta antagonists & inhibitors
- Abstract
Prostate cancer is a leading cause of cancer-related death in males worldwide. Indeed, advanced and metastatic disease characterized by androgen resistance and often associated with neuroendocrine (NE) differentiation remains incurable. Using the spontaneous prostate cancer TRAMP model, we have shown that mast cells (MCs) support in vivo the growth of prostate adenocarcinoma, whereas their genetic or pharmacologic targeting favors prostate NE cancer arousal. Aiming at simultaneously targeting prostate NE tumor cells and MCs, both expressing the cKit tyrosine kinase receptor, we have tested the therapeutic effect of imatinib in TRAMP mice. Imatinib-treated TRAMP mice experience a partial benefit against prostate adenocarcinoma, because of inhibition of supportive MCs. However, they show an unexpected outgrowth of prostate NE tumors, likely because of defective signaling pathway downstream of cKit receptor. Also unexpected but very effective was the inhibition of epithelial-stromal tumors of the seminal vesicles achieved by imatinib treatment. These tumors normally arise in the seminal vesicles of TRAMP mice, independently of the degree of prostatic glandular lesions, and resemble phyllodes tumors found in human prostate and seminal vesicles, and in breast. In both mice and in patients, these tumors are negative for cKit but express PDGFR-β, another tyrosine kinase receptor specifically inhibited by imatinib. Our results imply a possible detrimental effect of imatinib in prostate cancer patients but suggest a promising therapeutic application of imatinib in the treatment of recurrent or metastatic phyllodes tumors. Mol Cancer Ther; 16(2); 365-75. ©2016 AACR., (©2016 American Association for Cancer Research.)
- Published
- 2017
- Full Text
- View/download PDF
27. Mesenchymal Transition of High-Grade Breast Carcinomas Depends on Extracellular Matrix Control of Myeloid Suppressor Cell Activity.
- Author
-
Sangaletti S, Tripodo C, Santangelo A, Castioni N, Portararo P, Gulino A, Botti L, Parenza M, Cappetti B, Orlandi R, Tagliabue E, Chiodoni C, and Colombo MP
- Subjects
- Animals, Antigen Presentation, Antineoplastic Agents pharmacology, Breast Neoplasms drug therapy, Breast Neoplasms genetics, Breast Neoplasms pathology, Celecoxib pharmacology, Cell Line, Tumor, Doxorubicin analogs & derivatives, Doxorubicin pharmacology, Epithelial-Mesenchymal Transition genetics, Extracellular Matrix pathology, Female, Gene Expression, Humans, Mast Cells drug effects, Mast Cells immunology, Mast Cells pathology, Mice, Mice, Inbred BALB C, Mice, Knockout, Myeloid Cells drug effects, Myeloid Cells pathology, Myeloid-Derived Suppressor Cells drug effects, Myeloid-Derived Suppressor Cells pathology, Neoplasm Grading, Osteonectin deficiency, Polyethylene Glycols pharmacology, T-Lymphocytes, Regulatory drug effects, T-Lymphocytes, Regulatory immunology, T-Lymphocytes, Regulatory pathology, Xenograft Model Antitumor Assays, Breast Neoplasms immunology, Epithelial-Mesenchymal Transition immunology, Extracellular Matrix immunology, Myeloid Cells immunology, Myeloid-Derived Suppressor Cells immunology, Osteonectin genetics
- Abstract
The extracellular matrix (ECM) contributes to the biological and clinical heterogeneity of breast cancer, and different prognostic groups can be identified according to specific ECM signatures. In high-grade, but not low-grade, tumors, an ECM signature characterized by high SPARC expression (ECM3) identifies tumors with increased epithelial-to-mesenchymal transition (EMT), reduced treatment response, and poor prognosis. To better understand how this ECM3 signature is contributing to tumorigenesis, we expressed SPARC in isogenic cell lines and found that SPARC overexpression in tumor cells reduces their growth rate and induces EMT. SPARC expression also results in the formation of a highly immunosuppressive microenvironment, composed by infiltrating T regulatory cells, mast cells, and myeloid-derived suppressor cells (MDSCs). The ability of SPARC to induce EMT depended on the localization and suppressive function of myeloid cells, and inhibition of the suppressive function MDSCs by administration of aminobisphosphonates could revert EMT, rendering SPARC-overexpressing tumor cells sensitive to Doxil. We conclude that that SPARC is regulating the interplay between MDSCs and the ECM to drive the induction of EMT in tumor cells., (Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
28. Ultrasound-guided intra-tumor injection of combined immunotherapy cures mice from orthotopic prostate cancer.
- Author
-
Mauri G, Chiodoni C, Parenza M, Arioli I, Tripodo C, and Colombo MP
- Subjects
- Adenoviridae genetics, Animals, Antibodies, Monoclonal pharmacology, CD8-Positive T-Lymphocytes immunology, Cell Proliferation, Combined Modality Therapy, Fluorescent Antibody Technique, Humans, Immunoenzyme Techniques, Injections, Intralesional, Interleukin-10 immunology, Interleukin-10 metabolism, Male, Mice, Mice, Inbred C57BL, Prostatic Neoplasms diagnostic imaging, Prostatic Neoplasms immunology, Tumor Cells, Cultured, Chemokines, CC genetics, Genetic Therapy, Immunotherapy, Interleukin-10 antagonists & inhibitors, Oligodeoxyribonucleotides genetics, Prostatic Neoplasms therapy, Ultrasonography
- Abstract
Intra-tumor injection of immunotherapeutic agents is often the most effective, likely because of concomitant modification of tumor microenvironment. We tested an immunotherapeutic regimen consisting of CpG oligonucleotides and of adenovirus-mediated gene delivery of CCL16 chemokine directly into orthotopically implanted prostate tumors by ultrasound-guided injection, followed by systemic administration of an anti-IL-10R antibody. This combination treatment induced rapid stromal rearrangement, characterized by massive leukocyte infiltration and large areas of necrosis, a scenario that eventually led to complete tumor rejection and systemic immunity in 75 % of the treated mice. In vivo T lymphocyte depletion experiments demonstrated that the efficacy of CCL16/CpG/anti-IL-10R combination treatment relies upon CD8 T lymphocytes whereas CD4 T cells are dispensable. The results underlie the feasibility of echo-guided local immunotherapy of tumors located in visceral organs that are not easily accessible.
