Molecular Diagnostics for Malignant Effusion

描述(由申请人提供)：长期目标是确定应用创新的分子方法来改进渗出标本中癌症诊断的可行性。腹水(腹膜)或胸腔积液是肿瘤和各种良性疾病患者常见的临床表现。这些积液标本的诊断对于临床肿瘤学家最好地处理患者是至关重要的。传统上，细胞学评估是根据细胞的形态特征来诊断恶性和良性积液，并在恶性积液中确定癌的原发部位。不幸的是，这种形态检查本质上是主观的，往往无法发现恶性细胞或在恶性积液中确定癌的来源。我们假设：1)检测肿瘤释放的DNA中与肿瘤相关的遗传变化(等位基因失衡和DNA完整性增加)和分泌的蛋白标记物(HLA-G、激肽释放酶5和激肽释放酶8)单独或联合提供新的分子方法，以提高诊断恶性腹水的敏感性和特异性；2)可以使用基于基因表达的癌症分类图精确预测恶性积液的原始部位。为了验证这两种假设，我们建议检测肿瘤释放的DNA中的多个分子遗传变化，并检测腹水上清液中分泌的HLA-G、激肽释放酶5和激肽释放酶8的水平。我们将建立一个基于有限一组基因表达的癌症分类图，以预测恶性胸腔积液中的癌症类型。 由于传统的诊断积液标本中肿瘤的形态学方法的局限性，临床肿瘤学迫切需要针对肿瘤分子诊断的转译研究。这项建议的总体意义是开发创新的分子分析方法，以帮助出现积液的患者进行癌症诊断，积液是癌症患者的常见临床表现。虽然这些患者通常出现在晚期，但准确的诊断和针对不同类型癌症的适当治疗方案可以对预后产生积极影响。这一建议的结果可能会为分子细胞学引入新的范例，并为肿瘤学家提供管理患者的新视角。

DESCRIPTION (provided by applicant): The long-term objective is to determine the feasibility of applying innovative molecular approaches to improve cancer diagnosis in effusion specimens. Ascites (peritoneal) or pleural effusion is a common clinical presentation in patients with neoplastic and a variety of benign diseases. Diagnosis of these effusion specimens has paramount importance for clinical oncologists to best manage patients. Traditionally, cytological evaluation based on morphological features of cells is performed to diagnose malignant versus benign effusions and to determine the primary sites of carcinomas in malignant effusions. Unfortunately, such morphological examination is subjective in nature and often fails to detect malignant cells or identify the origin of the carcinoma in a malignant effusion. We hypothesize that 1) detection of tumor-associated genetic alterations (allelic imbalance and increased DNA integrity) in tumor-released DNA and the secreted protein markers (HLA-G, kallikrein 5 and kallikrein 8) alone or in combination provide novel molecular approaches to increase the sensitivity and specificity in diagnosing malignant ascites and 2) the primary site of a malignant effusion can be precisely predicted using a gene expression based cancer classification map. To test both hypotheses, we propose to detect multiple molecular genetic changes in tumor-released DNA and measure the levels of secretory HLA-G, kallikrein 5 and kallikrein 8 in the ascites supernatant. We will establish a cancer classification map based on the expression of a limited set of genes to predict the cancer types in malignant effusions. Translational researches aiming at the molecular diagnostics for cancer are urgently needed in clinical oncology because of the limitations in traditional morphological approaches to diagnose cancer in effusion specimens. The overall significance of this proposal is to develop innovative molecular assays to assist cancer diagnosis in patients who present effusion, a common clinical presentation in cancer patients. Although these patients often present at a late stage, outcome can be positively affected by accurate diagnoses followed by appropriate therapeutic regimens specific to different types of carcinoma. The results from this proposal will likely introduce new paradigms to molecular cytology and provide oncologists a new perspective in managing their patients.