The role of osteoblast progenitors in response to bone anabolic agents

成骨细胞祖细胞对骨合成代谢剂的反应的作用

• 批准号: 10404415
• 负责人: HENRY M. KRONENBERG
• 金额: $ 92.4万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10404415
• 关键词: Accounting; Address; Agonist; Anabolic Agents; Anabolism; Antibodies; Antibody Therapy; Antigens; Apoptosis; Bioinformatics; Biopsy; Bone Formation Stimulation; Bone Marrow; Bone Marrow Aspiration; Bone Resorption; Cell Separation; Cells; Center for Translational Science Activities; Clinical; Clinical Research; Color; Complement; Core Biopsy; Data; Disease; Early treatment; Equilibrium; Female; Flow Cytometry; Forteo; Fracture; Future; Goals; Heterogeneity; Hip Fractures; Human; Marrow; Mesenchymal; Modeling; Molecular; Molecular Target; Mus; Osteoblasts; Osteoclasts; Osteocytes; Osteogenesis; Osteoporosis; Osteoporotic; Parathyroid Hormone Receptor; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Population; Postmenopause; Pre-Clinical Model; Prevention; Proliferating; Public Health; Role; Sampling; Stromal Cells; Subgroup; Surface; Testing; Time; Translational Research; Woman; aged; aging population; antibody detection; aspirate; bone; bone mass; cell bank; cellular targeting; exhaustion; fracture risk; fragility fracture; gene conservation; gene network; gene regulatory network; next generation; novel; optimal treatments; osteoblast differentiation; osteogenic; osteoprogenitor cell; peripheral blood; progenitor; prospective; response; side effect; single-cell RNA sequencing; skeletal; skeletal stem cell; stem cell biology; stem cells; transcriptomics; treatment effect

Project Summary – Project 1 The goal of this project is to understand the effects of osteoporosis anabolic therapies on osteoblast progenitors. This project aligns with the overall goal of the CORT in that skeletal stem cells (including osteoblast progenitors) represent an important, yet poorly understood, target of osteoporosis anabolic therapy. Specifically, this project will focus on determining why the bone anabolic effect of romosozumab (anti-sclerostin antibody) wanes so quickly over time. To accomplish this translational research goal, complementary approaches will be pursued using patient and mouse samples. First, in Aim 1, in a prospective clinical study, we will obtain bone-relevant bio-samples (trans-iliac crest core biopsies, bone marrow aspirates, and peripheral blood) from post-menopausal women with osteoporosis during the course of romosozumab therapy. These newly-collected samples will be analyzed along with bone marrow samples previously obtained from post-menopausal women treated with teriparatide. Therefore, we will compare and contrast the effects of these two distinct osteoporosis anabolic agents. In this project, bone marrow aspirates will be analyzed by multi-color flow cytometry, and non-hematopoietic stromal cells will be studied using a cocktail of antibodies that detects surface-expressed antigens that sub-divide marrow stroma in order to define treatment effects on human marrow stromal fractions. In addition, non-hematopoietic stromal cells will be prospectively isolated for single cell RNA-sequencing. This analysis will afford unbiased perspectives on the molecular heterogeneity of marrow stroma, effects of osteoporosis anabolic treatments, and key information to explain the waning efficacy of romosozumab over time. To complement these studies using human-derived samples, in Aim 2 we will combine lineage tracing with single cell RNA-sequencing in mice to assess the effects of sclerostin antibody on osteoblast precursors over time. Incorporating lineage tracing into our approach will prove that putative progenitor populations become osteoblasts, and build confidence in cross-species, conserved gene regulatory networks that are responsible the effects of osteoporosis anabolic agents over time. Results from this project will be compared to comparable approaches in Project 2 which focus on osteocytes. The Bioinformatics Core will play a crucial role in analysis of cross-species single cell transcriptomic data. Ultimately, this merged analysis will generate powerfully-informed, novel hypotheses regarding bone stem cell biology and responses to bone anabolic agents.

项目概要 – 项目 1 该项目的目标是了解骨质疏松症合成代谢疗法对 该项目与 CORT 在该骨骼中的总体目标一致。 干细胞（包括成骨细胞祖细胞）代表了一种重要但知之甚少的细胞， 具体来说，该项目将重点确定原因。 romosozumab（抗硬化素抗体）的骨合成代谢作用随着时间的推移而迅速减弱。 为了实现这一转化研究目标，将采取互补的方法 首先，在目标 1 中，我们将在一项前瞻性临床研究中获得患者和小鼠样本。 骨相关生物样本（经髂嵴核心活检、骨髓抽吸物和 外周血）来自患有骨质疏松症的绝经后妇女 这些新收集的样本将与骨骼一起进行分析。 先前从接受特立帕肽治疗的绝经后妇女身上获得的骨髓样本。 因此，我们将比较和对比这两种不同的骨质疏松合成代谢的影响 在这个项目中，骨髓抽吸物将通过多色流式细胞术进行分析， 和非造血基质细胞将使用检测的抗体混合物进行研究 表面表达的抗原可细分骨髓基质以确定治疗效果 此外，还将对人骨髓基质细胞进行非造血基质细胞的检测。 前瞻性分离用于单细胞 RNA 测序，该分析将提供公正的结果。 骨髓基质分子异质性、骨质疏松合成代谢影响的观点 治疗方法以及解释 romosozumab 疗效随时间减弱的关键信息。 使用人类样本补充这些研究，在目标 2 中，我们将结合谱系 在小鼠中使用单细胞 RNA 测序进行追踪，以评估硬化蛋白抗体对 随着时间的推移，将成骨细胞前体纳入我们的方法将证明这一点。 假定的祖细胞群变成成骨细胞，并建立跨物种的信心， 负责骨质疏松合成代谢影响的保守基因调控网络 随着时间的推移，该项目的结果将与类似方法进行比较。 项目 2 重点关注骨细胞，生物信息学核心将在分析中发挥至关重要的作用。 最终，这种合并分析将生成跨物种单细胞转录组数据。 关于骨干细胞生物学和骨反应的强有力的新颖假设 合成代谢剂。