- Published
- 2013
- Full Text
- View/download PDF
29. Neutrophil extracellular traps mediate transfer of cytoplasmic neutrophil antigens to myeloid dendritic cells toward ANCA induction and associated autoimmunity.
- Author
-
Sangaletti S, Tripodo C, Chiodoni C, Guarnotta C, Cappetti B, Casalini P, Piconese S, Parenza M, Guiducci C, Vitali C, and Colombo MP
- Subjects
- Animals, Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis pathology, Apoptosis, Autoantigens, Blotting, Western, Cell Differentiation, Cell Proliferation, Cytosol metabolism, Dendritic Cells metabolism, Enzyme-Linked Immunosorbent Assay, Female, Flow Cytometry, Humans, Immunization, Immunoenzyme Techniques, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Myeloid Cells metabolism, Neutrophils metabolism, RNA, Messenger genetics, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Skin immunology, Skin metabolism, Skin pathology, Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis etiology, Antibodies, Antineutrophil Cytoplasmic blood, Antibodies, Antineutrophil Cytoplasmic immunology, Autoimmunity immunology, Cytosol immunology, Dendritic Cells immunology, Myeloid Cells immunology, Neutrophils immunology
- Abstract
Antineutrophil cytoplasmic antibodies (ANCAs) target proteins normally retained within neutrophils, indicating that cell death is involved in the autoimmunity process. Still, ANCA pathogenesis remains obscure. ANCAs activate neutrophils inducing their respiratory burst and a peculiar form of cell death, named NETosis, characterized by formation of neutrophil extracellular traps (NETs), decondensed chromatin threads decorated with cytoplasmic proteins endorsed with antimicrobial activity. NETs have been consistently detected in ANCA-associated small-vessel vasculitis, and this association prompted us to test whether the peculiar structure of NET favors neutrophil proteins uploading into myeloid dendritic cells and the induction of ANCAs and associated autoimmunity. Here we show that myeloid DCs uploaded with and activated by NET components induce ANCA and autoimmunity when injected into naive mice. DC uploading and autoimmunity induction are prevented by NET treatment with DNAse, indicating that NET structural integrity is needed to maintain the antigenicity of cytoplasmic proteins. We found NET intermingling with myeloid dendritic cells also positive for neutrophil myeloperoxidase in myeloperoxidase-ANCA-associated microscopic poliangiitis providing a potential correlative picture in human pathology. These data provide the first demonstration that NET structures are highly immunogenic such to trigger adaptive immune response relevant for autoimmunity.
- Published
- 2012
- Full Text
- View/download PDF
30. SPARC oppositely regulates inflammation and fibrosis in bleomycin-induced lung damage.
- Author
-
Sangaletti S, Tripodo C, Cappetti B, Casalini P, Chiodoni C, Piconese S, Santangelo A, Parenza M, Arioli I, Miotti S, and Colombo MP
- Subjects
- Animals, Bleomycin toxicity, Bone Marrow Cells metabolism, Chimera, Collagen metabolism, Down-Regulation, Leukocytes physiology, Mice, Mice, Inbred BALB C, Osteonectin metabolism, Transforming Growth Factor beta metabolism, Tumor Necrosis Factor-alpha biosynthesis, Fibroblasts metabolism, Macrophages physiology, Osteonectin physiology, Pneumonia chemically induced, Pulmonary Fibrosis chemically induced, Transforming Growth Factor beta physiology
- Abstract
Fibrosis results from inflammatory tissue damage and impaired regeneration. In the context of bleomycin-induced pulmonary fibrosis, we demonstrated that the matricellular protein termed secreted protein acidic and rich in cysteine (SPARC) distinctly regulates inflammation and collagen deposition, depending on its cellular origin. Reciprocal Sparc(-/-) and wild-type (WT) bone marrow chimeras revealed that SPARC expression in host fibroblasts is required and sufficient to induce collagen fibrosis in a proper inflammatory environment. Accordingly, Sparc(-/-) >WT chimeras showed exacerbated inflammation and fibrosis due to the inability of Sparc(-/-) macrophages to down-regulate tumor necrosis factor production because of impaired responses to tumor growth factor-β. Hence, the use of bone marrow cells expressing a dominant-negative form of tumor growth factor-β receptor type II under the monocyte-specific CD68 promoter, as a decoy, phenocopied Sparc(-/-) donor chimeras. Our results point to an unexpected dual role of SPARC in oppositely influencing the outcome of fibrosis., (Copyright © 2011 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)
- Published
- 2011
- Full Text
- View/download PDF
31. Mast cell targeting hampers prostate adenocarcinoma development but promotes the occurrence of highly malignant neuroendocrine cancers.
- Author
-
Pittoni P, Tripodo C, Piconese S, Mauri G, Parenza M, Rigoni A, Sangaletti S, and Colombo MP
- Subjects
- Adenocarcinoma enzymology, Animals, Carcinoma, Neuroendocrine enzymology, Cell Degranulation physiology, Cell Line, Tumor, Disease Progression, Epithelial-Mesenchymal Transition, Humans, Male, Mast Cells enzymology, Matrix Metalloproteinase 9 biosynthesis, Mice, Mice, Inbred C57BL, Mice, Transgenic, Prostatic Neoplasms enzymology, Proto-Oncogene Proteins c-kit biosynthesis, Receptors, Tumor Necrosis Factor, Member 25, Tumor Microenvironment, Adenocarcinoma pathology, Carcinoma, Neuroendocrine pathology, Mast Cells pathology, Prostatic Neoplasms pathology
- Abstract
Mast cells (MC) are c-Kit-expressing cells, best known for their primary involvement in allergic reactions, but recently reappraised as important players in either cancer promotion or inhibition. Here, we assessed the role of MCs in prostate tumor development. In prostate tumors from both tumor-prone transgenic adenocarcinoma of the mouse prostate (TRAMP) mice and human patients, MCs are specifically enriched and degranulated in areas of well-differentiated (WD) adenocarcinoma but not around poorly differentiated (PD) foci that coexist in the same tumors. We derived novel TRAMP tumor cell lines, representative of WD and PD variants, and through pharmacologic stabilization or genetic ablation of MCs in recipients mice, we showed that MCs promote WD adenocarcinoma growth but are dispensable for PD tumors. WD tumors rely on MCs for matrix metalloprotease 9 (MMP-9) provision, as reconstitution of MC-deficient mice with wild-type but not MMP-9(-/-) MCs was sufficient to promote their growth. In contrast, PD tumors are MMP-9 self-competent, consistently with epithelial-to-mesenchymal transition. Such a dual source of MMP-9 was confirmed in human tumors, suggesting that MCs could be a good target for early-stage prostate cancer. Interestingly, in testing whether MC targeting could block or delay tumorigenesis in tumor-prone TRAMP mice, we observed a high incidence of early and aggressive tumors, characterized by a neuroendocrine (NE) signature and c-Kit expression. Taken together, these data underscore the contribution of MCs in tumor progression and uncover a new, opposite role of MCs in protecting against the occurrence of aggressive NE variants in prostate cancer.
