Immunologic Mechanisms,Biomarkers and Subsets in CFS/ME

描述(申请人提供)：传统的慢性疲劳综合症/肌肉痛脑脊髓炎(CFS/ME)治疗方法未能有效地治疗与CFS/ME相关的潜在功能障碍。迈阿密CFS/ME研究小组与Broderick计算实验室(阿尔伯塔大学)合作，开发了一种基于系统生物学的方法来探索CFS/ME的潜在机制，这是R01赠款的目标，题为“CFS中的免疫机制、生物标记物和亚群”。这种“好日子不好日子”的纵向方案包括在18个月的时间里从CFS/ME患者身上提取四个血样。除了基线和18个月的临床评估和抽血外，患者在症状相对稳定的时候看一次(“好日子”)，在症状严重程度恶化或复发的时期看一次(“糟糕的一天”)。在这个项目中，我们将进一步研究潜在的疾病机制，驱动我们观察和发表的免疫信号改变模式，目标是设计一种基于多重ELISA的强大的分析方法，捕获与临床最相关的相互作用，连接免疫、内分泌和神经系统功能的标记物。我们将通过基因表达谱调查我们的细胞库中与先前测量的细胞因子、趋化因子和神经激素的表达有关的基因。我们将使用计算生物学的方法来整合这些结果，以识别基于路径的疾病特定基因集，以便使用NanoString平台进行验证。通过与蛋白质表达结果的比对，这些将被进一步提炼。我们将测试这个定制的基因表达板，将其作为区分CFS/ME与其他复杂的多症状疾病和健康个体的生物标记物。为了努力了解慢性免疫激活和疾病持久性的机制，我们将利用我们之前的发现和我们的生物库样本来进一步表征与疾病进展和复发有关的生物标记物。众所周知，CFS患者自然杀伤细胞(NK)功能低下，部分原因是细胞内裂解酶(穿孔素和颗粒酶)和细胞因子(IL-15)减少。因此，我们将基于先前NK细胞功能和信号转导缺陷的结果进行临床前研究，以评估靶向免疫治疗对IL-15的影响。我们将使用我们的生物库样本来研究体外暴露于CFS/ME患者和健康对照组的PBMC和分离的NK细胞的IL-15对NK功能和细胞内裂解蛋白(穿孔素和颗粒酶)的影响。我们相信，这种细胞因子可能有资格在CFS/ME中进行临床试验，因为它在改变潜在的疾病机制方面具有潜在的RLE。

DESCRIPTION (provided by applicant): Conventional Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME) treatments have failed to effectively treat the underlying dysfunction associated with CFS/ME. The Miami CFS/ME research group in collaboration with the Broderick computational laboratory (University of Alberta) developed an approach based on systems biology to probe the underlying mechanisms of CFS/ME, which was the goal of the R01 grant entitled "Immunologic mechanisms, biomarkers and subsets in CFS". This "good day bad day" longitudinal protocol involves drawing four blood samples from CFS/ME patients over an 18-month period. In addition to the baseline and an 18 month clinical evaluations and blood draws, patients were seen once when symptomatology was relatively stable ("good day"), and once during a period of worsening symptom severity or relapse ("bad day"). In this project extension, we will further investigate the potential illness mechanisms that drive the altered patterns of immune signaling that we observed and published, with the objective of designing a robust multiplex ELISA-based assay that captures the most clinically relevant interactions linking markers of immune, endocrine and nervous system function. We will survey our cell bank for genes involved in the expression of previously measured cytokines, chemokines, and neurohormones through gene expression profiling. We will use a computational biology approach to integrate these results to identify pathway-based illness specific gene sets for validation with the NanoString platform. These will be further refined by alignment with protein expression results. We will test this custom gene expression panel as a biomarker assay for distinguishing CFS/ME from other complex multi symptom illnesses and from healthy individuals in an effort to understand the mechanism of chronic immune activation and disease persistence, we will leverage our prior findings and our bio-bank samples to further characterize biomarkers involved in disease progression and relapse. It is well understood that natural killer (NK) cell function is poor in patients with CFS, partly due to reduced intracellular lytic enzymes (perforin and granzymes) and cytokines (IL-15). Therefore, we will conduct preclinical studies based on previous results of deficient NK cell function and signaling to assess the impact of targeting immunotherapy to IL-15. We will use our bio-bank samples to study the effects of ex vivo exposure to IL-15 of PBMC and isolated NK cells from CFS/ME cases and healthy controls in terms of changes in NK function and intracellular lytic proteins (perforin and granzyme). We believe that this cytokine will likely qualify for clinical trials in CFS/ME due to its potential rle in modifying underlying disease mechanisms.