CGRP-CLR mediated regulation of bone healing

CGRP-CLR 介导的骨愈合调节

• 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
• 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

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，高度血管化且密集 支配。骨断裂的研究主要集中于生长因子和血管的作用 在康复过程中，但我们假设感觉神经信号是调节的关键部分 启动断裂愈伤组织所需的茎/祖细胞反应的机制。许多 证据线指向感觉神经或与感觉神经相关的信号作为启动子 骨头口音和康复。例如，化学感官神经治疗后，骨折愈合受损， 但是，尚未定义的感官神经衍生的信号促进了康复。我们假设损害 peroste骨中的感觉神经通过降钙素基因相关的骨裂层序列 肽（CGRP） - 降钙素喜欢受体（CLR）信号传导。考虑到多个CGRP，这很重要 最近，FDA批准了抑制剂用于预防和治疗偏头痛。 在AIM 1中，我们将评估CGRP抑制剂对骨愈合的影响。鉴于CGRP扮演 在骨转换和潜在愈合中的作用，了解CGRP抑制对 断裂愈合。在AIM 2中，我们将确定对CGRP信号响应的细胞谱系或谱系 在使用靶向缺失的CLR受体删除过程中进行愈合。早期骨折愈合中的CLR缺失将是 使用诱导CRE的针对以下谱系：MPC（αSMA-CREER），软骨细胞（ACAN）-CREER， 骨折愈合过程中使用Col2.3Creer和内皮（CDH5-CREER）中的成骨细胞。影响 将确定愈伤组织形成和力量以及愈伤组织中的分化。我们将检查 使用体内方法研究MPC扩展，分化的CGRP/CLR作用的细胞机制 和血管化。我们建议通过区分配体的效应来剖析CLR信号传导。主 配体是通过CLR和坡道起作用的CGRP和肾上腺素蛋白，其中Clr-ramp1是主要复合物 对于CGRP信号传导，而Clr-ramp2和ramp3负责ADM信号激活。在目标3中，我们将 使用ADMFL/FL小鼠评估ADM缺失对间充质种群的影响。我们还将定义 使用10倍基因组学中，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有关CGRP新批准抑制剂的影响的关键信息 信号传导在骨骼愈合上展示以及哪些细胞机制最终通过CLR受体影响愈合， 调节CGRP/CLR感觉单的测试方法可能会导致一种治疗策略以增强 骨骼愈合。