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

描述（由申请人提供）：最先进成像技术的常规使用增加导致偶然发现的良性和恶性肾脏小肿块的发生率较高。很大一部分患有这种肿块的患者是老年人，更虚弱的人群，对于肾细胞癌（RCC），60-70%的患者现在无症状。在目前的干预措施包括微创到标准外科手术的疾病中，需要对此类患者提出降低发病率的治疗建议。最近，针吸活检的基于形态学的诊断对治疗决策的影响已被确定为具有两个主要局限性：无法获得足够数量和质量的诊断材料用于形态学检查，以及无法在形态学上区分特定的RCC亚型。在目前的I期/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学，而是基于肾皮质肿瘤的固有差异遗传学，由Cancer Genetics，Inc.该试验的应用将允许诊断肾皮质肿瘤的四种主要亚型（三种恶性，一种良性），与形态学检查一起将影响该疾病临床管理的所有阶段。第一阶段;具体目标1：基于BAC的FISH探针组在肾组织芯活检中鉴别肾皮质肿瘤四种主要亚型的回顾性验证由于使用CGI设计的基于BAC的探针组的初步研究显示了四种组织学亚型的良好鉴别诊断，因此将在从切除标本中获得的145个活检样本的队列中验证探针组的灵敏度、特异性、阳性和阴性预测值，其中切除标本具有已知但通过标准病理学设盲的形态。阶段II;具体目标1：开发单拷贝FISH探针组。为了在不侵犯Cot-1 DNA用于杂交的专利的情况下商业化销售基于FISH的检测，将设计、繁殖、标记和验证一组单拷贝探针，用于I期使用的肾穿刺活检和手术标本的子集。第二阶段;具体目标2：前瞻性验证单拷贝FISH探针组作为一种潜在的检测方法，在针吸活检中诊断RCC。在IRB批准的研究中，将在从可疑肾脏肿块患者前瞻性获得的图像引导经皮肾活检中评价单拷贝探针组。将根据预期在2年内采集和分析的300份活检样本中已知但设盲的标准病理学形态评价分析的准确度。该项目的总体目标是开发、生产和销售一种稳健、灵敏和特异的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，影响患者治疗建议和护理成本。公共卫生相关性：随着先进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，但目前还不清楚这些肿块是恶性的，需要治疗，还是良性的。在本申请中开发和验证的基于基因的肾癌检测试剂盒可用于对此类患者获得的肿块进行针吸活检，并发症很少，将有助于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.