Development of a diagnostic platform for PTSD blood biomarkers

开发 PTSD 血液生物标志物诊断平台

• 批准号: 10455593
• 负责人: Michael S Breen
• 金额: $ 21.13万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10455593
• 关键词: Binding Sites; Biological; Biological Markers; Blood specimen; CD14 gene; CD19 gene; CD8B1 gene; Cells; Clinical; DNA Binding; Data; Development; Dexamethasone; Diagnostic; Discrimination; Disease; Etiology; Exposure to; Flow Cytometry; Future; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Glean; Glucocorticoid Receptor; Glucocorticoids; Goals; Hematological Disease; Heterogeneity; Hormones; Immune; Immune system; Impairment; Individual; Individual Differences; Inflammatory; Laboratory Finding; Light; Measures; Mediating; Modeling; Molecular; NCAM1 gene; Neurosecretory Systems; Outcome; Participant; Pathway interactions; Patients; Pattern; Peripheral; Peripheral Blood Mononuclear Cell; Post-Traumatic Stress Disorders; Predisposition; Production; Recovery; Regulation; Regulator Genes; Regulatory Element; Research; Resolution; Signal Transduction; Site; Stress; System; Techniques; Technology; Therapeutic; Tissues; Translating; Trauma; Variant; Veterans; Work; biological adaptation to stress; biomarker development; case control; cell type; chromatin immunoprecipitation; combat veteran; cytokine; density; diagnostic biomarker; diagnostic platform; diagnostic screening; differential expression; gene expression variation; genome-wide; hypothalamic-pituitary-adrenal axis; in vitro Model; indexing; insight; liquid biopsy; novel; post-trauma; receptor binding; receptor expression; resilience; response; stress reactivity; transcriptome; transcriptome sequencing; transcriptomics; trauma exposure; traumatic event; Binding Sites; Biological; Biological Markers; Blood specimen; CD14 gene; CD19 gene; CD8B1 gene; Cells; Clinical; DNA Binding; Data; Development; Dexamethasone; Diagnostic; Discrimination; Disease; Etiology; Exposure to; Flow Cytometry; Future; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Glean; Glucocorticoid Receptor; Glucocorticoids; Goals; Hematological Disease; Heterogeneity; Hormones; Immune; Immune system; Impairment; Individual; Individual Differences; Inflammatory; Laboratory Finding; Light; Measures; Mediating; Modeling; Molecular; NCAM1 gene; Neurosecretory Systems; Outcome; Participant; Pathway interactions; Patients; Pattern; Peripheral; Peripheral Blood Mononuclear Cell; Post-Traumatic Stress Disorders; Predisposition; Production; Recovery; Regulation; Regulator Genes; Regulatory Element; Research; Resolution; Signal Transduction; Site; Stress; System; Techniques; Technology; Therapeutic; Tissues; Translating; Trauma; Variant; Veterans; Work; biological adaptation to stress; biomarker development; case control; cell type; chromatin immunoprecipitation; combat veteran; cytokine; density; diagnostic biomarker; diagnostic platform; diagnostic screening; differential expression; gene expression variation; genome-wide; hypothalamic-pituitary-adrenal axis; in vitro Model; indexing; insight; liquid biopsy; novel; post-trauma; receptor binding; receptor expression; resilience; response; stress reactivity; transcriptome; transcriptome sequencing; transcriptomics; trauma exposure; traumatic event

