Models and Gene Therapies for AAT Deficiency

AAT 缺乏症的模型和基因疗法

• 批准号: 10270088
• 负责人: Terence R. Flotte
• 金额: $ 278.91万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-09 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10270088
• 关键词: Address; Adenine; Affinity; Alleles; Animal Model; Antibody Response; Antigen Receptors; Biodiversity; Biological Assay; Biological Markers; CRISPR/Cas technology; Capsid; China; Chronic Obstructive Airway Disease; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Complex; Cryoelectron Microscopy; Deaminase; Densitometry; Disease; Disease model; Doctor of Philosophy; Elastin; Environment; Enzymes; European; Exhibits; Faculty; Ferrets; Future; Gene Silencing; Gene Transduction Agent; Genes; Genetic; Genetic Diseases; Homozygote; Human; Human Genetics; Hybrids; Immune Evasion; Immune response; Immunology; Impairment; Interferon Type II; Laboratories; Lead; Libraries; Life; Liver; Lung; Lung CAT Scan; Lung Inflammation; Lung diseases; Measurement; Measures; Mechanics; Mediating; Modeling; Molecular; Mus; Muscle; Mutation; Other Genetics; Outcome; Pharmaceutical Preparations; Phenotype; Physiological; Physiology; Population; Property; Proteins; Pulmonary Emphysema; RNA-Directed DNA Polymerase; Recombinant adeno-associated virus (rAAV); Regulatory T-Lymphocyte; Research Personnel; Role; Safety; Serotyping; Serum; Site; Structure-Activity Relationship; System; Technology; Testing; Time; Transducers; Transgenes; Transgenic Animals; Transgenic Organisms; Variant; Veterinary Medicine; Veterinary Schools; Wild Type Mouse; adeno-associated viral vector; airway obstruction; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; base; cell mediated immune response; clinically relevant; collaborative environment; comparative efficacy; design; enzyme activity; enzyme linked immunospot assay; gene delivery system; gene replacement; gene therapy; immunogenic; immunoregulation; improved; innovation; mouse model; mutant; neutralizing antibody; neutralizing monoclonal antibodies; new technology; next generation; nonhuman primate; novel; programs; reconstitution; response; screening; success; tool; vector

Project Summary (OVERALL) Alpha-1 antitrypsin deficiency (AATD) is caused by mutations in the SERPINA1 gene. The E342K (PI*Z) mutant allele is very common among those of European ancestry, and E342K homozygotes encode a protein with impaired secretion, resulting in deficient AAT serum levels. Since AAT normally protects elastin in the lung from degradation, loss of effective AAT triggers lung inflammation, airways obstruction and emphysema, which is the primary life-limiting manifestation of AATD. The projects within this proposal seek to pursue numerous parallel strategies to develop a gene therapy for AATD. Most of these strategies revolve around the use of recombinant adeno-associated virus (rAAV)-based vectors, a platform technology that has been very successful for other genetic diseases. In Project 1, optimized rAAV vectors will be studied in genetically defined animal models (including mice and ferrets) in comparison with transgenic reconstitution studies using a regulated conditional transgenic system to compare two relevant potential target replacement levels (11µM and 25µM) and clinically relevant endpoints will be studied. In Project 2, novel CRISPR variants will be used for gene editing, base editing and prime editing strategies to treat AATD. In Project 3, we will screen naturally occurring AAV capsid libraries obtained from remote populations in Western China to identify capsids with enhanced efficacy and safety for AATD gene therapy. Finally, in project 4, we will use novel Treg and CAR-Treg strategies to selectively modulate anti-vector immune responses. There will also be two cores. Core A will provide each project with important Vector Immunology assays, which can identify limitations due to host immune responses to AAV capsids, the AAT transgene or to Cas9-derived proteins. Core B will provide animal models and physiologic measurements in the animal models for testing of optimized rAAV vectors, gene editing tools and immune modulation approaches. Program investigators have a track record of interactions and collaborations that we anticipate will continue in future years.

项目摘要（总体） α-1抗胰蛋白酶缺乏症（AATD）是由SERPINA1基因突变引起的。这 E342K（PI*Z）突变等位基因在欧洲血统中很常见，E342K 纯合子编码具有分泌受损的蛋白质，导致AAT血清水平不足。 由于AAT通常保护肺中的弹性蛋白免受降解，因此有效AAT触发损失 肺部炎症，气道异议和肺气肿，这是主要的生命限制 AATD的表现。该提案中的项目寻求追求无数的平行 开发AATD基因疗法的策略。这些策略中的大多数围绕使用 基于重组腺相关病毒（RAAV）的载体，一种平台技术 对于其他遗传疾病，我们非常成功。在项目1中，优化的RAAV矢量将是 与一般定义的动物模型（包括小鼠和雪貂）相比，研究 转基因重建研究使用受管制的有条件转基因系统进行比较 两个相关的潜在目标置换水平（11µm和25µm）和临床相关 端点将进行研究。在项目2中，新颖的CRISPR变体将用于基因编辑， 基础编辑和主要的编辑策略来治疗AATD。在项目3中，我们将自然筛选 发生从中国西部偏远人群获得的AAV CAPSID图书馆以确定 Capsids具有提高的AATD基因治疗效率和安全性。最后，在项目4中，我们将 使用新颖的Treg和Car-Treg策略来选择性调节抗媒介免疫反应。 还将有两个核心。核心A将为每个项目提供重要的向量 免疫学评估，可以确定由于宿主对AAV的免疫反应而引起的局限性 衣壳，AAT转换或Cas9衍生的蛋白质。核心将提供动物模型，并 动物模型中的生理测量，用于测试优化的RAAV载体，基因 编辑工具和免疫调节方法。计划调查人员有一个记录 我们预期的互动和合作将在未来的几年中继续进行。