Metabolomics of cGVHD

cGVHD 的代谢组学

基本信息

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
项目类别：
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

项目摘要

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，慢性 GVHD 是导致晚期发病和死亡的主要原因。同种异体移植后的死亡率。我们做出了重要的观察：T 细胞：B 细胞协同性和类别转换 Ig 组织沉积引起多器官系统cGVHD，伴有闭塞性细支气管炎（BO），一种非感染性气道阻塞性和上皮重塑疾病预示着患者 5 年生存率极低。从证明- cGVHD 模型中的概念，我们提供了 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）对于一线类固醇治疗失败的患者。 CGVHD 可由生发中心 (GC) T 细胞和 B 细胞协同作用诱导，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) 在在 cGVHD/BO 早期阶段，糖酵解随着时间的推移而减少，与衰竭一致。 cGVHD 致病性 Tfh 和抑制 GC 的滤泡调节细胞 (Tfr) 必须在具有高活性氧和高活性氧的 GC 中适应并茁壮成长。能源有限。我们的中心假设是，cGVHD 对 GC 细胞施加独特的代谢需求 cGVHD 发病机制和对受损肺上皮祖细胞的独特需求，阻碍了修复和再生。选择最佳的单一或联合药物疗法来最佳治疗已确定的 cGVHD/BO 需要靶向治疗致病性（Tfhs、GC B 细胞）和保留 Tfrs 和肺干/祖细胞。我们将快速检验这个假设支持 GC 的增殖 Tfh 依赖于多种能源（谷氨酰胺分解、糖酵解、脂肪酸合成） (FAS)，而异常 GC B 细胞依赖糖酵解和 FAS。相关地，我们将检验以下假设： 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药物靶标。目标 1A 建议：(1) 询问 GC 子集以获取用于能量的精确途径；（2）。测试在 cGVHD/BO 小鼠的谱系限制 GC 子集中诱导单一代谢基因缺失是否会改善肺功能和免疫参数；和（3）。测试所选候选药物体内代谢逆转情况正在进行的 cGVHD/BO。在目标 1B 中，我们假设一个集中的 CRISPR 引导 RNA 代谢文库约为 40-50 来自 Target 1A 药物结果、U-[13C]-底物（葡萄糖、谷氨酰胺棕榈酸酯）标记和 RNA-seq 的基因靶点转入第 0 天输注的 Cas9 转基因供体细胞的数据将识别未被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药物所抵消对肺损伤修复/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气道上皮细胞和间充质细胞中配体-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识别受 cGVHD/BO 不利影响的细胞。在目标 2B，我们假设目标 1 药物可有效靶向 GC，目标 2 药物可有效支持肺细胞再生将指导药物选择以抑制cGVHD/BO发病机制并修复肺损伤。优越的 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阐明发病机制并导致新疗法。