Epigenetic Profiling of circulating cell-free DNA for the Monitoring of Graft-Versus-Host Disease after Hematopoietic Cell Transplantation

异基因造血细胞移植(HCT)是一种广泛用于治疗多种恶性肿瘤和非霍奇金淋巴瘤的方法。 非恶性血液病。在美国，每年有超过8500名患者接受HCT移植。 然而，高达50%的患者在第一年因移植物抗宿主病(GVHD)而出现并发症 HCT。当供者免疫细胞攻击患者自身组织时，就会发生移植物抗宿主病。GVHD可以在任何地方发生， 皮肤、肠道和肝脏最常受到影响。GVHD仍然是更广泛传播的主要障碍之一 同种异体红细胞移植的应用。 AGVHD的早期诊断对于指导治疗决定和预防严重的器官损伤和死亡至关重要。 不幸的是，几乎没有信息丰富的诊断方法可用。在目前的临床实践中，诊断为 AGVHD几乎完全依赖于临床症状的存在，需要通过侵入性活检进行确认 手术，如皮肤活检、结肠镜检查、上消化道内窥镜检查或肝脏活检。HCT患者迫切需要一个 更好的选择。 这项提案的目标是开发和应用血液测试来检测急性移植物抗宿主病(AGVHD)，并 预测长期的HCT结果。这项测试使用了全基因组的甲基化标记图谱，包括 在血液中循环中的无细胞DNA(CfDNA)，以追踪其组织起源，并量化组织特异性 HCT后损伤。这一概念得到了重要的试点数据的支持。 我们将进行一项回顾研究，探讨cfDNA在单器官aGVHD HCT受者分层中的作用。 (目标1)，并对cfDNA用于检测GVHD早期发病和预测红细胞压积的应用进行了前瞻性研究 风险(目标2)。这些目标共同探索了高度创新的概念，即对组织的测量 血浆中cfDNA的来源能够检测aGVHD，ii)量化aGVHD的严重程度并确定其器官 参与，iii)预测HCT后死亡率，以及iv)预测aGVHD治疗的结果。 这项建议直接解决了一个迫切的、尚未得到满足的医疗需求：aGVHD的早期和非侵入性诊断 在HCT之后。因此，这些研究具有很强的翻译性。一种信息量很大的非侵入性监控工具可以 告知新的治疗方式和移植策略。认识到普遍预防措施的局限性 针对aGVHD的治疗，cfDNA检测可以发现未来用于指导预防性治疗策略，从而 减少移植后长期免疫抑制治疗的需要。此外，早期检测到的 AGVHD可以缩短治疗免疫抑制的持续时间，改善患者的预后。

Allogeneic Hematopoietic Cell Transplantation (HCT) is a widely used therapy for a variety of malignant and nonmalignant hematologic diseases. More than 8,500 patients, receive HCT transplants in the US each year. Yet, up to 50% of patients suffer complications due to graft-versus-host disease (GVHD) in the first year after HCT. GVHD occurs when donor immune cells attack the patient’s own tissues. GVHD can occur anywhere, with the skin, gut and liver most frequently affected. GVHD remains one of the major barriers to a more widespread application of allogeneic HCT. Early diagnosis of aGVHD is critical to inform treatment decisions and prevent serious organ injury and death. Unfortunately, few informative diagnostic approaches are available. In current clinical practice, diagnosis of aGVHD relies almost entirely on the presence of clinical symptoms and requires confirmation via invasive biopsy procedures, such as skin biopsy, colonoscopy, upper endoscopy or liver biopsy. HCT patients urgently need a better alternative. The goal of this proposal is to develop and apply a blood test to detect acute GVHD (aGVHD) and to predict long-term HCT outcomes. This test employs genome-wide profiling of methylation marks comprised within circulating cell-free DNA (cfDNA) in blood to trace their tissues-of-origin, and to quantify tissue-specific injury after HCT. This concept is supported by significant pilot data. We will perform a retrospective study of the utility of cfDNA to stratify HCT recipients with single organ aGVHD (Aim 1), and a prospective study of the utility of cfDNA to detect the early onset of GVHD and to predict HCT risk (Aim 2). Together these aims explore the highly innovative concept that a measurement of the tissues-of- origin of cfDNA in plasma enables to i) detect aGVHD, ii) quantify the severity of aGVHD and determine its organ involvement, iii) predict post-HCT mortality, and iv) predict the outcome of aGVHD treatments. This proposal directly addresses an urgent, unmet medical need: the early and noninvasive diagnosis of aGVHD after HCT. These studies are therefore highly translational. A highly informative, noninvasive monitoring tool may inform new treatment modalities and transplant strategies. Recognizing the limitations of universal prophylactic therapy against aGVHD, cfDNA assays could find future use in guiding pre-emptive treatment strategies, thereby reducing the need for prolonged immunosuppressive therapy after transplant. Furthermore, earlier detection of aGVHD could lead to shorter durations of therapeutic immunosuppression and improved patient outcomes.