Epigenetic and chemical approaches to generate function beta cells

描述（由申请人提供）：胰岛素产生β细胞的再生以及胰岛移植是1型糖尿病（T1 D）治疗最有前途的长期解决方案。这种方法成功的主要障碍是器官供体的短缺和同种异体移植后终身免疫抑制的必要性。两个最近的进展，成功地将体细胞重编程为多能ES细胞样IPS（诱导多能干细胞）细胞和成功地将人ES细胞分化为功能性胰岛样簇（ILC），已经打开了通过将体细胞重编程为iPS细胞随后引导分化来产生患者特异性胰岛素分泌ILC的可能性。事实上，在一项概念验证研究中，我们最近已经证明，使用这种方法可以从人类皮肤成纤维细胞中产生胰岛素分泌细胞。尽管这一令人鼓舞的结果，几个障碍必须克服之前，这种技术的价值可以实现临床。主要障碍包括：1）使用携带病毒载体的iPS细胞的安全性问题; 2）当前方案在产生功能性β细胞方面的效率低; 3）移植的β细胞的自身免疫破坏。鉴于大量的iPS细胞群体正在解决第一个问题，许多免疫学实验室正在试图解决第三个问题，目前的提案计划使用表观遗传，化学和ECM方法来解决第二个问题。具体目标是： 1)胎儿和成熟人β细胞表观遗传特征的表征; 2)定形内胚层、胰腺祖细胞和内分泌祖细胞的表观遗传特征的表征; 3)鉴定促进β细胞分化的化学分子; 4)细胞外基质和其他环境因素对β细胞分化的影响的表征;

DESCRIPTION (provided by applicant): Regeneration of insulin-producing beta-cells as well as islet transplantation is the most promising long-term solutions for Type 1 diabetes (T1D) treatment. A major obstacle to the success of this approach is the shortage of organ donors and the necessity for life-long immunosupression after allograft transplantation. Two recent advances, the success in reprogramming somatic cells into pluripotent ES cell-like IPS (induced pluripotent stem) cells and the success in differentiation of human ES cells into functional islet-like clusters (ILCs), have opened the possibility of generating patient-specific insulin-secreting ILCs through the reprogramming of somatic cells into iPS cells followed by guided differentiation. Indeed, in a proof-of-concept study, we have recently demonstrated that insulin-secreting cells can be generated from human skin fibroblasts using this approach. Despite this encouraging result, several obstacles have to be overcome before the value of this technique can be realized clinically. The major obstacles include: 1) safety concerns over the use of iPS cells that harbor viral vectors; 2) low efficiency of current protocols in generating functional beta-cells; 3) autoimmune destruction of the transplanted beta-cells. Given that the large iPS cell community is attacking the first problem and many immunology labs are trying to solve the third problem, the current proposal plans to use epigenetic, chemical, and ECM approaches to attack the second issue. The Specific Aims are: 1) Characterization of epigenetic features of fetal and mature human beta-cells; 2) Characterization of epigenetic features of definitive endoderm, pancreatic progenitors, and endocrine progenitors; 3) Identification of chemical molecules that promote beta-cell differentiation; 4) Characterization of the effect of extracellular matrix and other environmental factors on beta-cell differentiation;