Metabolomics of cGVHD

cGVHD 的代谢组学

• 批准号: 10493800
• 负责人: Bruce R Blazar
• 金额: $ 59.42万
• 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/10493800
• 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; Epithelial; 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; Lead; Leucine; Libraries; Ligands; Lung; Maintenance; Mesenchymal; Mesenchyme; Metabolic; Metabolic Pathway; Metabolism; Metformin; Modeling; Morbidity - disease rate; Mus; Natural regeneration; Non-Insulin-Dependent Diabetes Mellitus; Nutrient; Nutrient Depletion; Organoids; Outcome; Oxidation-Reduction; Oxidative Phosphorylation; Palmitates; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Predictive Value; Proliferating; RNA library; RNA metabolism; Reactive Oxygen Species; Regulatory T-Lymphocyte; Resistance; SLC2A1 gene; Sampling; Steroids; Structure of germinal center of lymph node; Supporting Cell; System; T-Lymphocyte; Testing; Therapeutic Index; Time; Tissues; Trachea; Transgenic Organisms; airway epithelium; allotransplant; cell regeneration; cell type; chronic graft versus host disease; conditional knockout; drug candidate; drug metabolism; exhaustion; fatty acid oxidation; gene discovery; 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; single-cell RNA sequencing; stem; stem cell proliferation; 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慢性GVHD（cGVHD）的新疗法，慢性GVHD是晚期发病的主要原因， 同种异体移植后死亡率我们发现T细胞：B细胞协同性和类别转换的IG 组织沉积引起的多器官系统cGVHD伴闭塞性细支气管炎（BO），一种非感染性气道 阻塞性和上皮重塑疾病，预示着患者的5年生存率很低。从证据- 在cGVHD模型中，我们提供了导致6种新疗法临床试验的关键数据（其中2种现已获得FDA批准） 一线类固醇治疗失败的病人CGVHD可由生发中心（GC）T细胞和B细胞协同性诱导， 产生抗宿主抗体和纤维化虽然我们观察到GC T滤泡辅助细胞（Tfh）增加了糖酵解， 在早期cGVHD/BO阶段，糖酵解随时间减少，与衰竭一致。cGVHD致病性Tfh和 抑制GC的Tfr必须适应高活性氧的GC并在其中生长， 有限的能源。我们的中心假设是cGVHD对GC细胞施加了独特的代谢需求， cGVHD发病机制和对受损肺上皮祖细胞的不同需求，阻碍修复和再生。 选择最佳的单一或联合药物疗法来治疗已建立的cGVHD/BO需要靶向 致病性（Tfhs，GC B细胞）和保留Tfrs和肺干/祖细胞。我们将检验这个假设， 支持GC的增殖的Tfh依赖于多种能量来源（氨解、糖酵解、脂肪酸合成 (FAS)，而异常GC B细胞依赖于糖酵解和FAS。与此相关，我们将测试假设， GC亚群（目的1）和再生肺干/祖细胞（目的2）所需的代谢途径将导致 新的药物靶点目标1A提出：（1）询问GC子集用于能量的精确途径;（2）。 测试在cGVHD/BO小鼠的谱系限制性GC亚群中诱导单个代谢基因缺失是否会改善 肺功能和免疫参数;和（3）.在体内测试选定的代谢候选药物，以逆转 持续cGVHD/BO。在目标1B中，我们假设约40-50 μ g/ml的聚焦CRISPR指导RNA代谢文库可以用于治疗癌症。 来自目标1A药物结果、U-[13 C]-底物（葡萄糖、谷氨酰胺棕榈酸酯）标记和RNA-seq的基因靶点 数据，转导到第0天输注的Cas9转基因供体细胞中将鉴定未发现的代谢基因 对于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， 远端和近端气道上皮和间充质细胞中的配体-受体相互作用。肺细支气管， 肺泡和气管球类器官培养物将用于鉴定受cGVHD/BO不利影响的细胞。在 aim 2B，我们假设aim 1药物有效靶向GC，aim 2药物支持肺细胞增殖， 再生将指导药物选择，以抑制cGVHD/ BO发病机制和修复肺损伤， 预测上级cGVHD/BO结局的价值。这些新颖的方法提供了前所未有的机制 数据来阐明发病机制并导致新的治疗方法。