- Published
- 2011
- Full Text
- View/download PDF
32. Macrophage-derived SPARC bridges tumor cell-extracellular matrix interactions toward metastasis.
- Author
-
Sangaletti S, Di Carlo E, Gariboldi S, Miotti S, Cappetti B, Parenza M, Rumio C, Brekken RA, Chiodoni C, and Colombo MP
- Subjects
- Animals, Base Sequence, DNA Primers, Fibronectins metabolism, Flow Cytometry, Gene Silencing, Immunohistochemistry, Integrin beta Chains genetics, Mice, Mice, Inbred BALB C, Mice, Knockout, Osteonectin genetics, RNA, Small Interfering, Extracellular Matrix metabolism, Macrophages metabolism, Neoplasm Metastasis, Osteonectin physiology
- Abstract
Other than genetic imprinting and epithelial to mesenchymal transition, cancer cells need interaction with the nearby stroma toward metastasis. Secreted protein acidic and rich in cysteine (SPARC) is a matricellular protein known to regulate extracellular matrix (ECM) deposition and cell-ECM interaction. Gene expression profiles associate SPARC to malignant progression. Using reciprocal bone marrow chimeras between SPARC knockout and wild-type mice, we show that SPARC produced by inflammatory cells is necessary for spontaneous, but not experimental, i.v. metastasis. Macrophage-derived SPARC induces cancer cell migration and enhances their migration to other ECM proteins at least through alpha(v)beta(5) integrin. Indeed, RNA interference knockdown of beta(5) integrin expression reduces cell migration in vitro and metastasis in vivo. Together these results show that macrophage-derived SPARC takes part in metastasis, acting at the step of integrin-mediated migration of invasive cells.
- Published
- 2008
- Full Text
- View/download PDF
33. In Ewing's sarcoma CCN3(NOV) inhibits proliferation while promoting migration and invasion of the same cell type.
- Author
-
Benini S, Perbal B, Zambelli D, Colombo MP, Manara MC, Serra M, Parenza M, Martinez V, Picci P, and Scotlandi K
- Subjects
- Animals, Cell Adhesion, Cell Line, Cell Proliferation, Clone Cells metabolism, Clone Cells pathology, Connective Tissue Growth Factor, Female, Gene Expression Regulation, Neoplastic, Immediate-Early Proteins genetics, Intercellular Signaling Peptides and Proteins genetics, Matrix Metalloproteinases metabolism, Mice, Mice, Nude, Neoplasm Invasiveness, Nephroblastoma Overexpressed Protein, Receptor, IGF Type 1 metabolism, Receptor, Platelet-Derived Growth Factor alpha metabolism, Receptor, Platelet-Derived Growth Factor beta metabolism, Sarcoma, Ewing genetics, Transfection, Cell Movement, Immediate-Early Proteins metabolism, Intercellular Signaling Peptides and Proteins metabolism, Sarcoma, Ewing metabolism, Sarcoma, Ewing pathology
- Abstract
Altered expression of CCN3 has been observed in a variety of musculoskeletal tumours, including Ewing's sarcoma (ES). Despite its widespread distribution, very little is known about its biological functions and molecular mechanisms of action. We transfected CCN3 gene into a CCN3-negative ES cell line and analysed the in vitro and in vivo behaviours of stably transfected clones. Forced expression of CCN3 significantly reduced cell proliferation in vitro, growth in anchorage-independent conditions, and tumorigenicity in nude mice. Despite the antiproliferative effect, CCN3-transfected ES cells displayed increased migration and invasion of Matrigel. The decreased expression of alpha2beta1 integrin receptor and the increased amount of cell surface-associated matrix metalloproteinase (MMP)-9 following the expression of CCN3 may be the basis for the increased migratory abilities of transfected cells. Cells lacking alpha2beta1 are less facilitated to have stable anchorage since the predominant collagen extracted from ES tissue is indeed type I collagen, and proMMP-9 was recently found to provide a cellular switch between stationary and migratory ES cell phase. Our findings are in line with those recently obtained in glioblastoma. However, the underlying molecular mechanisms appear to be different, further highlighting the importance of the cellular context in the regulation of function of CCN proteins.
- Published
- 2005
- Full Text
- View/download PDF
34. Reconstitution of human telomerase reverse transcriptase expression rescues colorectal carcinoma cells from in vitro senescence: evidence against immortality as a constitutive trait of tumor cells.
