Department of Hepatobiliary and Pancreatic Surgery, Affiliated Hospital of Nantong University, Medical School of Nantong University, Nantong, 226001, China; Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, 226001, China.
Center of Gallbladder Disease, Shanghai East Hospital, Institute of Gallstone Disease, Tongji University School of Medicine, Shanghai, 200000, China.
Biomaterials. 2022 Dec;291:121882. doi: 10.1016/j.biomaterials.2022.121882. Epub 2022 Oct 31.
The biomimetic pancreatic microenvironment improves the differentiation efficiency and function of human embryonic stem cell-derived β-cells (SC-β cells). Thus, a laminin subunit alpha 2-gelatin methacrylate (LAMA2-GelMA) hybrid hydrogel as a bionics carrier for the formation and maturation of endocrine lineage was developed in our research, based on pancreas proteomics analysis of postnatal mice. Pancreatic endocrine cells cultured on the hybrid hydrogel in vitro, which was composed of 0.5 μg/mL LAMA2 protein and 4% GelMA, the expression of transcription factors (TFs), including NKX6.1, NKX6.2, and NEUROD1 were upregulated. Single-cell transcriptomics was performed after LAMA2 knockdown during the early differentiation of pancreatic progenitor (PP) cells, a marked decrease in the forkhead box protein A2 (FOXA2)/GATA-binding factor 6 (GATA6) cluster was detected. Also, we clarified that as a receptor of LAMA2, integrin subunit alpha 7 (ITGA7) participated in Integrin-AKT signaling transduction and influenced the protein levels of FOXA2 and PDX1. In vivo experiments showed that, PP cells encapsulated in the LAMA2-GelMA hydrogel exhibited higher serum C-peptide levels compared to the GelMA and Matrigel groups in nude mice and reversed hyperglycemia more quickly in STZ-induced diabetic nude mice. Taken together, our findings highlighted the feasibility of constructing a pancreas-specific microenvironment based on proteomics and tissue engineering for the treatment of diabetes.
仿生胰腺微环境可提高人胚胎干细胞源性β细胞（SC-β 细胞）的分化效率和功能。因此，我们基于对出生后小鼠胰腺蛋白质组学的分析，开发了一种层粘连蛋白亚单位 α2-明胶甲基丙烯酰（LAMA2-GelMA）杂化水凝胶作为内分泌谱系形成和成熟的仿生载体。在体外，胰腺内分泌细胞在由 0.5μg/mL LAMA2 蛋白和 4% GelMA 组成的杂化水凝胶上培养，其转录因子（TFs），包括 NKX6.1、NKX6.2 和 NEUROD1 的表达上调。在胰腺祖细胞（PP）分化早期敲低 LAMA2 后进行单细胞转录组学分析，发现叉头框蛋白 A2（FOXA2）/GATA 结合因子 6（GATA6）簇明显减少。此外，我们还阐明了作为 LAMA2 的受体，整合素亚单位α7（ITGA7）参与整合素-AKT 信号转导，并影响 FOXA2 和 PDX1 的蛋白水平。体内实验表明，与 GelMA 和 Matrigel 组相比，包封在 LAMA2-GelMA 水凝胶中的 PP 细胞在裸鼠中表现出更高的血清 C 肽水平，并在 STZ 诱导的糖尿病裸鼠中更快地逆转高血糖。总之，我们的研究结果强调了基于蛋白质组学和组织工程构建胰腺特异性微环境治疗糖尿病的可行性。
