264 results on '"Kajaste-Rudnitski A"'
Search Results
2. Removal of innate immune barriers allows efficient transduction of quiescent human hematopoietic stem cells
- Author
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Valeri, Erika, Unali, Giulia, Piras, Francesco, Abou-Alezz, Monah, Pais, Giulia, Benedicenti, Fabrizio, Lidonnici, Maria Rosa, Cuccovillo, Ivan, Castiglioni, Ilaria, Arévalo, Sergio, Spinozzi, Giulio, Merelli, Ivan, Behrendt, Rayk, Oo, Adrian, Kim, Baek, Landau, Nathaniel R., Ferrari, Giuliana, Montini, Eugenio, and Kajaste-Rudnitski, Anna
- Published
- 2024
- Full Text
- View/download PDF
3. In vivo macrophage engineering reshapes the tumor microenvironment leading to eradication of liver metastases
- Author
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Kerzel, Thomas, Giacca, Giovanna, Beretta, Stefano, Bresesti, Chiara, Notaro, Marco, Scotti, Giulia Maria, Balestrieri, Chiara, Canu, Tamara, Redegalli, Miriam, Pedica, Federica, Genua, Marco, Ostuni, Renato, Kajaste-Rudnitski, Anna, Oshima, Masanobu, Tonon, Giovanni, Merelli, Ivan, Aldrighetti, Luca, Dellabona, Paolo, Coltella, Nadia, Doglioni, Claudio, Rancoita, Paola M.V., Sanvito, Francesca, Naldini, Luigi, and Squadrito, Mario Leonardo
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- 2023
- Full Text
- View/download PDF
4. Genetic engineering meets hematopoietic stem cell biology for next-generation gene therapy
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Ferrari, Samuele, Valeri, Erika, Conti, Anastasia, Scala, Serena, Aprile, Annamaria, Di Micco, Raffaella, Kajaste-Rudnitski, Anna, Montini, Eugenio, Ferrari, Giuliana, Aiuti, Alessandro, and Naldini, Luigi
- Published
- 2023
- Full Text
- View/download PDF
5. Protocol to differentiate monolayer human induced pluripotent stem cells into inflammatory responsive astrocytes
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Anna Maria Sole Giordano, Monah Abou Alezz, Ivan Merelli, and Anna Kajaste-Rudnitski
- Subjects
Cell Biology ,Cell Differentiation ,Immunology ,Molecular Biology ,Neuroscience ,Stem Cells ,Science (General) ,Q1-390 - Abstract
Summary: Glia, and in particular astrocytes, are one of the major players in neurological and neuroinflammatory disorders. Here, we present a protocol to efficiently generate inflammatory responsive astrocytes from human induced pluripotent stem cells in a monolayer culture. We describe steps for neural differentiation to reach a homogeneous population of neural progenitor cells, followed by their differentiation into neural/glial progenitors. Finally, we detail enrichment to a 90% pure inflammatory responsive astrocyte population.For complete details on the use and execution of this protocol, please refer to Giordano et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
- Published
- 2023
- Full Text
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6. AAV vectors trigger DNA damage responses and STING-dependent inflammation in human CNS cells
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Kajaste-Rudnitski, Anna, primary, Costa-Verdera, Helena, additional, Meneghini, Vasco, additional, Fitzpatrick, Zachary, additional, Alezz, Monah Abou, additional, Fabyanic, Emily, additional, Huang, Xin, additional, Dzhashiashvili, Yulia, additional, Ahiya, Avantika, additional, Mangiameli, Elisabeth, additional, Valeri, Erika, additional, Crivicich, Giovanni, additional, Cuccovillo, Ivan, additional, Caccia, Roberta, additional, Bertin, Berangere, additional, Ronzitti, Giuseppe, additional, Engel, Esteban, additional, Merelli, Ivan, additional, Mingozzi, Federico, additional, Gritti, Angela, additional, and Kuranda, Klaudia, additional
- Published
- 2024
- Full Text
- View/download PDF
7. Antiviral immunity and nucleic acid sensing in haematopoietic stem cell gene engineering
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Piras, Francesco and Kajaste-Rudnitski, Anna
- Published
- 2021
- Full Text
- View/download PDF
8. Laboratory-Scale Lentiviral Vector Production and Purification for Enhanced Ex Vivo and In Vivo Genetic Engineering
- Author
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Monica Soldi, Lucia Sergi Sergi, Giulia Unali, Thomas Kerzel, Ivan Cuccovillo, Paola Capasso, Andrea Annoni, Mauro Biffi, Paola Maria Vittoria Rancoita, Alessio Cantore, Angelo Lombardo, Luigi Naldini, Mario Leonardo Squadrito, and Anna Kajaste-Rudnitski
- Subjects
Lentiviral vectors ,purification process ,manufacturing ,gene therapy ,ex vivo ,in vivo ,Genetics ,QH426-470 ,Cytology ,QH573-671 - Abstract
Lentiviral vectors (LVs) are increasingly employed in gene and cell therapy. Standard laboratory production of LVs is not easily scalable, and research-grade LVs often contain contaminants that can interfere with downstream applications. Moreover, purified LV production pipelines have been developed mainly for costly, large-scale, clinical-grade settings. Therefore, a standardized and cost-effective process is still needed to obtain efficient, reproducible, and properly executed experimental studies and preclinical development of ex vivo and in vivo gene therapies, as high infectivity and limited adverse reactions are important factors potentially influencing experimental outcomes also in preclinical settings. We describe here an optimized laboratory-scale workflow whereby an LV-containing supernatant is purified and concentrated by sequential chromatographic steps, obtaining biologically active LVs with an infectious titer and specific activity in the order of 109 transducing unit (TU)/mL and 5 × 104 TU/ng of HIV Gag p24, respectively. The purification workflow removes >99% of the starting plasmid, DNA, and protein impurities, resulting in higher gene transfer and editing efficiency in severe combined immunodeficiency (SCID)-repopulating hematopoietic stem and progenitor cells (HSPCs) ex vivo, as well as reduced activation of inflammatory responses ex vivo and in vivo as compared to TU-matched, laboratory-grade vectors. Our results highlight the value of accessible purified LV production for experimental studies and preclinical testing.
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- 2020
- Full Text
- View/download PDF
9. D-mannose suppresses macrophage IL-1β production
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Simone Torretta, Alessandra Scagliola, Luisa Ricci, Francesco Mainini, Sabrina Di Marco, Ivan Cuccovillo, Anna Kajaste-Rudnitski, David Sumpton, Kevin M. Ryan, and Simone Cardaci
- Subjects
Science - Abstract
Mannose is present at trace levels in blood and regulates cancer growth. Here the authors show that supraphysiological levels of mannose can also regulate macrophages, limiting their production of IL-1β and increasing resistance of mice to LPS-induced endotoxemia and DSS-induced colitis.
