Using Embryos to Understand the Chromatin State of Mesoderm Induction

描述（由申请人提供）：出生缺陷，包括神经管、眼睛和肢体的缺陷，是由于胚胎发育过程中细胞规格不当造成的。形成这些特化组织所必需的主要胚胎学事件之一是中胚层的诱导。我们的目标是确定驱动中胚层形成的基因组和生化过程，长期目标是能够产生更复杂的组织类型。虽然介导中胚层诱导和许多其他胚胎过程的信号通路已经被很好地理解，但下游转录因子如何与染色质接口和通信仍然是一个谜。这种并置是正常细胞规范的核心，并且正在成为细胞重编程的关键因素。因此,

DESCRIPTION (provided by applicant): Birth defects, including those of the neural tube, eye and limb, are caused by improper cellular specification during embryogenesis. One of the primary embryological events necessary for the formation of these specialized tissues is the induction of mesoderm. Our aim is to identify the genomic and biochemical processes that drive the formation of mesoderm with the long-term goal of being able to generate more complex tissue types. While the signaling pathways that mediate mesoderm induction and many other embryological processes are well understood, how downstream transcription factors interface and communicate with chromatin is still a mystery. This juxtaposition is central for normal cellular specification, and is emerging as a critical element of cellular reprogramming. Therefore, inroads into this problem are important for understanding errors during development and will enable the establishment of better protocols for regenerative medicine. With the advent of genomic sequencing technologies, we can now ask fundamental questions about how signaling pathways interface with chromatin, whether they are permissive or active players in generating open chromatin structures and how these signals are communicated between neighboring cells. In this grant, we use Xenopus laevis and Xenopus tropicalis to address the involvement of the chromatin state and its interface with the Nodal signaling factor, smad2/3, during in vivo mesoderm induction. The wealth of embryological resources present in these species, the deep knowledge of their fate maps, combined with newly available genomic tools, presents a prime opportunity to revisit mechanisms underlying classic cell fate and inductive interactions using emerging modern technologies. The central hypothesis of this grant is that the interplay between chromatin state and smad2/3 underlie mesoderm induction and patterning. PUBLIC HEALTH RELEVANCE: Birth defects, including those of the neural tube, eye and limb, are caused by improper cellular specification during embryogenesis. The molecular signals underlying many of these disorders are understood, but how they interact with the genome is largely unknown. In this grant, we use the frog embryo to examine how signaling between neighboring cells influences their genomes, both in terms of chromatin signatures and in transcription factor occupancy. Our central hypothesis is that chromatin and its associated transcriptional states underlie the competency of tissues to communicate for normal embryonic development.