Choline metabolites as biomarkers in rheumatoid arthritis

 描述(由申请人提供)：本提案描述了一项为期三年的研究提案，旨在帮助首席研究员实现和扩大她的K08奖项的目标，同时它将有助于收集足够的初步数据来申请联邦NIH R01拨款。这位首席研究员目前持有由Firestein、Karin和Kavanaugh博士指导的职业发展指导奖K08 AR064834-01，该奖项题为“胆碱激酶：类风湿性关节炎的新靶点”。在她的职业发展中，重点之一是培养在风湿病领域以患者为导向的研究中进行独立调查所需的技能，更准确地说，是获得使用代谢组学研究进行生物标记物分析的经验。Her K08旨在通过磁共振光谱分析探讨胆碱代谢产物及其下游产物作为类风湿关节炎关节损害的诊断或预后生物标志物的作用。初步数据表明，将这一目标扩展到更详细的质谱学研究中，并实现对成纤维细胞样滑膜细胞(FLS)和类风湿关节炎(RA)患者血清中代谢物的无靶向研究将有助于实现这一目标。代谢组学是生物标志物研究中的一个新兴领域。基于可以利用代谢组的生理信息追踪疾病的假设，代谢物水平的变化可以用于生物标记物的研究。在类风湿关节炎(RA)的诊断或预测对不同治疗的反应方面有用的生物标志物仍然是一个未得到满足的医疗需求，特别是在对肿瘤坏死因子阻滞剂无效的患者或进行性关节损害的患者中。我们最近发现胆碱激酶是RA的一个潜在的治疗靶点。胆碱代谢在FLS功能中起着重要作用，而FLS功能有助于它们的攻击行为和关节破坏。值得注意的是，胆碱代谢是肿瘤学中潜在的预后标记物。我们的长期目标将是确定胆碱代谢物或其他代谢物是否适合作为FLS激活和关节损害的生物标志物。为了解决这个问题，作为一个特定的目标，我们将利用磁共振波谱(MRS)和超性能液相色谱质谱(UPLC-MS)更准确地分析静息FLS和RA相关细胞因子刺激后的胆碱代谢产物和下游效应分子。代谢物将在细胞和培养上清液中进行分析，以确定最合适的代谢物作为生物标志物。非靶向分析也将有助于确定其他潜在的生物标记物。我们还将确定在FLS中检测到的I代谢物在RA患者的血清或尿样中可检测到，与正常和其他炎症性疾病患者相比。选定的代谢物、血清阳性、腐蚀性疾病和DAS28所确定的疾病活动性之间的关系将被确定。如果成功，我们的研究将转化为重要的诊断和预后测试，将识别FLS激活和关节损伤的高危患者，并确定最合适的治疗方法。

 DESCRIPTION (provided by applicant): This proposal describes a three-year research proposal that aims to help the principal investigator to achieve and expand the aims of her K08 award, and at the same time it will help to gather enough preliminary data to apply for federal NIH R01 grant. The principal investigator currently holds a mentored career development award, K08 AR064834-01, entitled "Choline kinase: a novel target for rheumatoid arthritis" and mentored by Drs. Firestein, Karin and Kavanaugh. One of the points in her career development is to develop skill sets required to conduct independent investigation in patient-oriented research in the field of Rheumatology, and more precisely to get experience in biomarker analysis using metabolomics studies. Her K08 aims to explore the role of choline metabolites and its downstream products as diagnostic or prognostic biomarker of joint damage in rheumatoid arthritis by magnetic resonance spectrometry. Preliminary data suggest that expanding this aim into a more detailed study by mass spectrometry and realizing an untargeted study of the metabolites in both fibroblast-like synoviocytes (FLS) and serum from patients with rheumatoid arthritis (RA) will help to achieve this aim. Metabolomics is a new emerging field in biomarker research. Based on the assumption that diseases can be traced using physiological information from the metabolome, the changes in metabolite levels can be used for biomarker research. Biomarkers useful in diagnosis or predicting responses to different treatments in rheumatoid arthritis (RA) remain an unmet medical need, especially in patients not responsive to TNF blockers or in patients with progressive joint damage. We have recently identified choline kinase as a potential therapeutic target in RA. Choline metabolism plays an important role in FLS functions that contribute to their aggressive behavior and joint destruction. Of interest, choline metabolism is a potential prognostic marker in oncology. Our long-term objective would be to determine suitability of choline metabolites or other metabolites as a biomarker of FLS activation and joint damage. To address this question, and as a specific goal, we will analyze more precisely choline metabolites and downstream effectors in resting FLS and after stimulation with cytokines involved in RA by magnetic resonance spectrometry (MRS) and ultra-performance liquid chromatography mass spectrometry (UPLC-MS). Metabolites will be analyzed in cells and supernatants to determine the most suitable metabolites as biomarkers. Untargeted analysis will also help to identify other potential biomarkers. We will also determine i metabolites detected in FLS are detectable in serum or urine samples of RA patients compared to normal and patients with other inflammatory diseases. The relationship between chosen metabolites, seropositivity, erosive disease, and disease activity as determined by DAS28 would be determined. If successful, our studies could translate into important diagnostic and prognostic tests that will identify patients at high risk for FLS activation and joint damage, and identify the most appropriate therapy.