- Published
- 2020
- Full Text
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10. Efficient gene editing of human long-term hematopoietic stem cells validated by clonal tracking
- Author
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Ferrari, Samuele, Jacob, Aurelien, Beretta, Stefano, Unali, Giulia, Albano, Luisa, Vavassori, Valentina, Cittaro, Davide, Lazarevic, Dejan, Brombin, Chiara, Cugnata, Federica, Kajaste-Rudnitski, Anna, Merelli, Ivan, Genovese, Pietro, and Naldini, Luigi
- Published
- 2020
- Full Text
- View/download PDF
11. Genetic engineering meets hematopoietic stem cell biology for next-generation gene therapy
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Samuele Ferrari, Erika Valeri, Anastasia Conti, Serena Scala, Annamaria Aprile, Raffaella Di Micco, Anna Kajaste-Rudnitski, Eugenio Montini, Giuliana Ferrari, Alessandro Aiuti, and Luigi Naldini
- Subjects
Genetics ,Molecular Medicine ,Cell Biology - Published
- 2023
12. Understanding and Tackling Immune Responses to Adeno-Associated Viral Vectors
- Author
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Costa-Verdera, Helena, primary, Unzu, Carmen, additional, Valeri, Erika, additional, Adriouch, Sahil, additional, González Aseguinolaza, Gloria, additional, Mingozzi, Federico, additional, and Kajaste-Rudnitski, Anna, additional
- Published
- 2023
- Full Text
- View/download PDF
13. Corrigendum: Generation of Powerful Human Tolerogenic Dendritic Cells by Lentiviral-Mediated IL-10 Gene Transfer
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Michela Comi, Giada Amodio, Laura Passeri, Marta Fortunato, Francesca Romana Santoni de Sio, Grazia Andolfi, Anna Kajaste-Rudnitski, Fabio Russo, Luca Cesana, and Silvia Gregori
- Subjects
dendritic cells ,IL-10 ,cell therapy ,immune tolerance ,allogeneic transplantation ,Immunologic diseases. Allergy ,RC581-607 - Published
- 2021
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14. D-mannose suppresses macrophage IL-1β production
- Author
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Torretta, Simone, Scagliola, Alessandra, Ricci, Luisa, Mainini, Francesco, Di Marco, Sabrina, Cuccovillo, Ivan, Kajaste-Rudnitski, Anna, Sumpton, David, Ryan, Kevin M., and Cardaci, Simone
- Published
- 2020
- Full Text
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15. Generation of Powerful Human Tolerogenic Dendritic Cells by Lentiviral-Mediated IL-10 Gene Transfer
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Michela Comi, Giada Amodio, Laura Passeri, Marta Fortunato, Francesca Romana Santoni de Sio, Grazia Andolfi, Anna Kajaste-Rudnitski, Fabio Russo, Luca Cesana, and Silvia Gregori
- Subjects
dendritic cells ,IL-10 ,cell therapy ,immune tolerance ,allogeneic transplantation ,Immunologic diseases. Allergy ,RC581-607 - Abstract
The prominent role of dendritic cells (DC) in promoting tolerance and the development of methods to generate clinical grade products allowed the clinical application of tolerogenic DC (tolDC)-based therapies for controlling unwanted immune responses. We established an efficient method to generate tolerogenic human DC, producing supra-physiological levels of IL-10, by genetically engineering monocyte-derived DC with a bidirectional Lentiviral Vector (bdLV) encoding for IL-10 and a marker gene. DCIL−10 are mature DC, modulate T cell responses, promote T regulatory cells, and are phenotypically and functionally stable upon stimulation. Adoptive transfer of human DCIL−10 in a humanized mouse model dampens allogeneic T cell recall responses, while murine DCIL−10 delays acute graft-vs.-host disease in mice. Our report outlines an efficient method to transduce human myeloid cells with large-size LV and shows that stable over-expression of IL-10 generates an effective cell product for future clinical applications in the contest of allogeneic transplantation.
- Published
- 2020
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16. Constitutive IL-1RA production by modified immune cells protects against IL-1–mediated inflammatory disorders
- Author
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Colantuoni, Mariasilvia, primary, Jofra Hernandez, Raisa, additional, Pettinato, Emanuela, additional, Basso-Ricci, Luca, additional, Magnani, Laura, additional, Andolfi, Grazia, additional, Rigamonti, Chiara, additional, Finardi, Annamaria, additional, Romeo, Valentina, additional, Soldi, Monica, additional, Sergi Sergi, Lucia, additional, Rocchi, Martina, additional, Scala, Serena, additional, Hoffman, Hal M., additional, Gregori, Silvia, additional, Kajaste-Rudnitski, Anna, additional, Sanvito, Francesca, additional, Muzio, Luca, additional, Naldini, Luigi, additional, Aiuti, Alessandro, additional, and Mortellaro, Alessandra, additional
- Published
- 2023
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17. Lentiviral vectors escape innate sensing but trigger p53 in human hematopoietic stem and progenitor cells
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Francesco Piras, Michela Riba, Carolina Petrillo, Dejan Lazarevic, Ivan Cuccovillo, Sara Bartolaccini, Elia Stupka, Bernhard Gentner, Davide Cittaro, Luigi Naldini, and Anna Kajaste‐Rudnitski
- Subjects
gene therapy ,hematopoietic stem and progenitor cells ,innate sensing ,lentiviral vectors ,p53 signaling ,Medicine (General) ,R5-920 ,Genetics ,QH426-470 - Abstract
Abstract Clinical application of lentiviral vector (LV)‐based hematopoietic stem and progenitor cells (HSPC) gene therapy is rapidly becoming a reality. Nevertheless, LV‐mediated signaling and its potential functional consequences on HSPC biology remain poorly understood. We unravel here a remarkably limited impact of LV on the HSPC transcriptional landscape. LV escaped innate immune sensing that instead led to robust IFN responses upon transduction with a gamma‐retroviral vector. However, reverse‐transcribed LV DNA did trigger p53 signaling, activated also by non‐integrating Adeno‐associated vector, ultimately leading to lower cell recovery ex vivo and engraftment in vivo. These effects were more pronounced in the short‐term repopulating cells while long‐term HSC frequencies remained unaffected. Blocking LV‐induced signaling partially rescued both apoptosis and engraftment, highlighting a novel strategy to further dampen the impact of ex vivo gene transfer on HSPC. Overall, our results shed light on viral vector sensing in HSPC and provide critical insight for the development of more stealth gene therapy strategies.
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- 2017
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18. Efficient Ex Vivo Engineering and Expansion of Highly Purified Human Hematopoietic Stem and Progenitor Cell Populations for Gene Therapy
- Author
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Erika Zonari, Giacomo Desantis, Carolina Petrillo, Francesco E. Boccalatte, Maria Rosa Lidonnici, Anna Kajaste-Rudnitski, Alessandro Aiuti, Giuliana Ferrari, Luigi Naldini, and Bernhard Gentner
- Subjects
Medicine (General) ,R5-920 ,Biology (General) ,QH301-705.5 - Abstract
Summary: Ex vivo gene therapy based on CD34+ hematopoietic stem cells (HSCs) has shown promising results in clinical trials, but genetic engineering to high levels and in large scale remains challenging. We devised a sorting strategy that captures more than 90% of HSC activity in less than 10% of mobilized peripheral blood (mPB) CD34+ cells, and modeled a transplantation protocol based on highly purified, genetically engineered HSCs co-infused with uncultured progenitor cells. Prostaglandin E2 stimulation allowed near-complete transduction of HSCs with lentiviral vectors during a culture time of less than 38 hr, mitigating the negative impact of standard culture on progenitor cell function. Exploiting the pyrimidoindole derivative UM171, we show that transduced mPB CD34+CD38− cells with repopulating potential could be expanded ex vivo. Implementing these findings in clinical gene therapy protocols will improve the efficacy, safety, and sustainability of gene therapy and generate new opportunities in the field of gene editing. : In this article, Gentner and colleagues undertake a comprehensive strategy to advance ex vivo genetic engineering of HSCs for gene therapy. They experimentally define an optimal strategy to purify HSCs, which allows uncoupling long-term from short-term hematopoietic reconstitution, and implement ex vivo conditions that best preserve their biological properties applying novel transduction-enhancing compounds and pyrimidoindole derivatives to support HSC expansion. Keywords: HSC gene therapy, purified HSCs, HSC expansion, lentiviral vector transduction, prostaglandin E2, UM171
