Translational Studies of Paracrine CV Gene Transfer

旁分泌 CV 基因转移的转化研究

• 批准号: 8795690
• 负责人: H. Kirk Hammond
• 金额: --
• 依托单位: VA SAN DIEGO HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8795690
• 关键词: Apoptotic; Award; Biodistribution; Blood Circulation; CCI-779; Cardiac; Cardiovascular Diseases; Cardiovascular system; Clinical Trials; Congestive Heart Failure; Dependovirus; Disease; Distant; Dose; Doxycycline; Family suidae; Funding; Gene Transfer; Goals; Heart; Heart Diseases; Heart failure; Hormones; Immune response; Injection of therapeutic agent; Insulin-Like Growth Factor I; Intramuscular Injections; Intravenous; Laboratories; Length of Stay; Mediating; Methods; Modeling; Morbidity - disease rate; Myocardium; Office Visits; Patients; Peptides; Population; Primates; Rattus; Regulation; Research Design; Rodent; Safety; Serotyping; Serum; Site; Skeletal Muscle; System; Testing; Tetracyclines; Therapeutic; Toxic effect; Transgenes; Veterans; Virus; Water Supply; adeno-associated viral vector; base; economic cost; improved functioning; intravenous injection; novel; paracrine; promoter; response; selective expression; therapeutic transgene; transgene expression; translational study; vector

DESCRIPTION (provided by applicant): Gene transfer for the treatment of cardiovascular diseases is conceptually attractive, but difficulty in obtaining high yield transgene expression in the heart in a manner that can be easily and safely applied has been a chief impediment to progress. Current methods of gene transfer for heart disease include intramuscularinjection into heart muscle or intracoronary delivery, approaches that typically provide limited expression or are cumbersome to apply. Consequently, we have considered the usefulness of a vector encoding a paracrine-type transgene. In this approach, the transgene acts as a hormone, having cardiac effects after being released to the circulation from a distant site. This approach would circumvent the problem of attaining high yield cardiac gene transfer and enable patients to be treated by a systemic injection during an office visit. Furthermore, the approach proposed would eliminate the need for intravenous delivery of therapeutic peptides and thereby circumvent repeated and prolonged hospital stays, high morbidity, and enormous economic costs. The most suited vector to achieve these goals is the adeno-associated virus (AAV), which provides long term and extensive expression after intravenous delivery in rodents, pigs, and primates. Finally, by using regulated expression of the transgene in the AAV vector, one can achieve control of the level of transgene expression. To develop and optimally refine this approach is the focus of this proposal. Insulin-like growth factor-I (IGFI) is a peptide with protean favorable cardiovascular effects (inotrope, angiogenic, anti-apoptotic). Proof-of-concept studies in our laboratory recently showed that skeletal muscle injection of an AAV5 vector encoding IGFI improved function of the failing rat heart. IGFI will be an ideal therapeutic transgene in the proposed studies. To develop and refine such an approach, we propose to determine: a) mechanisms by which IGFI gene transfer increases contractile function; b) the optimal AAV vector and promoter for intravenous delivery that will provide maximal transgene expression with minimal immune response and off-target effects in rats; c) the optimal regulated transgene expression system to enable fine-tuning of serum transgene levels, and turning expression off and on as needed; d) the safety, efficacy, and survival advantage of IGFI gene transfer, using this optimal, paracrine-based approach in a rat model of congestive heart failure; and e) e) the minimally effective doses of AAV and activators of transgene expression following intravenous delivery of this optimal vector in normal pigs, using serum IGFI and immune response as end-points. These mechanistic and proof-of-concept studies are designed to be sufficient in scope to launch, under separate funding, a study in pigs with CHF, and subsequently to file an IND application to the FDA for the penultimate study: a clinical trial in veterans with congestive heart failure.

描述（由申请人提供）： 用于治疗心血管疾病的基因转移在概念上很有吸引力，但难以以容易且安全应用的方式在心脏中获得高产转基因表达一直是进展的主要障碍。目前用于心脏病的基因转移方法包括肌肉注射到心肌或冠状动脉内递送，这些方法通常提供有限的表达或应用起来很麻烦。因此，我们考虑了编码旁分泌型转基因的载体的有用性。在这种方法中，转基因充当激素，从远处释放到循环中后具有心脏作用。这种方法将规避获得高产量心脏基因转移的问题，并使患者能够在就诊期间接受全身注射治疗。此外，所提出的方法将消除静脉内输送治疗肽的需要，从而避免重复和长期住院、高发病率和巨大的经济成本。最适合实现这些目标的载体是腺相关病毒（AAV），它在啮齿动物、猪和灵长类动物中静脉注射后可提供长期和广泛的表达。最后，通过利用AAV载体中转基因的调节表达，可以实现转基因表​​达水平的控制。开发和优化这种方法是本提案的重点。 胰岛素样生长因子-I (IGFI) 是一种具有多种有利心血管作用（正性肌力、血管生成、抗细胞凋亡）的肽。我们实验室最近的概念验证研究表明，骨骼肌注射编码 IGFI 的 AAV5 载体可以改善衰竭大鼠心脏的功能。 IGFI 将成为拟议研究中理想的治疗性转基因。 为了开发和完善这种方法，我们建议确定：a) IGFI 基因转移增强收缩功能的机制； b) 用于静脉内递送的最佳 AAV 载体和启动子，将在大鼠中提供最大的转基因表达，同时具有最小的免疫反应和脱靶效应； c) 最佳调节转基因表达系统，能够微调血清转基因水平，并根据需要关闭和开启表达； d) 在充血性心力衰竭大鼠模型中使用这种基于旁分泌的最佳方法，IGFI 基因转移的安全性、有效性和生存优势； e) e) 在正常猪中静脉注射该最佳载体后，AAV 和转基因表达激活剂的最低有效剂量，使用血清 IGFI 和免疫反应作为终点。这些机制和概念验证研究的设计范围足以在单独的资助下启动一项针对患有 CHF 的猪的研究，并随后向 FDA 提交倒数第二项研究的 IND 申请：针对退伍军人的临床试验患有充血性心力衰竭。