- Author
-
Dalerba P, Guiducci C, Poliani PL, Cifola I, Parenza M, Frattini M, Gallino G, Carnevali I, Di Giulio I, Andreola S, Lombardo C, Rivoltini L, Schweighoffer T, Belli F, Colombo MP, Parmiani G, and Castelli C
- Subjects
- Animals, Cellular Senescence physiology, Colorectal Neoplasms genetics, DNA-Binding Proteins, Female, Genetic Vectors, Humans, Mice, Mice, SCID, Neoplasm Transplantation, RNA, Messenger biosynthesis, RNA, Messenger genetics, Retroviridae genetics, Telomerase genetics, Telomerase metabolism, Telomere physiology, Transduction, Genetic, Transplantation, Heterologous, Tumor Cells, Cultured, Colorectal Neoplasms enzymology, Colorectal Neoplasms pathology, Telomerase biosynthesis
- Abstract
Although in vitro establishment of new colorectal carcinoma (CRC) cell lines is an infrequent event, we have observed that primary cultures of CRC can be repeatedly and reproducibly initiated following in vitro plating of tumor-derived epithelial cells. These cultures, however, usually display a short life span as they undergo a limited number of cell passages before entering a state of irreversible growth arrest. In this study, we show that short-lived CRC primary cultures lack constitutive telomerase activity and undergo a senescence process characterized by progressive telomere shortening. Moreover, transduction of these cells with a retroviral vector encoding human telomerase reverse transcriptase (hTERT) is sufficient to reconstitute telomerase activity and allow immortalization. Detailed molecular characterization of hTERT-immortalized CRC cell lines confirms their individual tumor origin by showing expression of colonic epithelial differentiation markers, such as cytokeratin-20 (CK20), full match with class I and class II human leukocyte antigen genotyping of autologous B-lymphoblastoid cells, and presence of somatic mutations in key cancer genes (KRAS2, APC) identical to those of the corresponding autologous original tumor tissues. Moreover, functional characterization of hTERT-immortalized CRC cell lines shows that they have a transformed phenotype, being able to form colonies in soft agar and tumors in severe combined immunodeficient mice. Most interestingly, immunohistochemical analysis of original tumor tissues indicates that short-lived CRC primary cultures, although hTERT-negative in vitro, derive from hTERT-positive tumors. Taken together, our data show that, in a least subset of CRC, biochemical pathways involved in maintenance of telomere length, such as telomerase, are not activated in a constitutive way in all tumor cells.
- Published
- 2005
- Full Text
- View/download PDF
35. Intralesional injection of adenovirus encoding CC chemokine ligand 16 inhibits mammary tumor growth and prevents metastatic-induced death after surgical removal of the treated primary tumor.
- Author
-
Guiducci C, Di Carlo E, Parenza M, Hitt M, Giovarelli M, Musiani P, and Colombo MP
- Subjects
- Adenocarcinoma immunology, Adenocarcinoma secondary, Animals, Antigens, Neoplasm immunology, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes pathology, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes pathology, Cell Line, Tumor, Chemokines, CC genetics, Combined Modality Therapy methods, Combined Modality Therapy mortality, Dendritic Cells immunology, Dendritic Cells pathology, Epitopes, T-Lymphocyte immunology, Female, Genetic Vectors, Graft Rejection immunology, Graft Rejection mortality, Growth Inhibitors genetics, Humans, Inflammation genetics, Inflammation immunology, Inflammation pathology, Injections, Intralesional, Lung Neoplasms immunology, Lung Neoplasms mortality, Lung Neoplasms prevention & control, Lymph Nodes immunology, Lymph Nodes pathology, Lymphatic Metastasis immunology, Lymphatic Metastasis prevention & control, Mammary Neoplasms, Experimental immunology, Mammary Neoplasms, Experimental mortality, Mammary Neoplasms, Experimental therapy, Mice, Mice, Inbred BALB C, Mice, Knockout, Mice, Nude, T-Lymphocyte Subsets immunology, T-Lymphocyte Subsets pathology, Adenocarcinoma mortality, Adenocarcinoma surgery, Adenoviridae genetics, Antineoplastic Agents administration & dosage, Chemokines, CC administration & dosage, Growth Inhibitors administration & dosage, Lung Neoplasms secondary, Mammary Neoplasms, Experimental surgery
- Abstract
The CC chemokine ligand (CCL)16 exerts chemotactic activity on human monocytes and lymphocytes. Although no murine homologous has been defined, the TSA mouse adenocarcinoma cells engineered to express human CCL16 are rapidly rejected by syngenic mice. An adenovirus encoding CCL16 (AdCCL16) was generated using a Cre-Lox-based system and was used to determine whether this chemokine might also block pre-existing tumors. Both recombinant and viral CCL16 showed in vitro chemotactic activity for murine CD4(+) and CD8(+) lymphocytes and dendritic cells (DC). AdCCL16, but not the control empty vector, when injected in established nodules significantly delayed tumor growth. Immunohistochemistry revealed accumulation of CD4(+) and CD8(+) T cells and DC in the treated tumors as well as in draining lymph nodes. DC from such lymph nodes stimulated IFN-gamma by a T cell clone specific for the known TSA tumor-associated Ag (TAA), suggesting the tumor origin of these cells. Lymphocytes from the same nodes showed specific CTL activity against TSA tumor cells and their immunodominant TAA peptide. Antitumor activity required CD4, CD8, and IFN-gamma production, as shown using subset-depleted and knockout mice. Despite the robust and rapid immune response triggered by intratumoral injection of AdCCL16, the lesions were not completely rejected; however, the same treatment given before surgical excision of primary lesions prevented metastatic spread and cured 63% of mice bearing the 4T1 mammary adenocarcinoma, which is perhaps the most compelling model of spontaneous metastasis.
- Published
- 2004
- Full Text
- View/download PDF
36. Enhanced efficacy of tumor cell vaccines transfected with secretable hsp70.
- Author
-
Massa C, Guiducci C, Arioli I, Parenza M, Colombo MP, and Melani C
- Subjects
- Animals, Antigen-Presenting Cells physiology, Cell Line, Tumor, Cytotoxicity, Immunologic, Female, HSP70 Heat-Shock Proteins genetics, Immunologic Memory, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, T-Lymphocytes immunology, Transfection, HSP70 Heat-Shock Proteins metabolism, Immunotherapy, Adoptive, Neoplasms therapy
- Abstract
Tumor immunotherapy has exploited the ability of heat shock proteins to chaperone precursors of antigenic peptides to antigen-presenting cells and to activate efficiently an immune response against tumor-associated antigens. The most common strategy is based on the purification of heat shock protein-peptide complexes from tumor cell lines or from tumor surgical samples for in vivo administration. In this article, we have modified the murine-inducible hsp70 into a secreted protein and engineered tumor cells to secrete constitutively their antigenic repertoire associated with the hsp70 protein. In vitro studies showed that the relocalization of hsp70 from the cytoplasm to the secretory pathway did not modify the ability of hsp70 to interact with peptides derived either from natural tumor-associated antigens or model antigens, and that antigen-presenting cells specifically took up the secreted hsp70 and presented the chaperoned epitopes to T cells. In vivo studies showed that tumors secreting hsp70 displayed increased immunogenicity, with induction of a strong and specific CTL response. Mice injected with hsp70-secreting tumors showed increased survival and impaired tumor take compared with mice bearing parental tumors. More than 70% of mice rejected tumor cells secreting hsp70 through mechanisms that involve T lymphocytes and natural killer cells, with the induction of a memory response in the case of T lymphocytes. Moreover, hsp70 secretion increased the immunogenic potential of tumor cell vaccines.