- Published
- 2017
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19. Interferon‐inducible phospholipids govern IFITM3‐dependent endosomal antiviral immunity
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Unali, Giulia, primary, Crivicich, Giovanni, additional, Pagani, Isabel, additional, Abou‐Alezz, Monah, additional, Folchini, Filippo, additional, Valeri, Erika, additional, Matafora, Vittoria, additional, Reisz, Julie A, additional, Giordano, Anna Maria Sole, additional, Cuccovillo, Ivan, additional, Butta, Giacomo M, additional, Donnici, Lorena, additional, D'Alessandro, Angelo, additional, De Francesco, Raffaele, additional, Manganaro, Lara, additional, Cittaro, Davide, additional, Merelli, Ivan, additional, Petrillo, Carolina, additional, Bachi, Angela, additional, Vicenzi, Elisa, additional, and Kajaste‐Rudnitski, Anna, additional
- Published
- 2023
- Full Text
- View/download PDF
20. Towards improved yet regulated gene therapy for X-CGD
- Author
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Kajaste-Rudnitski, Anna, primary and Aiuti, Alessandro, additional
- Published
- 2023
- Full Text
- View/download PDF
21. Protocol to differentiate monolayer human induced pluripotent stem cells into inflammatory responsive astrocytes
- Author
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Giordano, Anna Maria Sole, primary, Abou Alezz, Monah, additional, Merelli, Ivan, additional, and Kajaste-Rudnitski, Anna, additional
- Published
- 2023
- Full Text
- View/download PDF
22. Interferon‐inducible phospholipids govern <scp>IFITM3</scp> ‐dependent endosomal antiviral immunity
- Author
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Giulia Unali, Giovanni Crivicich, Isabel Pagani, Monah Abou‐Alezz, Filippo Folchini, Erika Valeri, Vittoria Matafora, Julie A Reisz, Anna Maria Sole Giordano, Ivan Cuccovillo, Giacomo M Butta, Lorena Donnici, Angelo D'Alessandro, Raffaele De Francesco, Lara Manganaro, Davide Cittaro, Ivan Merelli, Carolina Petrillo, Angela Bachi, Elisa Vicenzi, and Anna Kajaste‐Rudnitski
- Subjects
IFITM ,SARS-CoV2 ,innate immunity ,phosphatidylinositol 3,4,5-trisphosphate (PIP3) ,viral entry ,General Immunology and Microbiology ,General Neuroscience ,phosphatidylinositol 3 ,Settore BIO/19 - Microbiologia Generale ,5-trisphosphate (PIP3) ,Molecular Biology ,General Biochemistry, Genetics and Molecular Biology - Published
- 2023
23. Laboratory-Scale Lentiviral Vector Production and Purification for Enhanced Ex Vivo and In Vivo Genetic Engineering
- Author
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Alessio Cantore, Monica Soldi, Lucia Sergi Sergi, Thomas Kerzel, Anna Kajaste-Rudnitski, Paola Capasso, Luigi Naldini, Mario Leonardo Squadrito, Andrea Annoni, Ivan Cuccovillo, Giulia Unali, Mauro Biffi, Paola M.V. Rancoita, Angelo Lombardo, Soldi, Monica, Sergi Sergi, Lucia, Unali, Giulia, Kerzel, Thoma, Cuccovillo, Ivan, Capasso, Paola, Annoni, Andrea, Biffi, Mauro, Rancoita, Paola Maria Vittoria, Cantore, Alessio, Lombardo, Angelo, Naldini, Luigi, Squadrito, Mario Leonardo, and Kajaste-Rudnitski, Anna
- Subjects
0301 basic medicine ,lcsh:QH426-470 ,Genetic enhancement ,Viral vector ,Cell therapy ,03 medical and health sciences ,0302 clinical medicine ,Plasmid ,In vivo ,Genetics ,medicine ,Progenitor cell ,lcsh:QH573-671 ,innate immunity ,Molecular Biology ,Severe combined immunodeficiency ,Chemistry ,lcsh:Cytology ,Lentiviral vectors ,medicine.disease ,gene therapy ,hematopoietic stem cells ,3. Good health ,Cell biology ,manufacturing ,in vivo ,lcsh:Genetics ,030104 developmental biology ,030220 oncology & carcinogenesis ,ex vivo ,Molecular Medicine ,purification process ,Original Article ,Ex vivo - Abstract
Lentiviral vectors (LVs) are increasingly employed in gene and cell therapy. Standard laboratory production of LVs is not easily scalable, and research-grade LVs often contain contaminants that can interfere with downstream applications. Moreover, purified LV production pipelines have been developed mainly for costly, large-scale, clinical-grade settings. Therefore, a standardized and cost-effective process is still needed to obtain efficient, reproducible, and properly executed experimental studies and preclinical development of ex vivo and in vivo gene therapies, as high infectivity and limited adverse reactions are important factors potentially influencing experimental outcomes also in preclinical settings. We describe here an optimized laboratory-scale workflow whereby an LV-containing supernatant is purified and concentrated by sequential chromatographic steps, obtaining biologically active LVs with an infectious titer and specific activity in the order of 109 transducing unit (TU)/mL and 5 × 104 TU/ng of HIV Gag p24, respectively. The purification workflow removes >99% of the starting plasmid, DNA, and protein impurities, resulting in higher gene transfer and editing efficiency in severe combined immunodeficiency (SCID)-repopulating hematopoietic stem and progenitor cells (HSPCs) ex vivo, as well as reduced activation of inflammatory responses ex vivo and in vivo as compared to TU-matched, laboratory-grade vectors. Our results highlight the value of accessible purified LV production for experimental studies and preclinical testing., Graphical Abstract, Lentiviral vectors (LVs) are powerful gene-transfer tools routinely exploited for distinct research and clinical applications. LVs produced in most research laboratories contain contaminants that can generate confounding effects in experimental studies. Soldi et al. describe a laboratory-scale workflow for purified LV production, highlighting enhanced gene-editing efficiency and diminished inflammatory responses.
- Published
- 2020
24. Towards improved yet regulated gene therapy for X-CGD
- Author
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Anna Kajaste-Rudnitski and Alessandro Aiuti
- Subjects
Immunology ,Cell Biology ,Hematology ,Biochemistry - Published
- 2023
25. Choice of template delivery mitigates the genotoxic risk and adverse impact of editing in human hematopoietic stem cells
- Author
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Ferrari, Samuele, primary, Jacob, Aurelien, additional, Cesana, Daniela, additional, Laugel, Marianne, additional, Beretta, Stefano, additional, Varesi, Angelica, additional, Unali, Giulia, additional, Conti, Anastasia, additional, Canarutto, Daniele, additional, Albano, Luisa, additional, Calabria, Andrea, additional, Vavassori, Valentina, additional, Cipriani, Carlo, additional, Castiello, Maria Carmina, additional, Esposito, Simona, additional, Brombin, Chiara, additional, Cugnata, Federica, additional, Adjali, Oumeya, additional, Ayuso, Eduard, additional, Merelli, Ivan, additional, Villa, Anna, additional, Di Micco, Raffaella, additional, Kajaste-Rudnitski, Anna, additional, Montini, Eugenio, additional, Penaud-Budloo, Magalie, additional, and Naldini, Luigi, additional
- Published
- 2022
- Full Text
- View/download PDF
26. Abstract 3297: IFNalpha by in vivo-engineered macrophages abates liver metastases and triggers counter regulatory responses limiting efficacy
- Author
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Kerzel, Thomas, primary, Beretta, Stefano, additional, Scamardella, Eloise, additional, Balestrieri, Chiara, additional, Canu, Tamara, additional, Pedica, Federica, additional, Norata, Rossana, additional, Sergi, Lucia Sergi, additional, Genua, Marco, additional, Renato, Ostuni, additional, Kajaste-Rudnitski, Anna, additional, Esposito, Antonio, additional, Oshima, Masanobu, additional, Tonon, Giovanni, additional, Sanvito, Francesca, additional, Squadrito, Mario Leonardo, additional, and Naldini, Luigi, additional
- Published
- 2022
- Full Text
- View/download PDF
27. Efficient gene editing of human long-term hematopoietic stem cells validated by clonal tracking
- Author
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Ferrari, S, Jacob, A, Beretta, S, Unali, G, Albano, L, Vavassori, V, Cittaro, D, Lazarevic, D, Brombin, C, Cugnata, F, Kajaste-Rudnitski, A, Merelli, I, Genovese, P, Naldini, L, Ferrari S., Jacob A., Beretta S., Unali G., Albano L., Vavassori V., Cittaro D., Lazarevic D., Brombin C., Cugnata F., Kajaste-Rudnitski A., Merelli I., Genovese P., Naldini L., Ferrari, S, Jacob, A, Beretta, S, Unali, G, Albano, L, Vavassori, V, Cittaro, D, Lazarevic, D, Brombin, C, Cugnata, F, Kajaste-Rudnitski, A, Merelli, I, Genovese, P, Naldini, L, Ferrari S., Jacob A., Beretta S., Unali G., Albano L., Vavassori V., Cittaro D., Lazarevic D., Brombin C., Cugnata F., Kajaste-Rudnitski A., Merelli I., Genovese P., and Naldini L.
