Girk3 in bone biology and disease

Girk3 在骨生物学和疾病中的作用

• 批准号: 10663602
• 负责人: Samantha R Weaver
• 金额: $ 10.49万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10663602
• 关键词: Adult; Affect; Age; Aging; Applied Skills; Award; Bioinformatics; Biological Assay; Biological Markers; Biology; Blood; Bone Density; Bone Development; Bone Diseases; Bone Marrow; Bone Marrow Cells; Bone Resorption; Bone remodeling; Calvaria; Cell Lineage; Cells; Cellularity; Clinic; Continuous Infusion; Cytometry; Data; Development; Development Plans; Disease; Educational workshop; Endocrinology; Enzyme-Linked Immunosorbent Assay; Female; Foundations; Fracture; GTP-Binding Proteins; Goals; Harvest; Hematopoietic; IL3 Gene; In Vitro; Inflammation; Inflammatory; Interleukin-1; Interleukin-1 beta; Ion Channel; Ion Channel Gating; Ions; Knowledge; Laboratories; Lead; Learning; Macrophage; Measures; Mechanics; Membrane; Mentors; Mentorship; Messenger RNA; Methods; Modeling; Molecular; Mus; Musculoskeletal; Osteoblasts; Osteoclasts; Osteocytes; Osteogenesis; Osteoporosis; Ovariectomy; Phase; Physiological; Physiological Processes; Population; Positioning Attribute; Postdoctoral Fellow; Postmenopausal Osteoporosis; Potassium; Potassium Channel; Process; Production; Publications; Publishing; Research; Research Personnel; Role; Safety; Serum; Signal Pathway; Signal Transduction; Skeleton; Testing; Time; Training; Wild Type Mouse; Woman; Work; aged; bone; bone cell; bone fragility; bone loss; bone mass; career; career development; cytokine; experimental study; improved; innovation; inward rectifier potassium channel; male; meetings; men; microCT; monocyte; mouse model; novel; osteoporosis with pathological fracture; pre-doctoral; prevent; programs; research and development; single-cell RNA sequencing; skeletal; skills; success; symposium; synergism; tool; transcriptome; transcriptomics

PROJECT SUMMARY / ABSTRACT The goals of this proposal are to: 1) obtain experimental skills and career training necessary to develop an independent research program investigating mechanisms of bone development and disease, and 2) characterize a novel target of bone remodeling called G protein-gated inwardly-rectifying K+ channel 3 (Girk3), a key regulator of potassium flux and physiological processes. Our data demonstrate that Girk3-/- mice develop high bone mass after skeletal maturity and have low interleukin-1 beta (IL-1β) and other circulating cytokines. The overall objective of this proposal is to test the hypothesis that Girk3 deletion enhances bone density in adult skeletons by altering the secretion of IL-1β and other monocyte-derived cytokines that modify bone resorption. During the mentored K99 phase, the specific aims are to identify how deletion of Girk3 in monocytes affects bone mass (Aim 1) and to determine how Girk3 regulates cytokine production by blood and bone marrow monocytes (Aim 2). Methods used to evaluate the role of Girk3 in bone remodeling will include dynamic and static histomorphometry, osteoblast and osteoclast differentiation assays, cytometry by time-of-flight (CyTOF), single cell RNA-sequencing (scRNA-seq), and bioinformatics. Completion of this aim will facilitate my transition into the independent R00 phase when the specific aim (Aim 3) will be to determine if Girk3 deletion can prevent bone loss in a murine model of osteoporosis via inhibition of IL-1β. The K99 phase will be conducted at Mayo Clinic and will focus on obtaining mentored training in professional development through meetings with a curated mentorship team, regular attendance and presentations at research seminars and national research conferences, participating in workshops on grantsmanship and responsible conduct in research, and seeking out networking opportunities, as well as conducting the experiments in Aims 1 and 2 and publishing the results. The R00 phase will be conducted in my independent laboratory and will focus on completion and publication of Aim 3 and developing an R01 application based on the results. The proposed plan synergizes new skills in osteoclast and ion channel biology, unbiased spectometry, single cell RNA sequencing and bioinformatics, and bone histomorphometry with prior expertise in endocrinology and osteoblast biology to create my own unique research trajectory. The career development plan and research strategy outlined in this application will produce a robust foundation for an independent research career in musculoskeletal biology.

项目摘要 /摘要 该建议的目标是：1）获得开发必要的实验技能和职业培训 独立研究计划调查骨骼发育和疾病的机制，以及2）表征 一个新型的骨重塑靶标，称为G蛋白门控的内部矫正K+通道3（GIRK3），一种钥匙调节剂 钾通量和物理过程。我们的数据表明，girk3 - / - 小鼠会发展高骨头 骨骼成熟后，白细胞介素-1β（IL-1β）和其他循环细胞因子具有低的白介素-1β（IL-1β）。总体 该提议的目的是检验以下假设：girk3缺失增强了成人骨骼的骨密度 通过改变IL-1β和其他单核细胞衍生的细胞因子的分泌，这些细胞因子改变了骨骼的分辨率。在 指导K99阶段，具体目的是确定单核细胞中GIRK3的缺失如何影响骨骼 （AIM 1）并确定GIRK3如何调节血液和骨髓单核细胞的细胞因子产生（AIM 2）。用于评估GIRK3在骨骼重塑中的作用的方法将包括动态和静态 组织形态计量学，成骨细胞和破骨细胞分化测定法，飞行时间（Cytof），单个细胞仪，单个 细胞RNA-sequencing（SCRNA-SEQ）和生物信息学。这个目标的完成将有助于我过渡到 当特定目标（AIM 3）是确定GIRK3缺失是否可以防止骨头 通过抑制IL-1β，骨质疏松症的鼠模型损失。 K99阶段将在Mayo诊所进行 并将专注于通过与精心策划的会议获得专业发展的培训 识别团队，定期出勤和研究下水道和国家研究的演讲 会议，参加研究授予技巧和负责任的行为的研讨会，并寻求 消除网络机会，以及在目标1和2中进行实验并发布结果。 R00阶段将在我的独立实验室中进行，并将集中于完成和发布 AIM 3并根据结果开发R01应用程序。拟议的计划使新技能协同 破骨细胞和离子通道生物学，无偏见，单细胞RNA测序和生物信息学以及 具有内分泌学和成骨细胞生物学的先前专业知识的骨骼组织形态计量学可以创建我自己的独特 研究轨迹。本申请中概述的职业发展计划和研究策略将产生 为肌肉骨骼生物学独立研究职业的强大基础。