Novel Diagnostic Tests for Renal Cell Carcinoma

肾细胞癌的新型诊断测试

• 批准号: 9031600
• 负责人: John A. Petros
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9031600
• 关键词: AQP1 gene; Affect; Air; Award; Base Sequence; Bathing; Benign; Biological Assay; Blood; Blood Tests; Blood typing procedure; CLIA certified; Camping; Caring; Centers for Disease Control and Prevention (U.S.); Cessation of life; Chemicals; Chronic Kidney Failure; Clinic; Clinical; Complement; Creatinine; Cystic Nephroma; DNA; DNA Sequence; DNA Sequence Alteration; Data; Detection; Development; Diagnosis; Diagnostic tests; Disease; Early Diagnosis; Enrollment; Enzyme-Linked Immunosorbent Assay; Evaluable Disease; Evaluation; Exposure to; Family; Fatal Outcome; Fibrous capsule of kidney; Future; Genes; Goals; Gold; Health; Health system; Histologic; Image; Individual; Kidney; Kidney Diseases; Kidney Failure; Kidney Neoplasms; Laboratories; Laboratory Research; Laws; Lesion; Link; Literature; Magnetic Resonance Imaging; Malignant - descriptor; Malignant Neoplasms; Marines; Measurement; Measures; Medical; Medical center; Metals; Methods; Military Personnel; Morbidity - disease rate; Mutate; Nephrectomy; Obesity; Occupational Exposure; Operative Surgical Procedures; Outcome; Outpatients; Oxyphilic Adenoma; Patient Selection; Patients; Peptides; Performance; Plasma; Population; Populations at Risk; Proteins; Renal Cell Carcinoma; Renal Mass; Renal carcinoma; Reporting; Research; Risk; Risk Factors; Scanning; Schedule; Sensitivity and Specificity; Site; Smoker; Solid; Solvents; Somatic Mutation; Specimen; Stage at Diagnosis; Staging; System; Testing; Tetrachloroethylene; Time; Tissues; Tobacco smoking; Toxin; Translations; Trichloroethylene; Tumor Tissue; Tumor-Derived; Ultrasonography; University Hospitals; Urine; Veterans; Water; Woman; Work; X-Ray Computed Tomography; base; blood group; cancer diagnosis; clinical application; clinically relevant; cohort; contaminated water; dechlorination; deep sequencing; drinking; evidence based guidelines; follow-up; high risk; lifetime risk; lymph nodes; male; men; mortality; novel; novel diagnostics; patient population; performance tests; prospective; public health relevance; screening; sound; tumor; tumor DNA; urinary; volatile organic compound

 DESCRIPTION (provided by applicant): Objectives. The objective of this proposal is to evaluate, in patients, two novel tests developed in our research laboratory for the detection of renal cell carcinoma (RCC). The primary objective is to rigorously determine sensitivity and specificity of the tests so that when the proposed work has been accomplished, evidence-based recommendations can be made for the use of the tests in specific patient populations and scenarios such as screening a healthy population, screening a high risk group such as patients with chronic kidney disease (CKD) or acquired cystic disease (ACD) and in the evaluation of radiologically detected renal masses. Research Plan. Two specific aims are proposed. In the first aim we will deploy the tests (quantitative determination of aquaporin-1 (AQP-1) protein in the urine and DNA-sequencing based detection of tumor- associated somatic mutations in the blood) in patients with known renal cancer (pre-op surgical cohort) and controls (healthy ambulatory population and individuals with CT scans showing no renal masses). In this way we will compare the new diagnostic tests' performance to gold standard diagnostic tests (tissue diagnosis for the positive controls and negative CT for the negative controls). In the second aim we will use the tests to screen patients seen in the Atlanta VA Renal Clinic that have CKD and abnormal screening ultrasounds, a high risk group. Methods. The urine AQP-1 test has been fully developed in a CLIA lab (Emory University Hospital clinical laboratory) and is a novel quantitative ELISA assay that is performed on proteins purified from urine. The analytic measurement range (AMR) is 0.25-16.0 ng/ml and is normalized to urine creatinine levels post- analytically. Preliminary data presented in this application demonstrate that all RCC urines tested to date are robustly positive while all healthy negative controls are below the AMR. The detection of tumor-derived DNA in the blood is performed by isolating DNA from patient plasma and then PCR amplifying the complete exonic sequence of 14 genes known to be mutated in RCC followed by deep sequencing using the Illumina platform, also established in a CLIA certified laboratory. Preliminary data demonstrates that all RCC tumor tissue sequenced in this manner demonstrates somatic mutations in at least one of the 14 genes and that 100% of evaluable patients had somatic mutations detected in the blood. Clinical Relevance. There is no laboratory test for RCC. Clinical outcome of RCC is essentially completely dependent on the stage at which the disease is detected with survival in excess of 95% for those tumors diagnosed at the earliest stage (small cancer confined to the renal capsule) compared to 5% or less once metastatic. The ultimate clinical translation of these tests would be to drastically alter the stage at which RCC is diagnosed thereby significantly decreasing morbidity and mortality from this disease that currently claims the lives of over 13,000 patients per year. This is especially relevant to veterans' health since hundreds of thousands of Marines and their families were exposed to TCE, a chemical degreasing agent, at Camp Lejeune (and many other Air Force and Marine bases) and the CDC has determined that these individuals are at significantly increased risk of developing RCC and that the Veterans Affairs health system is responsible for caring for these individuals. The development of screening tests for RCC within the VA system could be seen as particularly responsive to this congressionally directed mandate.