- Published
- 2004
- Full Text
- View/download PDF
37. Autologous and MHC class I-negative allogeneic tumor cells secreting IL-12 together cure disseminated A20 lymphoma.
- Author
-
Curti A, Parenza M, and Colombo MP
- Subjects
- Animals, Bystander Effect immunology, Cytokines biosynthesis, Cytokines metabolism, Immunity, Cellular, Interleukin-12 biosynthesis, Interleukin-12 genetics, Interleukin-12 metabolism, Lymphocyte Activation, Lymphoma immunology, Mice, Mice, Inbred BALB C, Neoplasms, Experimental immunology, Neoplasms, Experimental pathology, Neoplasms, Experimental therapy, Receptors, OX40, T-Lymphocytes, Cytotoxic immunology, Transduction, Genetic, Treatment Outcome, Tumor Cells, Cultured, Tumor Necrosis Factor Receptor Superfamily, Member 7 metabolism, Cancer Vaccines administration & dosage, Cancer Vaccines immunology, Cancer Vaccines pharmacology, Histocompatibility Antigens Class I immunology, Lymphoma pathology, Lymphoma therapy, Receptors, Tumor Necrosis Factor
- Abstract
Cytokine gene-modified tumor cells have increased immunogenicity and retain the antigenic repertoire of a particular neoplasia. However, practical concerns have led to an increased interest in allogeneic gene-transduced bystander cells as a broader source of cytokines for autologous tumor cell-based vaccines. Here, we show that allogeneic B78H1 major histocompatibility complex (MHC) class I-negative and -positive (H-2K(b)- and D(b)-transfected) cells induced cytotoxic T lymphocytes (CTLs) and protection in BALB/c mice at comparable levels in response to a challenge with C26 (H-2(d)) colon carcinoma cells sharing the tumor-associated antigen envelope glycoprotein 70 (env-gp70) with both cell lines. Class I-negative B78H1 cells transduced to express interleukin-12 (IL-12) and mixed with autologous A20 tumor cells led to eradication of preestablished A20 lymphoma in 50% or 100% of treated mice after 3 or 4 vaccinations, respectively, whereas A20 cells alone or mixed with nontransduced B78H1 cured none or 50% of mice after 3 or 4 vaccinations, respectively. Immunization with the IL-12-producing bystander cell line increased tumor-specific proliferation and type 1 cytokine production by CD4(+) T cells. By contrast, CD4 T-cell function appeared impaired after immunization with A20 cells alone or mixed with B78H1 cells. Indeed, only CD4(+) T cells from IL-12-treated mice could be restimulated with anti-OX40 monoclonal antibody (mAb) in place of a fourth cellular boost. Moreover, the IL-12-based tumor vaccine induced expansion of tumor-specific interferon-gamma (IFN-gamma)-producing CD8(+) T cells. These results are clinically relevant for the development of feasible IL-12 cancer vaccines based on engineered class I-negative bystander cells.
- Published
- 2003
- Full Text
- View/download PDF
38. Dendritic cells infiltrating tumors cotransduced with granulocyte/macrophage colony-stimulating factor (GM-CSF) and CD40 ligand genes take up and present endogenous tumor-associated antigens, and prime naive mice for a cytotoxic T lymphocyte response.
- Author
-
Chiodoni C, Paglia P, Stoppacciaro A, Rodolfo M, Parenza M, and Colombo MP
- Subjects
- Animals, Antigens, Neoplasm immunology, CD40 Ligand, Colonic Neoplasms immunology, Female, Flow Cytometry, Mice, Mice, Inbred BALB C, Tumor Cells, Cultured, Antigens, Neoplasm metabolism, Dendritic Cells immunology, Granulocyte-Macrophage Colony-Stimulating Factor genetics, Membrane Glycoproteins genetics, T-Lymphocytes, Cytotoxic immunology
- Abstract
We transduced BALB/c-derived C-26 colon carcinoma cells with granulocyte/macrophage colony-stimulating factor (GM-CSF) and CD40 ligand (CD40L) genes to favor interaction of these cells with host dendritic cells (DCs) and, therefore, cross-priming. Cotransduced cells showed reduced tumorigenicity, and tumor take was followed by regression in some mice. In vivo tumors were heavily infiltrated with DCs that were isolated, phenotyped, and tested in vitro for stimulation of tumor-specific cytotoxic T lymphocytes (CTLs). BALB/c C-26 carcinoma cells express the endogenous murine leukemia virus (MuLV) env gene as a tumor-associated antigen. This antigen is shared among solid tumors of BALB/c and C57BL/6 mice and contains two epitopes, AH-1 and KSP, recognized in the context of major histocompatibility complex class I molecules H-2Ld and H-2K(b), respectively. DCs isolated from C-26/GM/CD40L tumors grown in (BALB/c x C57BL/6)F1 mice (H-2d x b) stimulated interferon gamma production by both anti-AH-1 and KSP CTLs, whereas tumor-infiltrating DCs (TIDCs) of BALB/c mice stimulated only anti-AH-1 CTLs. Furthermore, TIDCs primed naive mice for CTL activity as early as 2 d after injection into the footpad, whereas double-transduced tumor cells required at least 5 d for priming; this difference may reflect direct DC priming versus indirect tumor cell priming. Immunohistochemical staining indicated colocalization of DCs and apoptotic bodies in the tumors. These data indicate that DCs infiltrating tumors that produce GM-CSF and CD40L can capture cellular antigens, likely through uptake of apoptotic bodies, and mature in situ to a stage suitable for antigen presentation. Thus, tumor cell-based vaccines engineered to favor the interaction with host DCs can be considered.
