Phenotyping, Human Tissue and Biomarkers Core

表型、人体组织和生物标志物核心

• 批准号: 10621248
• 负责人: ISSAM A AWAD
• 金额: $ 30.53万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10621248
• 关键词: ADAMTS; Acute; Affect; American; Anticoagulants; Articulation; Biological Markers; Biological Specimen Banks; Blinded; Blood Vessels; Blood capillaries; Brain; CD14 gene; Caenorhabditis elegans; Cavernous Hemangioma; Cells; Chicago; Chronic; Clinical; Clonal Expansion; Collaborations; Collection; Cytoprotection; DNA analysis; Data; Densitometry; Development; Diet; Discipline; Disease; Dissection; Dose; Endothelium; Ensure; Environmental Risk Factor; Epilepsy; Evaluation; Familiarity; Gene Silencing; Genes; Genotype; Germ Lines; Hand; Hemangioma; Hemorrhage; Human; Human Subject Research; Hypoxia; Infiltration; Inflammatory; Institution; Intestines; Investigational Therapies; Iron; Knowledge; Laboratories; Lasers; Lesion; Libraries; Link; MicroRNAs; Modeling; Molecular Probes; Molecular Profiling; Monitor; Mucins; Mus; Mutate; Mutation Analysis; Ontology; Operative Surgical Procedures; Organism; Outcome; PIK3CG gene; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phenotype; Phosphotransferases; Plasma; Plasma Proteins; Proteins; Proteolysis; Protocols documentation; RNA analysis; Research Personnel; Resected; Rho-associated kinase; Role; Sample Size; Sampling; Services; Severity of illness; Signal Transduction; Site; Somatic Mutation; Specimen; Stains; Statistical Methods; Stimulus; TLR4 gene; Techniques; Thrombomodulin; Thrombospondin 1; Thrombospondins; Translating; Translations; Universities; Validation; Work; activated Protein C; adjudication; atorvastatin; biobank; biomarker discovery; biomarker validation; candidate marker; cell free DNA; cerebral cavernous malformations; clinical translation; clinically relevant; cohort; data structure; dietary supplements; differential expression; effective therapy; exosome; experimental study; human subject; human tissue; immunohistochemical markers; innovation; lifetime risk; metabolomics; microCT; microbiome; mind control; molecular marker; mouse model; neurovascular unit; next generation; novel; novel therapeutics; peripheral blood; placebo controlled trial; prevent; primary outcome; programs; prospective; protein expression; risk stratification; secondary outcome; statistics; stroke risk; synergism; tool; transcriptomics; trial readiness; versican