 描述（由申请人提供）： 目标.本提案的目的是在患者中评估我们研究实验室开发的两种用于检测肾细胞癌（RCC）的新型测试。主要目标是严格确定检测的灵敏度和特异性，以便在完成拟议工作后，可以提出基于证据的建议，以便在特定患者人群和场景中使用检测，例如筛查健康人群，筛查高危人群，如慢性肾病（CKD）或获得性囊性疾病（ACD）患者以及在放射学检测的肾肿块的评价中。研究计划。提出了两个具体目标。在第一个目标中，我们将在已知患有肾癌的患者（术前手术队列）和对照（健康的流动人群和CT扫描显示无肾脏肿块的个体）中部署测试（尿液中水通道蛋白-1（AQP-1）蛋白的定量测定和血液中肿瘤相关体细胞突变的基于DNA测序的检测）。通过这种方式，我们将比较新诊断测试与金标准诊断测试的性能（阳性对照的组织诊断和阴性对照的阴性CT）。在第二个目标中，我们将使用这些测试来筛查在亚特兰大VA肾脏诊所就诊的患有CKD和异常筛查超声的患者，这是一个高危人群。方法.尿液AQP-1测试已经在CLIA实验室（埃默里大学医院临床实验室）中完全开发，并且是一种对从尿液中纯化的蛋白质进行的新型定量ELISA测定。分析测量范围（AMR）为0.25-16.0 ng/ml，并标准化为分析后的尿肌酐水平。本申请中提供的初步数据表明，迄今为止检测的所有RCC尿液均为强阳性，而所有健康阴性对照均低于AMR。通过从患者血浆中分离DNA，然后PCR扩增已知在RCC中突变的14个基因的完整外显子序列，然后使用Illumina平台进行深度测序，检测血液中肿瘤来源的DNA，该平台也在CLIA认证的实验室中建立。初步数据表明，以这种方式测序的所有RCC肿瘤组织均在14个基因中的至少一个中显示体细胞突变，并且100%的可评估患者在血液中检测到体细胞突变。临床相关性。RCC没有实验室检测。RCC的临床结果基本上完全取决于检测到疾病的阶段，对于在最早阶段诊断的那些肿瘤（局限于肾囊的小癌症），生存率超过95%，而一旦转移则为5%或更低。这些测试的最终临床转化将大大改变RCC诊断的阶段，从而显著降低这种疾病的发病率和死亡率，目前每年有超过13，000名患者死亡。这与退伍军人的健康特别相关，因为数十万海军陆战队及其家人在勒琼营（以及许多其他空军和海军基地）接触了TCE，一种化学毒剂，CDC已经确定这些人患RCC的风险显着增加，退伍军人事务部卫生系统负责照顾这些人。在VA系统内开发RCC筛查测试可以被视为对国会指示的任务特别敏感。