Improved diagnostic and treatment methods are needed for chronic graft-versus-host disease (cGVHD), the leading cause of late nonrelapse mortality (NRM) in long-term survivors of allogenic hematopoietic cell transplantation. Validated biomarkers that facilitate disease diagnosis and classification generally are lacking in cGVHD. Here, we conducted whole serum proteomics analysis of a well-established murine multiorgan system cGVHD model. We discovered 4 upregulated proteins during cGVHD that are targetable by genetic ablation or blocking antibodies, including the RAS and JUN kinase activator, CRKL, and CXCL7, CCL8, and CCL9 chemokines. Donor T cells lacking CRK/CRKL prevented the generation of cGVHD, germinal center reactions, and macrophage infiltration seen with wild-type T cells. Whereas antibody blockade of CCL8 or CXCL7 was ineffective in treating cGVHD, CCL9 blockade reversed cGVHD clinical manifestations, histopathological changes, and immunopathological hallmarks. Mechanistically, elevated CCL9 expression was present predominantly in vascular smooth muscle cells and uniquely seen in cGVHD mice. Plasma concentrations of CCL15, the human homolog of mouse CCL9, were elevated in a previously published cohort of 211 cGVHD patients compared with controls and associated with NRM. In a cohort of 792 patients, CCL15 measured at day +100 could not predict cGVHD occurring within the next 3 months with clinically relevant sensitivity/specificity. Our findings demonstrate for the first time the utility of preclinical proteomics screening to identify potential new targets for cGVHD and specifically CCL15 as a diagnosis marker for cGVHD. These data warrant prospective biomarker validation studies.

中文翻译：

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小鼠慢性移植物抗宿主病蛋白质组分析发现 CCL15 作为患者的新型生物标志物


慢性移植物抗宿主病（cGVHD）需要改进的诊断和治疗方法，这是同种异体造血细胞移植长期幸存者晚期非复发死亡率（NRM）的主要原因。 cGVHD 通常缺乏有助于疾病诊断和分类的经过验证的生物标志物。在这里，我们对成熟的小鼠多器官系统 cGVHD 模型进行了全血清蛋白质组学分析。我们发现了 4 种在 cGVHD 期间上调的蛋白质，这些蛋白质可通过基因消融或阻断抗体来靶向，包括 RAS 和 JUN 激酶激活剂、CRKL 以及 CXCL7、CCL8 和 CCL9 趋化因子。缺乏 CRK/CRKL 的供体 T 细胞可阻止野生型 T 细胞所见的 cGVHD、生发中心反应和巨噬细胞浸润的产生。虽然 CCL8 或 CXCL7 的抗体阻断对治疗 cGVHD 无效，但 CCL9 阻断却逆转了 cGVHD 的临床表现、组织病理学变化和免疫病理学特征。从机制上讲，CCL9 表达升高主要存在于血管平滑肌细胞中，并且在 cGVHD 小鼠中独特可见。在先前发表的 211 名 cGVHD 患者队列中，与对照组相比，CCL15（小鼠 CCL9 的人类同源物）的血浆浓度升高，并且与 NRM 相关。在 792 名患者的队列中，+100 天测量的 CCL15 无法预测未来 3 个月内发生的 cGVHD，具有临床相关的敏感性/特异性。我们的研究结果首次证明了临床前蛋白质组学筛选可用于确定 cGVHD 的潜在新靶点，特别是 CCL15 作为 cGVHD 的诊断标志物。这些数据值得进行前瞻性生物标志物验证研究。