Mechanistic Studies of MUC4 and NGAL in Pancreatic Adenocarcinoma

MUC4和NGAL在胰腺癌中的作用机制研究

• 批准号: 8256488
• 负责人: Michael James Baine
• 金额: $ 2.22万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-16 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8256488
• 关键词: Adenocarcinoma; Biochemical; Biochemical Pathway; Biological Markers; Biological Models; Cancer Biology; Cancer Patient; Cancer Prognosis; Cancer cell line; Carcinoma; Carcinoma in Situ; Cell Line; Development; Diagnosis; Diagnostic; Disease; Doctor of Philosophy; Down-Regulation; ERBB2 gene; Early Diagnosis; Future; Gelatinase A; Genes; Human; Immunocompetent; Immunohistochemistry; In Vitro; Investigation; Knowledge; Lesion; Link; Literature; Malignant Neoplasms; Malignant neoplasm of pancreas; Membrane; Methods; Modeling; Mucins; Mus; Neoplasm Metastasis; Outcome; PI3K/AKT; Pancreas; Pancreatic Adenocarcinoma; Pancreatic Intraepithelial Neoplasia; Pathway interactions; Patients; Pattern; Physicians; Plasma; Play; Prognostic Marker; Proteins; Regulation; Reporting; Role; Scientist; Screening procedure; Staging; Students; Surrogate Markers; Symptoms; Testing; Therapeutic; Time; Tissue Microarray; Tissues; Training; Translational Research; Treatment Efficacy; Tumor Tissue; Tumorigenicity; Western Blotting; Work; base; career; clinical decision-making; design; expectation; high risk; human disease; immunocytochemistry; improved; interest; man; neoplastic; novel; outcome forecast; overexpression; pancreatic cancer cells; pancreatic neoplasm; peripheral blood; prognostic; research study; secretory protein; tumor; tumor growth; tumor progression; tumorigenic

DESCRIPTION (provided by applicant): Pancreatic cancer remains the most lethal cancer known to man, owing primarily to its lack of early symptom resulting in late diagnosis and lack of consistent treatment efficacy. MUC4 is a large-sized membrane- anchored mucin which is aberrantly overexpressed by most human carcinomas. We have previously reported that MUC4 promotes pancreatic tumor growth and metastasis by multiple mechanisms. In parallel to these studies, using immunohistochemistry in tissue microarrays containing well characterized tumors tissues, we observed elevated expression of neutrophil gelatinase associated lipocalin (NGAL), also known as Lipocalin 2 or uterocalin, during pancreatic cancer progression. Interestingly, the expression of NGAL correlated with MUC4 expression in these tissues. Immunocytochemistry showed co-expression of NGAL and MUC4 in pancreatic cancer tissue sections. Recent studies by our group have shown that NGAL is a potential diagnostic biomarker and its expression is upregulated as early as the preneoplastic PanIN-2 stage. Additionally, preliminary studies have shown that, in the pancreatic cancer cell line CaPan-1, MUC4 knockdown leads to decreased expression of NGAL. In the present study, I will further investigate the link between NGAL and MUC4 in pancreatic cancer using the cell lines CoLo-357, CaPan-1, MiaPaCa, and PANC-1 as well as healthy and neoplastic pancreatic tissue from both humans and murine models in pursuit of the hypothesis that MUC4, acting through HER2, regulates NGAL expression in pancreatic cancer and that this connection will prove highly relevant to the prognostic abilities of NGAL as a pancreatic cancer surrogate biomarker. In pursuit of this investigation I have formulated three specific aims. In the first specific aim, I will use knockdown and overexpression experiments as well as Western blotting, Quantitative-Real Time PCR, and immunohistochemistry to determine if expression of NGAL and MUC4 are reliably linked in pancreatic cancer and if this link is present at the translational and/or transcriptional level. In the second aim, I will elucidate the pathway by which NGAL and MUC4 expression are linked using currently-known upstream and downstream proteins and pathways as a starting-point. Specifically, based on literature and my preliminary results, I will begin by assessing if MUC4 regulates NGAL expression through HER2. In the third aim, I will use an immunocompetent murine model genetically manipulated to spontaneously produce pancreatic cancer that has been developed in our lab to assess the correlation between tissue and plasma NGAL and tissue MUC4 levels throughout the development of and advancement of pancreatic cancer. Through these aims, I expect to conclusively elucidate the biochemical connection between MUC4 and NGAL while improving on current evidence potentiating NGAL's utility as both an early diagnostic and prognostic surrogate biomarker for pancreatic cancer. PUBLIC HEALTH RELEVANCE: Pancreatic cancer remains the most lethal human malignancy, owing mostly to a lack of early diagnosis and abysmal treatment efficacy in late-stage disease. Preliminary evidence suggests that MUC4, one of the earliest and most differentially upregulated genes in pancreatic cancer and which positively correlates with metastatic and tumorigenic potential, is upstream of NGAL, a secreted protein also found to be overexpressed in pancreatic tumor tissues as well as in the peripheral blood of pancreatic cancer patients. This study aims to define the mechanism by which MUC4 and NGAL expressions are linked, thereby bolstering the evidence for peripheral blood NGAL's use as a potential early diagnostic and prognostic marker in pancreatic cancer.

