Protein and proteolytic activity biomarkers of early stage pancreatic cancer

早期胰腺癌的蛋白质和蛋白水解活性生物标志物

• 批准号: 10090577
• 负责人: PAUL TEMPST
• 金额: $ 62.35万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-03 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10090577
• 关键词: Address; Age; Award; Benign; Biological; Biological Assay; Biological Markers; Biology; Body Weight decreased; Cachexia; Cancer Biology; Cancer Etiology; Carcinoma in Situ; Cessation of life; Characteristics; Chemistry; Clinical; Clinical Course of Disease; Colorectal Cancer; Deltastab; Detection; Development; Diagnosis; Disease; Ductal Epithelium; Early Diagnosis; Enzymes; Fluorescence; Fractionation; Gender; Genetic; Grant; Human; Isotopes; Label; Liquid substance; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Mass Spectrum Analysis; Methods; Minority; Modeling; Mutation; Neoplasm Metastasis; Operative Surgical Procedures; Organ; Pancreas; Pancreatic Diseases; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Pattern; Peptide Hydrolases; Performance; Phosphoproteins; Play; Prognosis; Proteins; Proteolysis; Proteome; Proteomics; Public Health; Recurrence; Reference Standards; Resected; Risk; Role; Sampling; Screening procedure; Sensitivity and Specificity; Serum; Serum Proteins; Symptoms; Testing; Tumor Suppression; Tumor Tissue; base; biomarker identification; biomarker panel; cancer cell; cancer type; candidate marker; cell type; chronic pancreatitis; clinical application; clinical biomarkers; cohort; detection method; differential expression; early detection biomarkers; improved; innovation; insight; instrumentation; malignant breast neoplasm; mortality; neoplastic cell; pancreatic ductal adenocarcinoma cell; patient screening; potential biomarker; premalignant; prognostic model; protein biomarkers; protein degradation; screening; tumor; tumor progression

ABSTRACT Identification of very early stage disease, including cancer in situ, is currently the most promising approach to reduce pancreatic ductal adenocarcinoma (PDAC) mortality. Development of serum biomarkers for early diagnosis is particularly significant for tumors located in the pancreas, an organ that is relatively inaccessible to examination. The search for clinically useful biomarkers in PDAC has been challenging. The well-defined genetic pathway from normal ductal epithelium to invasive PDAC suggests that genetic alterations predate development of invasive and incurable cancer. As such, there is the potential to detect the resulting protein and PTM changes before the tumor has progressed to an incurable stage. Unfortunately, all biomarkers investigated to date lack the sensitivity and specificity to serve as a clinically useful screening test. This proposal therefore addresses a major unmet clinical need: currently, no strategy has been developed to effectively detect early stage PDAC. We have recently developed two complementary approaches to identify potential biomarkers. The first seeks to identify and quantify PDAC-derived protein biomarkers in serum using an isotopic mass-labeled proteome, produced and/or secreted by cultured PDAC cells, as an internal standard. The second approach quantifies activities of selected proteases, a class of enzymes that plays an important role in cancer biology. We propose to expand upon the serum protein and enzymatic biomarkers that we have discovered, by improving the analytical sensitivity and by analyzing carefully collected, statistically relevant and gender- and age-matched sample sets (PDAC; pre-cancerous; chronic pancreatitis; etc.). Central to our efforts will be (1) mass spectrometry-based protein quantitation and fluorescence-based protease activity assays, (2) deep and quantitative proteome and phosphoproteome analysis of cancer cells, secretomes and minute amounts of PDAC tumor tissues, and (3) tumor cell type-specific cellular and secreted protein reference standards, termed Super-SILAC and SEC-super-SILAC, that enable accurate relative quantitation of selected proteins in biological fluids and tumor tissues. We propose to test these hypotheses under the following three highly integrated study aims: (1) We will characterize panels of cellular, secreted, and tumor tissue proteins specific for PDAC as well as panels of serum proteases and prioritize candidate biomarkers for further development based on four criteria met. (2) We will then study protein and protease biomarker candidates in human serum. We hypothesize a subset of these biomarker candidates are differentially expressed or active in serum from early PDAC versus benign pancreatic disease. (3) Biomarkers that show promise will be subjected to verification by using larger sample cohorts to build mixed (i.e., proteins and protease activities) multivariate PDAC prognostic models. Performance characteristics of this model will be determined and validated for clinical use. Development of effective, noninvasive biomarkers for screening patients at risk for PDAC, where none currently exist, will have a significant impact on the treatment and clinical course of this disease.

摘要 识别非常早期的疾病，包括原位癌，是目前最有希望的方法 降低胰腺导管腺癌(PDAC)死亡率。早期血清生物标志物的研究进展 诊断对位于胰腺的肿瘤尤其重要，胰腺是一种相对难以接近的器官 考试。在PDAC中寻找临床有用的生物标记物一直是具有挑战性的。定义明确的 从正常导管上皮到侵袭性PDAC的遗传途径表明基因改变早于 发展为浸润性和不可治愈的癌症。因此，有可能检测到产生的蛋白质和 在肿瘤进展到无法治愈的阶段之前，PTM会发生变化。不幸的是，所有的生物标志物 到目前为止，研究缺乏作为临床有用的筛查试验的敏感性和特异性。这 因此，该提案解决了一个主要的未得到满足的临床需求：目前，还没有制定战略来 有效检测早期PDAC。我们最近开发了两种互补的方法来确定 潜在的生物标志物。第一个目的是识别和定量血清中PDAC衍生的蛋白生物标志物 一种同位素质量标记的蛋白质组，由培养的PDAC细胞产生和/或分泌，作为内部标准。 第二种方法量化选定的蛋白酶的活性，这是一类发挥重要作用的酶 在癌症生物学中的作用。我们建议对我们已有的血清蛋白和酶生物标志物进行扩展 发现，通过提高分析灵敏度和分析仔细收集的统计相关性和 性别和年龄匹配的样本集(PDAC；癌前病变；慢性胰腺炎等)。对我们的努力至关重要 将会有(1)基于质谱学的蛋白质定量和基于荧光的蛋白酶活性分析，(2) 癌细胞、分泌体和微小蛋白质组的深层定量蛋白质组和磷蛋白质组分析 PDAC肿瘤组织的数量，以及(3)肿瘤细胞类型特定的细胞和分泌蛋白参考 标准，称为Super-SILAC和SEC-Super-SILAC，能够准确地相对定量选定的 生物液和肿瘤组织中的蛋白质。我们建议在以下三个方面检验这些假设 高度集成的研究目标：(1)我们将表征细胞、分泌和肿瘤组织蛋白质组 针对PDAC以及血清蛋白水解酶的面板，并优先选择候选生物标记物 基于符合四个标准的发展。(2)然后，我们将研究蛋白质和蛋白酶生物标记物的候选 人血清。我们假设这些候选生物标记物的子集在 早期PDAC的血清与良性胰腺疾病的对比。(3)有希望的生物标记物将受到影响 通过使用更大的样本队列来建立混合(即蛋白质和蛋白酶活性)多变量进行验证 PDAC预后模型。将确定并验证此型号的性能特征 临床应用。开发有效的、非侵入性的生物标志物用于筛查有PDAC风险的患者，其中 目前都不存在，将对这种疾病的治疗和临床病程产生重大影响。