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In Vivo Gene Editing for HIV-1 Cure

体内基因编辑治疗 HIV-1

基本信息

批准号：
10331787
负责人：
IRVIN S.Y. CHEN
金额：
$ 67.8万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-11 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10331787
关键词：
AIDS therapy Ablation Address Anti-Retroviral Agents Antibody Therapy BLT mice Bar Codes Binding Biodistribution Bioinformatics Biological Products Blood Circulation Brain CCR5 gene CD4 Positive T Lymphocytes CRISPR/Cas technology Cell surface Cells Charge Chemical Engineering Chemical Structure Clinical Research Complex Crosslinker DNA Disease Encapsulated Engineering Freeze Drying Gene Delivery Gene-Modified Genes Genetic Engineering Genetic Translation Guide RNA HIV Genome HIV-1 Hematopoietic Hematopoietic stem cells Homing Immune Immune system In Situ In Vitro Individual Infection Intravenous Journals Knock-out Lentivirus Vector Ligands Liver Lung Lymphoid Mainstreaming Modeling Mutate Nanotechnology Nucleic Acids Organ Patients Pharmaceutical Preparations Polymers Positioning Attribute Property Proteins Proviruses Publications Reagent Resistance Rest Role Scientist Site Small Interfering RNA Surface Properties System T-Lymphocyte Testing The Sun Therapeutic Thinness Time Tissues Transduction Gene Viral Vector Virion Work clinical practice delivery vehicle endonuclease experience gene therapy immunogenicity in vivo knock-down macromolecule macrophage monomer mouse model nanocapsule nanotechnology platform neoplastic cell novel nucleic acid-based therapeutics particle polymerization preclinical study reactivation from latency receptor small molecule tool transcription activator-like effector nucleases vector virtual zinc finger nuclease

项目摘要

PROJECT SUMMARY The overall hypothesis to be tested in this proposal is that a novel class of nanocapsules can effectively deliver gene editing components into the two primary HIV-1 target cells, T-cells and macrophages, and mutagenize the HIV-1 provirus such that replication and/or reactivation from latency is aborted. While gene modification is challenging, the advantage over small molecule drugs is that the HIV-1 provirus or genes necessary for HIV-1 expression and/or infection can be directly knocked down or knocked out without the need to kill the infected cells. Efficient gene-modification activity has been achieved by a number of systems including zinc-finger nucleases (ZNFs), transcription activator-like effector nucleases (TALENs), homing endonucleases, and most recently, the CRISPR/Cas9 system. Despite the promise of these new gene editing tools, therapeutic nucleic acids and proteins are rapidly lost from circulation and delivery vehicles cannot deliver gene modifying reagents by effective means to impact HIV-1 reservoirs. Thus, to date, all applications of gene modification for HIV-1 disease are currently practiced on cells removed from the body and transduced ex vivo. From our past experience with engineered lentiviral vectors, we recognize the difficult challenges of developing tools for in vivo gene editing, but also the promise and potential of bringing gene therapy into mainstream clinical practice. Our prior experience teaches us that viral vectors suffer from limitations in titer, adequate biodistribution, poor transduction of resting T-cells, complex genetic engineering, and immunogenicity. Recently, we developed a nanotechnology platform whereby individual macromolecules, protein, siRNA, gRNA, or DNA, are encapsulated and protected within a thin polymer shell by in situ polymerization of monomers and stabilized by environmentally responsive crosslinkers. In many respects, these “nanocapsules” are similar to virion particles, being of similar size and, like virions, protect the single encased gene. However, they have the advantage of simple manufacturing to higher “titer”, storage by freeze-dry, and, most importantly, the ability to easily alter the surface properties of chemical structure, charge, and ligand conjugation which determines factors such as biodistribution, cell binding, and entry. Since the properties of the nanocapsule are conferred by the shell which shields the cargo, virtually any nucleic acid or protein cargo can be interchanged. By judicious choice of polymer shell and crosslinkers, we successfully engineered nanocapsules which enhance biodistribution to reservoir sites, release a model cargo in time release fashion, and target specific cells in vivo through ligand recognition of cell surface molecules. Furthermore, these nanocapsules themselves are relatively non- immunogenic and shield the cargo from the immune system. These proof of principle studies begin to overcome the challenges outlined above and thus provide the basis for our proposed studies.

项目总结