Metabolomics of cGVHD

cGVHD 的代谢组学

• 批准号: 10698171
• 负责人: Bruce R Blazar
• 金额: $ 56.98万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10698171
• 关键词: 3-Dimensional; Acetyl-CoA Carboxylase; Affect; Alveolar; B-Lymphocytes; Benchmarking; Biological Assay; Bronchiolitis Obliterans; Candidate Disease Gene; Cell Proliferation; Cells; Chronic; Chronic stress; Clinic; Clinical; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Combination Drug Therapy; Complex; Data; Defect; Deposition; Disease; Distal; Drug Targeting; Energy-Generating Resources; Enzymes; Epithelium; FDA approved; Fatty Acids; Fibrosis; Functional disorder; Gene Deletion; Gene Targeting; Genes; Gluconeogenesis; Glucose; Glucose Transporter; Glutaminase; Glutamine; Glycolysis; Goals; Guide RNA; Helper-Inducer T-Lymphocyte; Human; Hypoxia; Immune; Immune System Diseases; Immune response; Immunoglobulin Class Switching; Immunoglobulins; In Situ; Injury; Knock-out; Knockout Mice; Label; Libraries; Ligands; Lung; Maintenance; Mesenchymal; Mesenchyme; Metabolic; Metabolic Pathway; Metabolism; Metformin; Modeling; Morbidity - disease rate; Mus; Natural regeneration; Non-Insulin-Dependent Diabetes Mellitus; Norleucine; Nutrient; Nutrient Depletion; Organoids; Outcome; Oxidation-Reduction; Oxidative Phosphorylation; Palmitates; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Predictive Value; Proliferating; Pseudostratified Epithelium; RNA library; RNA metabolism; Reactive Oxygen Species; Resistance; SLC2A1 gene; Sampling; Steroids; Structure of germinal center of lymph node; Supporting Cell; T-Lymphocyte; Testing; Therapeutic Index; Time; Tissues; Trachea; Transgenic Organisms; airway epithelium; allotransplant; body system; candidate identification; cell regeneration; cell type; chronic graft versus host disease; conditional knockout; drug candidate; drug metabolism; exhaustion; fatty acid oxidation; gene discovery; glutamine analog; improved; in vivo; in vivo evaluation; inhibitor; injury and repair; insight; lipid biosynthesis; lung injury; lung regeneration; lung repair; metabolomics; mortality; new therapeutic target; novel strategies; novel therapeutics; phase III trial; preservation; prevent; progenitor; pulmonary function; receptor; repaired; restraint; single-cell RNA sequencing; stem; stem cell proliferation; stem cell self renewal; stem cells; therapy outcome; transcriptome sequencing

Abstract Our goal is to develop new therapies for chronic GVHD (cGVHD), the leading cause of late morbidity and mortality after allotransplant. We made the important observation that T cell:B cell cooperativity and class-switched Ig tissue deposition caused multi-organ system cGVHD with bronchiolitis obliterans (BO), a non-infectious airway obstructive and epithelial remodeling disorder that portends an abysmal 5 year survival for patients. From proof-of- concept in cGVHD models, we provided key data leading to clinical trials of 6 new therapies (2 now FDA approved) for patients failing first-line steroids. CGVHD can be induced by germinal center (GC) T- and B- cell cooperativity, to produce anti-host Abs and fibrosis. While we observed GC Tfollicular helper cells (Tfh) to have increased glycolysis in the early cGVHD/BO phase, glycolysis decreased over time, consistent with exhaustion. cGVHD pathogenic Tfh and Tfollicular regulatory cells (Tfr) that restrain GCs must adapt and thrive in GCs with high reactive oxygen species and limited energy sources. Our central hypothesis is that cGVHD imposes unique metabolic demands on GC cells for cGVHD pathogenesis and distinct demands on damaged lung epithelial progenitors, impeding repair and regeneration. Choosing the best single or combined drug therapies to treat established cGVHD/BO optimally requires targeting pathogenic (Tfhs, GC B cells) and sparing Tfrs and lung stem/progenitor cells. We will test the hypothesis that rapidly proliferating Tfh that support GCs depend on multiple energy sources (glutaminolysis, glycolysis, fatty acid synthesis (FAS), while aberrant GC B cells rely on glycolysis and FAS. Relatedly, we will test the hypothesis that knowing metabolic pathways required by GC subsets (aim 1) and regenerating lung stem/ progenitor cells (aim 2) will lead to new druggable targets. Aim 1A proposes to: (1) Interrogate GC subsets for the precise pathways used for energy; (2). Test if inducing single metabolism gene deletion in a lineage-restricted GC subset in cGVHD/BO mice will improve pulmonary function and immune parameters; and (3). Test selected metabolism drug candidates in vivo to reverse ongoing cGVHD/BO. In aim 1B, we hypothesize that a focused CRISPR guide RNA metabolism library of ~40-50 gene targets from aim 1A drug results, U-[13C]-substrate (glucose, glutamine palmitate) labeling and RNA-seq data, transduced into Cas9 transgenic donor cells infused on day 0 will identify undiscovered metabolism genes critical for cGVHD/BO pulmonary dysfunction. Drugs to treat immune dysfunction may be offset by detrimental effects on lung injury repair/regeneration. In aim 2A, we will use scRNA-seq to define altered cell states and ligand-receptor interactions in distal and proximal airway epithelial and mesenchymal cells. Lung bronchiolar, alveolar and tracheosphere organoid cultures will be used to identify cells adversely affected by cGVHD/BO. In aim 2B, we hypothesize that aim 1 drugs effective in targeting GCs and aim 2 drugs in supporting lung cell regeneration will guide drug selection to inhibit cGVHD/ BO pathogenesis and repair lung injury with a high predictive value for superior cGVHD/BO outcomes. These novel approaches provide unprecedented mechanistic data to elucidate pathogenesis and lead to new therapies.

