Molecular Diagnostics for Malignant Effusion

描述（由申请人提供）：长期目标是确定应用创新分子方法提高积液标本癌症诊断的可行性。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临床肿瘤学家最好地管理患者至关重要。传统上，基于细胞形态学特征的细胞学评估用于诊断恶性和良性积液，并确定恶性积液中癌的原发部位。不幸的是，这种形态学检查本质上是主观的，往往不能发现恶性细胞或在恶性积液中确定癌的起源。我们假设1)检测肿瘤释放的DNA和分泌的蛋白标记物（HLA-G， kallikrein 5和kallikrein 8）中的肿瘤相关遗传改变（等位基因失衡和DNA完整性增加）单独或联合为提高诊断恶性腹水的敏感性和特异性提供了新的分子方法；2)使用基于基因表达的癌症分类图可以精确预测恶性积液的原发部位。为了验证这两种假设，我们建议检测肿瘤释放DNA的多种分子遗传变化，并测量腹水上清中分泌HLA-G、钾化因子5和钾化因子8的水平。我们将建立一个基于有限基因表达的癌症分类图，以预测恶性积液中的癌症类型。

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.