Gene Therapy for Diabetes

糖尿病基因治疗

• 批准号: 10017960
• 负责人: Markus Grompe
• 金额: $ 72.91万
• 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/10017960
• 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.

项目摘要 在1型糖尿病中丢失的β细胞（T1D）中存在两种潜在方法。首先是 移植源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患者的原位生成新的β细胞，而无需细胞移植。这可以实现 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。基因疗法 向量用于传递重编程因子。 我们和其他人最近发现，可以通过逆行导管纠正小鼠中的糖尿病 注入重编程向量。导管内输送的优点是递送高剂量的基因 局部治疗载体，最大程度地减少全身副作用，并达到较高的局部浓度 重编程因素。此外，这种管理途径在人类中很容易可行，如ERCP （内窥镜逆行胆管造影）是临床胃肠病学中的常规程序。 啮齿动物中的临床前工作表明α细胞是重编程的主要目标，而 胰管也可以转化为功能性β样细胞。 在此提案中，我们将开发用于重新编程的α细胞的AAV矢量 导管内递送后，人类和非人类素数。我们正在建立进步 在当前的hirn UC4赠款中，我们开发了能够转导的新型AAV Capsids 人体内分泌细胞高效率。我们还发展了能够 仅限于β细胞的转化表达。 在AIM 1中，我们将产生新颖的AAV Capsids（变体），它们在转导胰腺方面高效 体内非人类素数逆行注射后α细胞和导管细胞。高度创新的Capsid 将使用进化方法。在AIM 2中，我们将生成专门引导转换表达式的CRE 到重编程靶标，即α细胞。细胞类型的特定启动子将与microRNA结合 识别要素以实现这一目标。最后，在AIM 3中，AIM 1和CRES产生的AAV CAPSIDS 在AIM 2中开发的将结合使用，以产生优化的AAV，能够将重编程因素传递给 α细胞及其重新编程的功能将在非人类隐私中进行评估。 这项工作的成功执行将生成确定是否是否这样的临床前数据 方法具有在人类中临床应用的潜力。