描述（由申请人提供）：胰腺癌仍然是人类已知的最致命的癌症，主要是由于其缺乏早期症状，导致晚期诊断和缺乏一致的治疗效果。MUC 4是一种大尺寸的膜锚定粘蛋白，其在大多数人类癌中异常过表达。我们以前曾报道MUC 4通过多种机制促进胰腺肿瘤的生长和转移。在这些研究的同时，使用免疫组织化学在含有良好表征的肿瘤组织的组织微阵列中，我们观察到在胰腺癌进展期间中性粒细胞明胶酶相关脂质运载蛋白（NGAL）（也称为脂质运载蛋白2或子宫运载蛋白）的表达升高。有趣的是，在这些组织中NGAL的表达与MUC 4的表达相关。免疫细胞化学显示NGAL和MUC 4在胰腺癌组织切片中共表达。我们小组最近的研究表明，NGAL是一种潜在的诊断生物标志物，其表达早在肿瘤前PanIN-2阶段就上调。此外，初步研究表明，在胰腺癌细胞系CaPan-1中，MUC 4敲低导致NGAL表达降低。在本研究中，我将使用细胞系科洛-357、CaPan-1、MiaPaCa和PANC-1以及来自人和鼠模型的健康和肿瘤胰腺组织进一步研究胰腺癌中NGAL和MUC 4之间的联系，以追求MUC 4通过HER 2起作用的假设，调节胰腺癌中NGAL的表达，并且这种联系将证明与NGAL作为胰腺癌替代生物标志物的预后能力高度相关。在进行这项调查时，我提出了三个具体目标。在第一个具体目标中，我将使用敲低和过表达实验以及Western印迹，定量实时PCR和免疫组织化学来确定NGAL和MUC 4的表达是否在胰腺癌中可靠地联系，以及这种联系是否存在于翻译和/或转录水平。在第二个目标中，我将使用目前已知的上游和下游蛋白质和途径作为起点，阐明NGAL和MUC 4表达的连接途径。具体而言，基于文献和我的初步结果，我将开始评估MUC 4是否通过HER 2调节NGAL表达。在第三个目标中，我将使用我们实验室开发的经遗传操作自发产生胰腺癌的免疫活性小鼠模型来评估胰腺癌发展和进展过程中组织和血浆NGAL与组织MUC 4水平之间的相关性。通过这些目标，我希望最终阐明MUC 4和NGAL之间的生化联系，同时改善目前的证据，增强NGAL作为胰腺癌早期诊断和预后替代生物标志物的效用。 公共卫生相关性：胰腺癌仍然是最致命的人类恶性肿瘤，主要是由于缺乏早期诊断和晚期疾病的治疗效果差。初步证据表明，MUC 4是胰腺癌中最早和最差异上调的基因之一，与转移和致瘤潜力正相关，是NGAL的上游，NGAL是一种分泌蛋白，也发现在胰腺肿瘤组织以及胰腺癌患者的外周血中过表达。本研究旨在确定MUC 4和NGAL表达相关的机制，从而支持外周血NGAL作为胰腺癌早期诊断和预后标志物的证据。