- Published
- 1999
- Full Text
- View/download PDF
39. IgG2a induced by interleukin (IL) 12-producing tumor cell vaccines but not IgG1 induced by IL-4 vaccine is associated with the eradication of experimental metastases.
- Author
-
Rodolfo M, Melani C, Zilocchi C, Cappetti B, Luison E, Arioli I, Parenza M, Canevari S, and Colombo MP
- Subjects
- Animals, Antibodies, Monoclonal pharmacology, Antibodies, Neoplasm biosynthesis, Female, Interleukin-4 metabolism, Lung Neoplasms secondary, Lung Neoplasms therapy, Mice, Mice, Inbred BALB C, Vaccination, Cancer Vaccines, Immunoglobulin G biosynthesis, Interleukin-12 metabolism, Neoplasm Metastasis
- Abstract
We evaluated whether antibody response correlates with tumor therapy by cytokine gene-modified tumor cell vaccines. To characterize the antibody (Ab) response against a known antigen, colon carcinoma C26 cells and C26 variants engineered to produce interleukin (IL) 12 or IL-4 were further transduced to express the human tumor-associated antigen gp38 folate receptor (FR) alpha. Irradiated IL-12- and IL-4-producing C26/FR alpha cell vaccines cured 50 and 30% of mice bearing C26/FR alpha lung micrometastases. Treatment induced a rapid, CD4-dependent Ab production dominated by IgG2a and IgG1 in response to the IL-12 or IL-4 vaccine, respectively. In contrast, untreated tumor-bearing mice showed a late serological response dominated by IgM. Anti-FR alpha IgG1 and IgG2a were able to suppress tumor metastases upon passive transfer in vivo. Sera from mice cured by the IL-12 vaccine displayed a higher binding activity, a higher anti-FR alpha IgG2a content, and a higher complement-mediated tumor cell lysis in vitro compared to the sera from nonresponder mice. Such a correlation was not found in the sera of mice treated with the IL-4 vaccine. These data indicate that cytokine-producing tumor cell vaccines strongly influence antibody response, and that in the case of the IL-12-based vaccine, the Ab titer correlates with the therapeutic response, thus suggesting its use for monitoring the outcome of vaccination in cancer patients.
- Published
- 1998
40. Interferon gamma-independent rejection of interleukin 12-transduced carcinoma cells requires CD4+ T cells and Granulocyte/Macrophage colony-stimulating factor.
- Author
-
Zilocchi C, Stoppacciaro A, Chiodoni C, Parenza M, Terrazzini N, and Colombo MP
- Subjects
- Animals, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes physiology, Chemokine CXCL10, Chemokines, CXC analysis, Colonic Neoplasms metabolism, Cytotoxicity, Immunologic genetics, Cytotoxicity, Immunologic immunology, Gene Expression Regulation, Neoplastic genetics, Immunohistochemistry, In Situ Hybridization, Interleukin-12 pharmacology, Mice, Mice, Inbred BALB C, Mice, Knockout, Neoplasms, Experimental genetics, Nitrites metabolism, T-Lymphocytes, Cytotoxic metabolism, Tumor Cells, Cultured, CD4-Positive T-Lymphocytes physiology, Cell Transformation, Neoplastic genetics, Granulocyte-Macrophage Colony-Stimulating Factor metabolism, Interferon-gamma genetics, Interleukin-12 genetics
- Abstract
We analyzed the ability of interferon (IFN)-gamma knockout mice (GKO) to reject a colon carcinoma transduced with interleukin (IL)-12 genes (C26/IL-12). Although the absence of IFN-gamma impaired the early response and reduced the time to tumor onset in GKO mice, the overall tumor take rate was similar to that of BALB/c mice. In GKO mice, C26/IL-12 tumors had a reduced number of infiltrating leukocytes, especially CD8 and natural killer cells. Analysis of the tumor site, draining nodes, and spleens of GKO mice revealed reduced expression of IFN- inducible protein 10 and monokine induced by gamma-IFN. Despite these defects, GKO mice that rejected C26/IL-12 tumor, and mice that were primed in vivo with irradiated C26/IL-12 cells, showed the same cytotoxic T lymphocyte activity but higher production of granulocyte/macrophage colony-stimulating factor (GM-CSF) as compared with control BALB/c mice. Treatment with monoclonal antibodies against GM-CSF abrogated tumor regression in GKO but not in BALB/c mice. CD4 T lymphocytes, which proved unnecessary or suppressive during rejection of C26/IL-12 cells in BALB/c mice, were required for tumor rejection in GKO mice. CD4 T cell depletion was coupled with a decline in GM-CSF expression by lymphocytes infiltrating the tumors or in the draining nodes, and with the reduction and disappearance of granulocytes and CD8 T cells, respectively, in tumor nodules. These results suggest that GM-CSF can substitute for IFN-gamma in maintaining the CD8-polymorphonuclear leukocyte cross-talk that is a hallmark of tumor rejection.
- Published
- 1998
- Full Text
- View/download PDF
41. Interleukin 12 gene therapy of MHC-negative murine melanoma metastases.
- Author
-
Nanni P, Rossi I, De Giovanni C, Landuzzi L, Nicoletti G, Stoppacciaro A, Parenza M, Colombo MP, and Lollini PL
- Subjects
- Animals, Cancer Vaccines immunology, Cytotoxicity, Immunologic, Genetic Therapy, Immunocompromised Host, Lung Neoplasms secondary, Male, Mice, Mice, Inbred C57BL, Mice, Nude, Neovascularization, Pathologic, Transduction, Genetic, H-2 Antigens, Interleukin-12 genetics, Major Histocompatibility Complex, Melanoma, Experimental therapy, Neoplasm Metastasis prevention & control
- Abstract
Immunological gene therapy of cancer relies heavily on the activation of T cells, but tumors with defects in MHC gene expression are not recognized by MHC-restricted T cells. To investigate the potential of cytokine genes for the therapy of MHC-negative tumors, we transduced B78H1, a class I-negative murine melanoma clone, with a polycistronic vector carrying murine interleukin (IL)-12 genes. The clones studied produced 400-25,000 pg/ml IL-12; their in vitro growth properties were similar to those of parental cells. A complete inhibition of growth was observed in vivo both after s.c. and i.v. administration of all IL-12 clones. IL-12-transduced cells were also used as a therapeutic vaccine in mice bearing micrometastases by nontransduced parental cells. A significant (80-90%) reduction in the number of lung nodules was obtained. Immunohistochemical analysis and studies in immunocompromised hosts showed that T cells and natural killer cells had a significant role in the elimination of IL-12-releasing cells. In situ hybridization with cytokine probes detected a strong increase in the proportion of leukocytes positive for IFN-gamma, tumor necrosis factor alpha, IL-1beta, and IFN-inducible protein 10 at the site of rejection of IL-12-engineered tumor cells. However, it was clear that the loss of in vivo growth was also due to T-cell- and natural killer cell-independent factors, possibly related to the antiangiogenic properties of IL-12. In conclusion, tumor therapy based on IL-12 gene transduction was effective on a MHC-negative metastatic tumor, suggesting a possible application to MHC-defective human neoplasms.