The cerebral cavernous malformation (CCM) is a common hemorrhagic vascular anomaly, caused by germ line and/or endothelial somatic mutations in three known genes. It affects more than a million Americans, predisposing them to a lifetime risk of stroke and epilepsy. A lesion is alarmingly more likely to rebleed after a prior symptomatic hemorrhage, and there is currently no therapy to prevent the genesis or clinical progression of lesions. In the past 5 years, our program project has clarified key mechanisms of CCM pathogenesis through exceptional collaboration and synergy among four laboratories. These included the identification of MEKK3/KLF2/4 signaling as a primary trigger of lesion development in the setting of Ccm loss, downstream activation of RhoA kinase (ROCK), and roles of microbiome driven TLR4/CD14 signaling, intestinal mucin loss, hypoxia, dietary supplements, thrombospondin-1, thrombomodulin and clonal expansion of mutated endothelium. The Scientific Core at U Chicago optimized the high throughput assessment of lesion burden using micro-CT, and the quantification of chronic hemorrhage using densitometry of non-heme iron Perls staining. We developed familiarity with data structure for optimal sample sizes and statistics, and a discipline of prospective articulation of outcomes and blinding in the various experiments. Protocols were harmonized, and models, specimens and data were shared across projects and sites. The Core produced transcriptomic libraries of mutated cells, C. elegans, and neurovascular units from human and murine lesions with various genotypes and at different stages of development. We streamlined the collection and distribution of genotyped human CCM lesions from excised surgical specimens to the project sites. And we assessed the peripheral blood of human subjects for protein levels and microRNAs related to the signaling aberrations. Yet critical knowledge gaps remain, as pilot data has motivated hypotheses about the roles of Ccm deficient endothelium in lesion genesis versus maturation, PIK3 signaling, ADAMTS proteolysis of versican, and activated protein C anticoagulant and cytoprotective pathways. Core services shall continue, further enhanced by new techniques aimed at facilitating single-cell RNA and DNA analyses from lesions, morphometric assessment of acute hemorrhage (as a clinically relevant phenotypic feature distinct from chronic bleeding), and micro-sampling and processing for plasma biomarker discovery in mice. Each of the projects will be assisted by and take advantage of the proposed Phenotyping, Human Tissue and Biomarkers Core, aimed at (1) phenotype assessment and human lesion dissection, and (2) biomarker discovery and validation. The new hypotheses being probed will identify novel therapies, and the resulting biomarkers will facilitate risk stratification and disease monitoring in patients. In a strategic sense, the proposed Core will continue to enhance collaboration and synergy among the project sites, ensure rigor, and facilitate innovation. The Core shall allow cross-validation between mouse and human samples, further leveraging the translation of discoveries to human subjects.

脑海绵状血管畸形（CCM）是一种常见的出血性血管畸形， 和/或三种已知基因中的内皮体细胞突变。它影响了超过一百万的美国人， 使他们一生都有中风和癫痫的危险。令人担忧的是， 既往症状性出血，目前尚无预防发生或临床进展的治疗方法 的病变。在过去的5年里，我们的项目已经阐明了CCM发病机制的关键机制， 四个实验室之间的出色合作和协同作用。其中包括查明 MEKK3/KLF2/4信号传导作为Ccm丢失背景下病变发展的主要触发因素，下游 RhoA激酶（ROCK）的活化，以及微生物组驱动的TLR4/CD14信号传导的作用，肠粘蛋白损失， 缺氧，膳食补充剂，血小板反应蛋白-1，血栓调节蛋白和突变的克隆扩增 内皮细胞芝加哥大学的科学核心优化了病变负荷的高通量评估， 显微CT，以及使用非血红素铁Perls染色的密度测定法定量慢性出血。我们 熟悉最佳样本量和统计的数据结构，以及前瞻性的学科 在各种实验中阐明结果和设盲。协议是统一的，模型， 标本和数据在各项目和地点之间共享。核心产生了转录组文库， 突变细胞、C. elegans和来自具有各种基因型的人和鼠病变的神经血管单位， 在不同的发展阶段。我们简化了基因型人类CCM的收集和分发 从切除的手术标本到项目现场的病变。我们评估了人类的外周血 受试者的蛋白质水平和microRNA相关的信号畸变。然而，关键的知识差距 由于试验数据激发了关于Ccm缺陷内皮细胞在病变发生中作用的假设， 与成熟、PIK3信号传导、多能蛋白聚糖的ADAMTS蛋白水解和活化的蛋白C抗凝剂， 细胞保护途径。核心服务将继续下去，并通过旨在促进 单细胞RNA和DNA分析病变，急性出血的形态学评估（作为临床 与慢性出血不同的相关表型特征），以及血浆的微量取样和处理 在小鼠中发现生物标志物。每个项目都将得到拟议的 表型，人类组织和生物标志物核心，旨在（1）表型评估和人类病变 解剖，和（2）生物标志物的发现和验证。正在探索的新假设将确定新的 这些生物标志物将有助于患者的风险分层和疾病监测。中 从战略意义上讲，拟议核心区将继续加强项目地点之间的合作和协同作用， 确保严谨，促进创新。核心应允许小鼠和人类之间的交叉验证 样本，进一步利用发现人类受试者的翻译。