Dissecting molecular mechanisms implicated in age- and osteoarthritis-related decline in anabolism in articular cartilage

剖析与年龄和骨关节炎相关的关节软骨合成代谢下降有关的分子机制

• 批准号: 10541847
• 负责人: DENIS EVSEENKO
• 金额: $ 33.83万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10541847
• 关键词: ATAC-seq; Acetylation; Acute; Address; Adult; Affect; Age; Aging; Agonist; American; Anabolism; Arthritis; Automobile Driving; Binding; Biological; Cartilage; Cartilage injury; Catabolism; Cell Differentiation process; Cell Proliferation; Cells; ChIP-seq; Chondrocytes; Chondrogenesis; Chromatin; Complex; Data; Degenerative polyarthritis; Development; Disease; Dominant-Negative Mutation; Equilibrium; Exhibits; Family; Genes; Genetic; Genetic Transcription; Goals; Growth; Heterodimerization; Heterogeneity; Hip Joint; Human; Human Development; IL6ST gene; In Vitro; Inflammation; Inflammatory; Insulin-Like Growth Factor I; Integrin alpha Chains; Integrin alpha4; Interleukin-6; Joints; Knee joint; LIFR gene; Laboratories; Life; Ligands; Link; MAP Kinase Gene; Mammals; Mediating; Mesenchymal Stem Cells; Metabolism; Modern Medicine; Modification; Molecular; Molecular Conformation; Morbidity - disease rate; Mus; Mutagenesis; Neoplasms; Outcome; Pathogenesis; Pathogenicity; Pathologic; Pathway interactions; Phenotype; Phosphorylation; Phosphorylation Inhibition; Population; Post-Translational Protein Processing; Process; Production; Proliferating; Proto-Oncogene Proteins c-akt; Regenerative capacity; Regenerative response; STAT3 gene; Signal Transduction; Skeletal Development; Source; Stat3 protein; Surface; Synovial joint; Testing; Therapeutic; Work; aged; articular cartilage; bone morphogenetic protein receptors; cartilage regeneration; cartilage repair; cell type; cytokine; cytokine receptor gp130; fetal; functional outcomes; gain of function; in vivo; innovation; loss of function; member; molecular phenotype; nanoproteomic; osteochondral tissue; osteogenic; permissiveness; preservation; progenitor; promoter; public health relevance; receptor; reparative capacity; response; restoration; senescence; single-cell RNA sequencing; small molecule; subchondral bone; transcription factor; transcriptome sequencing

ABSTRACT The reparative capacity in human articular cartilage is generally considered to be low or negligible, and this intrinsic capacity decreases with age. As a result, articular cartilage injuries often result in irreversible damage leading to osteoarthritis (OA). We and others have recently defined heterogeneity in articular chondrocytes at both the molecular and cellular levels. Work in mice and other mammals has implicated a subset of cells in the superficial layer of articular cartilage as the source of regenerative capacity; to date, these findings have not been extended to a specific population of chondrocytes in human ontogeny. Our previous studies have shown that unlike adult chondrocytes, fetal chondrocytes are highly proliferative and migratory, and exhibit high basal levels of phosphorylated signal transducer and activator of transcription 3 (pSTAT3). Our preliminary data also nominate cells expressing integrin α4 (ITGA4) and bone morphogenetic protein receptor (BMPR1B) as the most immature chondrocytes in human articular cartilage throughout human development. Moreover, we have shown at the molecular level that ITGA4+BMPR1B+ cells are enriched for active STAT3 (pSTAT3), which are known to drive proliferation, anabolism and preserve differentiation potential. Importantly, adult ITGA4+BMPR1B+ cells are localized to the superficial layer and also express the highest levels of SOX9, which is strongly identified with osteochondral progenitor identity and anabolism; indeed, ITGA4+BMPR1B+ cells are robustly chondro- and osteogenic in vitro. The percentage of ITGA4+BMPR1B+ cells and levels of pSTAT3 tightly correlate with biological age, decreasing from 20-30% in developing joints down to 1-2% in aged adult healthy cartilage. We hypothesize that active STAT3 is expressed in immature articular chondrocytes and is a permissive factor required for immature cell anabolism and differentiation in response to specific instructive signals in the niche. We propose to define the direct transcriptional targets of STAT3 in human articular chondrocytes at different ontogenic stages and under conditions similar to the pro-inflammatory state driven by IL-6 family cytokines in OA. To address how IL-6 family cytokines can drive varied biological and functional outcomes in a context- specific manner, we will employ nanoproteomics and targeted mutagenesis to determine how specific post- translational modifications in the core IL-6 family cytokine receptor gp130 differ in fetal vs. adult chondrocytes stimulated with IL-6 family cytokines. Finally, we will apply single cell RNA-Seq to further refine the molecular and cellular phenotype of immature articular chondrocytes. In parallel, we will assess the molecular and functional consequences of STAT3 gain and loss of function in articular chondrocytes. We propose that cells with higher levels of pSTAT3 will evidence broader differentiation potential in vivo, resulting from changes mediated by STAT3 in chromatin conformation. The overall impact of this highly innovative study is to define the cellular and molecular phenotype of immature articular chondrocytes throughout human ontogeny and to link this to the potential for cartilage repair and/or regeneration during aging.

