Micro-western array methodology for assessment of preanalytical variability in bi

用于评估双组分分析前变异性的微西方阵列方法

基本信息

批准号：
8848052
负责人：
KEVIN P. WHITE
金额：
$ 24.06万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-05-12 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8848052
关键词：
Address Adrenal Gland Neoplasms Age Anesthetics Antibodies Biological Bladder Neoplasm Blood Cells Chicago Clinic Clinical Collection Communities Core Facility Daughter Drug Industry Drug usage Esophageal Ethics Excision Exposure to Freezing Future Gene Expression Gene Proteins Genomics Head and Neck Cancer Head and Neck Neoplasms Head and neck structure Health Hemorrhage Hour Hypoxia Informatin Institution Intestines Ischemia Lung Malignant Neoplasms Masks Measurement Measures Medical center Methodology Methods Modeling Modification Monitor Operating Rooms Operative Surgical Procedures Perioperative Pharmaceutical Preparations Phase Post-Translational Protein Processing Postoperative Period Probability Procedures Process Prostatic Neoplasms Protein Analysis Protein Array Proteins Proteomics Proxy Race Regression Analysis Resources Sampling Services Signal Transduction Signaling Protein Slice Source Specimen Stomach Stress Surgeon Technology Temperature Testing The Cancer Genome Atlas Thyroid Gland Time Tissue Banks Tissues Universities Validation Warm Ischemia Work anticancer research biobank efficacy testing in vivo metabolomics novel nutrition pancreatic neoplasm personalized medicine protein expression rectal response sample collection scale up sex tumor

项目摘要

DESCRIPTION (provided by applicant): Biospecimens are an invaluable resource to the biomedical community. The objective of this proposal is to employ a combination of micro-western arrays and reverse phase protein arrays to dramatically increase the throughput of antibody validation as well as the comprehensiveness of proteins that can be examined in biospecimens. We will use this platform to examine the relationship of about 500 protein abundances and modification states with sources of in- and ex-vivo peri-operative sources of preanalytical variability. We will record standard sources of per-operative variability during the normal course of surgical resection of head-and- neck tumors and relate these variables to differential protein expression and modification. In parallel, we will measure changes in protein expression and modification following defined times of ex-vivo ischemia to mimic a standard source of ex-vivo peri-operative pre-analytical variability. In summary, we will develop standard operating procedures for surgical biospecimen removal and test the efficacy of a platform employing micro- western arrays and reverse phase protein arrays for measuring changes in the expression of about 500 cell signaling proteins in approximately 140 biospecimens subjected either to standard perioperative handling variables or to defined and controlled sources of preanalytical variability. Following surrogate variable analysis to control for known biological covariates (such as age, sec, etc.), we will use linear mixed effects modeling to identify baseline protein levels associated with preanalytical variability. Two-step, cubic regression will be used to identify proteins with significant ex-vivo temporal expression changes following resection. Our model will be used to establish metrics of tissue quality and to identify protein signatures indicative of biospecimen quality.

描述（由申请人提供）：生物测量是生物医学界的宝贵资源。该提案的目的是采用微西阵列和反相蛋白阵列的组合，以显着增加抗体验证的吞吐量以及可以在生物测量中检查的蛋白质的全面性。我们将使用此平台来检查大约500种蛋白质丰度和修饰状态的关系与体外和前围手术量源的来源的关系。我们将记录在头和颈部肿瘤的正常手术切除过程中均操作变异性的标准来源，并将这些变量与差异蛋白表达和修饰联系起来。同时，我们将根据定义的前缺血时间来衡量蛋白质表达和修饰的变化，以模仿前体术前分析变异性的标准来源。总而言之，我们将开发用于除手术生物概念的标准操作程序，并测试采用微阵列和反相蛋白阵列的平台的功效，以测量大约140个细胞信号蛋白在大约140个生物疗法中表达的变化，该蛋白在大约140个生物疗法中，对标准的毛皮变量或对定义的源自定义的源自定义的可变性。在替代变量分析以控制已知的生物协变量（例如年龄，SEC等）之后，我们将使用线性混合效应建模来识别与缩小性变异性相关的基线蛋白水平。两步，立方回归将用于鉴定切除后具有明显的外部时间表达变化的蛋白质。我们的模型将用于建立组织质量的指标，并确定指示生物质量质量的蛋白质特征。