- Abstract
Targeted gene editing in hematopoietic stem cells (HSCs) is a promising treatment for several diseases. However, the limited efficiency of homology-directed repair (HDR) in HSCs and the unknown impact of the procedure on clonal composition and dynamics of transplantation have hampered clinical translation. Here, we apply a barcoding strategy to clonal tracking of edited cells (BAR-Seq) and show that editing activates p53, which substantially shrinks the HSC clonal repertoire in hematochimeric mice, although engrafted edited clones preserve multilineage and self-renewing capacity. Transient p53 inhibition restored polyclonal graft composition. We increased HDR efficiency by forcing cell-cycle progression and upregulating components of the HDR machinery through transient expression of the adenovirus 5 E4orf6/7 protein, which recruits the cell-cycle controller E2F on its target genes. Combined E4orf6/7 expression and p53 inhibition resulted in HDR editing efficiencies of up to 50% in the long-term human graft, without perturbing repopulation and self-renewal of edited HSCs. This enhanced protocol should broaden applicability of HSC gene editing and pave its way to clinical translation.
- Published
- 2020
28. DNA damage contributes to neurotoxic inflammation in Aicardi-Goutières Syndrome astrocytes
- Author
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Anna Maria Sole Giordano, Marco Luciani, Francesca Gatto, Monah Abou Alezz, Chiara Beghè, Lucrezia Della Volpe, Alessandro Migliara, Sara Valsoni, Marco Genua, Monika Dzieciatkowska, Giacomo Frati, Julie Tahraoui-Bories, Silvia Clara Giliani, Simona Orcesi, Elisa Fazzi, Renato Ostuni, Angelo D’Alessandro, Raffaella Di Micco, Ivan Merelli, Angelo Lombardo, Martin A.M. Reijns, Natalia Gromak, Angela Gritti, and Anna Kajaste-Rudnitski
- Subjects
Inflammation ,Autoimmune Diseases of the Nervous System ,Astrocytes ,DNA Damage ,Humans ,Nervous System Malformations ,Immunology ,Immunology and Allergy - Abstract
Aberrant induction of type I IFN is a hallmark of the inherited encephalopathy Aicardi-Goutières syndrome (AGS), but the mechanisms triggering disease in the human central nervous system (CNS) remain elusive. Here, we generated human models of AGS using genetically modified and patient-derived pluripotent stem cells harboring TREX1 or RNASEH2B loss-of-function alleles. Genome-wide transcriptomic analysis reveals that spontaneous proinflammatory activation in AGS astrocytes initiates signaling cascades impacting multiple CNS cell subsets analyzed at the single-cell level. We identify accumulating DNA damage, with elevated R-loop and micronuclei formation, as a driver of STING- and NLRP3-related inflammatory responses leading to the secretion of neurotoxic mediators. Importantly, pharmacological inhibition of proapoptotic or inflammatory cascades in AGS astrocytes prevents neurotoxicity without apparent impact on their increased type I IFN responses. Together, our work identifies DNA damage as a major driver of neurotoxic inflammation in AGS astrocytes, suggests a role for AGS gene products in R-loop homeostasis, and identifies common denominators of disease that can be targeted to prevent astrocyte-mediated neurotoxicity in AGS.
- Published
- 2022
29. Choice of template delivery mitigates the genotoxic risk and adverse impact of editing in human hematopoietic stem cells
- Author
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Samuele Ferrari, Aurelien Jacob, Daniela Cesana, Marianne Laugel, Stefano Beretta, Angelica Varesi, Giulia Unali, Anastasia Conti, Daniele Canarutto, Luisa Albano, Andrea Calabria, Valentina Vavassori, Carlo Cipriani, Maria Carmina Castiello, Simona Esposito, Chiara Brombin, Federica Cugnata, Oumeya Adjali, Eduard Ayuso, Ivan Merelli, Anna Villa, Raffaella Di Micco, Anna Kajaste-Rudnitski, Eugenio Montini, Magalie Penaud-Budloo, and Luigi Naldini
- Subjects
Gene Editing ,Integrases ,DNA, Viral ,Genetics ,Molecular Medicine ,Humans ,Cell Biology ,CRISPR-Cas Systems ,Tumor Suppressor Protein p53 ,Hematopoietic Stem Cells ,DNA Damage - Abstract
Long-range gene editing by homology-directed repair (HDR) in hematopoietic stem/progenitor cells (HSPCs) often relies on viral transduction with recombinant adeno-associated viral vector (AAV) for template delivery. Here, we uncover unexpected load and prolonged persistence of AAV genomes and their fragments, which trigger sustained p53-mediated DNA damage response (DDR) upon recruiting the MRE11-RAD50-NBS1 (MRN) complex on the AAV inverted terminal repeats (ITRs). Accrual of viral DNA in cell-cycle-arrested HSPCs led to its frequent integration, predominantly in the form of transcriptionally competent ITRs, at nuclease on- and off-target sites. Optimized delivery of integrase-defective lentiviral vector (IDLV) induced lower DNA load and less persistent DDR, improving clonogenic capacity and editing efficiency in long-term repopulating HSPCs. Because insertions of viral DNA fragments are less frequent with IDLV, its choice for template delivery mitigates the adverse impact and genotoxic burden of HDR editing and should facilitate its clinical translation in HSPC gene therapy.
- Published
- 2022
30. Efficient gene editing of human long-term hematopoietic stem cells validated by clonal tracking
- Author
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Giulia Unali, Ivan Merelli, Stefano Beretta, Aurelien Jacob, Luisa Albano, Pietro Genovese, Samuele Ferrari, Anna Kajaste-Rudnitski, Davide Cittaro, Valentina Vavassori, Federica Cugnata, Luigi Naldini, Chiara Brombin, Dejan Lazarevic, Ferrari, S, Jacob, A, Beretta, S, Unali, G, Albano, L, Vavassori, V, Cittaro, D, Lazarevic, D, Brombin, C, Cugnata, F, Kajaste-Rudnitski, A, Merelli, I, Genovese, P, Naldini, L, Ferrari, S., Jacob, A., Beretta, S., Unali, G., Albano, L., Vavassori, V., Cittaro, D., Lazarevic, D., Brombin, C., Cugnata, F., Kajaste-Rudnitski, A., Merelli, I., Genovese, P., and Naldini, L.
