Transcriptome-based biomarkers in Human Osteoarthritis

人类骨关节炎中基于转录组的生物标志物

• 批准号: 6742957
• 负责人: Steven B Abramson
• 金额: $ 14.75万
• 依托单位: HOSPITAL FOR JOINT DISEASES ORTHO INST
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6742957
• 关键词: biomarker; biomechanics; cartilage disorder; clinical research; cooperative study; disease /disorder onset; gene expression; genetic susceptibility; human subject; interleukin 1; leukocytes; longitudinal human study; messenger RNA; microarray technology; osteoarthritis; pathologic process; patient oriented research; polymerase chain reaction; prostaglandin endoperoxide synthase; synovial membrane

DESCRIPTION(Provided by Applicant): Osteoarthritis (OA) is a disease characterized by altered biomechanics, genetic susceptibility and hormonal influences. The disease process affects the entire joint structure, including cartilage, synovial membrane, subchondral bone, which are active sites of growth factor, cytokine, protease and inflammatory mediator production. Reliable biomarkers remain elusive in this disease. Recent studies in our laboratory revealed the unexpected and novel observation that peripheral blood leukocytes (PBMCs) in OA are activated. Using gene expression array, OA-PBMCs were shown to differentially express clusters of transcripts that were distinct from normal controls. Among the upregulated transcripts was COX-2; cytokine-induced production of PGE2 was 5-fold higher in OA-PBMCs than in normal. A cluster of 54 transcripts sensitive to pharmacological intervention (COX-2 specific inhibitors) could also be identified in OA-PBMCs. Independently, human OA affected cartilage showed a specific (proliferation/neoplastic) gene expression signature in OA, which differed from normal and RA cartilage. These findings suggest that there will be "transcriptome-based biomarkers" in OA patients, in PBMCs and/or cartilage. We will utilize the HJD Arthritis Database and Tissue Bank of 500 OA patients to expand these studies. We will examine PBMCs to establish disease and stage specific gene expression signatures in early and late human OA. A cohort of 50 normals, 50 patients with early OA (OA-E) and 50 patients with late OA (OA-L) will be analyzed. RNA from these PBMCs will be stored. Using gene expression array technology we will identify common transcripts that are differentially expressed in early and/or late OA versus normal. These "transcriptome biomarkers" will be confirmed by Real time PCR on three occasions over a six-month study period. In a second specific aim we will compare gene expression profile changes in human OA PBMCs and OA-affected cartilage and synovium. We hypothesis that bioinformatic analysis will: (a) show OA-PBMC "transcriptome-based" biomarkers that distinguish OA vs. normal, as well as markers that identify early vs. late disease. (b) identify end stage OA specific "cartilage dysfunctional transcripts", involved in joint destruction, which may also be differentially expressed in early and/or late OA PBMCs. Markers of OA-cartilage that are expressed in early disease will be of particular interest as candidates that predict disease progression. In summary, this proposal will utilize genomic technology to identify transcriptome-based biomarkers in OA-PBMC, which we propose act as circulating "sensors" of complex metabolic and inflammatory processes within the diseased tissues of the OA joint. These studies will open the OA field to novel prognostic and pharmacogenomic future strategies.

描述（由申请人提供）： 骨关节炎 (OA) 是一种以生物力学、遗传易感性和激素影响改变为特征的疾病。疾病过程影响整个关节结构，包括软骨、滑膜、软骨下骨，这些都是生长因子、细胞因子、蛋白酶和炎症介质产生的活性位点。这种疾病的可靠生物标志物仍然难以捉摸。我们实验室最近的研究揭示了一个意想不到的新观察结果：OA 中的外周血白细胞 (PBMC) 被激活。使用基因表达阵列，OA-PBMC 被证明能够差异表达与正常对照不同的转录物簇。上调的转录本包括 COX-2； OA-PBMC 中细胞因子诱导的 PGE2 产量比正常情况高 5 倍。在 OA-PBMC 中还可以鉴定出对药物干预（COX-2 特异性抑制剂）敏感的 54 个转录物簇。独立地，受 OA 影响的人类软骨在 OA 中显示出特定的（增殖/肿瘤）基因表达特征，这与正常和 RA 软骨不同。这些发现表明，OA 患者、PBMC 和/或软骨中将存在“基于转录组的生物标志物”。我们将利用包含 500 名 OA 患者的 HJD 关节炎数据库和组织库来扩展这些研究。我们将检查 PBMC，以确定早期和晚期人类 OA 的疾病和阶段特异性基因表达特征。将分析 50 名正常人、50 名早期 OA 患者 (OA-E) 和 50 名晚期 OA 患者 (OA-L) 的队列。来自这些 PBMC 的 RNA 将被存储。使用基因表达阵列技术，我们将识别早期和/或晚期 OA 与正常情况下差异表达的常见转录本。这些“转录组生物标志物”将在六个月的研究期间通过实时 PCR 进行三次确认。在第二个具体目标中，我们将比较人类 OA PBMC 和受 OA 影响的软骨和滑膜的基因表达谱变化。我们假设生物信息学分析将： (a) 显示区分 OA 与正常的 OA-PBMC“基于转录组”的生物标志物，以及识别早期与晚期疾病的标志物。 (b) 鉴定终末期 OA 特异性“软骨功能障碍转录物”，参与关节破坏，其也可能在早期和/或晚期 OA PBMC 中差异表达。在早期疾病中表达的 OA 软骨标志物作为预测疾病进展的候选者将受到特别关注。总之，该提案将利用基因组技术来识别 OA-PBMC 中基于转录组的生物标志物，我们建议将其作为 OA 关节患病组织内复杂代谢和炎症过程的循环“传感器”。这些研究将为 OA 领域开辟新的预后和药物基因组学未来策略。