Characterizing the neural crest response to BMP signaling through gastrulation and neurulation

通过原肠胚形成和神经形成表征神经嵴对 BMP 信号的反应

• 批准号: 10448701
• 负责人: Michael Louis Piacentino
• 金额: $ 6.96万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10448701
• 关键词: Award; Binding; CHARGE syndrome; COVID-19 pandemic; Cartilage; Cephalic; Cleft Palate; Co-Immunoprecipitations; Congenital Abnormality; Craniofacial Abnormalities; Craniosynostosis; Defect; Dentin; Development; Dominant-Negative Mutation; Face; Funding; Ganglia; Genes; Goals; Image; Measures; Molecular; Neural Crest; Neural Crest Cell; Parents; Peripheral; Phase; Plant Roots; Play; Publishing; Research; Role; Signal Transduction; Therapeutic; Time; Tooth structure; Training; Universities; bone; bone morphogenetic protein receptor type I; career development; chromatin immunoprecipitation; comparative; craniofacial; craniofacial development; craniofacial structure; epithelial to mesenchymal transition; experimental study; gastrulation; in vivo; migration; novel; pandemic disease; prevent; programs; public health relevance; response; skills; transcriptome; translational approach

PROJECT SUMMARY The cranial neural crest (NC) contributes to the formation of many craniofacial structures including the bones and cartilage of the face, tooth dentin, and peripheral ganglia. Cell signaling regulates different aspects of cranial NC specification, epithelial-to-mesenchymal transition (EMT), and differentiation and disruptions in this developmental program result in many cranial NC-derived craniofacial birth defects including craniosynostosis, Treacher Collins and CHARGE syndromes, and cleft palate. BMP signaling plays a crucial role during the specification and differentiation of cranial NC, and more recently, BMP signaling was shown to control cranial NC EMT. A mechanistic understanding of the role of BMP signaling during cranial NC development is essential to develop novel preventative and therapeutic measures against craniofacial defects. Under the support of this parent award, I have made significant progress toward understanding the mechanisms of BMP signaling underlying cranial NC EMT and migration. My preliminary results suggest that BMP signaling peaks during cranial NC cells during EMT, whereas analysis of fixed sections suggest that signaling level are reduced at earlier (during specification) and later (during migration) stages (Piacentino et al., 2021). To understand the role of BMP signaling during this phase, I have inhibited BMP signaling using a dominant negative Type I BMP receptor (dnBMPR1A) and demonstrated that BMP signaling is essential for cranial NC migration, independent of specification or delamination. Furthermore, I have identified and validated novel targets of BMP signaling using a comparative transcriptome profiling approach (Piacentino et al., 2021). Unfortunately, the COVID-19 pandemic led to university closures and research restrictions that have severely delayed my career development. While I have remained productive and have published results toward the goals of the original proposal (Piacentino et al., 2021), and have identified a novel endocytic mechanism that regulates BMP signaling in cranial NC cells (Piacentino et al., 2020) [Preprint], pandemic setbacks have prevented me from completing the training outlined in my initial Aims. A funding extension will allow me to complete these projects and gain critical training in 1) live time-lapse imaging to carefully examine BMP signaling dynamics in vivo, 2) chromatin-immunoprecipitation (ChIP) experiments to identify direct versus indirect targets of BMP signaling, and 3) co-immunoprecipitation (co-IP) experiments essential to identify the binding partners of the BMP target genes, Id1/2/3/4. This training is essential for me to gain the skills necessary to establish a strong independent research program that will make lasting impacts on the field of BMP signaling in craniofacial development.

项目摘要 颅神经rest（NC）有助于形成许多颅面结构 包括脸部的骨头和软骨，牙齿牙本质和周围神经节。细胞信号传导 调节颅NC规范的不同方面，上皮到间质转变（EMT）， 该发展计划中的差异和干扰导致许多颅内NC衍生 颅面的先天缺陷，包括颅突，背叛者柯林斯和电荷综合症，以及 裂口味。 BMP信号在颅骨的规范和分化过程中起着至关重要的作用 NC以及最近的BMP信号被证明可以控制颅NC EMT。机械 了解BMP信号在颅NC开发过程中的作用对于发展至关重要 针对颅面缺陷的新型预防和治疗措施。 在该父母奖的支持下，我取得了重大进步 BMP信号传导的机制和迁移。我的初步结果 建议BMP信号传导在EMT期间颅NC细胞期间达到峰值，而对固定的分析 部分表明，在较早（规范）和更晚的早期（期间）（期间）降低了信号传导水平（在 迁移）阶段（Piacentino等，2021）。了解在此期间BMP信号的作用 阶段，我使用主要的I型BMP受体（DNBMPR1A）抑制了BMP信号传导 并证明BMP信号对于颅NC迁移至关重要，与 规格或分层。此外，我已经确定并验证了BMP的新颖目标 使用比较转录组分析方法的信号传导（Piacentino等，2021）。 不幸的是，共同199的大流行导致了大学的关闭和研究限制 严重延迟了我的职业发展。虽然我保持生产力 已发表的结果朝着原始提案的目标（Piacentino等，2021），并具有 确定了一种新型的内吞机制，该机制调节颅NC细胞中的BMP信号传导（Piacentino 等，2020）[预印]，大流行挫折阻止了我完成概述的训练 在我的最初目标中。资金扩展将使我能够完成这些项目并获得关键 训练1）实时延时成像以仔细检查体内BMP信号传导动力学，2） 染色质免疫沉淀（芯片）实验，以识别直接与间接靶标的BMP 信号传导和3）识别结合伙伴必不可少的共免疫沉淀（CO-IP）实验 BMP靶基因的ID1/2/3/4。这项培训对于我获得必要的技能至关重要 建立一个强大的独立研究计划，将对BMP领域产生持久影响 颅面发育中的信号传导。