- Subjects
G2 Phase ,Transcription, Genetic ,Genetic enhancement ,Transplantation, Heterologous ,Biomedical Engineering ,Bioengineering ,Biology ,Applied Microbiology and Biotechnology ,Stem-cell biotechnology ,Article ,S Phase ,03 medical and health sciences ,Mice ,Viral Proteins ,0302 clinical medicine ,Gene therapy ,Genome editing ,Animals ,Humans ,Cell Lineage ,Haematopoietic stem cell ,E2F ,030304 developmental biology ,Gene Editing ,0303 health sciences ,Base Sequence ,Targeted Gene Repair ,HEK 293 cells ,Recombinational DNA Repair ,Reproducibility of Results ,Dependovirus ,Hematopoietic Stem Cells ,Xenograft Model Antitumor Assays ,Cell biology ,Clone Cells ,Up-Regulation ,Transplantation ,Haematopoiesis ,Targeted gene repair ,HEK293 Cells ,Cell Tracking ,Molecular Medicine ,Stem cell ,Tumor Suppressor Protein p53 ,030217 neurology & neurosurgery ,Biotechnology - Abstract
Targeted gene editing in hematopoietic stem cells (HSCs) is a promising treatment for several diseases. However, the limited efficiency of homology-directed repair (HDR) in HSCs and the unknown impact of the procedure on clonal composition and dynamics of transplantation have hampered clinical translation. Here, we apply a barcoding strategy to clonal tracking of edited cells (BAR-Seq) and show that editing activates p53, which substantially shrinks the HSC clonal repertoire in hematochimeric mice, although engrafted edited clones preserve multilineage and self-renewing capacity. Transient p53 inhibition restored polyclonal graft composition. We increased HDR efficiency by forcing cell-cycle progression and upregulating components of the HDR machinery through transient expression of the adenovirus 5 E4orf6/7 protein, which recruits the cell-cycle controller E2F on its target genes. Combined E4orf6/7 expression and p53 inhibition resulted in HDR editing efficiencies of up to 50% in the long-term human graft, without perturbing repopulation and self-renewal of edited HSCs. This enhanced protocol should broaden applicability of HSC gene editing and pave its way to clinical translation. Transient p53 inhibition and induced cell-cycle progression increase clonal engraftment and homology-directed repair in hematopoietic stem cells.
- Published
- 2020
- Full Text
- View/download PDF
31. Lentiviral vectors escape innate sensing but trigger p53 in human hematopoietic stem and progenitor cells
- Author
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Piras, Francesco, Riba, Michela, Petrillo, Carolina, Lazarevic, Dejan, Cuccovillo, Ivan, Bartolaccini, Sara, Stupka, Elia, Gentner, Bernhard, Cittaro, Davide, Naldini, Luigi, and Kajaste‐Rudnitski, Anna
- Published
- 2017
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- View/download PDF
32. HIV-1 envelope-dependent restriction of CXCR4-using viruses in child but not adult untransformed CD4+ T-lymphocyte lines
- Author
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Mariani, Samanta A., Brigida, Immacolata, Kajaste-Rudnitski, Anna, Ghezzi, Silvia, Rocchi, Alessia, Plebani, Anna, Vicenzi, Elisa, Aiuti, Alessandro, and Poli, Guido
- Published
- 2012
- Full Text
- View/download PDF
33. DNA damage contributes to neurotoxic inflammation in Aicardi-Goutières syndrome astrocytes
- Author
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Giordano, Anna Maria Sole, primary, Luciani, Marco, additional, Gatto, Francesca, additional, Abou Alezz, Monah, additional, Beghè, Chiara, additional, Della Volpe, Lucrezia, additional, Migliara, Alessandro, additional, Valsoni, Sara, additional, Genua, Marco, additional, Dzieciatkowska, Monika, additional, Frati, Giacomo, additional, Tahraoui-Bories, Julie, additional, Giliani, Silvia Clara, additional, Orcesi, Simona, additional, Fazzi, Elisa, additional, Ostuni, Renato, additional, D’Alessandro, Angelo, additional, Di Micco, Raffaella, additional, Merelli, Ivan, additional, Lombardo, Angelo, additional, Reijns, Martin A.M., additional, Gromak, Natalia, additional, Gritti, Angela, additional, and Kajaste-Rudnitski, Anna, additional
- Published
- 2022
- Full Text
- View/download PDF
34. HIV-1 silencing mediated by TRIM22 inhibition of Sp1 binding to the promoter
- Author
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E. Vicenzi, F. Turrini, S.S. Marelli, A. Kajaste-Rudnitski, C. Van Lint, A.T. Das, B. Berkout, and G. Poli
- Subjects
Microbiology ,QR1-502 ,Public aspects of medicine ,RA1-1270 - Published
- 2015
- Full Text
- View/download PDF
35. Oncogene-induced maladaptive activation of trained immunity in the pathogenesis and treatment of Erdheim-Chester disease
- Author
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Mihai G. Netea, Riccardo Biavasco, Lorenzo Dagna, Simone Cenci, Travis Nemkov, Giulio Cavalli, Philippe Maksud, Claudio Doglioni, Gianfranco Distefano, Daniela Cesana, Charles A. Dinarello, Eleonora Cantoni, Anna Kajaste-Rudnitski, Corrado Campochiaro, Simone Cardaci, Daniela Gnani, Isak W. Tengesdal, Julien Haroche, Samuel Zambrano, Alessandro Tomelleri, Giacomo De Luca, Davide Stefanoni, Francesco Piras, Ivan Merelli, Angelo D'Alessandro, Maddalena Panigada, Davide Mazza, Rob J.W. Arts, Jorge Domínguez-Andrés, Alessia Loffreda, Laura Cassina, Raffaella Molteni, Eugenio Montini, Alessandra Boletta, Leo A. B. Joosten, Elisabetta Ferrero, Marina Ferrarini, Biavasco, Riccardo, Molteni, Raffaella, Stefanoni, Davide, Nemkov, Travi, Netea, Mihai G., Domínguez-Andrés, Jorge, Arts, Rob JW, Merelli, Ivan, Mazza, Davide, Zambrano, S, Panigada, Maddalena, Cantoni, Eleonora, Tengesdal, Isak, Maksud, Philippe, Piras, Francesco, De Luca, Giacomo, Cassina, Laura, Distefano, Gianfranco, Loffreda, Alessia, Gnani, Daniela, Doglioni, Claudio, Tomelleri, Alessandro, Campochiaro, Corrado, Joosten, Leo, Dinarello, Charles A, Kajaste-Rudnitski, Anna, Haroche, Julien, Cardaci, Simone, Cenci, Simone, Dagna, Lorenzo, Ferrarini, Marina, Ferrero, Elisabetta, Boletta, Alessandra, D'Alessandro, Angelo, Montini, Eugenio, and Cavalli, Giulio
- Subjects
Proto-Oncogene Proteins B-raf ,Erdheim-Chester Disease ,Myeloid ,medicine.medical_treatment ,Immunology ,lnfectious Diseases and Global Health Radboud Institute for Molecular Life Sciences [Radboudumc 4] ,Inflammation ,Biochemistry ,Myeloid Neoplasm ,Proinflammatory cytokine ,Epigenesis, Genetic ,Pathogenesis ,medicine ,Humans ,Point Mutation ,Protein kinase B ,Cells, Cultured ,business.industry ,Monocyte ,Macrophages ,Immunity ,Cell Biology ,Hematology ,Oncogenes ,Cytokine ,medicine.anatomical_structure ,Cancer research ,medicine.symptom ,business - Abstract
Trained immunity (TI) is a proinflammatory program induced in monocyte/macrophages upon sensing of specific pathogens and is characterized by immunometabolic and epigenetic changes that enhance cytokine production. Maladaptive activation of TI (ie, in the absence of infection) may result in detrimental inflammation and development of disease; however, the exact role and extent of inappropriate activation of TI in the pathogenesis of human diseases is undetermined. In this study, we uncovered the oncogene-induced, maladaptive induction of TI in the pathogenesis of a human inflammatory myeloid neoplasm (Erdheim-Chester disease, [ECD]), characterized by the BRAFV600E oncogenic mutation in monocyte/macrophages and excess cytokine production. Mechanistically, myeloid cells expressing BRAFV600E exhibit all molecular features of TI: activation of the AKT/mammalian target of rapamycin signaling axis; increased glycolysis, glutaminolysis, and cholesterol synthesis; epigenetic changes on promoters of genes encoding cytokines; and enhanced cytokine production leading to hyperinflammatory responses. In patients with ECD, effective therapeutic strategies combat this maladaptive TI phenotype; in addition, pharmacologic inhibition of immunometabolic changes underlying TI (ie, glycolysis) effectively dampens cytokine production by myeloid cells. This study revealed the deleterious potential of inappropriate activation of TI in the pathogenesis of human inflammatory myeloid neoplasms and the opportunity for inhibition of TI in conditions characterized by maladaptive myeloid-driven inflammation.
