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

描述（由申请人提供）：越来越多的常规使用最先进的成像技术导致偶然发现的小的肾脏肿块的发生率增加，无论是良性的还是恶性的。很大一部分有这种肿块的患者是年龄较大、身体较弱的人群，对于肾细胞癌（RCC）， 60-70%的患者现在无症状表现。对于这种疾病，目前的干预措施包括微创到标准外科手术，因此需要降低发病率的治疗建议。最近，基于形态学的针活检诊断对治疗决策的影响已被确定为两个主要局限性：无法获得足够数量和质量的形态学检查诊断材料，以及无法从形态学上区分特定的RCC亚型。在目前的I/ II期申请中，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cancer genetics， Inc.进行商业分销。应用该检查可以诊断肾皮质肿瘤的四种主要亚型（三种恶性，一种良性），结合形态学检查将影响该疾病临床管理的所有阶段。第一阶段;具体目的1：回顾性验证基于bac的FISH探针面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的探针板进行的初步研究显示对四种组织学亚型有良好的鉴别诊断，因此探针板的敏感性、特异性、阳性和阴性预测值将在145个活检标本的队列中得到验证，这些活检标本是由标准病理已知但不透明的切除标本。第二阶段;具体目标1：开发单拷贝FISH探针面板。为了在不侵犯杂交中使用的Cot-1 DNA专利的情况下商业化分销基于fish的测试，将设计、繁殖、标记和验证一套单拷贝探针，用于第一阶段使用的肾针活检和手术标本的子集。特异性目标2：前瞻性验证单副本FISH探针面板作为针活检中诊断RCC的潜在检测方法。在一项经irb批准的研究中，单拷贝探针将在图像引导下经皮肾活检中进行评估，这些活检来自可疑肾肿块的患者。测定的准确性将根据标准病理学的已知但盲法形态学进行评估，预计将在两年内收集和分析300个活检组织。该项目的总体目标是开发、生产和销售一种强大、敏感和特异性的FRCD试剂盒，该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。公共卫生相关性：由于常规使用复杂的成像技术，经常在老年患者中发现肾脏肿块，但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此应用中开发和验证，可用于从此类患者身上获得的肿块的针活检，几乎没有并发症，将帮助临床医生和外科医生做出更好的治疗决策，并防止不必要的昂贵手术。

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.