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.

摘要 Tristan Maerz博士（PI）是一名生物医学工程师和早期研究者，专注于 关节损伤和创伤后骨关节炎（PTOA）。安德烈阿尔福德，博士（合作PI）是一个矩阵生物学家专注于 关于细胞外基质蛋白质、细胞外基质和细胞之间的相互作用。他们一起拥有广泛的 补充专业知识，研究血小板反应蛋白-2在PTOA中的作用，使用创新的方法。 研究背景：PTOA约占骨关节炎病例的12%，构成约560万症状。 在美国的案例。滑膜炎现在越来越多地被认为是关节退行性变的永久因素， 它在风湿性疾病中的作用。伴随炎症的是血管生成增加，这促进了- 通过增加血管内炎症细胞和细胞因子的进入， 关节然而，没有研究表明调节血管生成的程度是否会影响关节内的血管生成。 关节损伤后最大炎症和下游PTOA严重程度。血小板反应蛋白-2（TSP 2）是一种内源性的 抗血管生成因子，该提案的初步数据表明，它是显着上调， 关节损伤后滑膜中，使其成为关节内血管生成和PTOA的潜在调节剂。 具体目标：1）。确定在损伤时暂时性TSP 2敲除增加的程度， 血管生成、滑膜炎和PTOA严重程度。2）。评估抗血管生成、抗炎和PTOA-miti- 关节内TSP 2基因转移的门控效应。 研究：在目标1中，单细胞RNAseq将用于分析TSP 2（+）滑膜细胞群， 相关的血管生成途径。然后，将使用全局的诱导型Rosa 26-CreERT 2; TSP 2flox小鼠来ab. 晚期TSP 2评估关节内血管生成增加影响滑膜炎、基质金属蛋白酶、 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。 基于分组的PTOA严重性。在目标2中，腺病毒TSP 2将关节内施用至WT和WT。 Rosa 26-CreERT 2; TSP 2flox小鼠关节损伤后，以及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TSP 2是否可以用作疾病修饰治疗剂。 创新：证明损伤后血管生成程度与PTOA严重程度之间的关系。 通过识别影响PTOA路径的潜在可修改急性过程，这将是一个高影响力的发现- 起源虽然TSP 2的抗血管生成作用在骨折愈合中已被广泛研究，但几乎没有文献报道。 已经阐明了其在滑膜和PTOA中的作用。最后，这项建议旨在利用七- 各种创新的成像方法，以表征关节catalysts和PTOA的严重程度，提供了一个潜在的， 使用创新的成像技术，为更早、更敏感的PTOA和OA诊断提供了一条新的平移路径。