- Published
- 2021
36. Abstract 3297: IFNalpha by in vivo-engineered macrophages abates liver metastases and triggers counter regulatory responses limiting efficacy
- Author
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Thomas Kerzel, Stefano Beretta, Eloise Scamardella, Chiara Balestrieri, Tamara Canu, Federica Pedica, Rossana Norata, Lucia Sergi Sergi, Marco Genua, Ostuni Renato, Anna Kajaste-Rudnitski, Antonio Esposito, Masanobu Oshima, Giovanni Tonon, Francesca Sanvito, Mario Leonardo Squadrito, and Luigi Naldini
- Subjects
Cancer Research ,Oncology - Abstract
The liver hosts an immune suppressive environment favouring metastatic seeding and proliferation of cancer cells. Pharmacological treatments, including immunotherapies, fail in the presence of liver metastases (LMS). Therefore, identifying new interventional tools and key targetable players involved in the immunosuppressive environment is of pivotal importance. We developed a lentiviral vector (LV)-based platform for selective genetic engineering of resident and tumour-associated macrophages enabling locally-sourced delivery of therapeutic molecules to LMS. Selective transgene expression is driven by a macrophage specific promoter and fine-tuned by microRNA target sequences. Upon systemic delivery of the LV, we observed enhanced transgene expression in macrophages located in areas surrounding LMS. We then equipped the LV with an IFNα-coding sequence, a cytokine with pleiotropic immune effects. Long term analysis in mice showed LV dose-dependent, sustained and well-tolerated IFNα expression. To investigate the therapeutic efficacy, we employed a colorectal cancer (CRC) organoid-based syngeneic mouse model of LMS containing molecular and histopathological hallmarks of the human disease. IFNα LV treatment significantly delayed LMS growth reaching a complete response in up to 50%. Single cell omics of LMS from IFNα LV-treated mice showed upregulation of IFNα-responsive genes, macrophage skewing to an antigen presenting (M1-like) polarization state, and expansion as well as reduced exhaustion of LMS-associated antigen specific CD8 T cells. Employing spatial transcriptomics, we found that the interface between LMS and liver parenchymal tissue was the major site of IFNα action, which was associated with enhanced immune activation and antigen presentation. Furthermore, we observed decreased angiogenesis and hypoxia in IFNα LV-treated LMS. When comparing LMS of treatment responsive to resistant mice, we found accumulation of activated CD8 T-cells in responsive whereas a high number of immunosuppressive T regulatory type 1 (TR1)-like cells in resistant mice. Molecular analyses suggest that TR1-like cell infiltration was associated with increased IL10 signaling in resistant mice. Furthermore, we found a positive correlation between IFNα and TR1-like cell signatures in human LMS and primary CRC thus supporting the link between IFNα activation and expansion of TR1-like cells in cancer. In summary, we developed an innovative gene-based platform that upon a single well-tolerated intravenous LV infusion rapidly promotes a protective therapeutic response against LMS through enabling immune activation. However, we also found that TR1-like cells might promote tumor immune evasion in presence of IFNα signaling in this setting, suggesting targeting of TR1-like cells when facing resistance to cancer immunotherapies that trigger IFNα signaling. Citation Format: Thomas Kerzel, Stefano Beretta, Eloise Scamardella, Chiara Balestrieri, Tamara Canu, Federica Pedica, Rossana Norata, Lucia Sergi Sergi, Marco Genua, Ostuni Renato, Anna Kajaste-Rudnitski, Antonio Esposito, Masanobu Oshima, Giovanni Tonon, Francesca Sanvito, Mario Leonardo Squadrito, Luigi Naldini. IFNalpha by in vivo-engineered macrophages abates liver metastases and triggers counter regulatory responses limiting efficacy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3297.
- Published
- 2022
37. Corrigendum: Generation of Powerful Human Tolerogenic Dendritic Cells by Lentiviral-Mediated IL-10 Gene Transfer
- Author
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Giada Amodio, Fabio Russo, Francesca Romana Santoni de Sio, Silvia Gregori, Michela Comi, Grazia Andolfi, Anna Kajaste-Rudnitski, Luca Cesana, Laura Passeri, and Marta Fortunato
- Subjects
lcsh:Immunologic diseases. Allergy ,immune tolerance ,Allogeneic transplantation ,Immunology ,Gene transfer ,Biology ,allogeneic transplantation ,Immune tolerance ,Cell therapy ,Interleukin 10 ,IL-10 ,Cancer research ,Immunology and Allergy ,dendritic cells ,cell therapy ,lcsh:RC581-607 - Published
- 2021
38. Oncogene-induced maladaptive activation of trained immunity in the pathogenesis and treatment of Erdheim-Chester disease
- Author
-
Biavasco, Riccardo, primary, Molteni, Raffaella, additional, Stefanoni, Davide, additional, Nemkov, Travis, additional, Netea, Mihai G., additional, Domínguez-Andrés, Jorge, additional, Arts, Rob JW, additional, Merelli, Ivan, additional, Mazza, Davide, additional, Zambrano, Samuel, additional, Panigada, Maddalena, additional, Cantoni, Eleonora, additional, Tengesdal, Isak, additional, Maksud, Philippe, additional, Piras, Francesco, additional, De Luca, Giacomo, additional, Cassina, Laura, additional, Distefano, Gianfranco, additional, Loffreda, Alessia, additional, Gnani, Daniela, additional, Doglioni, Claudio, additional, Tomelleri, Alessandro, additional, Campochiaro, Corrado, additional, Joosten, Leo, additional, Dinarello, Charles A, additional, Kajaste-Rudnitski, Anna, additional, Haroche, Julien, additional, Cardaci, Simone, additional, Cenci, Simone, additional, Dagna, Lorenzo, additional, Ferrarini, Marina, additional, Ferrero, Elisabetta, additional, Boletta, Alessandra, additional, D'Alessandro, Angelo, additional, Montini, Eugenio, additional, and Cavalli, Giulio, additional
- Published
- 2021
- Full Text
- View/download PDF
39. Oncogene-induced maladaptive activation of trained immunity in the pathogenesis and treatment of Erdheim-Chester disease.
- Author
-
Molteni, R., Biavasco, R., Stefanoni, D., Nemkov, T., Dominguez Andres, J., Arts, R.J.W., Merelli, I., Mazza, D., Zambrano, S., Panigada, M., Cantoni, E., Tengesdal, I.W., Maksud, P., Piras, F., Cesana, D., Cassina, L., Distefano, G., Loffreda, A., Gnani, D., Luca, G. De, Tomelleri, A., Campochiaro, C., Joosten, L.A.B., Dinarello, C.A., Kajaste-Rudnitski, A., Haroche, J., Cardaci, S., Cenci, S., Dagna, L., Doglioni, C., Ferrarini, M., Ferrero, E., Boletta, A., d'Alessandro, A., Montini, E., Netea, M.G., Cavalli, G.C., Molteni, R., Biavasco, R., Stefanoni, D., Nemkov, T., Dominguez Andres, J., Arts, R.J.W., Merelli, I., Mazza, D., Zambrano, S., Panigada, M., Cantoni, E., Tengesdal, I.W., Maksud, P., Piras, F., Cesana, D., Cassina, L., Distefano, G., Loffreda, A., Gnani, D., Luca, G. De, Tomelleri, A., Campochiaro, C., Joosten, L.A.B., Dinarello, C.A., Kajaste-Rudnitski, A., Haroche, J., Cardaci, S., Cenci, S., Dagna, L., Doglioni, C., Ferrarini, M., Ferrero, E., Boletta, A., d'Alessandro, A., Montini, E., Netea, M.G., and Cavalli, G.C.
