Mechanisms of intestine development

此NIDDK指导研究科学家发展奖申请描述了一个为期5年的培训计划，旨在让我获得额外的技能和知识，以便我可以过渡到一个独立的研究科学家。在进行拟议的研究，并按照我的职业发展计划，我将增加我的科学剧目，并获得在肠道发育生物学的专业知识。利用这一新获得的肠道发育专业知识，我将建立一个科学的利基，这将使我有别于我的导师，并铺平道路，以一个强大的，校外资助的研究计划。 本申请中提出的研究将集中于了解胚胎肠中的干/祖细胞是如何使用传统的小鼠遗传学和我们开发的一种新型体外系统来指定和调节的，该系统允许我们从人类胚胎和诱导多能干细胞培养中产生人类肠组织。在培养中产生人类肠道组织将允许对人类肠道发育、稳态和疾病进行前所未有的研究。具体的目的是测试一个模型，其中Sox 9调节早期肠祖细胞，因为它们被限制在发育中的肠的增殖绒毛间区域。具体目标1将使用谱系标记和细胞示踪实验研究人类和小鼠肠干细胞的胚胎发育。特异性目的2将确定Sox 9在调节人类和小鼠肠绒毛间区域的胚胎发育中的作用。具体目标3将研究Wnt信号传导在发育期间绒毛间祖细胞区建立中的抑制作用。 总的来说，这里提出的研究符合跨NIH报告“消化系统疾病研究的机遇和挑战：国家消化系统疾病委员会的建议”（2009年），该报告指出："揭示出生前和新生儿生活中控制消化道发育和分化的机制可以为再生疗法治疗消化道癌症和其他疾病提供新的见解，并为疾病的发病机制提供了新的见解。“这个指导研究科学家发展奖的目标是发展专业知识，使我能够运行一个独立的，资助的研究计划，这将有助于改善人类健康和疾病。 公共卫生相关性：影响产前和围产期消化道的疾病包括坏死性小肠结肠炎（NEC）和短肠综合征（SBS），并导致早产儿死亡率和发病率增加。旨在改善这些和其他疾病的结果的再生疗法需要深入了解胎儿肠道如何发育。这项研究计划使用一种新的系统来研究培养中的人类胎儿肠道发育，并旨在研究发育中肠道的增殖细胞是如何建立和调节的。从这项工作中获得的见解可能适用于改善影响肠道的早期儿童疾病。

DESCRIPTION (provided by applicant): This NIDDK Mentored Research Scientist Development Award application describes a 5-year training plan designed to allow me to gain additional skill and knowledge so that I can transition into an independent research scientist. In carrying out the proposed research, and by following my career development plan I will add to my scientific repertoire and acquire expertise in intestinal developmental biology. Using this newly acquired expertise in intestine development, I will establish a scientific niche that will set me apart from my mentors and pave the way to a robust, extramurally funded research program. The research proposed in this application will focus on understanding how the stem/progenitor cells in the embryonic intestine are specified and regulated using both traditional mouse genetics, and a novel in vitro system we have developed that allows us to generate human intestinal tissue in culture from human embryonic and induced pluripotent stem cells. Generation of human intestinal tissue in culture will allow for unprecedented studies of human intestinal development, homeostasis and disease. The specific aims are designed to test a model where Sox9 regulates early intestinal progenitor cells as they are restricted to the proliferative intervillus region of the developing intestine. Specific Aim 1 will investigate the embryonic development of human and mouse intestinal stem cells using lineage labeling and cell tracing experiments. Specific Aim 2 will determine the role of Sox9 in regulating embryonic development of the intervillus region in human and mouse intestine. Specific Aim 3 will investigate a suppressive role for Wnt signaling in establishment of the intervillus progenitor zone during development. Collectively, the research proposed here is in line with the Trans-NIH Report "Opportunities and Challenges in Digestive Diseases Research: Recommendations of the National Commission on Digestive Diseases" (2009) which states: "Uncovering the mechanisms that control development and differentiation of the digestive tract before birth and in neonatal life could generate new insights for regenerative therapies to treat digestive cancers and other diseases, as well as provide new insights into disease pathogenesis." The goal of this Mentored Research Scientist Development Award is to develop the expertise that will allow me to run an independent, funded research program that will contribute to improving human health and disease. PUBLIC HEALTH RELEVANCE: Diseases that affect the prenatal and perinatal digestive tract include Necrotizing Enterocolitis (NEC) and Short Bowel Syndrome (SBS), and lead to increased rates of mortality and morbididity in preterm infants. Regenerative therapies designed at improving the outcomes of these and other diseases require insights into how the fetal intestine develops. This research proposal uses a novel system to study human fetal intestinal development in culture, and is designed to investigate how the proliferative cells of the developing intestine are established and regulated. Insights gained from this work may be applied to improving early childhood diseases affecting the intestines.