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Phenotyping, Human Tissue and Biomarkers Core

表型、人体组织和生物标志物核心

基本信息

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
项目类别：
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

项目摘要

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.

脑海绵状血管瘤（CCM）是一种常见的出血性血管异常，由生殖系引起和/或三个已知基因的内皮体细胞突变。它影响了超过一百万美国人，使他们终生面临中风和癫痫的风险。令人担忧的是，病变在手术后更有可能再出血。既往有症状性出血，目前尚无治疗方法可以预防其发生或临床进展的病变。在过去的5年里，我们的项目通过以下方式阐明了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。其中包括识别 MEKK3/KLF2/4 信号传导是 Ccm 丢失情况下病变发展的主要触发因素，下游 RhoA 激酶 (ROCK) 的激活，以及微生物组驱动的 TLR4/CD14 信号传导的作用、肠道粘蛋白损失、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使用以下方法优化了病变负担的高通量评估显微 CT，以及使用非血红素铁 Perls 染色的光密度测定法对慢性出血进行量化。我们熟悉最佳样本量和统计数据的数据结构，以及前瞻性学科各种实验中结果的阐明和盲法。协议和模型得到了协调，样本和数据在项目和地点之间共享。核心产生的转录组文库来自人类和小鼠病变的突变细胞、线虫和神经血管单位，具有不同的基因型和处于不同的发展阶段。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与信号异常相关的蛋白质水平和 microRNA 的受试者。然而关键的知识差距仍然存在，因为试点数据激发了关于 Ccm 缺陷内皮在病变发生中的作用的假设与成熟、PIK3 信号传导、多能聚糖 ADAMTS 蛋白水解以及活化的蛋白 C 抗凝剂和细胞保护途径。核心服务将继续下去，并通过旨在促进病变的单细胞 RNA 和 DNA 分析、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），以及血浆的微量采样和处理小鼠生物标志物的发现。每个项目都将得到拟议的协助并利用表型、人体组织和生物标志物核心，旨在 (1) 表型评估和人类病变解剖，以及（2）生物标志物的发现和验证。正在探索的新假设将发现新颖的治疗方法以及由此产生的生物标志物将有助于患者的风险分层和疾病监测。在一个战略意义上，拟议的核心将继续加强项目地点之间的协作和协同，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，进一步利用将发现转化为人类受试者。