- Abstract
Item does not contain fulltext, Trained immunity (TI) is a proinflammatory program induced in monocyte/macrophages upon sensing of specific pathogens and is characterized by immunometabolic and epigenetic changes that enhance cytokine production. Maladaptive activation of TI (ie, in the absence of infection) may result in detrimental inflammation and development of disease; however, the exact role and extent of inappropriate activation of TI in the pathogenesis of human diseases is undetermined. In this study, we uncovered the oncogene-induced, maladaptive induction of TI in the pathogenesis of a human inflammatory myeloid neoplasm (Erdheim-Chester disease, [ECD]), characterized by the BRAFV600E oncogenic mutation in monocyte/macrophages and excess cytokine production. Mechanistically, myeloid cells expressing BRAFV600E exhibit all molecular features of TI: activation of the AKT/mammalian target of rapamycin signaling axis; increased glycolysis, glutaminolysis, and cholesterol synthesis; epigenetic changes on promoters of genes encoding cytokines; and enhanced cytokine production leading to hyperinflammatory responses. In patients with ECD, effective therapeutic strategies combat this maladaptive TI phenotype; in addition, pharmacologic inhibition of immunometabolic changes underlying TI (ie, glycolysis) effectively dampens cytokine production by myeloid cells. This study revealed the deleterious potential of inappropriate activation of TI in the pathogenesis of human inflammatory myeloid neoplasms and the opportunity for inhibition of TI in conditions characterized by maladaptive myeloid-driven inflammation.
- Published
- 2021
40. D-mannose suppresses macrophage IL-1β production
- Author
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Luisa Ricci, Sabrina Di Marco, S. Torretta, Ivan Cuccovillo, Francesco Mainini, Anna Kajaste-Rudnitski, Kevin M. Ryan, David Sumpton, Simone Cardaci, and Alessandra Scagliola
- Subjects
Lipopolysaccharides ,0301 basic medicine ,Regulatory T cell differentiation ,Science ,Interleukin-1beta ,General Physics and Astronomy ,Mannose ,Carbohydrate metabolism ,Article ,Monocytes ,General Biochemistry, Genetics and Molecular Biology ,Cell Line ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,In vivo ,Metabolomics ,Animals ,Humans ,Macrophage ,Monocytes and macrophages ,Inflammation ,Regulation of gene expression ,Mannosephosphates ,Multidisciplinary ,Chemistry ,Macrophages ,Cell Differentiation ,General Chemistry ,Macrophage Activation ,Colitis ,Hypoxia-Inducible Factor 1, alpha Subunit ,Cell biology ,030104 developmental biology ,Gene Expression Regulation ,Metabolic pathways ,Cell culture ,030220 oncology & carcinogenesis ,Metabolic Networks and Pathways ,Intracellular - Abstract
D-mannose is a monosaccharide approximately a hundred times less abundant than glucose in human blood. Previous studies demonstrated that supraphysiological levels of D-mannose inhibit tumour growth and stimulate regulatory T cell differentiation. It is not known whether D-mannose metabolism affects the function of non-proliferative cells, such as inflammatory macrophages. Here, we show that D-mannose suppresses LPS-induced macrophage activation by impairing IL-1β production. In vivo, mannose administration improves survival in a mouse model of LPS-induced endotoxemia as well as decreases progression in a mouse model of DSS-induced colitis. Phosphomannose isomerase controls response of LPS-activated macrophages to D-mannose, which impairs glucose metabolism by raising intracellular mannose-6-phosphate levels. Such alterations result in the suppression of succinate-mediated HIF-1α activation, imposing a consequent reduction of LPS-induced Il1b expression. Disclosing an unrecognized metabolic hijack of macrophage activation, our study points towards safe D-mannose utilization as an effective intervention against inflammatory conditions., Mannose is present at trace levels in blood and regulates cancer growth. Here the authors show that supraphysiological levels of mannose can also regulate macrophages, limiting their production of IL-1β and increasing resistance of mice to LPS-induced endotoxemia and DSS-induced colitis.
- Published
- 2020
41. Restriction factors of retroviral replication: the example of Tripartite Motif (TRIM) protein 5α and 22
- Author
-
Kajaste-Rudnitski, Anna, Pultrone, Cinzia, Marzetta, Flavia, Ghezzi, Silvia, Coradin, Tiziana, and Vicenzi, Elisa
- Published
- 2010
- Full Text
- View/download PDF
42. Assessing the Impact of Cyclosporin A on Lentiviral Transduction and Preservation of Human Hematopoietic Stem Cells in Clinically Relevant Ex Vivo Gene Therapy Settings
- Author
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Francesco Piras, Carolina Petrillo, Alessia Capotondo, Eugenio Montini, Andrea Calabria, Bernhard Gentner, Ivan Cuccovillo, Giulio Spinozzi, Anna Kajaste-Rudnitski, Luigi Naldini, Fabrizio Benedicenti, Alessandra Biffi, Petrillo, C., Calabria, A., Piras, F., Capotondo, A., Spinozzi, G., Cuccovillo, I., Benedicenti, F., Naldini, L., Montini, E., Biffi, A., Gentner, B., and Kajaste-Rudnitski, A.
- Subjects
Permissiveness ,engraftment and stemne ,Mice ,Transduction (genetics) ,0302 clinical medicine ,Transduction, Genetic ,Cyclosporin a ,Research Articles ,Mice, Knockout ,0303 health sciences ,Lentiviral transduction ,Cell Cycle ,Graft Survival ,Gene Transfer Techniques ,Hematopoietic Stem Cell Transplantation ,Chromosome Mapping ,Cell biology ,Haematopoiesis ,human hematopoietic stem cells ,engraftment and stemness ,lentiviral transduction ,transduction enhancers ,030220 oncology & carcinogenesis ,Cyclosporine ,Molecular Medicine ,Genetic Vector ,Stem cell ,Human ,Virus Integration ,Genetic Vectors ,Biology ,Lentiviru ,Viral vector ,Colony-Forming Units Assay ,03 medical and health sciences ,Genetics ,Animals ,Humans ,Progenitor cell ,human hematopoietic stem cell ,Molecular Biology ,Cell Proliferation ,030304 developmental biology ,Animal ,Lentivirus ,Hematopoietic Stem Cell ,Genetic Therapy ,Gene Transfer Technique ,Hematopoietic Stem Cells ,transduction enhancer - Abstract
Improving hematopoietic stem and progenitor cell (HSPC) permissiveness to lentiviral vector (LV) transduction without compromising their biological properties remains critical for broad-range implementation of gene therapy as a treatment option for several inherited diseases. This study demonstrates that the use of one-hit ex vivo LV transduction protocols based on either cyclosporin A (CsA) or rapamycin enable as efficient gene transfer as the current two-hit clinical standard into bone marrow–derived CD34+ cells while better preserving their engraftment capacity in vivo. CsA was additive with another enhancer of transduction, prostaglandin E2, suggesting that tailored enhancer combinations may be applied to overcome multiple blocks to transduction simultaneously in HSPC. Interestingly, besides enhancing LV transduction, CsA also significantly reduced HSPC proliferation, preserving the quiescent G0 fraction and the more primitive multipotent progenitors, thereby yielding the highest engraftment levels in vivo. Importantly, no alterations in the vector integration profiles could be detected between CsA and control transduced HSPC. Overall, the present findings contribute to the development of more efficient and sustainable LV gene therapy protocols, underscoring the benefits of scaling down required vector doses, as well as shortening the HSPC ex vivo culture time.
