Targeting medium chain fatty acid metabolism for the treatment of chronic Graft-versus-Host Disease

摘要 慢性移植物抗宿主病(Cgvhd)已成为发病率和死亡率的主要原因之一。 异基因造血细胞移植(HCT)后。然而，cGVHD的治疗选择有限。 预防和治疗。类固醇是cGVHD的标准初始治疗方法，在预防cGVHD方面往往失败。 永久性器官损伤，并与重大毒性有关。脂肪酸(FA)代谢 决定T细胞(TC)在慢性移植物抗宿主病(CGVHD)中的命运和功能。然而，中链脂肪酸(MCFA)如何 参与cGVHD的病理生物学还没有研究。GPR84被称为MCFA传感 受体。CGVHD患者外周血非核细胞(PBMC)和粪便与小鼠 GPR84信号通路在cGVHD病理生物学中的意义。科学的 目前研究建议的前提是了解GPR84介导的MCFA信号如何调节T细胞 因此，GPR84靶向是控制cGVHD的有效方法。在……里面 目的1、阐明GPR84介导的MCFA代谢在慢性移植物抗宿主病(CGVHD)中的生物学作用。我们会 使用GPR84基因敲除小鼠和富含MCFA的饮食来了解MCFA信号在 闭塞性细支气管炎(BO)和胸腺受损的cGVHD。在目标2中，我们将阐明机制 负责GPR84介导的MCFA信号在cGVHD病理生物学中的影响。利用稳定性 同位素分解代谢组学、高维流式细胞术和单细胞ATAC-SEQ，我们将验证如何 GPR84信号通过调节细胞代谢影响TC和卵泡辅助TC(TFH)的存活。 迁移、分化和功能。我们将阐明CD8+T滤泡细胞(TFC)的功能，这是我们发现的 并阐明GPR84信号如何在GVHD后调节这一新的细胞亚群 移植。在目标3中，我们研究GPR84靶向是否将成为cGVHD的一种治疗选择 同时保留了GVL效应。我们将使用药理拮抗剂的组合；CRISPR-Cas9 人到小鼠cGVHD模型的诱导基因沉默和饮食规划方法来测试OUR 假设。我们还将研究GPR84靶向在预防类固醇耐药方面是否有效 CGVHD。因为GPR84抑制剂GLPG1205已被发现在二期临床中安全有效；精确 目前的研究表明，饮食和微生物调节疗法在cGVHD的治疗中非常重要 提案具有很高的翻译性，非常重要。

Abstract Chronic graft versus host disease (cGVHD) has emerged as one of the major cause of morbidity and mortality after allogeneic hematopoietic cell transplantation (HCT). However, there is limited therapeutic option for cGVHD prevention and therapy. Steroid, the standard initial therapy of cGVHD, often is unsuccessful in preventing permanent organs damage and has been associated with significant toxicities. Fatty acid (FA) metabolism decides T cell (Tc) fate and function in cGVHD development. However, how medium chain fatty acid (MCFA) participates in cGVHD pathobiology has not been studied. GPR84 has been well known as the MCFA sensing receptor. Using peripheral blood nonnuclear cells (PBMCs) and fecals from cGVHD patients along with murine models of cGVHD, we elucidate the implication of GPR84 signaling in cGVHD pathobiology. The scientific premise of current research proposal is to understand how GPR84- mediated MCFA signaling regulates T cell metabolism in cGVHD and therefore validate GPR84 targeting as an effective approach to control cGVHD. In Aim 1, we will elucidate the biological function of GPR84-mediated MCFA metabolism in cGVHD. We will use GPR84 genetically knockout mice and MCFA- enriched diet to understand the role of MCFA signaling in bronchiolitis obliterans (BO)-, and thymus-damaged cGVHD. In Aim 2, we will elucidate mechanisms responsible for the impact of GPR84- mediated MCFA signaling in cGVHD pathobiology. Utilizing stable isotope-resolved metabolomics, high dimensional flow cytometry, and single cell ATAC-seq, we will validate how GPR84 signaling by regulating cellular metabolism would influence Tc and follicular helper Tc (TFH) survival, migration, differentiation, and function. We will elucidate the function CD8+T follicular cells (TFC), which is found enriched in cGVHD patient’s PBMCs and elucidate how GPR84 signaling regulates this novel cell subset post- transplant. In Aim 3, we investigate whether GPR84 targeting would be a therapeutic option for cGVHD while preserving the GVL effect. We will use a combination of pharmacological antagonist; CRISPR-cas9 induced gene silencing, and diet programming approaches with human-to-mouse cGVHD model to test our hypothesis. We will also examine if GPR84 targeting would be effective in the prevention steroid refractory cGVHD. Because GPR84 inhibitor GLPG1205 has been found safe and effective in phase 2 clinical; precise dietaries and microbiome- modulating therapies are important in the treatment of cGVHD, current research proposal is highly translational and critically important.