Gene Therapy for Diabetes

糖尿病基因治疗

• 批准号: 10239013
• 负责人: Markus Grompe
• 金额: $ 72.09万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10239013
• 关键词: 3' Untranslated Regions; Acinar Cell; Allogenic; Alpha Cell; Bar Codes; Beta Cell; Binding Sites; Biodistribution; Bioinformatics; Blood Glucose; Capsid; Cell Transplantation; Cells; Chemicals; Chronic; Clinical; Data; Databases; Dependovirus; Development; Diabetes Mellitus; Directed Molecular Evolution; Disease; Disease Management; Dose; Drug Kinetics; Duct (organ) structure; Ductal Epithelial Cell; Elements; Endocrine; Endoderm Cell; Endoscopic Retrograde Cholangiopancreatography; Enhancers; Evolution; Gastroenterology; Gene Delivery; Gene Expression; Gene Transduction Agent; General Population; Genetic Transcription; Goals; Grant; Human; In Situ; Individual; Injections; Insulin; Insulin-Dependent Diabetes Mellitus; Islet Cell; Islets of Langerhans; Libraries; Macaca mulatta; Mediating; Methods; MicroRNAs; Molecular; Mus; Natural regeneration; Pancreas; Pancreatic duct; Patients; Premature Mortality; Primates; Procedures; Property; Reagent; Regulatory Element; Reporter; Reporter Genes; Rodent; Route; Safety; Site; Specificity; Structure of alpha Cell of islet; System; Technology; Therapeutic; Transplantation; Tropism; Variant; Viral Vector; Virus; Work; adeno-associated viral vector; aptamer; base; beta cell replacement; cell type; clinical application; clinical translation; diabetes mellitus therapy; gene therapy; genetic payload; humanized mouse; in vivo; innovation; islet; mouse model; nonhuman primate; novel; pre-clinical; promoter; side effect; stem cells; transcription factor; transduction efficiency; transgene expression; unnatural amino acids; vector

PROJECT SUMMARY Two potential approaches exist for the replacement of the β-cells lost in type 1 diabetes (T1D). The first is to transplant new β-cells derived from either allogeneic pancreas donors or stem cells. The second approach is to generate new β-cells in situ in the T1D patient without any need for cell transplantation. This can be achieved by transcription factor mediated reprogramming of endodermal cell types related to β-cells. Gene therapy vectors are used to deliver the reprogramming factors. We and others have recently found that it is possible to correct diabetes in mice by retrograde ductal injection of reprogramming vectors. Intraductal delivery has the advantage of delivering a high dose of gene therapy vector locally, minimizing systemic side effects and achieving a high local concentration of reprogramming factors. Furthermore, this route of administration is readily feasible in humans, as ERCP (endoscopic retrograde cholangio-pancreatography) is a routine procedure in clinical gastroenterology. Preclinical work in rodents indicates that α-cells are the prime target for reprogramming, while pancreatic ducts may also be converted to functional β-like cells. In this proposal, we will develop AAV vectors that are optimized for reprogramming the α-cells of humans and non-human primates to the β-cell fate after intraductal delivery. We are building on the progress made in our current HIRN UC4 grant, in which we developed novel AAV capsids capable of transducing human endocrine cells with high efficiency. We also evolved cis-regulatory elements (CREs) capable of restricting transgene expression to only β-cells. In Aim 1, we will produce novel AAV capsids (variants) that are highly efficient in transducing pancreatic α-cells and duct cells after retrograde injection in non-human primates in vivo. Highly innovative capsid evolution methods will be used. In Aim 2, we will generate CREs that direct transgene expression specifically to the reprogramming target, i.e. α-cells. Cell-type specific promoters will be combined with microRNA recognition elements to achieve this goal. Finally, in Aim 3, AAV capsids generated by Aim 1 and CREs developed in Aim 2 will be combined to produce optimized AAV capable of delivering reprogramming factors to α-cells and its capability of reprogramming will be assessed in non-human primates. Successful execution of this work will generate the preclinical data needed to determine whether this approach has potential for clinical application in humans.

项目概要 有两种潜在的方法可以替代 1 型糖尿病 (T1D) 中丢失的 β 细胞。第一个是 移植来自同种异体胰腺供体或干细胞的新 β 细胞。第二种方法是 在 T1D 患者体内原位产生新的 β 细胞，无需任何细胞移植。这是可以实现的 通过转录因子介导的与 β 细胞相关的内胚层细胞类型的重编程。基因治疗 载体用于传递重编程因子。 我们和其他人最近发现可以通过逆行导管来纠正小鼠的糖尿病 注射重编程载体。导管内递送具有递送高剂量基因的优点 局部治疗载体，最大限度地减少全身副作用并实现高局部浓度 重编程因素。此外，这种给药途径在人类中很容易实现，因为 ERCP （内镜逆行胰胆管造影）是临床胃肠病学的常规手术。 啮齿动物的临床前工作表明 α 细胞是重编程的主要目标，而 胰管也可能转化为功能性β样细胞。 在本提案中，我们将开发 AAV 载体，该载体经过优化，可以对 α 细胞进行重编程 人类和非人类灵长类动物导管内分娩后β细胞的命运。我们正在取得进展 我们目前的 HIRN UC4 资助项目中，我们开发了能够转导的新型 AAV 衣壳 人体内分泌细胞效率高。我们还进化了顺式调控元件（CRE），能够 将转基因表达仅限于 β 细胞。 在目标 1 中，我们将生产能够高效转导胰腺的新型 AAV 衣壳（变体） 非人灵长类动物体内逆行注射后的α细胞和导管细胞。高度创新的衣壳 将使用进化方法。在目标 2 中，我们将生成专门指导转基因表达的 CRE 到重编程目标，即α细胞。细胞类型特异性启动子将与microRNA结合 实现这一目标的识别要素。最后，在目标 3 中，目标 1 和 CRE 生成的 AAV 衣壳 Aim 2中开发的AAV将被结合起来产生优化的AAV，能够将重编程因子传递给 α细胞及其重编程能力将在非人类灵长类动物中进行评估。 这项工作的成功执行将生成确定该药物是否有效所需的临床前数据。 该方法具有在人类临床应用的潜力。