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型的治疗性，用来证明原理的减少动脉粥样硬化危险因素LDL。由于不良的副作用或有限的反应有限，大约有10％的人口患心血管和周围血管疾病的风险降低了当前脂质降低疗法的治疗剂量。肥胖症和II型糖尿病的临床管理由于肝脏造成的肝脏并发症以及与脂质动员从肝脏转移到循环系统相关的动脉粥样硬化风险。长期认识到，长期认识到，编辑酶（APOBEC-1）基因疗法对于诱导人肝脏中载脂蛋白B（APOB）mRNA的诱导以及通过非常低密度的脂蛋白（VLDL）的表达和分泌减少血清LDL的理论值已得到认可。面临的挑战是如何通过治疗方法递送APOBEC-1，同时最大程度地降低了由于不管制的酶活性而导致的副作用风险。通过蛋白质转导使用TAT在APOBEC-1上，在原代肝细胞中已经实现了受控的编辑活性。已经设计了一种新型酶替代疗法的方法，涉及从生产细胞中分泌的Apobec-1嵌合体的产生，并仅在将APOB mRNA编辑转换为相邻的肝细胞后才激活APOB mRNA编辑。通过感染的肝细胞对APOBEC的分泌限制了其在生产细胞中的积累，从而消除了过量编辑中脱靶影响的风险。嵌合APOBEC-1构建体将在体外优化用于分泌转导的及其在急性动物研究中评估的治疗益处。具体目的是：（1）构建用于分泌，转导和诱导受体细胞活性的APOBEC嵌合体； （2）将优化的Tat-Apobec嵌合体转移到RAAV2载体中，以在体外和体内感染肝细胞，以及（3）证明实验动物中的Secretio转导策略及其安全性及其安全性及其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型的基因治疗方法，以安全有效地替代酶活性。拟议的研究用来证明通过递送酶Apobec-1在肝脏中诱导载脂蛋白B mRNA的诱导，以产生一种脂质携带的蛋白质，从而降低称为LDL胆固醇的血清中的动脉粥样硬化危险因素。此外，这种治疗方法将满足目前对可以减少脂肪肝病的干预措施的需求，而不会导致血清LDL升高和动脉粥样硬化风险。