Transcriptional regulation of progenitor cell fate in craniofacial ligament regeneration

颅面韧带再生中祖细胞命运的转录调控

• 批准号: 10709889
• 负责人: Troy Anderson
• 金额: $ 4.85万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10709889
• 关键词: ATAC-seq; Acute; Address; Adopted; Adult; Advisory Committees; Automobile Driving; Binding; Biological Assay; Biology; Biomechanics; Candidate Disease Gene; Cell Differentiation process; Cell Nucleus; Cells; Cephalic; Chromatin; Cicatrix; Clustered Regularly Interspaced Short Palindromic Repeats; Communication; Complex; Data; Degenerative polyarthritis; Deposition; Development; Enhancers; Epigenetic Process; Extracellular Matrix; Failure; Fellowship; Future; Gene Expression; Genetic; Genetic Transcription; Genomic Segment; Human; Injury; Institution; Joint Instability; Joint repair; Joints; Knock-out; Knockout Mice; Knowledge; Learning; Life; Ligaments; Mandible; Mentorship; Mesenchymal; Mesenchyme; Modeling; Molecular Target; Morphology; Mus; Mutation; Natural regeneration; Neural Crest; Operative Surgical Procedures; Oryctolagus cuniculus; Patient-Focused Outcomes; Periosteal Cell; Periosteum; Population; Postdoctoral Fellow; Productivity; Property; Recurrence; Risk; Scientist; Signal Transduction; Source; Specific qualifier value; Techniques; Temporomandibular Joint; Testing; Therapeutic; Time; Tissues; Training; Transcriptional Regulation; Translating; Transposase; Zebrafish; career; cell regeneration; cell type; craniofacial; differential expression; experimental study; gene regulatory network; healing; improved; in vivo; insight; ligament development; ligament injury; loss of function; multiple omics; mutant; novel; osteogenic; overexpression; progenitor; regenerative; regenerative therapy; repair model; repaired; scleraxis; single nucleus RNA-sequencing; skeletal; skills; stem cells; tendon development; transcription factor; translational study

PROJECT SUMMARY/ABSTRACT Ligament injuries are compounded by the drastically increased risks of reinjury and eventual osteoarthritis. These risks result from a failure of differentiation; torn mammalian ligaments reform with fibrous scar tissue rather than with true ligamentocytes, altering the biomechanical properties of the repaired ligament and destabilizing the nearby joint. While the current models (mice and rabbits) used to study ligament repair recapitulate ligament scarring, they are unlikely to yield novel insights into targets for regenerative therapeutics. To address this shortcoming, our lab has developed the highly regenerative zebrafish as a model for craniofacial ligament regeneration. Zebrafish have the remarkable capacity to regenerate their ligaments without scarring in under a month. Our preliminary data demonstrate that the jaw joint-supporting interopercle (IOP) ligament in zebrafish shows identical morphology to the uninjured ligament as soon as 28 days after transection. During this time, the injured ligament is replaced with a dense mesenchyme which deposits a complex extracellular matrix to remake a functional ligament. Additionally, preliminary lineage tracing experiments show that both the uninjured ligament and the regenerative mesenchyme are cranial neural crest-derived. This project aims to identify the source of the mesenchymal cells which reform the ligament, as well as the genetic and epigenetic changes which promote ligamentocyte fate. In Aim 1, I will use enhancer peaks with increased accessibility after ligament injury to firstly trace the lineage of the regenerative mesenchyme to the regenerated ligament, and secondly trace the lineage of the periosteum through regeneration. In Aim 2, I will analyze my single-nuclei multiomic (gene expression and chromatin accessibility for each cell) data from jaw joints through IOP ligament regeneration to generate a short list of candidate transcription factors using a selection funnel of motif accessibility, gene expression, and transcription factor binding. I will then assess in vivo if these transcription factors are expressed before ligamentocyte fate is adopted, and generate knockout zebrafish lines to assess if each transcription factor is necessary for ligamentocyte fate. Through these aims, we will unveil a novel population of cells capable of regenerating ligaments, as well as the transcription factors necessary for ligamentocyte differentiation. The training plan outlined through this proposal will develop the techniques, mentorship, and communication skills necessary to establish my future career as an independent scientist studying craniofacial regeneration. Through the mentorship of Dr. Joanna Smeeton (Sponsor), Dr. Stavros Thomopoulos (Co-sponsor), and my thesis advisory committee, I will be well prepared for my transition to a postdoctoral fellowship at a leading biomedical institution. Dr. Smeeton has extensive knowledge of the zebrafish as a model for craniofacial regeneration, and Dr. Thomopoulos is an expert in translational mouse tendon development with an outstanding mentorship record. Under their combined sponsorship, I will learn cutting-edge computational and in vivo genetics to develop a productive career in regenerative craniofacial biology.

项目总结/文摘