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.

项目总结/摘要 本建议的目标是：1）获得必要的实验技能和职业培训，以发展一个 独立的研究计划，调查骨骼发育和疾病的机制，以及2）表征 一种新的骨重建靶点，称为G蛋白门控内向整流K+通道3（Girk 3）， 钾的流动和生理过程。我们的数据表明，Girk 3-/-小鼠的骨量高， 在骨骼成熟后，具有低的白细胞介素-1 β（IL-1β）和其他循环细胞因子。整体 本研究的目的是检验Girk 3缺失增强成人骨骼骨密度的假设 通过改变IL-1β和其他单核细胞衍生的细胞因子的分泌来改变骨吸收。期间 指导K99期，具体目的是确定单核细胞中Girk 3的缺失如何影响骨量 (Aim 1）和确定Girk 3如何调节血液和骨髓单核细胞的细胞因子产生（Aim 2）。用于评价Girk 3在骨重建中的作用的方法将包括动态和静态方法。 组织形态计量学、成骨细胞和破骨细胞分化测定、飞行时间流式细胞术（CyTOF）、单 细胞RNA测序（scRNA-seq）和生物信息学。完成这一目标将有助于我过渡到 独立R 00阶段，具体目标（目标3）是确定Girk 3缺失是否可以防止骨 通过抑制IL-1β在骨质疏松症鼠模型中的损失。K99阶段将在马约诊所进行 并将专注于通过与策划的专业人士会面， 指导小组，定期出席研究研讨会和国家研究 会议，参加关于研究中的责任和负责任行为的讲习班，并寻求 此外，还将提供建立网络的机会，并开展目标1和2的实验，并公布结果。 R 00阶段将在我的独立实验室进行，重点是完成和出版 目标3，并根据结果开发R 01应用程序。拟议的计划协同新技能， 破骨细胞和离子通道生物学，无偏电泳，单细胞RNA测序和生物信息学， 骨组织形态计量学与内分泌学和成骨细胞生物学的专业知识，创造自己独特的 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 在肌肉骨骼生物学的独立研究生涯的坚实基础。