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

• 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
• 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

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）：生物标本是生物医学界的宝贵资源。本提案的目的是采用微western阵列和反相蛋白质阵列的组合，以显着提高抗体验证的吞吐量以及可以在生物标本中检测的蛋白质的全面性。我们将使用这个平台来检查大约500种蛋白质丰度和修饰状态与体内和离体围手术期分析前变异性来源的关系。我们将记录头颈部肿瘤手术切除正常过程中手术前变异的标准来源，并将这些变异与差异蛋白表达和修饰联系起来。同时，我们将测量离体缺血后蛋白质表达和修饰的变化，以模拟离体围手术期分析前变异性的标准来源。总之，我们将制定外科生物标本去除的标准操作程序，并测试使用微western阵列和反相蛋白质阵列的平台的有效性，以测量大约140个生物标本中约500个细胞信号蛋白的表达变化，这些生物标本受到标准围手术期处理变量或定义和控制的分析前变异性来源的影响。通过替代变量分析来控制已知的生物协变量（如年龄、年龄等），我们将使用线性混合效应模型来确定与分析前变异性相关的基线蛋白质水平。两步，三次回归将用于鉴定在切除后显着的离体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，并确定指示生物标本质量的蛋白质特征。