Defining mechanistic differences between embryonic and regenerative organogenesis

研究综述 成人组织再生与胚胎发育具有共同的关键遗传要求。然而，尽管 再生医学领域的浓厚兴趣，即允许受控使用的自然机制 关于胚胎因子在成人体内重新构建功能组织和器官的研究仍然知之甚少。这 建议将海葵海葵的咽部作为一种简单而遗传的载体 强大的模型系统，阐明发育和发育之间的机制异同 再生器官的形成。线虫咽部是唯一适合于解决这个问题的 在胚胎发育期间通过原肠形成的，功能器官能够 在成人完全切除后完全再生。在初步实验中，我已经确定 叉头盒A(FOXA)，一种进化上保守的转录因子，在两个咽中表达 发展和再生。此外，通过功能基因分析，我已经证明了 FOXA在胚胎发育过程中的咽部形成。这项研究计划的目的是：1)确定 FOXA在再生有机体咽部细胞测定中的作用，2)定义FOXA依赖的转录 胚胎咽形成所需的程序，以及3)阐明再生特有的咽部生长 由FOXA指导的计划。利用转基因成年动物品系、系统RNAi、 嵌合体基因的表达将评估FOXA在咽部发育和再生中的作用。另外， RNA测序(RNAseq)和染色质免疫沉淀DNA测序(ChIPseq)分析将 确定胚胎发生特异性和再生特异性FOXA基因靶点和结合部位。这些FOXA- 依赖基因和调控结合位点将通过遗传扰动进一步询问使用 系统RNAi和CRISPR介导的基因组编辑机械地比较胚胎和后胚胎 同一种动物的胚胎器官发生程序。旨在了解它们之间的相似之处和 发育和再生过程中复杂器官形成的差异将揭示对 作为整个器官再生基础的自然机制，可以提供概念上的见解来指导 未来再生疗法的发展。 好了！

RESEARCH SUMMARY Adult tissue regeneration shares key genetic requirements with embryonic development. However, despite intense interest from the field of regenerative medicine, the natural mechanisms that allow for the controlled use of embryonic factors to construct functional tissue and organs de novo in adults remains poorly understood. This proposal focuses on the pharynx of the starlet sea anemone Nematostella vectensis as a simple and genetically powerful model system to elucidate the mechanistic similarities and differences between developmental and regenerative organ formation. The Nematostella pharynx is uniquely suited to address this question as it is formed through gastrulation during embryonic development and the functional organ is capable of being regenerated in its entirety following complete removal in the adult. In preliminary experiments, I have identified forkhead box A (foxA), an evolutionarily conserved transcription factor, as expressed in both pharynx development and regeneration. Additionally, through functional genetic analysis I have shown a requirement for foxA in pharynx formation during embryogenesis. This research proposal aims to 1) determine the precise role of foxA in pharyngeal cell determination in a regenerative organism, 2) define the foxA-dependent transcriptional program required for embryonic pharynx formation, and 3) elucidate a regeneration-specific pharyngeal growth program directed by foxA. Functional genetic experiments using transgenic adult animal lines, systemic RNAi, and mosaic gene expression will assess the role of foxA in pharynx development and regeneration. Additionally, RNA sequencing (RNAseq) and chromatin immunoprecipitation DNA sequencing (ChIPseq) analyses will determine embryogenesis-specific and regeneration-specific foxA gene targets and binding sites. These foxA- dependent genes and regulatory binding sites will be further interrogated through genetic perturbation using systemic RNAi and CRISPR-mediated genome editing to mechanistically compare embryonic and post- embryonic organogenesis programs in the same animal. Studies aimed at understanding the similarities and differences of complex organ formation through development and regeneration will reveal deep insight into the natural mechanisms that underlie whole-organ regeneration and could provide conceptual insights to guide the development of future regenerative therapeutics. !