Single-cell analysis of the craniofacial skeletal stem cell niche

颅面骨骼干细胞生态位的单细胞分析

• 批准号: 10040381
• 负责人: Peter Fabian
• 金额: $ 10.31万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10040381
• 关键词: Adult; Advisory Committees; Age; Aging; Alkaptonuria; Arthritis; Biology; Bone Diseases; Buffers; Cartilage; Catabolism; Cell Maintenance; Cell physiology; Cells; Cephalic; Chemicals; Collagen Fiber; Communities; Consultations; Country; Craniofacial Abnormalities; DNA Sequence Alteration; Data; Defect; Development; Ensure; Enzymes; Face; Faculty; Failure; Fertilization; Fishes; Future; Genes; Genetic; Genetic Transcription; Genomics; Glycogen; Goals; Growth; Head; Health; Homeostasis; Human; Image; In Situ; In Situ Hybridization; Injury; Institution; Label; Lead; Liver; Maintenance; Mammals; Maternal Phenylketonuria; Mechanics; Mediating; Mentors; Mentorship; Mesenchymal; Metabolic; Metabolic Diseases; Metabolic Pathway; Molecular Profiling; Mosaicism; Mus; Mutant Strains Mice; Mutate; Mutation; Neural Cell Adhesion Molecule L1; Neural Crest; Organ; Pattern; Periosteal Cell; Periosteum; Phase; Phenylalanine; Phenylalanine Metabolism Pathway; Phenylketonurias; Population; Positioning Attribute; Publishing; Reporting; Research; Robinow syndrome; Role; Scientist; Series; Signal Transduction; Skeleton; Source; Testing; Time; Tissues; Training; Validation; WNT5A gene; Work; Yang; Zebrafish; adult stem cell; blastomere structure; bone; career; career development; cartilage repair; cell type; craniofacial; craniofacial bone; craniofacial repair; cranium; design; experience; high resolution imaging; imaging approach; innovation; insight; member; mutant; novel; paracrine; premature; professor; regenerative therapy; repaired; self-renewal; single cell analysis; single-cell RNA sequencing; skeletal; skeletal tissue; skeletogenesis; stem cell homeostasis; stem cell niche; stem cells; tenure track; tissue repair; transcriptomics

Stem cells reside in specialized niches that support their long-term maintenance. Disruption of the niche, for example due to aging, injury, or genetic mutations, can lead to declines in stem cells that result in an ability to maintain and repair tissues. Compared to our understanding of the stem cells that maintain and repair skeletal tissues, we know much less about the cell types that constitute the niche for skeletal stem cells. In this proposal, I have taken an innovative single-cell transcriptomic approach to characterize potential niche cells in the zebrafish face. Preliminary data have uncovered molecular signatures of putative niche cells, including homologs of genes that when mutated cause craniofacial malformations in humans: WNT5A (Robinow Syndrome), HGD (Alkaptonuria, aka “black bone disease”), and PAH (maternal Phenylketonuria). This analysis suggests a role for Wnt5a as a niche-derived paracrine factor for skeletal stem cell maintenance, and an unexpected local role for phenylalanine catabolism, outside the liver, in protecting craniofacial skeletal tissues. The aims outlined in this proposal leverage powerful genomic, genetic, and high-resolution imaging approaches to test that niche cells are located in the outer periosteum in both fish and mammals (Aim 1), that niche cells and Wnt5a secretion act to maintain skeletal stem cells in the inner periosteum (Aim 2), and that efficient breakdown of phenylalanine in niche cells is critical for craniofacial skeletal health (Aim 3). Completion of these aims will inform how defects in the niche result in both developmental defects in the face (such as in Robinow Syndrome) and a failure to maintain the facial skeleton (such as with the severe arthritis seen in Alkaptonuria). These findings may also lead to future therapies aimed at better maintaining and repairing the skeleton through modulating the niche. The project and mentorship plan outlined in this proposal were designed to lay the groundwork for my career goal of obtaining a position as a tenure-track Assistant Professor at a top-tier academic research institution. During the K99 phase, I will receive mentorship in zebrafish biology from Gage Crump and crucial mouse training from my co-mentor Yang Chai. Regular interactions with clinician-scientists Shoji Yano and Kathryn Moseley, experts in Phenylketonuria and metabolic diseases and members of my advisory committee, will allow me to connect findings to human health. Career development activities at USC will prepare me for the transition to an independent faculty position during the R00 phase. As USC hosts one of the most experienced communities of craniofacial and skeletal biologists in the country, there are few better places to conduct this research and acquire the training to achieve my career goals.

干细胞居住在支持其长期维护的专门壁nikes中。利基的破坏，因为 由于衰老，损伤或遗传突变，例如，干细胞的下降会导致能力下降 维护和修复组织。与我们对维护和修复骨骼的干细胞的理解相比 组织，我们对构成骨骼干细胞利基市场的细胞类型的了解少得多。在这个 提案，我采用了一种创新的单细胞转录组方法来表征潜在的小众细胞 斑马鱼的脸。初步数据发现了假定的小众细胞的分子特征，包括 当突变引起人类颅面畸形时的基因同源物：wnt5a（robinow 综合征），HGD（Alkaptonuria，又名“黑骨病”）和PAH（母型苯酮尿症）。这个分析 提出WNT5A作为骨骼干细胞维持的小众旁分泌因子的作用，一个 肝内苯丙氨酸分解代谢的意外作用，在保护颅面骨骼组织中。 该提案中概述的目的利用强大的基因组，遗传和高分辨率成像 测试小众细胞位于鱼类和哺乳动物的外部per骨（AIM 1）的方法， 利基细胞和Wnt5a分泌作用以维持内部骨膜中的骨骼干细胞（AIM 2），并且 小众细胞中苯丙氨酸的有效分解对于颅面骨骼健康至关重要（AIM 3）。完成 这些目的中的目标将告知利基市场缺陷如何导致两种发育缺陷（例如 罗宾诺综合症）和未能维持面部骨骼的（例如， 碱）。这些发现也可能导致未来的疗法，以更好地维护和维修 骨骼通过调节利基市场。 该提案中概述的项目和精明计划旨在为我的职业生涯奠定基础 获得一流学术研究机构的终身助理助理教授的目标。 在K99阶段，我将获得斑马生物学的指标，从斑马和关键的小鼠中 我的Yang Chai的培训。与临床科学家Shoji Yano和Kathryn的定期互动 Moseley是苯酮尿和代谢疾病的专家，我的咨询委员会成员将 请允许我将发现与人类健康联系起来。南加州大学的职业发展活动将为我做好准备 在R00阶段过渡到独立的教师职位。作为美国南加州大学拥有最有经验的人之一 该国颅面和骨骼生物学家的社区，几乎没有更好的地方可以进行此事 研究并获得培训以实现我的职业目标。