Determining the Role of Plastin-3 in Osteoblast Differentiation and Mineralization

确定 Plastin-3 在成骨细胞分化和矿化中的作用

• 批准号: 10469332
• 负责人: Samantha Meagan Chin
• 金额: $ 3.27万
• 依托单位: WASHINGTON UNIVERSITY
• 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 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10469332
• 关键词: 2 year old; Actins; Address; Adult; Affect; Alkaline Phosphatase; Biochemical; Biological Models; Biology; Bone Density; Bone Development; Bone Diseases; Bone Resorption; Bundling; Calcium; Cellular biology; Child; Childhood; Clinical; Collagen Diseases; Core Facility; Cytoskeleton; Defect; Development; Diagnosis; Disease; Elderly; F-Actin; Fishes; Fracture; Functional disorder; Future; Genes; Genetic study; Goals; In Vitro; Individual; Institution; Knock-in; Life; Light; Link; Mediating; Medical; Mentorship; Modeling; Molecular; Mus; Musculoskeletal; Mutation; Osteoblasts; Osteoclasts; Osteocytes; Osteogenesis; Osteoporosis; Osteoporotic; Pathogenicity; Pathway interactions; Patients; Physicians; Physiologic calcification; Physiological; Play; Process; Proteins; Recording of previous events; Recurrence; Regulation; Research; Research Personnel; Role; Scientist; Structure; Techniques; Testing; Therapeutic; Time; Training; Training Programs; Transgenic Organisms; Undifferentiated; Universities; Washington; Zebrafish; age related; bone; bone fragility; bone health; bone loss; bone mass; bone turnover; early onset; effective therapy; experimental study; genetic analysis; in vivo; in vivo Model; insight; knock-down; mineralization; multidisciplinary; mutant; new therapeutic target; novel; osteoblast differentiation; osteogenic; plastin; response; side effect; skills; therapeutic target; transcriptome sequencing; treatment response

Samantha Chin (PD/PI) PROJECT SUMMARY/ABSTRACT Osteoporosis is the most common bone disorder in the world and represents a significant clinical and societal burden due to related bone fractures. Although classically viewed as an age-related disorder, osteoporosis now more commonly describes a general condition of low bone mineral density that can arise in children as well as adults. The mainstay of osteoporosis treatment is anti-resorptive therapy, which seeks to curb bone resorption. This approach, however, is plagued by undesirable side-effects and concerns regarding long- term efficacy, particularly in children, underscoring the pressing need for alternative therapeutic targets and strategies to effectively treat osteoporosis. Plastin-3 (PLS3) is a calcium-sensitive actin-bundling protein that has recently been linked to the development of childhood-onset osteoporosis; however, the underlying pathophysiology is completely unknown. This is due in part to the fact that the role of PLS3 in bone health remains to be identified. This proposal will address these questions and build a basis to develop PLS3 as a novel anabolic anti-osteoporosis target. Genetic studies in mouse and zebrafish models suggest that PLS3 plays a role in osteoblast-mediated bone formation; however, it remains unclear how PLS3 mechanistically contributes to these processes. Aim 1 will employ pathogenic PLS3 mutants that are defective in either actin-bundling or calcium-regulation to elucidate how PLS3 promotes osteoblast differentiation and mineralization in cultured osteoblasts. In addition, RNA-seq analysis will also be used to identify novel pathways that contribute to mineralization in order to provide insight to the specific role of PLS3 and actin dynamics in osteoblast mineralization. To better understand how PLS3 contributes to regulation of bone development in vivo, Aim 2 is focused on developing a zebrafish model system to study the effect of pathogenic PLS3 mutations on bone formation. Taken together, this proposal will fill major gaps in our understanding of how PLS3 and its regulation of actin cytoskeleton dynamics contribute to bone health and the development of osteoporosis. The applicant has assembled a multi-disciplinary mentorship team with experts in the actin cytoskeleton, bone biology, cell biology, and zebrafish model systems that will support the applicant in completion of these aims. The proposal also takes advantage of the Washington University’s strengths in musculoskeletal and zebrafish research including the institution’s cutting-edge cores and facilities. Additionally, Washington University Medical Scientist Training Program has a rich history of supporting physician scientists at various stages of training that will also be invaluable to the applicant’s development. This training and mentorship will provide the applicant critical skills that will facilitate the transition to independent researcher and physician-scientist.

Samantha Chin（PD/PI） 项目总结/摘要 骨质疏松症是世界上最常见的骨疾病，代表了一个重要的临床和病理学问题。 由于相关骨折造成的社会负担。尽管传统上被视为一种与年龄相关的疾病， 骨质疏松症现在更普遍地描述了一种低骨矿物质密度的一般状况， 儿童以及成人。骨质疏松症治疗的支柱是抗吸收疗法，其旨在抑制 骨吸收然而，这种方法受到不期望的副作用和对长期使用的担忧的困扰。 长期疗效，特别是在儿童中，强调迫切需要替代治疗靶点， 有效治疗骨质疏松症的策略。Plastin-3（PLS 3）是一种钙敏感的肌动蛋白捆绑蛋白， 最近被认为与儿童期发病的骨质疏松症的发展有关;然而， 病理生理学完全未知。这部分是由于PLS 3在骨骼健康中的作用 仍有待确认。该建议将解决这些问题，并建立一个基础，发展PLS 3作为一种新的 合成代谢抗骨质疏松目标。 在小鼠和斑马鱼模型中的遗传研究表明，PLS 3在成骨细胞介导的骨形成中起作用。 然而，仍然不清楚PLS 3如何在机制上有助于这些过程。目标1将 使用致病性PLS 3突变体，这些突变体在肌动蛋白成束或钙调节方面都有缺陷，以阐明 PLS 3如何促进培养的成骨细胞中的成骨细胞分化和矿化。此外，RNA-seq 分析还将用于确定有助于矿化的新途径，以提供见解 PLS 3和肌动蛋白动力学在成骨细胞矿化中的特定作用。为了更好地了解PLS 3 有助于调节体内骨发育，目标2致力于开发斑马鱼模型系统 研究致病性PLS 3突变对骨形成的影响。总的来说，这项提案将填补主要的 我们对PLS 3及其对肌动蛋白细胞骨架动力学的调节如何促进骨形成的理解存在差距。 健康和骨质疏松症的发展。 申请人已经组建了一个多学科的导师团队，其中包括肌动蛋白细胞骨架方面的专家， 骨生物学，细胞生物学和斑马鱼模型系统，将支持申请人完成这些 目标。该提案还利用了华盛顿大学在肌肉骨骼和 斑马鱼研究，包括该机构的尖端核心和设施。此外，华盛顿大学 医学科学家培训计划有着丰富的历史，在各个阶段支持医生科学家， 培训，这也将是宝贵的申请人的发展。这种培训和指导将提供 申请人的关键技能，这将有助于过渡到独立的研究人员和医生，科学家。