CGRP-CLR mediated regulation of bone healing

CGRP-CLR 介导的骨愈合调节

基本信息

批准号：
10659882
负责人：
IVO Kalajzic
金额：
$ 41.72万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-05 至 2028-02-29
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10659882
关键词：
Affect Binding Biology Biomechanics Bone Injury Bone callus Calcitonin Calcitonin Gene-Related Peptide Calcitonin-Gene Related Peptide Receptor Cell Differentiation process Cell Lineage Cells Chemicals Chondrocytes Chondrogenesis Chronic Clinical Complex Data Denervation Endothelial Cells Endothelium Exhibits FDA approved Fracture Future Genes Genetic Transcription Genomics Growth Factor Histology Impaired healing Impairment Intervention Lead Ligands Long-Term Effects Mechanics Mediating Medical Care Costs Mesenchymal Mesenchymal Stem Cells Migraine Modeling Molecular Molecular Target Mus Neuropeptides Osteoblasts Osteogenesis Outcome Pathogenesis Peptide Signal Sequences Periosteal Cell Periosteum Pharmaceutical Preparations Phenotype Population Positioning Attribute Prevention Preventive treatment Process Proteins Quality of life RAMP1 RAMP2 RAMP3 Receptor Signaling Regulation Risk Role Sensory Sensory Receptors Signal Transduction Site Testing Therapeutic Tissues Undifferentiated Vascularization adrenomedullin afferent nerve aggrecan antagonist bone bone fracture repair bone healing bone turnover calcitonin receptor-like receptor healing improved in vivo in vivo evaluation inducible Cre inhibitor migraine treatment nerve supply neurotransmission novel pain signal promoter receptor response response to injury stem cell proliferation stem cells targeted treatment tomography translational approach

项目摘要

Periosteum, a primary site of the fracture healing response, is highly vascularized and densely innervated. Studies on bone fracture have primarily focused on the role of growth factors and vascularization in the healing process, but we postulate that sensory nerve signals are a critical part of the regulatory mechanism that initiate the stem/progenitor cell responses required for fracture callus formation. A number of lines of evidence point to sensory innervation, or signals associated with sensory nerves as promoters of bone accrual and healing. For example, fracture healing is impaired following chemical sensory denervation, but the sensory nerve-derived signals that promote healing as not yet defined. We hypothesize that damage to sensory nerves in the periosteum orchestrates the bone-healing cascade through calcitonin gene-related peptide (CGRP) – calcitonin like receptor (CLR) signaling. This is important given that multiple CGRP inhibitors were recently approved by the FDA for prevention and treatment of migraines. In Aim 1 we will evaluate the effects of CGRP inhibitors on bone healing. Given that CGRP plays a role in bone turnover and potentially healing, it is important to understand the impact of CGRP inhibition on fracture healing. In the Aim 2, we will determine which cell lineage or lineages responsive to CGRP signals during healing using targeted deletion of the CLR receptor. CLR deletion in early fracture healing will be targeted to the following lineages using inducible Cre’s: MPCs (αSMA-CreER), chondrocyte (Acan)-CreER, osteoblasts using Col2.3CreER and in endothelial (Cdh5-CreER) during fracture healing. The effects on callus formation and strength, as well as differentiation in the callus will be determined. We will examine cellular mechanisms of CGRP/CLR action utilizing in vivo approaches to study MPC expansion, differentiation and vascularization. We propose to dissect CLR signaling by distinguishing effects of the ligands. The main ligands are CGRP and Adrenomedullin that act via CLR and RAMP, of which CLR-RAMP1 is main complex for CGRP signaling while CLR-RAMP2 and RAMP3 is responsible for ADM signal activation. In Aim 3 we will evaluate effects of ADM deletion in mesenchymal population using ADMfl/fl mice. We will also define downstream signaling mechanism of CLR deletion in MSCs and endothelial cells using 10x genomics. Our study will also provide critical information on the effects that newly approved inhibitors of CGRP signaling exhibit on bone healing and what cellular mechanisms affect healing via CLR receptor Finally, testing approaches to modulate CGRP/CLR sensory singling may lead to a therapeutic strategy to enhance bone healing.

骨膜是骨折愈合反应的主要部位，血管高度丰富且致密对骨折的研究主要集中在生长因子和血管化的作用上。在愈合过程中，但我们假设感觉神经信号是调节的关键部分启动骨折愈伤组织形成所需的干/祖细胞反应的机制。有证据表明感觉神经支配，或与感觉神经相关的信号作为促进剂例如，化学感觉去神经支配后骨折愈合受到损害，但我们尚未确定促进愈合的感觉神经源信号。骨膜中的感觉神经通过降钙素基因相关协调骨愈合级联肽 (CGRP) – 降钙素样受体 (CLR) 信号传导，鉴于存在多个 CGRP，这一点很重要。抑制剂最近被 FDA 批准用于预防和治疗偏头痛。在目标 1 中，我们将评估 CGRP 抑制剂对骨愈合的影响，因为 CGRP 发挥着重要作用。由于 CGRP 抑制对骨转换和潜在愈合的影响，了解 CGRP 抑制对骨转换和潜在愈合的影响非常重要。在目标 2 中，我们将确定哪些细胞谱系对 CGRP 信号有反应。在骨折愈合过程中使用靶向删除CLR受体会导致早期骨折愈合。使用诱导型 Cre 靶向以下谱系：MPC (αSMA-CreER)、软骨细胞 (Acan)-CreER、骨折愈合过程中使用 Col2.3CreER 和内皮细胞 (Cdh5-CreER) 的成骨细胞的影响。我们将检查愈伤组织的形成和强度以及愈伤组织的分化。利用体内方法研究 CGRP/CLR 作用的细胞机制来研究 MPC 扩张、分化我们建议通过区分配体的主要作用来剖析 CLR 信号传导。配体是CGRP和肾上腺髓质素，通过CLR和RAMP发挥作用，其中CLR-RAMP1是主要复合物 CGRP 信号传导，而 CLR-RAMP2 和 RAMP3 负责 ADM 信号激活。使用 ADMfl/fl 小鼠评估 ADM 缺失对间充质群体的影响。使用 10x 基因组学研究 MSC 和内皮细胞中 CLR 缺失的下游信号传导机制。我们的研究还将提供关于新批准的 CGRP 抑制剂的影响的重要信息信号传导对骨愈合的影响以及哪些细胞机制通过 CLR 受体影响愈合最后，调节 CGRP/CLR 感觉信号传导的测试方法可能会导致一种增强治疗策略骨头愈合。