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.

抽象的 当前，包括癌症在内的非常早期疾病的鉴定是目前最有前途的方法 降低胰腺导管腺癌（PDAC）死亡率。早期开发血清生物标志物 诊断对于位于胰腺中的肿瘤特别重要，胰腺是一种相对无法接近的器官 考试。在PDAC中寻找临床上有用的生物标志物一直具有挑战性。定义明确的 从正常的导管上皮到入侵PDAC的遗传途径表明，遗传改变早于 侵入性和无法治愈的癌症的发展。因此，有可能检测所得蛋白质和 PTM在肿瘤发展到无法治愈的阶段发生变化。不幸的是，所有生物标志物 迄今为止，已调查缺乏作为临床有用筛查测试的敏感性和特异性。这 因此，提案解决了主要的未满足临床需求：目前，尚未制定策略 有效检测早期PDAC。我们最近开发了两种互补方法来识别 潜在的生物标志物。第一次试图使用血清中PDAC衍生的蛋白质生物标志物识别和量化血清中的蛋白质生物标志物 由培养的PDAC细胞产生和/或分泌的同位素质量标记的蛋白质组，作为内标。 第二种方法量化了选定蛋白酶的活动，这是一类发挥重要作用的酶 在癌症生物学中的作用。我们建议扩展我们拥有的血清蛋白和酶生物标志物 通过提高分析灵敏度并分析精心收集的统计相关和 性别匹配和年龄匹配的样本集（PDAC；前癌症；慢性胰腺炎；等）。我们努力的中心 将是（1）基于质谱的蛋白质定量和基于荧光的蛋白酶活性测定，（2） 癌细胞，分泌组和微小的深层蛋白质组和磷蛋白组分析 PDAC肿瘤组织的量和（3）肿瘤细胞类型特异性细胞和分泌蛋白参考 标准标准，称为超锡和Sec-super-silac，可以准确地相对定量 生物液和肿瘤组织中的蛋白质。我们建议在以下三个中检验这些假设 高度综合的研究目的：（1）我们将表征细胞，分泌和肿瘤组织蛋白的面板 针对PDAC和血清蛋白酶的面板特定，并优先考虑候选生物标志物以进一步 根据四个标准满足开发。 （2）然后我们将研究蛋白质和蛋白酶生物标志物候选物 人血清。我们假设这些生物标志物候选者的一个子集在 早期PDAC与良性胰腺疾病的血清。 （3）表现出承诺的生物标志物将受到约束 通过使用较大的样品队列构建混合（即蛋白质和蛋白酶活性）多变量来验证 PDAC预后模型。该模型的性能特征将得到确定和验证 临床用途。开发有效的无创生物标志物，用于筛查有PDAC风险的患者 目前不存在，将对这种疾病的治疗和临床过程产生重大影响。