Developing a robust native extracellular matrix to improve islet function with attenuated immunogenicity for transplantation

开发强大的天然细胞外基质，以改善胰岛功能，并减弱移植的免疫原性

• 批准号: 10596047
• 负责人: Mary Navarro
• 金额: $ 30万
• 依托单位: STEMBIOSYS, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-03 至 2025-01-02
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10596047
• 关键词: Address; Adopted; Agrin; Allogenic; Allografting; Amniotic Fluid; Apoptosis; Architecture; Attenuated; Basement membrane; Beta Cell; Biological Assay; Blood; Blood Glucose; Blood Vessels; Blood capillaries; Bone Marrow; Cell Proliferation; Cell physiology; Cells; Clinical Trials; Coculture Techniques; Collagen Type I; Collagen Type VI; Complications of Diabetes Mellitus; Data; Decontamination; Development; Diabetes Mellitus; Digestion; Extracellular Matrix; Foundations; Genes; Glucose; Goals; Good Manufacturing Process; Growth; HLA-DR Antigens; Hepatic; Human; Hypoglycemia; Immune; Immunofluorescence Microscopy; Immunosuppression; Implant; In Vitro; Inflammatory Response; Injections; Insulin; Insulin-Dependent Diabetes Mellitus; Intellectual Property; Islet Cell; Islets of Langerhans; Islets of Langerhans Transplantation; Laboratory Procedures; Laminin; Lymphocyte; Maintenance; Marketing; Mediating; Microscopic; Morphology; Natural regeneration; Non-Insulin-Dependent Diabetes Mellitus; Pancreas; Physiological; Pluripotent Stem Cells; Portal vein structure; Positioning Attribute; Procedures; Production; Proteins; Proteomics; Quality of life; Rattus; Reagent; Recovery; Regulation; Reporting; Research; Risk; Stromal Cells; System; T-Lymphocyte; Technology; Testing; Time; Tissue Donors; Transplantation; Vascular Endothelial Cell; Work; allograft rejection; attenuation; clinical application; collagenase; commercialization; curative treatments; humanized mouse; immunogenicity; improved; in vivo; innovation; insulin secretion; islet; islet allograft; manufacture; mechanical properties; novel strategies; perlecan; preservation; public health priorities; response; restoration; subcutaneous; tissue culture

Project Summary/Abstract Compared to daily multiple insulin injections, pancreatic islet transplantation for type 1 diabetes (loss of β-cells) or the latter stages of type 2 diabetes (β-cells fail to produce sufficient insulin) provides a near physiologic regulation of normal blood glucose levels and significantly improves quality of life by minimizing severe hypoglycemia and diabetic complications. However, a shortage of donors, loss of allograft function over time, and the need for lifelong immunosuppression must be resolved before this approach can be widely adopted. The overall goal of this proposal is to improve islet transplantation using a native extracellular matrix (ECM)- based culture system, which mimics the pancreatic microenvironment, to obtain large quantities of high-quality islets with attenuated immunogenicity. Previously, we reported that culture of rat pancreatic islets on native ECM, produced by bone marrow stromal cells, promoted growth of islet vascular endothelial cells (VECs), production of islet-bm associated proteins, improvement of β- cell function, and attenuation of islet immunogenicity (Appendix 1). Recently, we developed an ECM synthesized by human amniotic fluid (AF)-derived pluripotent stem cells. Proteomic analysis suggested that the protein composition of AF-ECM and pancreatic ECM were similar and this was validated in our preliminary studies. Moreover, human islets maintained on AF-ECM showed significantly improved insulin secretion in response to glucose stimulation. Here, we propose to explore AF-ECM, a surrogate pancreatic-like ECM (pI-ECM), for potential use in maintaining human islets. We will prepare pl-ECM employing Good Manufacturing Practices (GMP) and hypothesize that maintenance of islets on pl-ECM, prepared using GMP compliant conditions, will facilitate the recovery of large numbers of high-quality human islets with decreased immunogenicity. To test this hypothesis, we will manufacture pl-ECM with reagents and facilities that are GMP compliant and then compare its architecture, mechanical properties, and protein composition with our current research use only (RUO) version (Aim 1). We will also compare the viability, function and immunogenicity of human islets maintained on GMP-compliant versus RUO- pl-ECM (Aims 2&3). If the proposed work is successful, it will provide the necessary preliminary data and rationale for moving forward with a small-scale clinical trial and commercialize a new, GMP-grade version of our matrix product for use in the manufacture of cell products for clinical applications.

项目摘要/摘要 与每日多次注射胰岛素相比，胰岛移植治疗1型糖尿病 (β细胞丢失)或2型糖尿病后期(β细胞不能产生足够的胰岛素) 提供对正常血糖水平的近乎生理调节，并显著改善 通过最大限度地减少严重低血糖和糖尿病并发症，提高生活质量。然而，a 供者短缺，随着时间的推移同种异体移植功能丧失，需要终生 在这一方法被广泛采用之前，必须解决免疫抑制问题。整体而言 这项建议的目标是使用天然细胞外基质(ECM)改善胰岛移植。 模拟胰腺微环境的基础培养系统，以获得大量 免疫原性减弱的高质量胰岛。此前，我们报道了大鼠的培养 由骨髓基质细胞产生的天然ECM上的胰岛促进胰岛生长 血管内皮细胞，产生胰岛-BM相关蛋白，改善β- 细胞功能和胰岛免疫原性减弱(附录1)。最近，我们开发了一种 人羊水来源的多能干细胞合成细胞外基质。蛋白质组学分析 提示AF-ECM和胰腺ECM的蛋白质组成相似，这是 在我们的初步研究中得到了验证。此外，在AF-ECM上维持的人胰岛显示 显著改善对葡萄糖刺激的胰岛素分泌。在此，我们建议 探索AF-ECM，一种替代胰腺样ECM(PI-ECM)，用于维持 人类的小岛。我们将准备采用良好制造规范(GMP)的PL-ECM和 假设使用符合GMP的条件制备的PL-ECM上的胰岛的维护， 将促进大量减少的优质人类胰岛的回收 免疫原性。为了验证这一假设，我们将用试剂和设备制造pl-ecm。 然后比较其结构、机械性能和蛋白质 作文与我们目前的研究用途(RUO)版本(目标1)。我们还将比较 人胰岛活性、功能和免疫原性维持在GMP依从性和RuO-R基础上 PL-ECM(目标2和3)。如果拟议的工作成功，它将提供必要的初步 推进小规模临床试验并将一种新的、 我们的基质产品的GMP级版本，用于生产临床细胞产品 申请。