Targeting Pathological Angiogenesis after Joint Injury

针对关节损伤后的病理性血管生成

• 批准号: 10214725
• 负责人: ANDREA I ALFORD
• 金额: $ 37.75万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10214725
• 关键词: Acute; Address; Adenoviruses; Adult; Affect; Angiogenesis Inhibitors; Angiogenic Factor; Anti-Inflammatory Agents; Attenuated; Biological Availability; Biological Process; Biomedical Engineering; Blood Vessels; Cartilage; Catabolism; Cell Survival; Cells; Chimera organism; Chondrocytes; Chronic; Collagen; Data; Degenerative polyarthritis; Development; Diagnosis; Disease; Disease Progression; Doctor of Philosophy; Endothelial Cells; Exhibits; Extracellular Matrix; Fibroblasts; Fracture Healing; Future; Gene Transfer; Health; Histology; Human; Hyperplasia; Hypertrophy; Image; Inflammation; Inflammatory; Injury; Joints; Knock-out; Literature; Lymphoid; Matrix Metalloproteinases; Measures; Mesenchymal; Methodology; Minerals; Molecular; Mus; Pain; Pathogenesis; Pathologic; Pathologic Neovascularization; Pathway interactions; Patients; Peptide Hydrolases; Physiologic Ossification; Population; Process; Property; Proteins; Research; Research Personnel; Rheumatism; Rheumatoid Arthritis; Role; Severities; Structure; Synovial Cell; Synovial Membrane; Synovitis; THBS2 gene; Testing; Therapeutic; Time; Tissues; Up-Regulation; Vascular Endothelial Growth Factors; Vascularization; Work; angiogenesis; arthropathies; base; blood vessel development; bone; cell type; chemokine; cytokine; effective therapy; experimental study; histological image; injured; innovation; joint destruction; joint inflammation; joint injury; molecular imaging; novel; osteochondral tissue; overexpression; prevent; response; single-cell RNA sequencing; thrombospondin 2; transcriptome sequencing

ABSTRACT INVESTIGATORS: Tristan Maerz, PhD (PI) is a biomedical engineer and early-stage investigator focused on joint injury and post-traumatic osteoarthritis (PTOA). Andrea Alford, PhD (Co-PI) is a matrix biologist focused on the interactions of matricellular proteins, the extracellular matrix, and cells. Together, they have extensive complementary expertise to study the role of thrombospondin-2 in PTOA using innovative methodologies. RESEARCH CONTEXT: PTOA accounts for ~12% of osteoarthritis cases, constituting ~5.6 million sympto- matic cases in the US. Synovitis is now increasingly recognized as a perpetuator of joint degeneration, analo- gous to its role in rheumatic diseases. Alongside inflammation is increased angiogenesis, which promotes in- flammation and tissue catabolism by increasing vascular access to inflammatory cells and cytokines within the joint. However, no study has demonstrated whether modulating the degree of angiogenesis affects intraarticu- lar inflammation and downstream PTOA severity after joint injury. Thrombospondin-2 (TSP2) is an endogenous anti-angiogenic factor, and this proposal’s preliminary data demonstrates that it is markedly upregulated in the synovium following joint injury, making it a potential regulator of intraarticular angiogenesis and PTOA. SPECIFIC AIMS: 1). Determine the extent to which temporal TSP2 knockout at the time of injury increases an- giogenesis, synovitis, and PTOA severity. 2). Assess the anti-angiogenic, anti-inflammatory, and PTOA-miti- gating effect of intraarticular TSP2 gene transfer. RESEARCH PLAN: In Aim 1, single-cell RNAseq will be used to profile TSP2(+) synovial cell populations and associated angiogenic pathways. A global, inducible Rosa26-CreERT2;TSP2flox mouse will then be used to ab- late TSP2 to assess the extent to which increased intraarticular angiogenesis affects synovitis, matrix metallo- proteinase (MMP) activity (assessed using molecular imaging), pain and function, and histological and imag- ing-based PTOA severity. In Aim 2, adenoviral TSP2 will be administered intraarticularly to both WT and Rosa26-CreERT2;TSP2flox mice after joint injury, and the downstream effect on intraarticular angiogenesis, synovitis, MMP activity, pain and function, chondrocyte hypertrophy, and PTOA severity will be assessed to elucidate whether TSP2 can be employed as a disease-modifying therapeutic. INNOVATION: Demonstrating a relationship between the degree of post-injury angiogenesis and PTOA sever- ity would be a high-impact finding by identifying a potentially-modifiable acute process that affects PTOA path- ogenesis. While TSP2’s anti-angiogenic role has been studied extensively in fracture healing, little to no litera- ture has elucidated its role in the synovium and in the context of PTOA. Lastly, this proposal aims to use sev- eral innovative imaging methodologies to characterize joint catabolism and PTOA severity, providing a poten- tial translational path for earlier, more sensitive PTOA and OA diagnosis using innovative imaging.

抽象的 研究人员：Tristan Maerz 博士（PI）是一位生物医学工程师和早期研究人员，专注于 Andrea Alford 博士（Co-PI）是一位专注于关节损伤和创伤后骨关节炎 (PTOA) 的基质生物学家。 关于基质细胞蛋白、细胞外基质和细胞的相互作用，它们具有广泛的作用。 使用创新方法研究血小板反应蛋白-2 在 PTOA 中的作用的补充专业知识。 研究背景：PTOA 约占骨关节炎病例的 12%，约占 560 万症状。 在美国，滑膜炎现在被越来越多地认为是关节退化、类似的原因。 除了炎症之外，它还促进了血管生成，从而促进了风湿性疾病。 通过增加血管内炎症细胞和细胞因子的通路来抑制炎症和组织分解代谢 然而，没有研究证明调节血管生成的程度是否会影响关节内。 血小板反应蛋白-2 (TSP2) 是一种内源性炎症，与关节损伤后下游 PTOA 的严重程度有关。 抗血管生成因子，该提案的初步数据表明其在 关节损伤后的滑膜，使其成为关节内血管生成和 PTOA 的潜在调节剂。 具体目标： 1).确定损伤时 TSP2 暂时敲除增加的程度。 评估抗血管生成、抗炎和 PTOA 缓解作用。 关节内 TSP2 基因转移的门控效应。 研究计划：在目标 1 中，将使用单细胞 RNAseq 来分析 TSP2(+) 滑膜细胞群并 相关的血管生成途径。 晚期 TSP2 评估关节内血管生成增加影响滑膜炎、基质金属的程度 蛋白酶（MMP）活性（使用分子成像评估）、疼痛和功能以及组织学和影像学 在目标 2 中，将对 WT 和 WT 进行关节内注射腺病毒 TSP2。 Rosa26-CreERT2;TSP2flox小鼠关节损伤后，及其对关节内血管生成的下游影响， 将评估滑膜炎、MMP 活性、疼痛和功能、软骨细胞肥大和 PTOA 严重程度 阐明 TSP2 是否可以用作疾病缓解疗法。 创新：证明损伤后血管生成程度与 PTOA 严重程度之间的关系 通过确定影响 PTOA 路径的潜在可改变的急性过程，这将是一个具有高影响力的发现。 虽然 TSP2 在骨折愈合中的抗血管生成作用已被广泛研究，但很少有文献报道。 最后，该提案旨在使用 sev- 来阐明其在滑膜和 PTOA 中的作用。 时代创新的成像方法来表征关节分解代谢和 PTOA 严重程度，提供了潜在的 使用创新成像进行更早、更灵敏的 PTOA 和 OA 诊断的最终转化路径。