- Published
- 1998
42. Amount of interleukin 12 available at the tumor site is critical for tumor regression.
- Author
-
Colombo MP, Vagliani M, Spreafico F, Parenza M, Chiodoni C, Melani C, and Stoppacciaro A
- Subjects
- Adenocarcinoma pathology, Animals, Base Sequence, CD4-Positive T-Lymphocytes immunology, Colonic Neoplasms pathology, DNA Primers chemistry, Immunologic Memory, Interleukin-12 metabolism, Mice, Mice, Inbred BALB C, Molecular Sequence Data, Recombinant Proteins, Adenocarcinoma immunology, Colonic Neoplasms immunology, Interleukin-12 administration & dosage
- Abstract
The C26 colon carcinoma is resistant to systemic recombinant interleukin 12 (rIL-12) therapy. Transduction of C26 with genes encoding the two subunits of murine IL-12 to release 30-80 pg/ml resulted in delayed tumor onset after injection of 5 x 10(4) cells into syngeneic BALB/c mice and in 40% tumor regression after injection into CD4-depleted mice. Here, we analyzed the activity of rIL-12 (1 microgram/day) against C26 grown into CD4-depleted mice. Like in mice given injections of interleukin 12 (IL-12) gene-transduced C26 cells, depletion of CD4+ cells led to tumor regression in 6 of 14 mice, and immumocytochemical characterization of tumor-infiltrating leukocytes showed abundant infiltration by CD8+ T cells and asialoGM1+ natural killer cells, which were scanty in tumors from nondepleted mice. On the basis of the percentage of tumor regression and leukocyte infiltration we can conclude that, in the C26 system, systemic rIL-12 (1 pmicrogramg/day) produces the same results as 30-80 pg/ml IL-12 released at the tumor site. A new polycistronic retroviral vector was then used to increase the amount of IL-12 produced by C26-transduced cells. C26 cells releasing 5 ng/ml IL-12, nearly 100 times more than the above-mentioned transduced cells, were tumorigenic in less than 50% of the mice given injections of 5 x 10(4) cells. In mice given injections of 5 x 10(5) cells, an initial tumor take of 100% followed by a complete tumor regression. Tumor regression was associated with infiltration of CD8+ and asialoGM1+ cells, and mice that remained tumor free were immune to a rechallenge of nontransduced C26 cells. The results indicate that the amount of IL-12 made available at the tumor site may determine both the type and number of infiltrating leukocytes and the events leading to tumor regression as well as it may overcame host immunosuppression.
- Published
- 1996
43. Hypoxic tumor cell death and modulation of endothelial adhesion molecules in the regression of granulocyte colony-stimulating factor-transduced tumors.
- Author
-
Colombo MP, Lombardi L, Melani C, Parenza M, Baroni C, Ruco L, and Stoppacciaro A
- Subjects
- Adenocarcinoma blood supply, Adenocarcinoma ultrastructure, Animals, Cell Death, Cell Hypoxia, Colonic Neoplasms blood supply, Colonic Neoplasms ultrastructure, E-Selectin metabolism, Immunohistochemistry, Interleukin-1 physiology, Mice, Mice, Inbred BALB C, Microscopy, Electron, Necrosis pathology, Neoplasm Transplantation, RNA, Messenger biosynthesis, Remission Induction, Tumor Necrosis Factor-alpha physiology, Vascular Cell Adhesion Molecule-1 metabolism, Adenocarcinoma pathology, Cell Adhesion Molecules metabolism, Colonic Neoplasms pathology, Granulocyte Colony-Stimulating Factor genetics, Transfection
- Abstract
C-26 colon adenocarcinoma cells transduced with the granulocyte colony-stimulating factor (G-CSF) gene form large tumors when injected into sublethally irradiated mice. These tumors regress when leukocyte function is reconstituted. Electron microscopy and immunocytochemical analysis of regressing C-26/G-CSF nodules indicates that tumor destruction is due mainly to hypoxia resulting from the functional loss of tumor vasculature and is only marginally due to direct cytolysis. Desegregation of basal lamina, cell swelling, and loss of junctions characterized the vessels within regressing tumors. Tumor cells were necrotic or filled with lipid vacuoles regardless of the distance from nearby vessels. Damage of tumor vasculature was dependent on the infiltrating leukocytes and the cytotoxic cytokines they produced. Locally produced interleukin-1 and tumor necrosis factor-alpha (TNF-alpha) induced vascular cellular adhesion molecule-1 (VCAM-1) and E-selectin on tumor vessels. Treatment with monoclonal antibodies to interferon-gamma (IFN-gamma) or TNF-alpha blocked tumor regression by inhibiting VCAM-1 and E-selectin expression on tumor-associated endothelial cells resulting in a reduced number of infiltrating leukocytes. Thus, C-26/G-CSF tumor regression presents features typical of hemorrhagic necrosis that occurs through the cytokines produced by infiltrating leukocytes in response to G-CSF.
