Pancreatic Cancer Protein Biomarkers for Early Detection

描述（由申请人提供）：胰腺癌是一种致命的疾病，因为大多数患者在癌症扩散到无法手术之前没有任何症状。此外，一旦癌症形成，化疗对生存的改善微乎其微。2厘米或更小的肿瘤比那些更大的有更好的生存结果。我们建议开发一种基于血清的胰腺癌早期检测方法。几乎所有的血清癌症生物标志物检测都是基于蛋白质的检测。因此，我们将使用蛋白质组学技术来鉴定胰腺癌和癌前病变特异性的蛋白质。我们的初步数据是令人兴奋的：1)我们已经在胰腺癌组织中鉴定并量化了656个蛋白，其中90个蛋白在癌症中上调，61个蛋白下调，至少发生了2倍的变化。2)迄今为止的验证研究表明，蛋白质组学测量是稳健和准确的。对这些蛋白质进行了功能分析，近四分之一是细胞外/膜蛋白-因此是胰腺癌生物标志物开发的绝佳候选者（膜蛋白脱落到血液中）。3)发现的两种蛋白生物标志物，Annexin II和Integrin B1，已被进一步研究；这两种蛋白在癌细胞中均有过表达，但在正常胰管或非胰管细胞中均无过表达。4)其中一种蛋白，Galectin 1，已通过western blotting验证，然后开发成ELISA。在初步试验中，凝集素1与正常血清鉴别胰腺癌的准确率为100%。5)我们开发了一种MALDI-TOF-TOF方法，用于直接从血清中测量蛋白质，因此最终不需要ELISA进行生物标志物检测。在本提案中，我们描述了综合开发候选胰腺癌生物标志物的方法，以及用于确定哪些生物标志物最适合开发为临床可用测试的算法。这里提出的研究有可能显著改变这种疾病的结果，这种疾病在过去100年里进展甚微。蛋白质组学技术的应用具有巨大的潜力。我们提出的研究可以导致：早期诊断-改善这种致命疾病的预后；加深对胰腺癌发生机制的认识用于未来治疗干预的靶向蛋白，包括新药/疫苗开发；疾病反应标志物的发展。

DESCRIPTION (provided by applicant): Pancreatic cancer is a uniformly lethal disease because most patients have no symptoms until the cancer has spread and become inoperable. Moreover, once the cancer has formed, chemotherapy offers only minimal improvement in survival. Cancers that are 2 cm or smaller have a much better survival outcome than those that are larger. We propose to develop a serum based early detection test for pancreatic cancer. Nearly all serum cancer biomarker tests are based on the detection of proteins. Therefore, we will use proteomics technology to identify the proteins that are specific to pancreatic cancer and pre-cancer. Our preliminary data is exciting: 1) we have already identified and quantified 656 proteins in pancreatic cancer tissue, of which 90 proteins were up-regulated and 61 were down-regulated in cancer by at least a 2-fold change. 2) Validation studies to date suggest that the proteomics measurements are robust and accurate. The proteins have been analyzed with regard to function, nearly one quarter are extracellular/membrane proteins- thus excellent candidates for pancreatic cancer biomarker development (membrane proteins are shed into the blood). 3) Two of the discovered protein biomarkers, Annexin II and Integrin B1, have been further investigated; both of these proteins have marked over-expression in cancer cells, but not in normal pancreatic ducts or in non-ductal pancreatic cells when evaluated by tissue array and immunohistochemistry. 4) One of the proteins, Galectin 1, has been validated by western blotting and then developed into an ELISA. Galectin 1 could distinguish pancreatic cancer versus normal sera with 100% accuracy in preliminary testing. 5) We have developed a MALDI-TOF-TOF methodology for measuring proteins directly from serum, so that ELISA's would eventually not be necessary for biomarker detection. In this proposal, we describe our methods for the comprehensive development of candidate pancreatic cancer biomarkers and the algorithm that will be used to determine which biomarkers are best suited for development into a clinically useable test. The studies proposed here have the potential to significantly change the outcome in a disease that has made minimal headway in the past 100 years. The use of proteomics technology has enormous potential. Our proposed studies can lead to: earlier diagnosis- improving the prognosis of this deadly disease; better understanding of tumorigenesis in pancreatic cancer; targeted proteins for future therapeutic interventions including new drug/vaccine development; development of disease response markers.