Genetic and physiologic regulation of pig islet development and function

摘要 啮齿类动物的研究从根本上提高了我们对胰岛细胞的认识， 发育和功能，但了解人类胰腺的啮齿动物模型的局限性 形成和疾病已经加强了对实验系统的兴趣， 人类胰腺发育和胰岛功能。我们的研究表明， 研究猪的胰岛发育，包括多种保守特征的证据， 在猪和人中的胰岛发育，在小鼠中未观察到。这包括表达 SIX 2和SIX 3等转录因子控制人胰岛β细胞命运的标志性特征 和功能我们已经开发出一种可靠的获取猪胰腺和胰岛的框架 并描述了分子、细胞、信号和遗传特征 从胎儿到新生儿阶段的发育猪α，β和δ细胞，包括第一个详细的 这些细胞的发育转录组。此外，我们的团队还制作了一些 第一个使用CRISPR/Cas9靶向的基因编辑猪。这些进步促使以下方面 目的： 目标1。阐明猪胰岛细胞发育和功能成熟的遗传结构 目标二。鉴定猪胰岛SIX 2和SIX 3表达的天然调节因子 目标3：猪HNF 1A杂合突变表型的研究 为了实现我们的目标，我们组建了一支由合作调查人员组成的优秀团队， 在发育生物学、猪遗传学、胰岛生物学、基因组学和 糖尿病研究这项工作应该允许以前无法实现的遗传和遗传学研究。 调节猪胰岛细胞的生理机制。这包括确定监管 连接染色质动力学和基因转录以控制胰岛α、β和δ细胞的特征 命运和功能，以及创建更好地再现人类疾病的MODY新模型 遗传学、发病机理和治疗学。我们的工作应该创造新的灵活的实验 从生理学角度研究胰岛细胞的发育和功能成熟的范例 和病理阶段，一个突出的优势，扩大了我们的建议的影响， 人体健康

Abstract Studies in rodents have fundamentally advanced our understanding of pancreatic islet cell development and function, but limitations of rodent models for understanding human pancreas formation and disease have intensified interest in experimental systems that more closely reflect human pancreas development and islet function. Our studies have revealed advantages for investigating islet development in pigs, including evidence for multiple conserved features of islet development in pigs and humans, not observed in mice. This includes expression of transcription factors like SIX2 and SIX3 which govern hallmark features of human islet β cell fate and function. We have developed a reliable framework for procuring pig pancreata and islets from any developmental stage, and described molecular, cellular, signaling and genetic features of developing pig α, β, and δ cells from fetal to neo-natal stages, including the first detailed developmental transcriptome of these cells. In addition, our team has generated some of the first gene-edited pigs using CRISPR/Cas9 targeting. These advances motivate the following Aims: Aim 1. Elucidate the genetic architecture of pig islet cell development and functional maturation Aim 2. Identify native regulators of pig islet SIX2 and SIX3 expression Aim 3. Investigate phenotypes of HNF1A heterozygous mutation in pigs To achieve our aims, we assembled a superb team of collaborating investigators with complementary expertise in developmental biology, pig genetics, islet biology, genomics and diabetes research. This work should allow previously unattainable investigation of genetic and physiological mechanisms regulating pig islet cells. This includes identification of regulatory features that connect chromatin dynamics and gene transcription to control islet α, β and δ cell fate and function, and creation of new models of MODY that better recapitulate human disease genetics, pathogenesis and therapeutics. Our work should create new flexible experimental paradigms to investigate development and functional maturation of islet cells from physiological and pathological stages, a striking advantage that broadens the impact of our proposal on human health.