- Published
- 1996
44. Interleukin 12 potentiates the curative effect of a vaccine based on interleukin 2-transduced tumor cells.
- Author
-
Vagliani M, Rodolfo M, Cavallo F, Parenza M, Melani C, Parmiani G, Forni G, and Colombo MP
- Subjects
- Adenocarcinoma metabolism, Animals, Cell Division physiology, Chemotherapy, Adjuvant, Colonic Neoplasms metabolism, Combined Modality Therapy, DNA, Complementary genetics, Drug Synergism, Granulocyte-Macrophage Colony-Stimulating Factor genetics, Granulocyte-Macrophage Colony-Stimulating Factor immunology, Granulocyte-Macrophage Colony-Stimulating Factor metabolism, Humans, Interleukin-2 genetics, Interleukin-2 immunology, Interleukin-2 metabolism, Killer Cells, Natural immunology, Lymphocytes, Tumor-Infiltrating immunology, Mice, Mice, Inbred BALB C, T-Lymphocytes immunology, Vaccines immunology, Adenocarcinoma immunology, Colonic Neoplasms immunology, Immunotherapy, Adoptive, Interleukin-12 pharmacology, Transduction, Genetic, Vaccines pharmacology
- Abstract
The purpose of these studies was to determine whether systemic administration of recombinant interleukin 12 (rIL-12) is able to potentiate an initial, but insufficient T-cell antitumor response. Mice challenged with carcinoma cells engineered to release interleukin 2 (IL-2) and displaying such a response received single or multiple i.p. injections of rIL-12. This combination of systemic rIL-12 and local IL-2 increased the percentage of mice that rejected two different IL-2 gene-transduced tumors. In another set of experiments more closely resembling a clinical situation, IL-2 gene-transduced tumors were used as vaccines in an attempt to cure mice bearing wild-type parental tumors. The combination of these vaccines with systemic rIL-12 cured mice more effectively than rIL-12 and IL-2 gene-transduced tumor vaccines alone.
- Published
- 1996
45. Alloreactivity and tumor antigens: generation of syngeneic antilymphoma killer lymphocytes by alloimmunization of mice with normal cells.
- Author
-
Sensi ML, Parenza M, and Parmiani G
- Subjects
- Animals, Antilymphocyte Serum pharmacology, Ascitic Fluid immunology, Cross Reactions, H-2 Antigens immunology, Mice, Mice, Inbred BALB C, Mice, Inbred DBA, Neoplasms, Experimental immunology, Antigens, Neoplasm immunology, Cytotoxicity, Immunologic, Killer Cells, Natural immunology, Lymphoma immunology
- Abstract
The possibility of obtaining a syngeneic antitumor effect by immunization with normal allogeneic cells was investigated by tests of the lytic activity of peritoneal exudate cells (PEC) of BALB/c mice immunized with lymphoid cells of either a single strain or a pool of six different allogeneic strains on the syngeneic Moloney virus-induced lymphoma YC8 target and on one of its clones designated YC8-D1. Significant cytotoxicity on both targets but not on two other BALB/c lymphomas was obtained with PEC of BALB/c mice singly immunized to the non-H-2-incompatible but H-2-compatible DBA/2 or B10.D2 lymphoid cells. The lack of lysis of YC8 cells by PEC of BALB/c mice immune to B10.A (H-2k,d) suggests that the in vitro killing was restricted by Kd-IEd region products of the major histocompatibility complex. Pool immunization was effective in generating antitumor cytotoxic lymphocytes only when DBA/2 lymphoid cells were included in the pool. The pattern of reactivity of effectors elicited in (BALB/c x DBA/2)F1 and in (BALB/c x B10.D2)F1 mice by immunization with DBA/2 and B10.D2 cells showed that at least two sets of antigens are recognized on YC8 targets, one shared by DBA/2 and B10.D2 tissues and the other expressed by DBA/2 cells only. Cold target blocking experiments indicated that the same effectors recognized non-H-2 antigens of DBA/2 and the cross-reacting YC8 determinants. The antitumor effect was mediated by T-cells, since it was abrogated by treatment of effectors with anti-Thy 1.2 serum plus complement. These data indicate that determinants defined by cytotoxic T-lymphocytes are expressed on the BALB/c lymphoma YC8 and cross-react with non-H-2 antigens of DBA/2 and B10.D2 strains.
- Published
- 1983
46. Adoptive immunotherapy of a BALB/c lymphoma by syngeneic anti-DBA/2 immune lymphoid cells: characterization of the effector population and evidence for the role of the host's non-T cells.
- Author
-
Colombo MP, Parenza M, and Parmiani G
- Subjects
- Animals, Cell Line, Hypersensitivity, Delayed immunology, Immunization, Passive, Immunologic Memory, Immunotherapy, Lymphocyte Activation, Lymphocyte Transfusion, Lymphoma immunology, Mice, Mice, Inbred BALB C, Mice, Inbred DBA, Cytotoxicity, Immunologic, Immunity, Cellular, Lymphocytes immunology, Lymphoma therapy
- Abstract
It has been previously shown that the BALB/c lymphoma YC8 is susceptible to lysis by syngeneic anti-DBA/2 lymphocytes and that YC8-bearing BALB/c mice can be cured by adoptive transfer of such immune effectors. In this study in vivo and in vitro functions of the curative immune lymphocytes have been evaluated together with the role of the host immune system in the mechanism of tumor eradication. It was found that the curative anti-DBA/2 lymphocytes were not directly cytotoxic to YC8 cells although they developed into YC8-lytic cells after in vitro restimulation with YC8. In vivo, the immune lymphocytes were able to mediate a tumor-specific delayed type hypersensitivity reaction against YC8 but had a low tumor-neutralizing activity in the Winn assay. Proliferation of infused BALB/c anti-DBA/2 lymphocytes was necessary for the in vivo therapeutic effect, since irradiation of effector cells or treatment of the donor immune lymphocytes with vinblastine abolished their curative capacity. Immunodepression of the T cell compartment of the prospective tumor-bearing animals by thymectomy plus irradiation or its abrogation in B mice (thymectomized, lethally irradiated, and reconstituted with fetal liver cells) did not interfere with the therapeutic effect of the transferred anti-DBA/2 lymphocytes. Blocking the macrophage functions of the host by carrageenan, however, abolished the therapeutic effect of immune lymphocytes. These data indicate that a radiation-resistant, non-T cell is involved in the tumor eradication induced by anti-DBA/2 lymphocytes. It was also shown that cured mice, tested 90 days after therapy, become resistant to 5 X 10(3) LD80 YC8 cells and that this resistance was due to the presence of memory cells derived from the transferred and not from the host lymphocyte population.
- Published
- 1985
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.