A longitudinal immunological and virological study for ME CFS biomarker discovery (Renewal)

这是对RO1研究的更新应用：ME/CFS生物标记物的纵向免疫学研究 “发现”。我们将招募和跟踪新病例和英国ME/CFS队列的病例样本，重点是 关于详细的免疫学和临床表型及其相互关系的分析。 参与者和纵向评估：110例ME/CFS病例(一半为重度，一半为轻度-中度)满足所有 疾控中心-19942、加拿大1和IOM4标准将每6至12个月评估一次，为期5个时间点，使 将生物标记物表达的变化映射到临床参数(反之亦然)；以精确分层 不同的病例类型；以及准确分析疾病进展不同阶段的生物标志物及其 与临床参数的时间关系。 活动和目标：在一项前瞻性队列研究中，我们将：i)招募和跟踪总共110名ME/CFS 病例，其中至少100例将拥有完整的数据和样本集；二)根据 症状严重程度的趋势，a)“改善”，b)“稳定”或c)“恶化”，使用与疼痛、疲劳和 功能状态39。对于深入的免疫学分析，我们将：i)进行详细的体外表型鉴定 流式细胞仪检测PBMC亚群；II)体外培养后测定NK细胞和T细胞的功能反应 流式细胞术刺激；III)分析刺激的PBMC培养上清液中分泌的细胞因子 多重珠阵列；以及，iv)MHC Class1和KIR的基因型供体，用于流式细胞术分析 数据。集成分析并将免疫学数据映射到临床数据，并确定 免疫参数先于、跟踪或预测临床轨迹，我们将：i)量化 免疫生物标记物和ii)确定与ME/CFS临床状态变化相关的生物标记物。我们 将在所有阶段考虑患者的观点、伦理、法律和社会问题。 为了确保质量和最大限度的研究产出，我们将继续使用英国ME/CFS队列程序和 对患者进行纵向随访时的方案。我们将遵循标准化的临床、数据和样本 对符合特定临床标准的ME/CFS患者进行随访的处理和实验室程序。 创新和战略需求：据我们所知，这是对ME/CFS进行的第一项长期前瞻性研究 纳入门诊和重症病例，并纳入全面的临床数据和深入的 免疫学分析。这项研究因其严格的纳入标准和详细的临床表型而独一无二， 为我们的分析增添了特定性和有效性。从长远来看，识别强大的生物标记物将使 ME/CFS表型与疾病严重程度和预后的相关性，并可能揭示新的治疗方案 干预措施。因为有100-250万美国人患有ME/CFS4，这项研究有可能改善 美国大量患者的生活，以及推动美国和 在国际上。患者和利益相关者都参与了研究的方方面面。

This is a renewal application for the RO1 study ‘A longitudinal immunological study for ME/CFS biomarker discovery’. We will recruit and follow-up new cases and a sample of cases of the UK ME/CFS cohort, focusing on detailed immunological and clinical phenotypes and analyses of their inter-relationships. Participants and longitudinal assessments: 110 ME/CFS cases (½ severe, ½ mild-moderate) meeting all of the CDC-19942, Canadian1 and IOM4 criteria will be assessed every 6 to 12 months for 5 time-points, enabling the mapping of changes in biomarker expression onto clinical parameters (and vice versa); to precisely stratify different case types; and to accurately analyse biomarkers at different stages of disease progression and their time-relationship with clinical parameters. Activities and objectives: In a prospective cohort study, we will: i) recruit and follow up a total of 110 ME/CFS cases, of which at least 100 cases will have complete data and sample sets; ii) stratify patients according to trends in symptom severity, a) ‘improving’, b) ‘stable’ or c) ‘worsening’, using scores related to pain, fatigue and functional status39. For in-depth immunological profiling, we will: i) conduct detailed ex vivo phenotyping of PBMC populations by flow cytometry; ii) measure the functional response of NK and T cells after in vitro stimulation by flow cytometry; iii) analyze secreted cytokines in supernatants of stimulated PBMC cultures by multiplex bead array; and, iv) genotype donors for MHC Class1 and KIR to inform analysis of flow cytometry data. To integrate analyses and map immunological data onto clinical data and determine whether changes in immune parameters precede, follow, or predict the clinical trajectory, we will: i) quantify correlations between immunological biomarkers and ii) identify biomarkers associated with changes in ME/CFS clinical status. We will consider patients’ perspectives, ethical, legal, societal issues at all stages. To ensure quality and maximum research output we will continue using the UK ME/CFS cohort procedures and protocols during the longitudinal follow up of patients. We will follow standardised clinical, data and samples handling and laboratory procedures in the follow up of ME/CFS patients meeting specific clinical criteria. Innovation and Strategic Need: This is, to our knowledge, the first long-term prospective study of ME/CFS to incorporate both ambulatory and severe cases and to include comprehensive clinical data and in-depth immunological profiling. The study is unique for its strict inclusion criteria and detailed clinical phenotyping, adding specificity and validity to our analyses. In the long term, identification of robust biomarkers will allow the correlation of ME/CFS phenotype with disease severity and prognosis and may reveal new options for interventions. Because 1- 2.5 million Americans have ME/CFS4, this study has the potential to improve the lives of a large patient population in the U.S. as well as advance the state of the field in the U.S. and internationally. Patients and stakeholders are involved in all aspects of the research.