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: 10313727
• 负责人: Samantha Meagan Chin
• 金额: $ 3.2万
• 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/10313727
• 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) 项目摘要/摘要 骨质疏松症是世界上最常见的骨骼疾病，具有重要的临床和 相关骨折造成的社会负担。尽管传统上被认为是一种与年龄相关的疾病， 骨质疏松症现在更常见地描述的是一种普遍的低骨密度状况，可发生在 无论是孩子还是成年人。骨质疏松症治疗的支柱是抗吸收疗法，它试图遏制 骨吸收。然而，这种方法受到不良副作用的困扰，以及对长期- 长期疗效，特别是在儿童中，强调迫切需要替代治疗靶点和 有效治疗骨质疏松症的策略。纤溶酶原蛋白-3(PLS3)是一种钙敏感的肌动蛋白捆绑蛋白，具有 最近被认为与儿童期骨质疏松症的发展有关；然而，潜在的 病理生理学完全未知。这在一定程度上是由于PLS3在骨骼健康中的作用 仍有待确认。这项提议将解决这些问题，并为将《PLS3》发展为小说奠定基础 合成代谢抗骨质疏松症靶点。 小鼠和斑马鱼模型的遗传学研究表明PLS3在成骨细胞介导的骨中发挥作用 然而，目前尚不清楚PLS3如何在机械上促进这些过程。目标1将 利用在肌动蛋白捆绑或钙调节方面有缺陷的致病PLS3突变体来阐明 PLS3是如何促进成骨细胞分化和矿化的。此外，RNA-seq 分析还将用于确定有助于成矿的新途径，以便提供洞察力 PLS3和肌动蛋白动力学在成骨细胞矿化中的具体作用。为了更好地了解PLS3如何 有助于体内骨骼发育的调节，目标2专注于开发斑马鱼模型系统 目的：研究致病基因PLS3突变对骨形成的影响。综上所述，这项提案将填补主要 我们对PLS3及其对肌动蛋白细胞骨架动力学的调节如何有助于骨骼的理解上的差距 健康与骨质疏松症的发展。 申请者已经组建了一个由肌动蛋白细胞骨架专家组成的多学科指导团队， 支持申请者完成这些课程的骨生物学、细胞生物学和斑马鱼模型系统 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在支持不同阶段的医学科学家方面有着丰富的历史 培训对申请者的发展也将是无价的。这一培训和指导将提供 申请者的关键技能将有助于向独立研究人员和内科科学家的转变。