Coupled Gene Delivery and Protein Transduction for the Reduction of Atheroscleros

联合基因传递和蛋白质转导可减少动脉粥样硬化

• 批准号: 7612615
• 负责人: John Linforth Daiss
• 金额: $ 17.58万
• 依托单位: FIRST WAVE TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2010-05-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7612615
• 关键词: Acute; Adenovirus Vector; Adverse effects; Affect; Albumins; Animals; Apolipoproteins B; Blood; Blood Circulation; C-terminal; Cardiovascular system; Cell Culture System; Cell Nucleus; Cells; Chimera organism; Chimeric Proteins; Cholesterol; Clinical Management; Clinical Trials; Coculture Techniques; Costs and Benefits; Coupled; Coupling; Cytoplasm; Development; Dose; Elements; Engineering; Enzymes; Evaluation; Face; Fatty Liver; Foundations; Gene Delivery; Gene Transfer; Goals; Hamsters; Hepatic; Hepatocyte; Histopathology; Human; In Situ; In Vitro; Infection; Inflammation; Intervention; LDL Cholesterol Lipoproteins; Lead; Length; Lipid Mobilization; Lipids; Lipoproteins; Liver; Liver diseases; Low-Density Lipoproteins; Malignant Neoplasms; Mediating; Messenger RNA; Methods; Modeling; Mus; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Pathology; Patients; Peripheral Vascular Diseases; Phase; Population; Populations at Risk; Positioning Attribute; Preparation; Production; Proteins; RNA Editing; Rattus; Research; Risk; Risk Factors; Safety; Serum; Serum Albumin; Stream; System; Terminator Codon; Therapeutic; Therapeutic Uses; Tissues; Transfer RNA; Translating; Vendor; Very low density lipoprotein; Work; apoB mRNA editing catalytic subunit; apolipoprotein B-48; base; cell type; cost; design; design and construction; enzyme activity; enzyme replacement therapy; gene therapy; hepatoma cell; human TYRP1 protein; improved; in vivo; intravenous administration; longitudinal animal study; novel; novel strategies; novel therapeutics; particle; prospective; public health relevance; research study; response; therapeutic gene; therapeutic protein; transduction efficiency; uptake; vector

DESCRIPTION (provided by applicant): The proposed research will develop a novel therapeutic using as proof of principle the reduction of the atherogenic risk factor, LDL. Approximately 10% of the population at risk for developing cardiovascular and peripheral vascular diseases is unable to take therapeutic doses of current lipid lowering therapies due to undesirable side effects or limited response. Clinical management of obesity and type II diabetes faces complications due to hepatosteatosis and the atherogenic risk associated with lipid mobilization from the liver into the circulation. The theoretical value of editing enzyme (APOBEC-1) gene therapy for the induction of apolipoprotein B (apoB) mRNA editing in human liver and reduction of serum LDL through the expression and secretion of apoB48-containing very low-density lipoproteins (VLDL) has long been recognized. The challenge has been how to deliver APOBEC-1 therapeutically while minimizing the risk of side effects due to unregulated enzyme activity. Controlled editing activity has been achieved in primary hepatocytes through protein transduction using TAT on APOBEC-1. A novel approach for enzyme replacement therapy has been devised involving production of APOBEC-1 chimeras in situ that are secreted from producer cells and activate apoB mRNA editing only after they transduce into neighboring hepatocytes. Secretion of APOBEC by infected hepatocytes limits its accumulation in the producing cells, eliminating the risk of off-target effects from excess editing. Chimeric APOBEC-1 constructs will be optimized in vitro for secretion-transduction and their therapeutic benefit assessed in acute animal studies. The Specific Aims are: (1) construction of a APOBEC chimera optimized for secretion, transduction and induction of activity in recipient cells; (2) transfer of the gene for the optimized TAT-APOBEC chimera into an rAAV2 vector for infection of hepatocytes in vitro and in vivo, and (3) demonstration of the secretio transduction strategy in experimental animals and its safety and affect on serum lipoproteins. Public Health Relevance: The biomedical impact of the proposed research is the development of a novel gene therapeutic approach for the safe and effective replacement of enzyme activity. The proposed research uses as proof- of-principle the induction of apolipoprotein B mRNA-editing in liver by the delivery of the enzyme APOBEC-1 in order to produce a lipid carrying protein that will reduce the atherogenic risk factor in serum known as LDL cholesterol. In addition, this therapeutic approach will satisfy the current need for interventions that can reduce fatty liver disease without resulting in elevated serum LDL and atherogenic risk.

描述（由申请人提供）：拟议的研究将开发一种新的治疗方法，使用降低致动脉粥样硬化风险因子LDL作为原理证明。大约10%的处于发生心血管和外周血管疾病风险的人群由于不良副作用或有限的反应而不能服用治疗剂量的当前降脂疗法。肥胖和II型糖尿病的临床管理面临着由于脂肪肝和与脂质从肝脏动员到循环中相关的致动脉粥样硬化风险引起的并发症。编辑酶（APOBEC-1）基因治疗通过表达和分泌含apoB 48的极低密度脂蛋白（VLDL）来诱导人肝脏中载脂蛋白B（apoB）mRNA编辑和降低血清LDL的理论价值早已被认识。挑战是如何在治疗上递送APOBEC-1，同时最大限度地减少由于酶活性不受调节而引起的副作用的风险。通过达特对APOBEC-1进行蛋白转导，在原代肝细胞中实现了受控编辑活性。已经设计了一种用于酶替代疗法的新方法，其涉及原位产生APOBEC-1嵌合体，所述嵌合体从生产细胞分泌并仅在它们进入相邻肝细胞后激活apoB mRNA编辑。受感染肝细胞分泌APOBEC限制了其在生产细胞中的积累，消除了过量编辑的脱靶效应风险。嵌合APOBEC-1构建体将在体外优化用于分泌-转导，并在急性动物研究中评估其治疗益处。具体目标是：（1）构建优化的APOBEC嵌合体，用于在受体细胞中分泌、转导和诱导活性;（2）将优化的TAT-APOBEC嵌合体的基因转移到rAAV 2载体中，用于体外和体内感染肝细胞;和（3）在实验动物中证明分泌转导策略及其安全性和对血清脂蛋白的影响。 公共卫生相关性：拟议研究的生物医学影响是开发一种新的基因治疗方法，用于安全有效地替代酶活性。所提出的研究使用通过递送酶APOBEC-1诱导肝脏中的载脂蛋白B mRNA编辑作为原理证明，以产生将降低血清中的致动脉粥样硬化风险因子（称为LDL胆固醇）的脂质携带蛋白。此外，这种治疗方法将满足目前对可以减少脂肪肝疾病而不会导致血清LDL升高和致动脉粥样硬化风险的干预措施的需求。