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改善渗出液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的常见临床表现。这些渗出液标本的诊断对于临床肿瘤学家最好地管理患者至关重要。传统上，基于细胞的形态学特征进行细胞学评价以诊断恶性与良性积液，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，并且经常不能检测恶性细胞或识别恶性积液中癌的起源。我们假设：1）检测肿瘤相关的遗传变异（等位基因不平衡和DNA完整性增加）在肿瘤释放的DNA和分泌的蛋白质标志物（HLA-G，激肽释放酶5和激肽释放酶8）单独或组合提供了新的分子方法来提高诊断恶性腹水的灵敏度和特异性，以及2）可以使用基于基因表达的癌症分类图谱精确预测恶性积液的原发部位。为了检验这两种假设，我们建议检测肿瘤释放DNA中的多个分子遗传变化，并测量腹水上清液中分泌型HLA-G、激肽释放酶5和激肽释放酶8的水平。我们将建立一个癌症分类图的基础上表达的一组有限的基因来预测恶性积液中的癌症类型。 由于传统的形态学方法在诊断渗出液中的肿瘤方面存在局限性，临床肿瘤学迫切需要针对肿瘤分子诊断的转化研究。该提案的总体意义是开发创新的分子检测方法，以协助存在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，但准确的诊断以及针对不同类型癌症的适当治疗方案可能会对结果产生积极影响。这项提议的结果可能会为分子细胞学引入新的范式，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.