Enhancement of Conditional Reprogramming Technology for Production of Functional Human Beta-Cells

项目摘要 供体胰岛供应不足严重限制了移植需求 胰岛素治疗不足的1型糖尿病(T1D)患者的治疗选择 控制他们的疾病。虽然最近导演的电影取得了很大成功 人胚胎干细胞和人诱导多能干细胞的分化 细胞(HU-IPSCs)转化为胰岛β细胞的过程仍然很不完善，效率也很低。至 为此，我们已经证明了条件重编程(CR)细胞培养技术可以 支持将人胰岛来源的β细胞显著扩增到至少一百万倍 金额。同样重要的是，我们的初步数据表明，CR-繁殖的人类β细胞 在体外可以很容易地形成成熟的分化标记和功能表型。 将这项前景看好的技术发展成人类β细胞替代的实用选择 治疗，我们寻求通过增强CR生长的人β-细胞的增殖能力 使用我们在其他CR反应患者中成功使用的重点筛查方法 单元类型。CR后扩展差异化步骤的优化也是一个组成部分 我们的建议，使用了其他实验室关于细胞表面相互作用如何 有助于推动人类β细胞的成熟和功能。使用CR技术的终极考验 扩增人类β细胞将使用这些细胞在T1D动物模型中并评估其 调节疾病病理的能力；恢复正常血糖并产生人类C- 接受移植的小鼠体内的多肽将是成功的积极标志。成功 完成这些具体的研究目标将使我们能够提出第二阶段SBIR 应用重点是将CR技术的临床前开发转化为GLP疗效和 安全测试和符合cGMP的电池制造。

Project Abstract Insufficient supply of donor pancreatic islets to meet the demand for transplants severely limits therapeutic options for Type 1 diabetic (T1D) patients in whom insulin therapy is insufficient to control their disease. And while there has been much recent success with the directed differentiation of human embryonic stem cells (hu-ESCs) and human induced pluripotent stem cells (hu-iPSCs) into pancreatic beta cells, the process is still quite imperfect and inefficient. To this end, we have shown that Conditional Reprogrammed (CR) cell culture technology can support the significant expansion of human islet-derived β-cells to at least one million-fold amount. Just as importantly, our preliminary data indicates that CR-propagated human β-cells can readily develop mature differentiation markers and functional phenotype in vitro. To develop this promising technology into a practical option for human β-cell replacement therapy, we seek to enhance the proliferative capacity of CR-grown human β-cells through the use of a focused screening approach that we have used with success with other CR-responsive cell types. The optimization of the post-CR expansion differentiation step is also a component of our proposal, using reproduced findings from other labs on how cell surface interactions can help drive human β-cell maturation and functionality. The ultimate test of using CR technology to expand human β-cells will be to use these cells in an animal model of T1D and to assess their ability to modulate the disease pathology; a return to euglycemia and production of human C- peptide in the transplant-receiving mice will be the positive hallmark of success. Successful completion of these specific research aims will position us to propose a Phase II SBIR application focused on the preclinical development of CR technology into GLP efficacy and safety testing and cGMP-compliant cell manufacturing.