Development and Validation of a FISH-Based Renal Cancer Detection Assay

描述(由申请人提供)：常规使用最先进的成像技术增加了意外发现的肾小肿块的发生率，包括良性和恶性。有这种肿块的患者中，有很大一部分是在年龄更大、更虚弱的人群中，对于肾细胞癌(RCC)，现在有60%-70%的患者没有症状。对于这种疾病，目前的干预措施包括对标准外科手术程序进行微创治疗，因此需要降低发病率的治疗建议。目前，针吸活检基于形态的诊断对治疗决策的影响主要有两个局限性：无法获得足够数量和质量的诊断材料进行形态检查，以及无法从形态上区分特定的肾细胞癌亚型。在目前的第一阶段/第二阶段应用中，我们建议开发一种新的基于荧光原位杂交(FISH)的肾癌检测(FRCD)试剂盒，该试剂盒不是基于细胞形态，而是基于肾皮质肿瘤的固有差异遗传学，由癌症遗传公司进行商业分销。应用这一测试将允许诊断四种主要的肾皮质肿瘤亚型(三种恶性，一种良性)，与形态检查一起将影响这种疾病的临床治疗的所有阶段。第一阶段；具体目标1：回顾验证基于BAC的FISH探针板在肾核心活检中区分肾皮质肿瘤的四种主要亚型。由于使用CGI设计的基于BAC的探针板的初步研究显示，该探针板对四种组织亚型有良好的鉴别诊断，因此该探针板的敏感性、特异性、阳性和阴性预测值将在145例活检中得到验证，这些活检标本来自标准病理学已知但盲目的形态切除的标本。第二阶段；具体目标1：开发单一副本鱼类探针板。为了在不侵犯杂交中Cot-1 DNA专利的情况下商业化分发基于FISH的检测，将设计、传播、标记一套单拷贝探针，并验证其用于第一阶段中使用的肾针活检和手术标本的子集；具体目标2：前瞻性地验证单拷贝FISH探针小组作为一种潜在的检测方法在针刺活检中诊断肾细胞癌的可能性。在IRB批准的一项研究中，单拷贝探针小组将在图像引导下经皮肾活检中进行评估，预期从可疑肾肿块患者中获得。该分析的准确性将根据标准病理学已知但盲目的形态进行评估，预计将在两年内收集和检测300例活检组织。该项目的总体目标是开发、生产和销售一种强大、敏感和特定的FRCD试剂盒，与标准病理学一起，将显著有助于肾肿块和肾癌患者的临床决策，影响患者的治疗建议和护理成本。与公共卫生相关：随着复杂的成像技术的常规使用，肾脏肿块经常在老年患者中被发现，但尚不清楚这些肿块是恶性的，需要治疗，还是良性的。基于基因的肾癌检测试剂盒将在这一应用中开发和验证，可用于对获得的肿块进行针刺活组织检查，而此类患者几乎没有并发症，这将帮助临床医生和外科医生做出更好的治疗决定，并防止不必要的昂贵外科手术。

DESCRIPTION (provided by applicant): Increased routine use of state-of-the-art imaging techniques has resulted in a higher incidence of small incidentally discovered renal masses, benign and malignant. A large proportion of patients with such masses, are within an older, more frail population and for renal cell carcinoma (RCC), 60-70% of patients now present asymptomatically. Treatment recommendations then with decreased morbidity are required for such patients, in a disease where current interventions include minimally invasive to standard surgical procedures. Recently, the impact of morphology-based diagnosis of needle biopsies on treatment decision making has been identified to have two main limitations: inability to obtain diagnostic material of adequate quantity and quality for morphologic examination, and inability to distinguish particular RCC subtypes morphologically. In the current Phase I/Phase II application, we propose to develop a novel fluorescence in situ hybridization (FISH)-based renal cancer detection (FRCD) kit that is based not on cell morphology, but on the inherent differential genetics of renal cortical neoplasms, for commercial distribution by Cancer Genetics, Inc. Application of this test will permit diagnosis of the four predominant subtypes of renal cortical neoplasms (three malignant, one benign) that together with morphologic examination would impact all phases of clinical management of this disease. Phase I; Specific Aim 1: Retrospective validation of a BAC-based FISH probe panel in discrimination of the four main subtypes of renal cortical neoplasms in renal core biopsies. Since preliminary studies using a BAC-based probe panel designed by CGI revealed good differential diagnosis of the four histologic subtypes, the sensitivity, specificity, positive and negative predictive values of the probe panel will be validated in a cohort of 145 biopsies obtained from resected specimens with known but blinded morphology by standard pathology. Phase II; Specific Aim 1: Development of a single-copy FISH probe panel. In order to commercially distribute a FISH-based test without patent infringement of Cot-1 DNA use in hybridizations, a set of single-copy probes will be designed, propagated, labeled, and validated for use in a subset of renal needle biopsies and surgical specimens used in Phase I. Phase II; Specific Aim 2: Prospective validation of the single-copy FISH probe panel as a potential assay in the diagnosis of RCC in needle biopsies. The panel of single-copy probes will be evaluated in image-guided percutaneous renal biopsies prospectively obtained from patients with suspicious renal masses in an IRB-approved study. The accuracy of the assay will be evaluated with respect to the known but blinded morphology by standard pathology, of the 300 biopsies expected to be collected and assayed over two years. The overall goal of the project then is to develop, produce, and market a robust, sensitive, and specific FRCD kit that together with standard pathology will significantly contribute to clinical decision making in patients with renal masses and kidney cancer, impacting patient treatment recommendations and cost of care. PUBLIC HEALTH RELEVANCE: With sophisticated imaging technologies routinely being used, kidney masses are often detected in older patients, but it unclear if these masses are malignant and need treatment, or are benign. The genetic-based kidney cancer detection kit to be developed and validated in this application that can be used on needle biopsies of the mass obtained with few complications from such patients, will help clinicians and surgeons to make better treatment decisions and prevent needless costly surgical procedures.