Human Genes that Influence HIV-1 Replication, Pathogenesis, and Immunity in IVDUs

影响 IVDU 中 HIV-1 复制、发病机制和免疫的人类基因

• 批准号: 8893936
• 负责人: JEREMY LUBAN
• 金额: $ 82.49万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8893936
• 关键词: Acquired Immunodeficiency Syndrome; Address; Alleles; Cells; DNA delivery; Development; Disease Progression; Experimental Models; Genes; HIV-1; Hematopoietic; Hematopoietic stem cells; Human; Immune system; Immunity; Infection; Lentivirus Vector; Mus; Pathogenesis; Pathology; RNA Interference; Reproducibility; Research; Resistance; Somatic Cell; Source; Substance of Abuse; System; Technology; Testing; Transfection; drug abuser; gene replacement; genetic variant; homologous recombination; human embryonic stem cell; immune function; in vivo; in vivo Model; intravenous drug user; knockout gene; nuclease; prevent; research study; reverse genetics; tool; transmission process; vaccination strategy

DESCRIPTION (provided by applicant): Despite 30 years of AIDS research, there is still no robust, reverse-genetic system for studying the in vivo function of human genes that influence HIV-1 replication, pathogenesis, and immunity. As the number of such genes skyrockets, including genes that determine rates of HIV-1 acquisition and disease progression among intravenous drug users, the need for such technology has never been greater. This proposal addresses technical hurdles that must be overcome before such an experimental system can be realized. RNAi transformed the functional assessment of human genes, but lack of reproducibility and inability to assess allelic variants limit utility. Mouse gene knockout technology is unsurpassed at unambiguous assignment of function to particular mammalian genes. Unfortunately, many human genes lack simple mouse orthologues, including APOBEC3G and TRIMS, two genes that restrict HIV-1 infection among intravenous drug users. Worse, HIV-1 does not replicate in mouse cells. The project proposed here will develop tools for targeted gene replacement by homologous recombination in cells of the human immune system, and for functional assessment of these modified cells within the context of an in vivo model of HIV-1 transmission, replication, immunity, and AIDS pathogenesis. Towards this end, ongoing, cutting-edge technical developments from several fields will be exploited, including human embryonic stem cells and somatic cell reprogramming, murine immune system substitution with human counterparts, lentiviral vectors that permit efficient delivery of DNA to transfection-resistant cells, and designer nucleases to stimulate homologous recombination. Development of a perpetual source of isogenic, human hematopoietic stem cells that can be genetically-modified in a controlled fashion, and used to generate an immune system within an in vivo experimental model, will permit us to draw firm conclusions concerning the function of particular human genes - or of particular alleles - in hematopoietic development, immune function, and HIV-1 replication and pathogenesis, all within a setting of substances of abuse.

描述（由申请人提供）：尽管进行了30年的艾滋病研究，但仍然没有可靠的反遗传系统来研究影响HIV-1复制，发病机理和免疫力的人类基因的体内功能。作为此类基因的数量，包括确定静脉吸毒者中HIV-1获取和疾病进展速率的基因，对这种技术的需求从未有所更大。该提案解决了在实现这种实验系统之前必须克服的技术障碍。 RNAi改变了人类基因的功能评估，但缺乏可重现性和无法评估等位基因变体的限制效用。小鼠基因基因敲除技术在功能向特定哺乳动物基因的明确分配中无与伦比。不幸的是，许多人类基因缺乏简单的鼠标 直系同源物，包括apobec3g和Trims，这两个基因限制了静脉吸毒者中的HIV-1感染。更糟糕的是，HIV-1不会在小鼠细胞中复制。此处提出的项目将开发用于人类免疫系统细胞中同源重组的靶向基因替代的工具，并在HIV-1传播，复制，免疫和AIDS发病机理的体内模型的背景下对这些修饰细胞的功能评估。 为此，来自多个领域的持续，尖端的技术发展将是 被剥削，包括人类胚胎干细胞和躯体细胞重编程，鼠 与人类对应物，慢病毒载体的免疫系统替代，允许有效 将DNA传递到抗转染的细胞和设计器核酸酶以刺激同源重组。可以以受控的方式进行遗传改性的等源性，人造血干细胞的永久来源的发展，并用于在体内实验模型中产生免疫系统，使我们能够在特定的人类基因的功能中得出有关特定人类基因的功能 - 或在所有人的疾病中，均具有免疫，并且在整个人类基因中的功能，并在整体上进行了免疫，并具有免疫，并构成免疫，并构成免疫功能，并在shement and sernita and sernice and in Vornice and Intical and Intical and Intical and Intical and Intical and Intical and Intical and iniv -1 nefive and iniv -1。