Regulation of Skeletal Development and Mechanosensitivity by the α2δ1 Auxiliary Voltage Sensitive Calcium Channel Subunit

α2β1 辅助电压敏感钙通道亚基对骨骼发育和机械敏感性的调节

• 批准号: 10188432
• 负责人: Christian Stephen Wright
• 金额: $ 1.71万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10188432
• 关键词: Adipocytes; Aging; Animal Experimentation; Animal Model; Antiepileptic Agents; Award; Binding; Bone Marrow; Bone Resorption; Bone remodeling; Calcium; Calcium Channel; Cell Lineage; Cell Surface Receptors; Cell membrane; Cells; Cellular biology; Chronic; Clinical; Complement; Complementary RNA; Data; Deterioration; Development; Dietary Fats; Dietary Intervention; Disease; Enterobacteria phage P1 Cre recombinase; Equilibrium; Event; Faculty; Fatty acid glycerol esters; Financial Support; Fostering; Funding; Genes; Goals; High Fat Diet; Hyperactivity; Impairment; In Vitro; Knock-out; Knockout Mice; Knowledge; Learning; Limb Bud; Measurable; Mechanics; Mediating; Mentors; Mesenchymal Stem Cells; Mesenchyme; Molecular; Molecular Biology; Molecular and Cellular Biology; Morphology; Mus; Musculoskeletal; Neurons; Nutritional; Obesity; Osteoblasts; Osteoclasts; Osteocytes; Osteogenesis; Osteopenia; Pharmaceutical Preparations; Pharmacology; Phenotype; Play; Prevalence; RNA; Regulation; Research; Research Proposals; Research Training; Role; Saline; Scientist; Skeletal Development; Skeletal bone; Skeleton; Stimulus; Technical Expertise; Techniques; Time; Training; Transgenic Mice; United States National Institutes of Health; Wild Type Mouse; Work; Writing; age related; biological research; bone; bone cell; bone loss; bone marrow mesenchymal stem cell; bone mass; bone quality; career; career development; cell type; design; disability; effective intervention; experience; fracture risk; gabapentin; in vivo; innovation; knock-down; lifestyle intervention; lipid biosynthesis; mechanical force; mechanical load; member; mouse model; novel; osteogenic; osteoprogenitor cell; painful neuropathy; receptor; response; skeletal; skeletal unloading; skills; stellate cell; stem cell fate; stem cells; trafficking; transcriptome; transcriptome sequencing; voltage

PROJECT SUMMARY/ABSTRACT The skeleton is highly sensitive to mechanical loading and unloading. Skeletal unloading increases bone marrow adiposity and accelerates the loss of bone quantity and quality. Conversely, skeletal loading decreases bone marrow adiposity and increases skeletal integrity. While this beneficial effect of skeletal loading is well- known, the specific cell types and the molecular mechanisms contributing to these effects are unclear. Preliminary data show that global deletion of the auxiliary α2δ1 voltage-sensitive calcium channel (VSCC) subunit results in osteopenia, impairing skeletal development, decreasing bone mass and bone formation, and increasing adiposity. As several cell types regulate bone formation and responses to skeletal loading, we hypothesize that the impaired activity of either osteocytes and/or bone marrow mesenchymal stem cells (MSCs) contribute to the decline in bone formation seen in global α2δ1 knockout mice. Thus, the proposed studies will examine the cell-specific mechanisms by which the auxiliary α2δ1 subunit regulates skeletal development and anabolic responses to loading. Using transgenic mouse models, α2δ1 will be selectively deleted in osteocytes and the limb-bud mesenchyme. Additionally, treatment with the neuropathic pain drug gabapentin, which binds α2δ1 will determine SA1) If deletion of α2δ1 in osteocytes or chronic GBP treatment impairs basal or load-induced bone formation with resultant loss in bone quality, and SA2) If deletion of α2δ1 in mesenchymal progenitors or chronic GBP treatment influences skeletal development, bone formation, and bone marrow adipogenesis. Additionally, Dr. Wright will conduct complementary RNA sequencing analyses and in vitro work to support in vivo results. Dr. Wright is a nutritional musculoskeletal scientist with considerable expertise in clinical dietary interventions, analytical analyses, and animal research whose long- term career goal is to become a NIH-funded, tenured faculty member who conducts innovative, translational musculoskeletal research. Dr. Wright is currently obtaining advanced training in molecular biology and animal modeling to complement his clinical background and develop the technical expertise needed to accomplish his career goals. Dr. Wright and his mentoring committee have developed a comprehensive training plan and research proposal that will build upon his previous research experience, and further expand his skills in basic biological research. Dr. Wright's primary training objectives include 1) Participating in professional development events; 2) Taking advanced didactic training courses; 3) Acquiring translational biomolecular research experience; 4) Learning new experimental techniques; and 5) Enhancing writing abilities. Collectively, these activities will expand Dr. Wright's knowledge and research abilities; providing him the crucial expertise necessary for a productive career. The F32 award will greatly aid in Dr. Wright's career development, providing the financial assistance and the protected time necessary to achieve these research and training objectives.

项目总结/摘要 骨骼对机械加载和卸载高度敏感。骨骼卸载增加骨骼 骨髓脂肪过多，加速骨数量和质量的损失。相反，骨骼负荷降低 骨髓肥胖和增加骨骼完整性。虽然骨骼负荷的这种有益效果是好的- 虽然已知这些效应，但具体的细胞类型和促成这些效应的分子机制尚不清楚。 初步数据显示，辅助性α2δ1电压敏感性钙通道（VSCC） 亚基导致骨质减少，损害骨骼发育，降低骨量和骨形成， 增加肥胖。由于几种细胞类型调节骨形成和对骨骼负荷的反应，我们 假设骨细胞和/或骨髓间充质干细胞的活性受损 骨髓间充质干细胞（MSC）导致在整体α2δ1敲除小鼠中观察到的骨形成下降。因此，拟议的 研究将检查辅助α2δ1亚单位调节骨骼肌的细胞特异性机制， 对负荷的发育和合成代谢反应。使用转基因小鼠模型，α2δ1将被选择性地 骨细胞和肢芽间充质缺失。此外，神经性疼痛药物治疗 结合α2δ1的加巴喷丁将决定SA 1）如果骨细胞中α2δ1缺失或慢性GBP治疗 损害基础或负荷诱导的骨形成，导致骨质量损失，SA 2）如果α2δ1缺失， 间充质祖细胞或慢性GBP治疗影响骨骼发育，骨形成， 骨髓脂肪生成此外，Wright博士将进行互补RNA测序分析， 并在体外工作以支持体内结果。Wright博士是一位营养肌肉骨骼科学家， 在临床饮食干预、分析和动物研究方面具有相当的专业知识， 长期职业目标是成为NIH资助的终身教职员工，进行创新，翻译 肌肉骨骼研究Wright博士目前正在接受分子生物学和动物医学方面的高级培训。 建模，以补充他的临床背景，并发展所需的技术专长，以完成他的 职业目标。赖特博士和他的指导委员会制定了一个全面的培训计划， 研究建议，将建立在他以前的研究经验，并进一步扩大他的基本技能， 生物学研究Wright博士的主要培训目标包括：1）参与专业培训 发展活动; 2）参加高级教学培训课程; 3）获得翻译生物分子 研究经验; 4）学习新的实验技术; 5）提高写作能力。总的来说， 这些活动将扩大赖特博士的知识和研究能力，为他提供关键的专业知识， 这是一个富有成效的职业生涯所必需的。F32奖将大大有助于赖特博士的职业发展， 实现这些研究和培训目标所需的财政援助和受保护的时间。