New Approaches to Gene Therapy for Alpha-1 Antitrypsin Deficiency

 描述(由申请人提供)：该翻译计划的总体目标是为因α-1抗胰蛋白酶(AAT)缺乏引起的肺部疾病开发一种明确的分子治疗方法，AAT基因突变是一种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S以来开发几种不同版本的重组腺相关病毒(RAAV)基因疗法治疗AAT缺乏症的经验基础上。该计划主任实验室之前由NHLBI资助的工作包括概念验证(POC)研究、正式的IND安全性研究，以及(在医生IND上)肌肉注射第一代rAAV2-AAT的第一阶段试验。随后，将相同的基因盒包装在rAAV1-AAT中用于IM注射(第二代rAAV1-AAT)，同样通过POC、IND使能以及NHLBI资金和一家小型生物技术公司(AGTC)资金之间的公私合作关系进行第一阶段和第二阶段研究。这些研究表明，血清AAT的表达持续数年，水平仅为治疗阈值的30倍，并证明了在这些患者中诱导调节性T细胞(Tregs)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的翻译研究以及一些新的方法。这些较新的方法包括基于RNA的创新疗法，如合成miRNAs(第三代)、CRISPR/Cas9方法(第四代)和新型AAV衣壳(第N代)。其他创新和加速翻译是由卓越的研究核心提供的，包括使用CRISPR/Cas9系统创造了AAT缺失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，以及将创新并为所有项目提供高质量载体的载体核心。通过这些连续几代载体，该计划寻求解决因α-1抗胰蛋白酶(AAT)缺乏而未得到满足的肺部疾病基因治疗需求，重点放在“节肝”系统基因治疗上，旨在治疗AAT肺部疾病而不加重AAT肝病。 叙述：这项提议寻求使用基因治疗中最尖端的科学技术来开发一种治疗遗传性肺部疾病的方法，称为AAT缺乏症。这种疾病导致一种形式的慢性阻塞性肺疾病(COPD)和肺气肿，在美国总共影响到1100多万患者。与非遗传性COPD相比，AAT缺乏症不太常见，据估计，在美国约有10,000至10,000例COPD病例，许多病例没有得到诊断。 项目1：肌肉导向rAAV1-AAT基因的临床试验和免疫学方面 治疗(Flotte，Terence R.，M.D.)

 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.)