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.

项目总结 颅神经脊(NC)在许多颅面结构的形成中起重要作用 包括面部的骨骼和软骨、牙本质和外周神经节。细胞信令 调节颅骨NC规范的不同方面，上皮向间充质转化(EMT)， 而这种发育程序的分化和中断导致了许多颅骨NC衍生 头面部出生缺陷，包括颅缝早闭、Treacher Collins和Charge综合征，以及 腭裂。BMP信号在颅骨的特化和分化过程中起着至关重要的作用 Nc，最近，BMP信号被证明控制颅骨Nc EMT。机械师 了解BMP信号在颅骨NC发育中的作用是发展的关键 新的预防和治疗头面部缺陷的措施。 在这个家长奖的支持下，我在理解 骨形态发生蛋白信号转导机制与颅骨NC、EMT和迁移的关系我的初步结果 提示在EMT期间，BMP信号在颅骨NC细胞中达到峰值，而分析固定的 部分建议在较早(规范期间)和较晚(在规范期间)降低信令级别 移民)阶段(Piacentino等人，2021年)。要了解BMP信号在这一过程中的作用 阶段，我使用显性负I型BMP受体(DnBMPR1A)抑制BMP信号转导 并证明了BMP信号对颅骨NC迁移是必不可少的，不依赖于 规格或分层。此外，我还确定并验证了BMP的新靶点 使用比较转录组图谱方法发出信号(Piacentino等人，2021年)。 不幸的是，新冠肺炎疫情导致大学关闭和研究限制 严重延缓了我的事业发展。虽然我一直保持着生产力，并 公布了实现原始提案目标的结果(Piacentino等人，2021年)，并已 发现了一种新的调节颅骨NC细胞BMP信号的内吞机制(Piacentino 等人，2020)[预印本]，大流行的挫折使我无法完成概述的培训 在我最初的目标中。资金延期将使我能够完成这些项目，并获得关键的 培训1)实时时间推移成像，以仔细检查体内BMP信号动力学，2) 染色质免疫沉淀(CHIP)实验鉴定BMP的直接和间接靶标 信号传递，以及3)共免疫沉淀(co-IP)实验对识别结合伙伴至关重要 在BMP靶基因中，ID1/2/3/4。这种培训对我获得必要的技能是必不可少的 建立一个强大的独立研究计划，对BMP领域产生持久的影响 头面部发育中的信号。