Lentivector Knockdown of CCR5 in Hematopoietic Stem and Progenitor Cells Confers Functional and Persistent HIV-1 Resistance in Humanized Mice
| Autor: | Annette Audigé, Patrick Salmon, Duo Li, Michael S. Pepper, Renier Myburgh, Stephan Regenass, Karl-Heinz Krause, Vincent Jaquet, Markus G. Manz, Mary-Aude Rochat, Roberto F. Speck, Sandra Ivic, Gustavo Gers-Huber |
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| Přispěvatelé: | University of Zurich, Speck, Roberto F |
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
1109 Insect Science
Genetic enhancement CD34 HIV Infections Mice SCID ddc:616.07 HIV Infections/immunology/virology 10234 Clinic for Infectious Diseases ddc:576.5 Genetic Therapy/methods Disease Resistance education.field_of_study Gene knockdown 2404 Microbiology virus diseases Viral Load Haematopoiesis Gene Knockdown Techniques Receptors CCR5 Immunology Population Genetic Vectors Virus Attachment 610 Medicine & health Biology Microbiology Gene Delivery Immune system Receptors HIV Virology HIV-1/physiology Animals Humans Progenitor cell education 2403 Immunology Transplantation Lentivirus/genetics Lentivirus Receptors CCR5/metabolism Genetic Therapy Hematopoietic Stem Cells Hematopoietic Stem Cells/immunology/virology Insect Science 10032 Clinic for Oncology and Hematology 10033 Clinic for Immunology 2406 Virology Cancer research HIV-1 Receptors HIV/antagonists & inhibitors |
| Zdroj: | Journal of Virology, Vol. 89, No 13 (2015) pp. 6761-72 |
| ISSN: | 0022-538X |
| Popis: | Gene-engineered CD34 + hematopoietic stem and progenitor cells (HSPCs) can be used to generate an HIV-1-resistant immune system. However, a certain threshold of transduced HSPCs might be required for transplantation into mice for creating an HIV-resistant immune system. In this study, we combined CCR5 knockdown by a highly efficient microRNA (miRNA) lentivector with pretransplantation selection of transduced HSPCs to obtain a rather pure population of gene engineered CD34 + cells. Low-level transduction of HSPCs and subsequent sorting by flow cytometry yielded >70% transduced cells. Mice transplanted with these cells showed functional and persistent resistance to a CCR5-tropic HIV strain: viral load was significantly decreased over months, and human CD4 + T cells were preserved. In one mouse, viral mutations, resulting presumably in a CXCR4-tropic strain, overcame HIV resistance. Our results suggest that HSPC-based CCR5 knockdown may lead to efficient control of HIV in vivo . We overcame a major limitation of previous HIV gene therapy in humanized mice in which only a proportion of the cells in chimeric mice in vivo are anti-HIV engineered. Our strategy underlines the promising future of gene engineering HIV-resistant CD34 + cells that produce a constant supply of HIV-resistant progeny. IMPORTANCE Major issues in experimental long-term in vivo HIV gene therapy have been (i) low efficacy of cell transduction at the time of transplantation and (ii) transduction resulting in multiple copies of heterologous DNA in target cells. In this study, we demonstrated the efficacy of a transplantation approach with a selection step for transduced cells that allows transplantation of an enriched population of HSPCs expressing a single (low) copy of a CCR5 miRNA. Efficient maintenance of CD4 + T cells and a low viral titer resulted only when at least 70% of the HIV target cells were genetically modified. These findings imply that clinical protocols of HIV gene therapy require a selective enrichment of genetically targeted cells because positive selection of modified cells is likely to be insufficient below this threshold. This selection approach may be beneficial not only for HIV patients but also for other patients requiring transplantation of genetically modified cells. |
| Databáze: | OpenAIRE |
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