Transformational platform for regenerating autologous transplantable endocrine tissue from human pancreatic matrix and pluripotent stem cells

从人胰腺基质和多能干细胞再生自体可移植内分泌组织的转化平台

• 批准号: 9169474
• 负责人: Jon S Odorico
• 金额: $ 22.31万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9169474
• 关键词: Address; Adult; Age; Anti-Inflammatory Agents; Anti-inflammatory; Autologous; Beta Cell; Biological; Biomedical Engineering; Blood Vessels; Blood flow; Cell Survival; Cell physiology; Cells; Child; Collaborations; Data; Deposition; Development; Diabetes Mellitus; Endocrine; Endocrine Glands; Endothelial Cells; Engineering; Engraftment; Exhibits; Extracellular Matrix; Family suidae; Future; Gel; Glucose; Goals; Health; Human; Hydrogels; Hypoxia; Immunosuppression; In Vitro; Inflammation; Insulin; Insulin-Dependent Diabetes Mellitus; Islets of Langerhans; Islets of Langerhans Transplantation; Kidney; Label; Life; Link; Liver; Mass Spectrum Analysis; Medical; Medicine; Mesenchymal; Methods; Modeling; Molds; Mus; Natural regeneration; Non-Insulin-Dependent Diabetes Mellitus; Nutrient; Organ; Organ Donor; Oxygen; Pancreas; Patients; Pharmacy Schools; Physiological; Pluripotent Stem Cells; Porifera; Portal vein structure; Procedures; Property; Proteins; Proteome; Proteomics; Regenerative Medicine; Replacement Therapy; Reporting; Research; Schools; Signal Transduction; Site; Staging; Stem cells; Stromal Cells; Techniques; Technology; Testing; Time; Tissue Engineering; Tissue Grafts; Tissue Sample; Tissue Transplantation; Tissues; Transplantation; allograft rejection; base; beta cell replacement; cell behavior; clinically relevant; design; diabetic; fetal; glycemic control; human embryonic stem cell; human tissue; implantation; improved; innovation; innovative technologies; insulin secretion; interest; intrahepatic; islet; minimally invasive; mouse model; new technology; novel; paracrine; prevent; regenerative; research study; scaffold; stem cell differentiation; subcutaneous

ABSTRACT Diabetes and its complications still claim the lives of millions of people despite continuing advances in insulin delivery technology primarily because insulin fails to achieve perfect glycemic control. On the other hand, beta cell replacement therapies including vascularized pancreas and isolated islet transplantation are able to fully restore normoglycemia, achieve insulin-independence and can delay end-organ complications. However, these latter therapies suffer from two key limitations, the shortage of organs and the need for life- long immunosuppression to prevent allograft rejection. Furthermore, the intrahepatic portal vein islet transplantation site used in humans is far from ideal and many islets are lost after implantation. An ideal beta cell replacement therapy strives towards both generating an abundant supply of functional beta cells and identifying a minimally invasive, well-vascularized, retrievable site for transplantation that is clinically applicable. After years of research it is now well established that human pluripotent stem cells (hPSCs) can be directed to differentiate into highly enriched physiological functional islet-like clusters (ILCs) in vitro that are capable of curing diabetes in mice. The extracellular matrix (ECM) is a critical component of the cellular niche that helps maintain cellular differentiation and provides tissue-specific signals to guide the fate and behavior of cells. Recent progress in the decellularization of organs has spurred great interest in using natural matrix for regenerative medical applications; yet, few studies have focused on the pancreas in general and the human pancreas to date has not been effectively decellularized and studied. Appreciating the importance of tissue-specific ECM, we have established effective techniques for the decellularization and delipidization of human pancreas tissue to produce several types of natural matrix constructs, including intact 3D matrix, molded sponge scaffolds and a spontaneous gelling hydrogel (hP-ECM). With the challenges of identifying a clinically applicable transplant site that provides for immediate and sufficient oxygen and nutrient delivery, we believe there is compelling rationale to take advantage of the proven proangiogenic and anti-inflammatory properties of ECs and MSCs. Thus, transplanting ILCs with hPSC-derived endothelial cells (ECs) and hPSC-derived mesenchymal stromal cells (MSCs), each providing essential properties, combined with hP-ECM into a prevascularized deviceless retrievable subcutaneous site might provide a more optimal transplant platform. Now, based on this innovative technology we aim to obtain a better understanding of the composition and function of natural hP-ECM in the context of hPSC differentiation to beta cells. The immediate objectives are to characterize human pancreatic extracellular matrix and to use this natural matrix in combination with stem cell-derived β cells, ECs and MSCs to reconstruct endocrine tissue capable of glucose- stimulated insulin-secretion after transplantation to mice. Our specific aims are to: 1) Comprehensively characterize the human pancreatic and islet ECM proteome, or matrixome, and compare the matrixome of different developmental ages using advanced quantitative mass spectrometry methods in collaboration with Dr. Linjun Li, 2) Construct a hP-ECM - cellular composite tissue graft combining hPSC-ILCs with ECs +/- MSCs and test its function in an immunodeficient murine diabetes model. Ultimately, we envision a bioengineered composite endocrine organ as a highly innovative regenerative medicine strategy for producing potentially autologous insulin-producing tissue for transplantation. These basic enabling studies are the first steps towards developing an effective, minimally invasive transplant platform that is available for all patients with diabetes.