抽象的 人类关节软骨的修复能力通常被认为很低或可以忽略不计，这 内在能力随着年龄的增长而下降，因此，关节软骨损伤通常会导致不可逆转的损伤。 我们和其他人最近定义了关节软骨细胞的异质性。 在小鼠和其他哺乳动物中的分子和细胞水平上的研究都涉及到细胞的一个子集。 迄今为止，这些发现还没有将关节软骨的表层作为再生能力的来源； 我们之前的研究已经扩展到人类个体发育中的特定软骨细胞群体。 与成人软骨细胞不同，胎儿软骨细胞具有高度增殖性和迁移性，并且表现出高基础能力 我们的初步数据还包括磷酸化信号转导子和转录激活子 3 (pSTAT3) 的水平。 提名表达整合素α4（ITGA4）和骨形态发生蛋白受体（BMPR1B）的细胞为最 此外，我们已经证明，在整个人类发育过程中，人类关节软骨中都存在未成熟的软骨细胞。 在分子水平上，ITGA4+BMPR1B+ 细胞富含活性 STAT3 (pSTAT3)，已知 重要的是，成年 ITGA4+BMPR1B+ 细胞是驱动增殖、合成代谢并保持分化潜力的。 定位于表层，并且还表达最高水平的 SOX9，这与 骨软骨祖细胞的身份和合成代谢；事实上，ITGA4+BMPR1B+ 细胞是强有力的软骨-和 ITGA4+BMPR1B+细胞的百分比和pSTAT3的水平与体外成骨密切相关。 生物年龄，从发育中关节的 20-30% 下降到老年健康软骨的 1-2%。 培养发现活性 STAT3 在未成熟的关节软骨细胞中表达，并且是一种许可因子 未成熟细胞合成代谢和分化所需的，以响应生态位中的特定指导信号。 我们建议定义 STAT3 在不同时间点的人关节软骨细胞中的直接转录靶标 个体发育阶段和类似于 IL-6 家族细胞因子驱动的促炎状态的条件下 OA。解决 IL-6 家族细胞因子如何在一定背景下驱动生物学和功能结果 - 具体方式，我们将采用纳米蛋白质组学和定向诱变来确定具体的后处理方式 核心 IL-6 家族细胞因子受体 gp130 的翻译修饰在胎儿与成人软骨细胞中存在差异 最后，我们将应用单细胞RNA-Seq进一步细化分子。 同时，我们将评估未成熟关节软骨细胞的分子和细胞表型。 我们提出关节软骨细胞中 STAT3 获得和功能丧失的功能后果。 pSTAT3 水平较高将证明体内变化引起的更广泛的分化潜力 这项高度创新的研究的总体影响是定义了染色质构象中的 STAT3 介导的。 整个人类个体发育过程中未成熟关节软骨细胞的细胞和分子表型并将其联系起来 这与衰老过程中软骨修复和/或再生的潜力有关。

项目成果

Author Correction: gp130/STAT3 signaling is required for homeostatic proliferation and anabolism in postnatal growth plate and articular chondrocytes.

作者更正：gp130/STAT3 信号传导对于出生后生长板和关节软骨细胞的稳态增殖和合成代谢是必需的。

• 发表时间: 2022-02-25
• 影响因子: 5.9
• 作者: Liu, Nancy Q;Lin, Yucheng;Li, Liangliang;Lu, Jinxiu;Geng, Dawei;Zhang, Jiankang;Jashashvili, Tea;Buser, Zorica;Magallanes, Jenny;Tassey, Jade;Shkhyan, Ruzanna;Sarkar, Arijita;Lopez, Noah;Lee, Siyoung;Lee, Youngjoo;Wang, Liming;Petrigliano
• 通讯作者: Petrigliano

Drug-induced modulation of gp130 signalling prevents articular cartilage degeneration and promotes repair.

药物诱导的 gp130 信号调节可防止关节软骨退化并促进修复。

• 发表时间: 2018
• 影响因子: 27.4
• 作者: Shkhyan, Ruzanna;Van Handel, Ben;Bogdanov, Jacob;Lee, Siyoung;Yu, Yifan;Scheinberg, Mila;Banks, Nicholas W;Limfat, Sean;Chernostrik, Arthur;Franciozi, Carlos Eduardo;Alam, Mohammad Parvez;John, Varghese;Wu, Ling;Ferguson, Gabriel B;Nsair, Al
• 通讯作者: Nsair, Al

Genetic ablation of adenosine receptor A3 results in articular cartilage degeneration.

腺苷受体 A3 的基因消融导致关节软骨退化。

• 发表时间: 2018
• 作者: Shkhyan, Ruzanna;Lee, Siyoung;Gullo, Francesca;Li, Lei;Peleli, Maria;Carlstrom, Mattias;Chagin, Andrei S;Banks, Nicholas W;Limfat, Sean;Liu, Nancy Q;Evseenko, Denis
• 通讯作者: Evseenko, Denis

A Single-Cell Culture System for Dissecting Microenvironmental Signaling in Development and Disease of Cartilage Tissue.

用于剖析软骨组织发育和疾病中微环境信号传导的单细胞培养系统。

• 发表时间: 2021
• 作者: Tassey, Jade;Sarkar, Arijita;Van Handel, Ben;Lu, Jinxiu;Lee, Siyoung;Evseenko, Denis
• 通讯作者: Evseenko, Denis

Cross-Communication Between Knee Osteoarthritis and Fibrosis: Molecular Pathways and Key Molecules.

膝骨关节炎和纤维化之间的交叉通讯：分子途径和关键分子。

• 发表时间: 2022
• 作者: Bolia, Ioanna K;Mertz, Kevin;Faye, Ethan;Sheppard, Justin;Telang, Sagar;Bogdanov, Jacob;Hasan, Laith K;Haratian, Aryan;Evseenko, Denis;Weber, Alexander E;Petrigliano, Frank A
• 通讯作者: Petrigliano, Frank A