Defining a therapeutic threshold in HSPC gene therapy for HIV-1 disease

定义 HIV-1 疾病 HSPC 基因治疗的治疗阈值

• 批准号: 9263831
• 负责人: Sanggu Kim
• 金额: $ 24.9万
• 依托单位: OHIO STATE UNIVERSITY
• 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 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9263831
• 关键词: Acquired Immunodeficiency Syndrome; Acute Myelocytic Leukemia; Alpha Cell; Animal Model; Anti-HIV Agents; Award; Behavior; Berlin; Biological Assay; Biology; Bone Marrow Transplantation; CCR5 gene; CD4 Positive T Lymphocytes; Cell Therapy; Cell Transplantation; Cells; Chronic Disease; Clinical; Clinical Research; Clinical Trials; Data; Development; Diagnostic; Diphtheria Toxin; Disease; Dose; Educational Background; Engineering; Engraftment; Environment; Evaluation; Financial Support; Future; Gene-Modified; Genes; Goals; Grant; HIV; HIV-1; Health; Hematopoietic stem cells; Highly Active Antiretroviral Therapy; Homologous Transplantation; Human; Immune system; Immunotherapy; Inbred Mouse; Infection; Institutes; Kinetics; Knowledge; Lentivirus Vector; Mentors; Mutation; Natural regeneration; Pathogenesis; Patients; Pharmacotherapy; Phase; Population; Positioning Attribute; Publishing; RNA Interference; Recovery; Regimen; Research; Resistance; Scientist; Stem cell transplant; Stem cells; System; T-Lymphocyte; Technology; Testing; Therapeutic; Toxic effect; Translational Research; antiretroviral therapy; attenuation; biomathematics; career; cell killing; chemokine receptor; design; expectation; experience; experimental study; gamma-Chemokines; gene therapy; high throughput screening; humanized mouse; improved; innovation; knock-down; macrophage; mathematical model; mouse model; nonhuman primate; novel; pandemic disease; preclinical study; preclinical trial; pressure; prevent; prognostic; receptor; reconstitution; small hairpin RNA; success; therapeutic gene; training opportunity

DESCRIPTION (provided by applicant): The success of hematopoietic stem progenitor cell (HSPC) genetic therapy for HIV-1 is dependent in part upon achieving sufficient levels of marking with therapeutic gene modification to protect the immune system from HIV-1 disease. I propose here to test the hypothesis that a certain threshold level of gene marking is required to effectively replace the HIV-damaged immune system with one that is resistant to HIV-1. Below this threshold, the immune system would not be protected; above the threshold, the immune system would be protected and effectively prevent AIDS. All past and current HSPC clinical trials - which are designed to achieve as high a level of gene marking as is safely possible, anticipating that HIV-1 will ultimately select for the HIV-protected cells - are moving forward without an understanding of this requirement. Herein, I propose to test the hypothesis using mixed bone marrow transplant murine models. I will use a sensitive and high- throughput clonal quantification assay I developed and mathematical modeling to investigate HSPC repopulation and T-cell dynamics above and below this threshold in the presence and absence of selection pressure. Dr. Irvin Chen, Director of the UCLA AIDS Institute and my primary mentor, and his research group were the first to develop lentivirus vectors for HIV-1 gene therapy using short hairpin RNAi against the HIV-1 co-receptor, C-C chemokine receptor type 5 (CCR5), and have demonstrated long-term and stable reduction of CCR5 in both non-human primates and humanized mice. As we prepare to introduce this shRNA into HSPC gene therapy for humans, I will use CCR5 knockdown as the population of T-cells protected from HIV-1 infection and analyze their repopulation kinetics. Knowledge gained from these experiments will help us to determine optimal conditions for efficient and safe selection of engineered cells and to create a mathematical model that can guide further experiments and preclinical trials of engineered HSPC transplantation. I am in an excellent position to pursue this important research thanks to the superior mentoring of Dr. Chen and Dr. Kenneth Lange of Biomathematics; my research and educational backgrounds in both engineering and basic biology; and UCLA's rich research environment. My long-term career goal is to become an independent and specialized translational research scientist in the stem cell gene therapy field. My current research goal is t help introduce the practice of quantitative assessment to better understand and improve treatment conditions and diagnostic/prognostic evaluation in both clinical and preclinical studies of HIV diseases. During the mentored phase of the K99/R00 award, I plan to expand my research expertise in both stem cell gene therapy and mathematical modeling. The research experience I will gain from this training opportunity, along with the financial support granted in the independent phase, will significantly help me to pursue my long-term career and research goals.

描述（由申请人提供）：HIV-1的造血茎祖细胞（HSPC）基因治疗的成功部分取决于通过治疗基因修饰获得足够的标记水平，以保护免疫系统免受HIV-1疾病的侵害。我在这里提出要检验以下假设：需要一定的基因标记阈值水平来有效地替代对HIV-1具有抗性的HIV损害免疫系统。在此阈值以下，免疫系统将不受保护。在阈值之上，免疫系统将受到保护并有效防止艾滋病。所有过去和当前的HSPC临床试验（旨在实现最高水平的基因标记），预计HIV -1最终将为受HIV保护的细胞选择 - 不了解这一要求。本文中，我建议使用混合骨髓移植鼠模型检验假设。在存在和不存在选择压力的情况下，我将使用我开发的灵敏且高吞吐量的克隆定量测定和数学建模来研究该阈值上方和下方的T细胞动力学。 UCLA AIDS研究所主任和我的主要导师伊尔文·陈（Irvin Chen）和他的研究小组是第一个使用短发夹RNAI对HIV-1共肽，C-C趋化因子受体5（CCR5）开发HIV-1基因疗法的慢病毒载体，并表明了长期和稳定的人类和稳定的人类和稳定的人类和稳定的人。当我们准备将此SHRNA引入人类HSPC基因治疗中时，我将使用CCR5敲低，因为T细胞的种群受到了HIV-1感染的影响，并分析了其重生动力学。从这些实验中获得的知识将有助于我们确定有效且安全选择工程细胞的最佳条件，并创建一个数学模型，该模型可以指导工程HSPC移植的进一步实验和临床前试验。由于陈（Chen）博士和生物护理学的肯尼斯·兰格（Kenneth Lange）博士的卓越指导，我能够进行这项重要的研究。我在工程和基本生物学方面的研究和教育背景；以及UCLA丰富的研究环境。我的长期职业目标是成为干细胞基因治疗领域的独立和专业的翻译研究科学家。我目前的研究目标是帮助介绍定量评估的实践，以更好地理解和改善HIV疾病临床和临床前研究中的治疗条件以及诊断/预后评估。在K99/R00奖的指导阶段，我计划扩大我在干细胞基因疗法和数学建模方面的研究专业知识。我将从这个培训机会中获得的研究经验，以及在独立阶段授予的财务支持，将极大地帮助我实现自己的长期职业和研究目标。