PROJECT SUMMARY Post-traumatic stress disorder (PTSD) is a disorder of stress reactivity with clinical manifestations that are evident when patients are triggered following exposure to a traumatic event. Impaired sensitivity to glucocorticoids (GCs) and alterations in peripheral blood mononuclear cell (PBMC) gene expression profiles, including genes implicated in GC signaling and inflammatory cytokine production, are at the crossroads of peripheral and central susceptibility pathways and represent promising PTSD biomarkers. However, their small case-control effect sizes have been a significant barrier to the development of actionable diagnostic biomarkers for the disorder. Notably, such studies have been performed post-trauma under baseline conditions and do not fully recapitulate the molecular response to adverse pathophysiological triggers, such as exposure to stress hormones. We developed a new in vitro model that sensitizes PBMCs to the synthetic GC, dexamethasone (DEX). Our model captures differences between individuals in sensitivity to GCs and significantly amplifies PTSD gene expression effect sizes into a range that may facilitate the development of actionable biomarkers. Still, cell type-specific differences in GC sensitivity have not been fully appreciated in PTSD and represent an important step forward for the development of translatable biomarkers and therapeutics. Here, we propose to combine our in vitro model with single-cell cellular indexing of transcriptomes and epitomes by sequencing (CITE-seq) and chromatin immunoprecipitation (ChIP)-seq to investigate the single-cell transcriptional responses to GC activation in PTSD. Specifically, we will investigate Aim 1) cell type-specific transcriptional differences using the CITE-seq approach on vehicle and DEX treated PBMCs derived from trauma-exposed combat veterans with PTSD(+; n=20) and without PTSD(-; n=20), as well as healthy trauma-free participants (HC; n=20). In doing so, we will study ~600,000 single cells (~5,000 cells per condition for each individual). Flow-cytometry will quantify the density and proportion of GR expression on five major immune cell types. Because the GR has site-specific DNA binding activity that differs between cells and conditions, we will Aim 2) perform anti-GR ChIP-seq data on five major immune cell subsets to identify genome-wide, cell type-specific GR binding sites. Integrating CITE-seq, GR expression and ChIP- seq across DEX exposed immune cell subsets will provide more nuanced and accurate insights into the cellular and molecular perturbations associated with altered GC sensitivity in PTSD. We have already measured genome-wide RNA-seq in bulk PBMCs treated with DEX and uncovered robust transcriptional differences distinguishing PTSD+ from PTSD-. Successful completion of this proposal will uncover cell type- specific transcriptional responses that will serve as definitive guides for the development of biomarkers for PTSD stress responsivity, vulnerability and resiliency.

项目摘要 创伤后应激障碍（PTSD）是一种应激反应性的疾病，临床表现是 暴露于创伤事件后会触发患者时，很明显。对敏感性受损 糖皮质激素（GC）和外周血单核细胞（PBMC）基因表达谱的改变， 包括与GC信号传导和炎性细胞因子产生有关的基因，位于 外围和中心敏感性途径，代表有希望的PTSD生物标志物。但是，他们的小 病例对照效应尺寸已成为开发可行诊断的重大障碍 该疾病的生物标志物。值得注意的是，在基线条件下创伤后已经进行了此类研究 并且不要完全概括对不良病理生理诱因的分子反应，例如暴露 压力激素。我们开发了一种新的体外模型，该模型将PBMC敏感到合成GC，即 地塞米松（Dex）。我们的模型捕获了个人对GC的敏感性和 显着将PTSD基因表达效应大小放大到可能有助于发展的范围 可操作的生物标志物。尽管如 PTSD并代表着开发可翻译生物标志物的重要一步 疗法。在这里，我们建议将我们的体外模型与单细胞细胞索引相结合 通过测序（Cite-Seq）和染色质免疫沉淀（CHIP）-Seq到TO TO TO的转录组和缩影 研究对PTSD中GC激活的单细胞转录响应。具体来说，我们将调查 AIM 1）使用cite-seq方法在车辆和DEX处理的情况 PBMC源自具有PTSD（+; n = 20）和没有PTSD（ - ; n = 20）的受创伤暴露的战斗退伍军人，AS 以及健康的无创伤参与者（HC; n = 20）。这样，我们将研究约600,000个单元格（约5,000个单元 每个条件的细胞）。流程仪将量化GR的密度和比例 对五种主要免疫细胞类型的表达。因为GR具有特定位点的DNA结合活性，不同 在细胞和条件之间，我们将瞄准2）对五个主要免疫细胞子集执行抗GR芯片seq数据 确定全基因组，细胞类型特异性的GR结合位点。整合cite-seq，gr表达和芯片 SEQ跨DEX暴露的免疫细胞子集将提供更细微和准确的见解 与PTSD中GC敏感性改变有关的细胞和分子扰动。我们已经 用DEX处理和未发现的稳健转录的大量PBMC中测量的全基因组RNA-SEQ 区分PTSD+与PTSD-的差异。该提案的成功完成将发现细胞类型 - 具体的转录响应将作为开发生物标志物的确定指南 PTSD压力响应性，脆弱性和弹性。