抽象的 糖尿病及其并发症仍然夺走了数百万人继续发展的生命 胰岛素输送技术主要是因为胰岛素无法获得完美的血糖控制。另一方面 手，β细胞替代疗法，包括血管化胰腺和孤立的胰岛移植是 有可能完全恢复正常血糖，达到胰岛素独立性并可能延迟末期并发症。 但是，这些后来的疗法遭受了两个关键局限性，器官短缺以及对生命的需求 - 长期免疫抑制以防止同种异体移植排斥。此外，e割门内静脉胰岛 人类使用的移植部位远非理想，植入后许多小岛丢失。理想的beta 细胞替代疗法旨在为产生大量的功能β细胞供应和 识别一个最小的侵入性，血管良好的，可检索的位点的移植，这是临床上的 适用的。 经过多年的研究，现在已经很好地确定了人类多能干细胞（HPSC）可以是 针对高度富集的物理功能胰岛样簇（ILC）的体外分化 能够在小鼠中治愈糖尿病。 细胞外基质（ECM）是细胞生态位的关键成分，有助于维持细胞 分化并提供组织特异性信号，以指导细胞的命运和行为。最近的进展 器官的脱细胞对使用天然基质进行再生医学引起了极大的兴趣 申请；然而，很少有研究集中在胰腺和迄今为止的人类胰腺上 没有有效的脱细胞和研究。欣赏组织特异性ECM的重要性，我们有 建立了用于人胰腺组织脱细胞和删除至的有效技术 生产几种类型的天然矩阵构建体，包括完整的3D矩阵，模制赞助商脚手架和一个 赞助胶凝水凝胶（HP-ECM）。 面对确定临床适用的移植部位的挑战，该网站提供立即和 充足的氧气和营养递送，我们认为有令人信服的理由可以利用这一验证 ECS和MSC的促血管生成和抗炎特性。那就是用HPSC衍生的ILC移植ILC 内皮细胞（ECS）和HPSC衍生的间质基质细胞（MSC），每个细胞提供必不可少的 与HP-ECM结合到一个无脱毛的无偏离检索皮下位点的特性可能 提供一个更最佳的移植平台。 现在，基于这种创新技术，我们旨在更好地了解组成和 天然HP-ECM在HPSC分化与β细胞的背景下的功能。直接的目标是 为了表征人类胰腺外基质并将这种天然矩阵组合使用 与干细胞衍生的β细胞，EC和MSC一起重建能够葡萄糖的内分泌组织 移植向小鼠后刺激的胰岛素分泌。 我们的具体目的是：1）全面表征人类胰腺和胰岛ECM蛋白质组， 或矩阵组，并使用高级定量质量比较不同发育年龄的基质组 光谱法与Linjun Li博士合作，2）构建HP -ECM-细胞复合组织 将HPSC-ILC与ECS +/- MSC结合的移植物，并在免疫缺陷的鼠糖尿病中测试其功能 模型。最终，我们将生物工程的复合内分泌器官视为高度创新的再生 医学策略，用于产生潜在的自体胰岛素组织进行移植。这些 基本启示研究是开发有效的，微创移植的第一步 适用于所有糖尿病患者的平台。