Mechanisms of Skeletal Morphogenesis During Digit Tip Regeneration

指尖再生过程中骨骼形态发生的机制

• 批准号: 10655300
• 负责人: Feini Qu
• 金额: $ 12.85万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10655300
• 关键词: Adult; Advisory Committees; Advocate; Affect; Alkaline Phosphatase; American; Amputation; Amputees; Animals; BMP2 gene; BMP7 gene; Basic Science; Bioinformatics; Biological; Biological Assay; Bone Morphogenetic Proteins; Bone Tissue; Calcium; Cell Culture System; Cell Lineage; Cell Shape; Cell Transplantation; Cells; Cellular Morphology; Cicatrix; Communities; Complex; Confocal Microscopy; Deposition; Development; Developmental Biology; Digit structure; Disease; Distal; Educational Activities; Embryo; Ensure; Environment; Eph Family Receptors; Ephrins; Feedback; Fluorescent in Situ Hybridization; Fostering; Gene Cluster; Gene Expression; Generations; Genes; Genetic; Goals; Growth; Hand; Histology; Homeobox Genes; Homeostasis; Human; Immunohistochemistry; In Vitro; Injury; Institution; Ligands; Limb Bud; Limb Development; Limb structure; LoxP-flanked allele; Measures; Mediating; Medical; Mentors; Methods; Minerals; Molecular; Monitor; Morphogenesis; Mus; Musculoskeletal; Natural regeneration; Osteoblasts; Osteogenesis; Pathway interactions; Patients; Pattern; Phosphorylation; Physical condensation; Positioning Attribute; Production; Prognosis; Quality of life; RNA; Regenerative Medicine; Regenerative capacity; Regulation; Regulator Genes; Regulatory Pathway; Research; Research Personnel; Research Project Grants; Resources; Role; Series; Signal Pathway; Signal Transduction; Signaling Protein; Site; Skeleton; Structure; System; Technology; Testing; Therapeutic; Tissues; Training; Training Activity; Training Programs; Transgenic Mice; Transplantation; Trauma; Universities; Up-Regulation; Washington; Western Blotting; Work; blastema; bone; bone fracture repair; career; career development; chromatin immunoprecipitation; clinically relevant; digit regeneration; digital; foot; gene function; gene regulatory network; genome editing; improved; in vitro testing; in vivo; in vivo regeneration; induced pluripotent stem cell; innovation; lentiviral-mediated; limb loss; limb regeneration; microCT; mouse model; musculoskeletal injury; novel; osteogenic; osteoprogenitor cell; overexpression; programs; regeneration potential; regenerative; regenerative biology; research faculty; second harmonic; single-cell RNA sequencing; skeletal; skeletal regeneration; skeletogenesis; skills; soft tissue; stem cell therapy; stem cells; success; tissue regeneration; transcriptome; transcriptomics; wound

Project Summary My long-term career goal is to become a successful independent investigator in the field of musculoskeletal regenerative medicine, developing therapeutics to promote the regeneration of complex tissues after injury or disease. The objective of this proposal is to help me transition to independence by providing me with critical scientific skills to investigate the cellular and molecular mechanisms of murine digit tip regeneration versus fibrotic scarring. To reach this objective, a thorough training plan has been established, including research aims and tailored training activities. The proposed research project will seek to develop a novel mouse model and an induced pluripotent stem cell (iPSC) in vitro culture system in order to dissect the functional role of regulatory genes involved in embryonic limb development and morphogenesis, including Hox genes. The central hypothesis of the proposal is that HoxA cluster genes, and specifically Hoxa13, are required during digit regeneration to coordinate osteogenic differentiation, outgrowth, and patterning via Eph/ephrin and bone morphogenetic protein (BMP) signaling. We will investigate this hypothesis by conditionally deleting HoxA cluster genes from osteoblast lineage cells in transgenic mice in Aim 1 and by modulating Hoxa13 gene expression in iPSCs in vitro using a lentiviral-mediated approach in Aims 2 and 3a. Finally, we will evaluate the therapeutic potential of Hoxa13-expressing cells delivered to the wound site of non-regenerative digits in Aim 3b. The project outlined in this application combines basic science with a clinically relevant in vivo platform and cutting-edge transcriptomic and bioinformatic technologies to query the gene regulatory networks and signaling pathways that lead to regeneration versus scarring after musculoskeletal injury. This proposal also includes a comprehensive series of educational activities that will prepare me for my independent research faculty position. The world-class institutional environment at Washington University in St. Louis provides a multitude of resources to ensure the successful completion of the proposed work, as well as ample opportunities for career development. Finally, the assembled Scientific and Career Advisory Committee, along with new mentoring relationships that I am fostering in the developmental and regenerative biology communities, will monitor research progress, provide constructive feedback, and advocate for my professional development as I begin my independent research career.

项目概要 我的长期职业目标是成为该领域的一名成功的独立调查员 肌肉骨骼再生医学，开发促进复杂组织再生的疗法 受伤或患病后。该提案的目的是通过提供我帮助我过渡到独立 具有关键的科学技能来研究小鼠指尖再生的细胞和分子机制 与纤维化疤痕相比。为了实现这一目标，制定了全面的培训计划，包括研究 目标和量身定制的培训活动。拟议的研究项目将寻求开发一种新型小鼠模型 和诱导多能干细胞（iPSC）体外培养系统，以剖析其功能作用 参与胚胎肢体发育和形态发生的调节基因，包括 Hox 基因。中央 该提案的假设是，HoxA 簇基因，特别是 Hoxa13，在数字化过程中是必需的。 通过 Eph/ephrin 和骨进行再生以协调成骨分化、生长和模式 形态发生蛋白（BMP）信号传导。我们将通过有条件删除 HoxA 簇来研究这个假设 目标 1 中转基因小鼠的成骨细胞谱系细胞的基因以及通过调节 Hoxa13 基因表达 在目标 2 和 3a 中使用慢病毒介导的方法体外 iPSC。最后，我们将评估治疗效果 目标 3b 中将表达 Hoxa13 的细胞递送至非再生手指伤口部位的潜力。项目 本申请中概述的结合了基础科学与临床相关的体内平台和尖端技术 转录组学和生物信息技术来查询基因调控网络和信号通路 导致肌肉骨骼损伤后的再生与疤痕形成。该提案还包括全面的 一系列的教育活动将使我为我的独立研究教员职位做好准备。世界一流的 圣路易斯华盛顿大学的制度环境提供了大量资源，以确保 成功完成拟议的工作，以及充足的职业发展机会。最后， 组建了科学和职业咨询委员会，以及我正在培养的新的指导关系 在发育和再生生物学界，将监测研究进展，提供建设性的 反馈，并在我开始独立研究生涯时倡导我的专业发展。