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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要与每日多次胰岛素注射相比，胰岛移植治疗 1 型糖尿病（β 细胞损失）或 2 型糖尿病后期（β 细胞无法产生足够的胰岛素）提供正常血糖水平的近乎生理调节并显着改善通过最大限度地减少严重低血糖和糖尿病并发症来提高生活质量。供体短缺、同种异体移植功能随着时间的推移而丧失以及需要终生在这种方法得以广泛采用之前，必须解决免疫抑制问题。该提案的目标是使用天然细胞外基质（ECM）改善胰岛移植 - 基于模拟胰腺微环境的培养系统，以获得大量之前，我们报道了大鼠培养物的免疫原性减弱的高质量胰岛。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 (VEC)、胰岛 bm 相关蛋白的产生、β- 的改善细胞功能和胰岛免疫原性减弱（附录 1）。由人羊水 (AF) 来源的多能干细胞合成的 ECM 蛋白质组分析。表明 AF-ECM 和胰腺 ECM 的蛋白质组成相似，这是此外，我们的初步研究证实了 AF-ECM 维持的人类胰岛。改善胰岛素分泌对葡萄糖刺激的显着反应。探索 AF-ECM，一种替代胰腺样 ECM (pI-ECM)，在维持我们将采用良好生产规范 (GMP) 来制备 pl-ECM 和坚持认为 pl-ECM 上的胰岛维护是使用符合 GMP 的条件制备的，将促进大量高质量人类胰岛的恢复为了检验这一假设，我们将使用试剂和设施制造 pl-ECM。符合 GMP 标准，然后比较其结构、机械性能和蛋白质与我们当前仅研究使用（RUO）版本（目标 1）进行比较。符合 GMP 与 RUO 的人类胰岛的活力、功能和免疫原性 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准备。推进小规模临床试验并将新的、我们的基质产品的 GMP 级版本，用于生产临床细胞产品应用程序。