Graft versus Host Disease Biomarkers: Prediction of Onset and Response to Therapy

描述（由申请人提供）：本申请涉及广泛的挑战领域（03）生物标志物发现和验证以及特定的挑战主题03-HL-101：鉴别和验证血液、血管、心脏和呼吸道功能障碍诊断和治疗反应的临床相关、可量化生物标志物。异基因造血干细胞移植是一种治疗多种恶性肿瘤的有效方法，但急性移植物抗宿主病（GVHD）阻碍了其临床应用。不幸的是，急性GVHD的诊断是基于临床标准，由于缺乏有效的诊断血液测试，必须通过三个靶器官（皮肤、胃肠道（GI）道或肝脏）之一的活检来确认。我们最近报道了一个四种生物标志物的小组，区分患者与非GVHD，预测长期生存的GVHD的严重程度无关。该分析已经扩展到鉴定特异性预测两个主要GVHD靶点（皮肤和胃肠道）中GVHD未来发展的生物标志物。在发现阶段，我使用完整蛋白质分析系统（IPAS）来鉴定在皮肤特异性GVHD或胃肠道特异性GVHD发作前14天收集的来自10名患者的合并患者血浆中存在的新的潜在生物标志物。我确定了16种候选蛋白质，这些蛋白质在继续发展GVHD的患者的血浆中显著升高，并且可以很容易地通过ELISA测定来测量。在这个奖项期间，我建议验证这些候选GVHD生物标志物的血浆样本从大约600异基因移植患者在密歇根大学治疗自2000年以来。此外，不到一半的急性GVHD标准一线治疗患者表现出完全缓解。因此，我建议将这项研究扩展到识别和验证预测GVHD治疗反应程度的生物标志物。使用IPAS，我将鉴定在治疗开始后4周从治疗耐药患者收集的合并血浆中升高的候选生物标志物，但在对治疗有反应的患者的血浆中未升高。然后，我将验证来自治疗前和治疗后4周的约300个匹配的GVHD患者样本的血浆样本中的候选生物标志物。GVHD的病理机制不仅涉及可溶性因素，还涉及细胞因素。因此，我建议作为一种补充和替代方法，评估细胞生物标志物，包括调节性T细胞，血液树突状细胞，血液单核细胞和循环T细胞亚群的水平，以预测GVHD风险和治疗反应性。具体目标1将验证全身、皮肤和胃肠道GVHD血浆候选生物标志物，鉴定和验证细胞生物标志物，并将这些发现整合到具有未来GVHD发生的最大预测潜力的凝聚性生物标志物组中。具体目标2将鉴定和验证血浆和细胞候选物，并将这些生物标志物整合到预测对GVHD治疗的反应性的小组中。 公共卫生相关性：异基因造血干细胞移植是一种潜在的治疗恶性肿瘤的方法，但其最严重的并发症急性移植物抗宿主病（GVHD）的发展阻碍了其应用。不幸的是，没有有效的诊断急性GVHD的血液测试。识别和验证GVHD的预测，诊断和治疗反应的生物标志物的策略将允许更好地利用许多血液癌症患者的治疗。

DESCRIPTION (provided by applicant): This application addresses the broad Challenge Area (03) Biomarker Discovery and Validation and the specific Challenge Topic, 03-HL-101: Identify and validate clinically relevant, quantifiable biomarkers of diagnostic and therapeutic responses for blood, vascular, cardiac, and respiratory tract dysfunction. Allogeneic hematopoietic stem cell transplantation is a potentially curative therapy for many malignant diseases but its clinical utility has been impeded by acute graft versus host disease (GVHD). Unfortunately, the diagnosis of acute GVHD is based on clinical criteria that must be confirmed by biopsy of one of three target organs (skin, gastrointestinal (GI) tract, or liver) due to a lack of a validated diagnostic blood test. We recently reported a four biomarker panel that discriminated between patients with and without GVHD that predicted long term survival independently of GVHD severity. This analysis has been extended to identify biomarkers that specifically predict the future development of GVHD in two major GVHD targets, the skin and the GI tract. In a discovery phase, I used an intact-protein-analysis-system (IPAS) to identify new potential biomarkers present in pooled patient plasma from 10 patients collected 14 days prior to the onset of skin- specific GVHD or GI tract-specific GVHD. I identified 16 candidate proteins that were significantly elevated in the plasma of patients that went on to develop GVHD and that could be readily measured by ELISA assays. During this award period, I propose to validate these candidate GVHD biomarkers in plasma samples from approximately 600 allogeneic transplant patients treated at the University of Michigan since 2000. Furthermore, less than half of patients treated with standard frontline treatment for acute GVHD exhibit complete responses. Therefore, I propose to extend this study to the identification and validation of biomarkers that predict the degree of response to GVHD therapy. Using IPAS, I will identify candidate biomarkers elevated in pooled plasma collected 4 weeks after treatment initiation from therapy-resistant patients but not in the plasma of patients who respond to therapy. I will then validate the candidate biomarkers in plasma samples from approximately 300 matched GVHD patient samples pre-treatment and 4 weeks post- treatment. GVHD pathology not only involves soluble factors but also cellular factors. Therefore, I propose as a complementary and alternative approach, to evaluate cellular biomarkers including levels of regulatory T cells, blood dendritic cells, blood monocytes, and subsets of circulating T cells for their capacity to predict GVHD risk and responsiveness to treatment. Specific aim 1 will validate general, skin, and GI-tract GVHD plasma candidate biomarkers, identify and validate cellular biomarkers, and integrate these findings into a cohesive biomarker panel with the greatest predictive potential for future GVHD occurrence. Specific aim 2 will identify and validate plasma and cellular candidates and integrate these biomarkers into a panel that predicts responsiveness to GVHD therapy. PUBLIC HEALTH RELEVANCE: Allogeneic hematopoietic stem cell transplantation is a potentially curative therapy for many malignant diseases whose applicability has been impeded by the development of its most serious complication, acute graft versus host disease (GVHD). Unfortunately there is no validated diagnostic blood test for acute GVHD. Strategies that identify and validate biomarkers of predictive, diagnostic and therapeutic responses for GVHD will allow for better harnessing of treatment in many patients with hematological cancers.