摘要我们的目标是开发治疗慢性移植物抗宿主病(CGVHD)的新疗法，慢性移植物抗宿主病是导致晚期发病率和 同种异体移植后死亡率。我们做了一个重要的观察：T细胞：B细胞的协同作用和类转换免疫球蛋白 组织沉积导致多器官系统cGVHD并闭塞性毛细支气管炎(BO)，这是一种非感染性呼吸道 梗阻和上皮重塑障碍，预示着患者糟糕的5年存活率。从证据中- 在cGVHD模型中的概念，我们提供了导致6种新疗法临床试验的关键数据(2种现已获得FDA批准) 对于服用一线类固醇失败的患者。生发中心(GC)T细胞和B细胞的协同作用可诱导cGVHD 产生抗宿主抗体和纤维化。虽然我们观察到GC毛囊辅助细胞(TFH)在体内糖酵解增加 在cGVHD/BO早期，糖酵解随着时间的推移而减少，与力竭一致。CGVHD致病TFH和 抑制GC的毛囊调节细胞(TFR)必须适应和生长在具有高活性氧物种和 有限的能源。我们的中心假设是cGVHD对GC细胞施加独特的代谢需求 CGVHD的发病机制和对受损的肺上皮祖细胞的不同要求，阻碍了修复和再生。 选择最佳的单一或联合药物疗法来治疗已确诊的cGVHD/BO需要靶向治疗 致病的(Tfhs、GC B细胞)和备用的TFR和肺干/祖细胞。我们将很快测试这一假设 支持GC的TFH的增殖依赖于多种能源(谷氨酰胺分解、糖酵解、脂肪酸合成 异常的GC B细胞依赖糖酵解和Fas。与此相关的是，我们将检验一个假设，即知道 GC亚群(目标1)和再生肺干/祖细胞(目标2)所需的代谢途径将导致 新的可下药靶点。目标1A建议：(1)询问GC子集以获得用于能量的精确路径；(2)。 测试在cGVHD/BO小鼠中诱导系谱限制性GC亚群中的单一代谢基因缺失是否会有所改善 肺功能和免疫指标；(3)在体内测试选定的代谢药物候选药物以逆转 正在进行的cGVHD/BO。在目标1B中，我们假设一个集中的CRISPR引导的RNA代谢文库~40-50 来自Aim 1A药物结果的基因靶点、U-[13C]底物(葡萄糖、谷氨酰胺棕榈酸酯)标记和RNA-SEQ 将数据转换到Cas9转基因供体细胞中，在第0天输注将识别未发现的代谢基因 对cGVHD/BO肺功能障碍至关重要。治疗免疫功能障碍的药物可能会被有害的 对肺损伤修复/再生的影响。在目标2A中，我们将使用scRNA-seq来定义改变的细胞状态和 远端和近端呼吸道上皮细胞和间充质细胞中配体-受体的相互作用。肺细支气管炎， 肺泡和气管球器官培养将用于鉴定受cGVHD/BO不利影响的细胞。在……里面 目标2B，我们假设目标1药物有效地靶向GC，目标2药物支持肺细胞 再生将指导药物的选择，以抑制cGVHD/BO的发病机制，并以较高的 对良好的cGVHD/BO结局的预测价值。这些新的方法提供了前所未有的机械性 为阐明发病机制和引导新的治疗方法提供数据。