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针对慢性GVHD（CGVHD）的新疗法，这是晚期发病率的主要原因 同种异体移植后的死亡率。我们做出了一个重要的观察结果，即T细胞：B细胞的协作性和类切换的Ig 组织沉积引起多器官系统CGVHD，其细支气管炎闭塞（BO），这是一种非感染气道 阻塞性和上皮重塑疾病预示了患者的5年生存期糟糕的疾病。从证明 CGVHD模型中的概念，我们提供了关键数据，导致6种新疗法的临床试验（现已批准了FDA） 对于一线类固醇失败的患者。 CGVHD可以由生发中心（GC）T-和B细胞协作诱导， 产生抗宿主ABS和纤维化。虽然我们观察到GC tfollicular辅助细胞（TFH）的糖酵解增加 随着时间的流逝，糖酵解的早期CGVHD/BO相逐渐减少，与疲惫一致。 CGVHD致病性TFH和 抑制GC的tfollicular调节细胞（TFR）必须适应高活性氧和高活性的GC和壮成长 能源有限。我们的中心假设是CGVHD对GC细胞施加了独特的代谢需求 CGVHD发病机理和对受损的肺上皮祖细胞的明显需求，阻碍修复和再生。 选择最佳的单一或合并的药物疗法来治疗已建立的CGVHD/BO最佳目标 致病性（TFHS，GC B细胞）和保留TFR和肺茎/祖细胞。我们将迅速检验的假设 支持GC的增殖TFH取决于多种能源（谷氨酰胺溶解，糖酵解，脂肪酸合成 （FAS），而异常的GC B细胞依赖于糖酵解和FAS。相关，我们将检验以下假设： GC子集（AIM 1）和再生肺茎/祖细胞所需的代谢途径（AIM 2）将导致 新的可药物目标。 AIM 1A提出：（1）询问用于能源的精确途径的GC子集； （2）。 测试如果在CGVHD/BO小鼠中诱导谱系限制的GC子集中的单一代谢基因缺失将改善 肺功能和免疫参数； （3）。测试选定的代谢候选药物在体内逆转 正在进行的CGVHD/BO。在AIM 1B中，我们假设一个重点的CRISPR指南RNA代谢库约40-50 AIM 1A药物结果的基因靶标U- [13C] - 基底（葡萄糖，谷氨酰胺）标记和RNA-Seq 在第0天注入的CAS9转基因供体细胞转导的数据将识别未发现的代谢基因 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 对肺损伤修复/再生的影响。在AIM 2A中，我们将使用SCRNA-SEQ定义改变的细胞状态，并且 远端和近端气道上皮和间质细胞中的配体 - 受体相互作用。肺支气管， 肺泡和气管层的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，并在支持肺部细胞中瞄准2种药物 再生将指导药物选择以抑制CGVHD/ BO发病机理和修复肺损伤 上级CGVHD/BO结果的预测价值。这些新颖的方法提供了前所未有的机械 数据以阐明发病机理并导致新疗法。