Chronic Graft-Versus-Host Disease Biomarkers: Prediction of Resistance to Therapy

慢性移植物抗宿主病（cGVHD）研究的主要障碍是无法预测 对cGVHD治疗的反应和随后的患者存活率。这种预测性生物标志物的缺乏部分地 由于cGVHD的复杂病理学，其涉及可溶性和细胞因子，但主要是由于 到目前为止，缺乏在开始特定的cGVHD治疗时收集的样本，特别是作为新的和 现在有更多针对cGVHD的治疗方法。因此，该领域仍然存在的主要问题是： 我们用一种药物靶向多个cGVHD通路？我们能检测生物标志物来预测未来吗？ 使用已经验证的cGVHD生物标志物治疗的耐药性？我们能不能发现更具体的标记 通过高通量蛋白质组学管道吗我们能否验证并利用这些预测性生物标志物， 为FDA批准这些生物标志物提供强有力的支持？在这个项目中，我们将使用样本（血浆 Pavletic博士试验检测Pacritanib（一种多激酶抑制剂， JAK 2和IRAK 1，在类固醇难治性/类固醇依赖性（SR/D）cGVHD患者中分析蛋白质组学 与cGVHD治疗无应答预测和无复发预后相关的特征 死亡率（NRM）。建议的标志物是基于以前的研究，将包括其他新的或假设的 因素我们将检验在cGVHD开始前测量的血浆蛋白质组学的假设， 用pacritinib（PAC）治疗将对HCT患者进行分层以预测耐药性，并在1年时对NRM进行分层。我们 将确定提供最佳灵敏度和特异性的不同生物标志物和组的阈值。 我们的总体假设通过解决两个具体目标（SA）来解决剩余的差距：SA 1： 先前鉴定的血浆cGVHD生物标志物[趋化因子（C-X-C基序）配体9（CXCL 9）、CXCL 10、基质 金属蛋白酶3（MMP 3）、Dickkopf相关蛋白3（DKK 3）和刺激2（ST 2;白细胞介素（IL）-33）。 受体）]预测对PAC治疗的抵抗？我们将使用ELISA从新鲜血液中测量它们 本合同期间收集的样本：治疗前和治疗后3个月和6个月的约35个样本。SA2： 是否可以通过我们的研究发现作为cGVHD抗性的关键生物驱动因素的其他血浆生物标志物？ 蛋白质组学管道比较反应者与非反应者？我们将用我们的好- 建立了蛋白质组学工作流程，可以识别和定量2000多种蛋白质。完成后，这些 研究将产生生物标志物组，可能有助于预测对治疗的反应和耐药性， 确定新治疗方法的候选者。

Major barriers to chronic graft-versus-host disease (cGVHD) research are the inability to predict the likelihood of response to cGVHD therapy and subsequent patient survival. This absence of predictive biomarkers is partly due to the complex pathology of cGVHD, which involves both soluble and cellular factors but mostly due to the paucity, so far, of samples collected when specific cGVHD treatments are initiated, particularly as novel and more targeted treatments for cGVHD are now available. Thus, major questions remaining in the field are: Can we target more than one cGVHD pathway with a single drug? Can we detect biomarkers to predict future resistance to treatment using already validated cGVHD biomarkers? Can we discover more specific markers through a high throughput proteomics pipeline? Can we validate and utilize these predictive biomarkers to provide strong support for FDA approval of these biomarkers? For this project, we will use samples (plasma and PBMCs) collected during Dr. Pavletic trial testing Pacritanib, a multi-kinase inhibitor with specificity for JAK2 and IRAK1, in patients with steroid-refractory/steroid-dependent (SR/D) cGVHD to analyze proteomic signatures associated with prediction of cGVHD therapy nonresponse, and with prognosis of nonrelapse mortality (NRM). Proposed markers are based on previous studies and will include other novel or hypothesized factors. We will test the hypothesis that plasma proteomic panels measured prior to the start of cGVHD treatment with pacritinib (PAC) will stratify HCT patients for prediction of resistance, and NRM at 1 year. We will determine the thresholds of different biomarkers and panels that provide best sensitivity and specificity. Our overarching hypothesis addresses gaps remaining by addressing two specific aims (SA): SA1: Are five previously identified plasma cGVHD biomarkers [chemokine (C-X-C motif) ligand 9 (CXCL9), CXCL10, matrix metalloproteinase 3 (MMP3), Dickkopf-related protein 3 (DKK3), and Stimulation 2 (ST2; the interleukin (IL)-33 receptor)] predictive of resistance to PAC therapy? We will measure them using ELISA from fresh blood samples collected during this award: ~35 samples pre-treatment and 3- and 6-months post-treatment. SA2: Can additional plasma biomarkers that are key biologic drivers of cGVHD resistance be discovered through our proteomics pipeline comparing responders vs. non responders? We will address this question using our well- established proteomic workflow that can identify and quantify more than 2000 proteins. Upon completion, these studies will result in biomarker panels that may facilitate prediction of response and resistance to therapy and identify candidates for new therapeutic approaches.