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

项目摘要/摘要 骨骼对机械加载和卸载高度敏感。骨骼卸载增加骨骼 骨髓肥大，加速骨量和质量的丧失。相反，骨骼负荷减少。 增加骨髓脂肪和增加骨骼的完整性。虽然骨骼负荷的这种有益效果很好- 已知，具体的细胞类型和导致这些效应的分子机制尚不清楚。 初步数据显示，辅助α2δ1电压敏感钙通道(VSCC)的全局缺失 亚单位导致骨量减少，损害骨骼发育，减少骨量和骨形成，以及 不断增加的肥胖症。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活性受损 (MSCs)有助于在全球α2δ1基因敲除小鼠中看到骨形成的下降。因此，拟议的 研究将检查辅助α2δ1亚单位调节骨骼的细胞特异性机制 发育和合成代谢对负荷的反应。利用转基因小鼠模型，α2δ1将被选择性地 在骨细胞和肢芽间充质中缺失。此外，使用神经性止痛药治疗 与α2δ1结合的加巴喷丁将决定1)骨细胞中α2δ1的缺失或慢性高血压治疗 损害基础或负荷诱导的骨形成，从而导致骨质量损失，以及2)如果α2δ1在 间充质祖细胞或慢性GBP治疗影响骨骼发育、骨形成和 骨髓成脂作用。此外，赖特博士还将进行互补的RNA测序分析 而体外工作则支持体内的研究结果。赖特博士是一位营养肌肉骨骼科学家， 在临床饮食干预、分析分析和动物研究方面拥有丰富的专业知识，其长期- 任期的职业目标是成为一名由NIH资助的终身教员，进行创新的、翻译的 肌肉骨骼研究。赖特博士目前正在接受分子生物学和动物方面的高级培训。 建模以补充他的临床背景，并发展完成HIS所需的技术专长 职业目标。赖特博士和他的指导委员会制定了一项全面的培训计划， 研究建议，将建立在他以前的研究经验的基础上，并进一步扩展他在基础 生物研究。赖特博士的主要培训目标包括：1)参与专业培训 发展事件；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。总而言之， 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 对于一个富有成效的职业生涯来说是必要的。F32奖项将极大地帮助赖特博士的职业发展，提供 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。

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.