- Published
- 2019
43. The lysine-rich intracellular loop and cell type-specific co-factors are required for IFITM3 antiviral immunity in hematopoietic stem cells
- Author
-
Unali, G., primary, Giordano, A.M.S, additional, Cuccovillo, I., additional, Alezz M., Abou, additional, Apolonia, L., additional, Merelli, I., additional, Malim, M. H., additional, Petrillo, C., additional, and Kajaste-Rudnitski, A., additional
- Published
- 2021
- Full Text
- View/download PDF
44. Corrigendum: Generation of Powerful Human Tolerogenic Dendritic Cells by Lentiviral-Mediated IL-10 Gene Transfer
- Author
-
Comi, Michela, primary, Amodio, Giada, additional, Passeri, Laura, additional, Fortunato, Marta, additional, Santoni de Sio, Francesca Romana, additional, Andolfi, Grazia, additional, Kajaste-Rudnitski, Anna, additional, Russo, Fabio, additional, Cesana, Luca, additional, and Gregori, Silvia, additional
- Published
- 2021
- Full Text
- View/download PDF
45. A novel STING variant triggers endothelial toxicity and SAVI disease
- Author
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Valeri, Erika, Breggion, Sara, Barzaghi, Federica, Abou Alezz, Monah, Crivicich, Giovanni, Pagani, Isabel, Forneris, Federico, Sartirana, Claudia, Costantini, Matteo, Costi, Stefania, Marino, Achille, Chiarotto, Eleonora, Colavito, Davide, Cimaz, Rolando, Merelli, Ivan, Vicenzi, Elisa, Aiuti, Alessandro, and Kajaste-Rudnitski, Anna
- Abstract
Gain-of-function mutations in STING cause STING-associated vasculopathy with onset in infancy (SAVI) characterized by early-onset systemic inflammation, skin vasculopathy, and interstitial lung disease. Here, we report and characterize a novel STING variant (F269S) identified in a SAVI patient. Single-cell transcriptomics of patient bone marrow revealed spontaneous activation of interferon (IFN) and inflammatory pathways across cell types and a striking prevalence of circulating naïve T cells was observed. Inducible STING F269S expression conferred enhanced signaling through ligand-independent translocation of the protein to the Golgi, protecting cells from viral infections but preventing their efficient immune priming. Additionally, endothelial cell activation was promoted and further exacerbated by cytokine secretion by SAVI immune cells, resulting in inflammation and endothelial damage. Our findings identify STING F269S mutation as a novel pathogenic variant causing SAVI, highlight the importance of the crosstalk between endothelial and immune cells in the context of lung disease, and contribute to a better understanding of how aberrant STING activation can cause pathology.
- Published
- 2024
- Full Text
- View/download PDF
46. Generation of Powerful Human Tolerogenic Dendritic Cells by Lentiviral-Mediated IL-10 Gene Transfer
- Author
-
Fabio Russo, Giada Amodio, Anna Kajaste-Rudnitski, Grazia Andolfi, Michela Comi, Marta Fortunato, Francesca Romana Santoni de Sio, Laura Passeri, Silvia Gregori, and Luca Cesana
- Subjects
lcsh:Immunologic diseases. Allergy ,Adoptive cell transfer ,immune tolerance ,T-Lymphocytes ,T cell ,Genetic Vectors ,Immunology ,Gene Expression ,Biology ,Monocytes ,Immunophenotyping ,Immune tolerance ,Viral vector ,Cell therapy ,Mice ,Immune system ,Transduction, Genetic ,medicine ,Animals ,Humans ,Immunology and Allergy ,dendritic cells ,Original Research ,Lentivirus ,Correction ,Interleukin-10 ,Cell biology ,allogeneic transplantation ,Interleukin 10 ,medicine.anatomical_structure ,Humanized mouse ,IL-10 ,Female ,cell therapy ,lcsh:RC581-607 - Abstract
The prominent role of dendritic cells (DC) in promoting tolerance and the development of methods to generate clinical grade products allowed the clinical application of tolerogenic DC (tolDC)-based therapies for controlling unwanted immune responses. We established an efficient method to generate tolerogenic human DC, producing supra-physiological levels of IL-10, by genetically engineering monocyte-derived DC with a bidirectional Lentiviral Vector (bdLV) encoding for IL-10 and a marker gene. DCIL−10 are mature DC, modulate T cell responses, promote T regulatory cells, and are phenotypically and functionally stable upon stimulation. Adoptive transfer of human DCIL−10 in a humanized mouse model dampens allogeneic T cell recall responses, while murine DCIL−10 delays acute graft-vs.-host disease in mice. Our report outlines an efficient method to transduce human myeloid cells with large-size LV and shows that stable over-expression of IL-10 generates an effective cell product for future clinical applications in the contest of allogeneic transplantation.
- Published
- 2020
47. Antiviral immunity and nucleic acid sensing in haematopoietic stem cell gene engineering
- Author
-
Anna Kajaste-Rudnitski and Francesco Piras
- Subjects
Genetic enhancement ,Genetic Vectors ,Review Article ,Computational biology ,Biology ,Antiviral Agents ,Viral vector ,03 medical and health sciences ,0302 clinical medicine ,Nucleic Acids ,Genetics ,Humans ,Progenitor cell ,Molecular Biology ,Gene ,030304 developmental biology ,Innate immunity ,0303 health sciences ,Innate immune system ,Haematopoietic stem cells ,Genetic Therapy ,Hematopoietic Stem Cells ,3. Good health ,Haematopoiesis ,030220 oncology & carcinogenesis ,Nucleic acid ,Molecular Medicine ,Stem cell - Abstract
The low gene manipulation efficiency of human hematopoietic stem and progenitor cells (HSPC) remains a major hurdle for sustainable and broad clinical application of innovative therapies for a wide range of disorders. Given that all current and emerging gene transfer and editing technologies are bound to expose HSPC to exogenous nucleic acids and most often also to viral vectors, we reason that host antiviral factors and nucleic acid sensors play a pivotal role in the efficacy of HSPC genetic manipulation. Here, we review recent progress in our understanding of vector–host interactions and innate immunity in HSPC upon gene engineering and discuss how dissecting this crosstalk can guide the development of more stealth and efficient gene therapy approaches in the future.
- Published
- 2020
- Full Text
- View/download PDF
48. Lipid-Based Nanoparticles as Nonviral Gene Delivery Vectors
- Author
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Pezzoli, Daniele, primary, Kajaste-Rudnitski, Anna, additional, Chiesa, Roberto, additional, and Candiani, Gabriele, additional
- Published
- 2013
- Full Text
- View/download PDF
49. A variant in the CD209 promoter is associated with severity of dengue disease
- Author
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Sakuntabhai, Anavaj, Turbpaiboon, Chairat, Casadémont, Isabelle, Chuansumrit, Ampaiwan, Lowhnoo, Tassanee, Kajaste-Rudnitski, Anna, Kalayanarooj, Sita Mint, Tangnararatchakit, Kanchana, Tangthawornchaikul, Nattaya, Vasanawathana, Sirijit, Chaiyaratana, Wathanee, Yenchitsomanus, Pa-thai, Suriyaphol, Prapat, Avirutnan, Panisadee, Chokephaibulkit, Kulkanya, Matsuda, Fumihiko, Yoksan, Sutee, Jacob, Yves, Lathrop, G Mark, Malasit, Prida, Desprès, Philippe, and Julier, Cécile
- Published
- 2005
- Full Text
- View/download PDF
50. Laboratory-Scale Lentiviral Vector Production and Purification for Enhanced Ex Vivo and In Vivo Genetic Engineering
- Author
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Soldi, Monica, primary, Sergi Sergi, Lucia, additional, Unali, Giulia, additional, Kerzel, Thomas, additional, Cuccovillo, Ivan, additional, Capasso, Paola, additional, Annoni, Andrea, additional, Biffi, Mauro, additional, Rancoita, Paola Maria Vittoria, additional, Cantore, Alessio, additional, Lombardo, Angelo, additional, Naldini, Luigi, additional, Squadrito, Mario Leonardo, additional, and Kajaste-Rudnitski, Anna, additional
- Published
- 2020
- Full Text
- View/download PDF
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