New Approaches to Gene Therapy for Alpha-1 Antitrypsin Deficiency

 描述（由申请人提供）：该翻译项目的总体目标是开发一种针对α-1抗胰蛋白酶（AAT）缺乏导致的肺部疾病的确定性分子疗法，这是一种由于AAT基因突变导致的相对常见的单基因疾病。该计划建立在研究人员自20世纪90年代末以来开发几种不同版本的基于重组腺相关病毒（rAAV）的AAT缺陷基因疗法的经验基础上。项目主任实验室先前由NHLBI资助的工作包括概念验证（POC）研究，正式的IND使能安全性研究，以及肌肉注射第一代rAAV 2- AAT（在医生IND上）的I期试验。随后，将相同的基因盒包装在用于IM注射的rAAV 1-AAT中（第二代rAAV 1-AAT），同样通过NHLBI资助和小型生物技术公司（AGTC）资助之间的公私合作伙伴关系中的POC，IND使能以及1期和2a期研究进行了研究。这些研究表明，血清AAT表达持续数年，仅低于治疗阈值30倍，并证明了在这些患者中诱导调节性T细胞（T细胞）的重要作用。在即将到来的提案中，UMMS Horae基因治疗中心（GTC）和RNA治疗研究所（RTI）的一组研究人员已经走到一起，推动第二代载体rAAV-AAT的翻译研究，沿着一些更新的方法。这些新方法包括基于RNA的创新疗法，如合成miRNA（第三代），CRISPR/Cas9方法（第四代）和新型AAV衣壳（第N代）。卓越的研究核心提供了额外的创新和加速翻译，包括使用CRISPR/Cas9系统创建AAT null小鼠的转基因动物核心，定义这些小鼠肺气肿表型的呼吸生理学核心，以及为所有项目创新并提供高质量载体的载体核心。通过这些连续几代载体，该计划旨在解决由于α-1抗胰蛋白酶（AAT）缺乏引起的肺部疾病基因治疗的未满足需求，重点关注“肝脏保留”系统性基因治疗，旨在治疗AAT肺部疾病而不会加重AAT肝脏疾病。 叙述：该提案旨在利用基因治疗中最前沿的科学技术来开发一种治疗遗传性肺病的方法，称为AAT缺乏症。这种疾病导致慢性阻塞性肺疾病（COPD）和肺气肿，在美国总共影响超过1100万患者。AAT缺乏症比非遗传性COPD不太常见，在美国估计约有10，000至100，000例，其中许多病例未被诊断。 项目1：肌肉定向rAAV 1-AAT基因的临床试验和免疫学方面 治疗（Flotte，Terence R.，（医学博士）

 DESCRIPTION (provided by applicant): The overall goal of this translational program is to develop a definitive molecular therapy for lung disease due to alpha-1 antitrypsin (AAT) deficiency, a relatively common single gene disease due to mutations in the AAT gene. The program builds on the experience of investigators in developing several different versions of recombinant adeno-associated virus (rAAV)-based gene therapies for AAT deficiency since the late 1990's. Previous NHLBI-funded work by the program director's laboratory included proof-of-concept (POC) studies, formal IND-enabling safety studies, and a phase 1 trial of intramuscular injection of a 1st generation rAAV2- AAT (on a physician IND). Subsequently, the same gene cassette was packaged in rAAV1-AAT for IM injection (2nd generation rAAV1-AAT) was likewise studied through POC, IND-enabling and phase 1 and phase 2a studies in a public-private partnership between NHLBI funding and funding by a small biotechnology company (AGTC). These studies showed serum AAT expression persisting for several years at level only 30-fold below the therapeutic threshold and demonstrated an important role for the induction of regulatory T cells (Tregs) in these patients. In the coming proposal, a group of investigators within the UMMS Horae Gene Therapy Center (GTC) and the RNA Therapeutics Institute (RTI) have come together to push translational studies of rAAV-AAT of the 2nd generation vector along with a number of newer approaches. These newer approaches include innovative RNA-based therapeutics, such as synthetic miRNAs (3rd generation), CRISPR/Cas9 approaches (4th generation), and novel AAV capsids (Nth generation). Additional innovation and accelerated translation is provided by superb research cores, including a transgenic animal core that has used the CRISPR/Cas9 system to create an AAT null mouse, respiratory physiology core to define the emphysema phenotype in these mice, and a vector core that will both innovate and provide high quality vector for all projects. With these successive generations of vectors, this program seeks to address the unmet need for gene therapy for lung disease due to alpha-1 antitrypsin (AAT) deficiency with an important focus on "liver-sparing" systemic gene therapy, designed to treat AAT lung disease without exacerbating AAT liver disease. Narrative: This proposal seeks to use the most cutting edge scientific techniques in gene therapy to develop a way to treat an inherited lung disease, called AAT deficiency. This disease leads to a form chronic obstructive pulmonary disease (COPD) and emphysema, which overall affect over 11 million patients in the US. AAT deficiency is less common than non-inherited forms of COPD, with estimates ranging from approximately 10,000 to 100,000 cases in the US, with many cases going undiagnosed. PROJECT 1: Clinical Trial and Immunologic aspects of muscle-directed rAAV1-AAT gene Therapy (Flotte, Terence R., M.D.)