TRGel微囊化结合全组织透明化和高分辨率成像技术解析胰岛再生动态演进过程及其分子机制

胰岛移植已成为治疗胰岛素缺乏型糖尿病的一种替代疗法。但由于临床上胰岛来源缺乏、分离困难以及移植排斥反应等，限制了胰岛移植的广泛应用。研究表明间充质干细胞可分化具有胰岛素分泌功能的细胞，通过微囊化包裹又能够保护胰岛素分泌细胞移植后的活性，降低免疫排斥。本课题拟应用骨髓、脂肪和脐带来源的间充质干细胞体外诱导分化为胰岛素分泌细胞并建立稳定的细胞分化、微小胰岛组织构建工艺；利用温度调控材料TRGel（poly(2,3)glucose）形成微囊并建立TRGel微囊化再生胰岛组织工艺；在糖尿病小鼠模型上检验再生微囊化胰岛组织的替代性治疗效果。借助单细胞测序和全组织透明化及高分辨率成像技术，综合免疫微环境分析，揭示间充质干细胞分化为胰岛素分泌细胞及微小胰岛组织再生过程多维度的表征和可视化以及关键的调控信号改变，探索移植体和周围环境的互作，从而为再生胰岛组织移植技术的应用提供分子依据。

Pancreatic islet transplantation has become a typical β-cell replacement therapy for the treatment of insulin-deficient diabetes. However, due to the lack of source of pancreatic islets, difficulty in isolation, and transplant rejection, the wide application of direct pancreatic islet transplantation is limited. Studies have shown that mesenchymal stem cells can differentiate into islet-like cells (or insulin producing cells, IPCs), and can protect the activity of IPCs after transplantation through microencapsulation and reduce immune rejection. In this project, we would like to use bone marrow, fat and umbilical cord-derived mesenchymal stem cells to induce IPCs in vitro and establish a stable cell differentiation and to construct encapsulation technology; to use temperature responsive material TRGel (poly(2,3)glucose) to form Microencapsulation and the establishment of TRGel microencapsulation regenerated islet tissue technology; the alternative treatment effect of regenerated microencapsulated islet tissue was tested on a diabetic mouse model. With the help of single-cell sequencing, whole-tissue clearing and high-resolution imaging technology, comprehensive immune microenvironment analysis, we aim to reveal the multi-dimensional characterization and visualization of the regeneration process of mesenchymal stem cells into IPCs and tiny islet tissues, as well as key regulatory signal changes. The interaction between the transplant and the surrounding environment provides a molecular basis for the application of regenerative islet tissue transplantation treatment technology for diabetes.