A cell-based liquid biopsy approach for early pancreatic cancer detection

用于早期胰腺癌检测的基于细胞的液体活检方法

• 批准号: 9288144
• 负责人: Neha Bhagwat
• 金额: $ 5.3万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9288144
• 关键词: Alpha Cell; Binding; Biological Assay; Blood; Cancer Detection; Cell Separation; Cells; Cohort Studies; Cystic Lesion; Data; Detection; Development; Devices; Diagnosis; Disease; Disease Progression; Early Diagnosis; Epithelial Cells; Flow Cytometry; Fluorescent in Situ Hybridization; General Population; Genes; Genetically Engineered Mouse; Goals; Growth; Guidelines; Hematopoietic; Human; Individual; Knowledge; Label; Lesion; Magnetic nanoparticles; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Microfluidic Microchips; Microfluidics; Mus; Mutation; Nature; Pancreas; Pancreatic Adenocarcinoma; Pancreatic Ductal Adenocarcinoma; Patients; Pilot Projects; Population; Predictive Value; Premalignant; Premalignant Cell; RNA; Sensitivity and Specificity; Staging; Stream; Techniques; Technology; Testing; Time; Tumor-Derived; Whole Blood; advanced disease; base; biomarker identification; cancer survival; cohort; high risk; high risk population; liquid biopsy; mouse model; neoplastic cell; next generation sequencing; novel marker; outcome forecast; pancreatic cell line; public health relevance; response; screening; transcriptome sequencing; tumor; tumor progression

 DESCRIPTION (provided by applicant): Pancreatic adenocarcinoma (PDA) is a lethal malignancy with a particularly poor prognosis due to the advanced nature of the disease at the time of diagnosis in most patients. Currently, there are no effective screening strategies for earl detection of PDA. Studies from our lab have demonstrated that pancreatic epithelial cells are shed into the blood stream at early stages of disease in genetically engineered mouse models as well as in patients with pre-malignant cystic lesions of the pancreas. These observations present a unique opportunity to use these cells as the basis for screening for PDA. The primary goal of this project is to develop a robust non-invasive test that can be used to detect early stage pancreatic ductal adenocarcinoma (PDA) based on the presence of circulating epithelial cells of pancreatic origin in blood. We will use magnetic nanoparticle technology and a microfluidic platform to enrich circulating epithelial cells (CECs) from whole blood followed by RNA-based detection, including RNA fluorescent in situ hybridization (RNA FISH) and quantitative PCR. We will validate this using a lineage-labeled genetically engineered mouse model of PDA in which CECs can be easily detected in the blood. Further, we will utilize the optimized device along with additional single cell isolation techniques such as the DEPArray(tm) technology or flow cytometry to perform next-generation sequencing on CECs in order to (i) identify novel biomarkers for more effective screening and diagnosis and (ii) investigate functional differences between CECs at different stages of pancreatic cancer progression. Finally we will implement a pilot study in a cohort of patients with known PDA to enrich and detect CECs in the blood using the assay developed in previous aims. We envision that this strategy could be adapted for a number of other applications including evaluating response to therapy, disease-staging and screening of high-risk populations, thereby leading to improvements in pancreatic cancer survival.

 描述（由申请人提供）：胰腺腺癌（PDA）是一种致命的恶性肿瘤，由于大多数患者在诊断时疾病已处于晚期，因此预后特别差。目前，尚无有效的早期检测PDA的筛查策略。我们实验室的研究表明，在基因工程小鼠模型以及患有胰腺癌前囊性病变的患者中，胰腺上皮细胞在疾病早期就会脱落到血流中。这些观察结果为使用这些细胞作为筛选 PDA 的基础提供了独特的机会。该项目的主要目标是开发一种强大的非侵入性测试，可用于根据血液中胰腺来源的循环上皮细胞的存在来检测早期胰腺导管腺癌（PDA）。我们将使用磁性纳米颗粒技术和微流体平台从全血中富集循环上皮细胞 (CEC)，然后进行基于 RNA 的检测，包括 RNA 荧光原位杂交 (RNA FISH) 和定量 PCR。我们将使用谱系标记的 PDA 基因工程小鼠模型来验证这一点，在该模型中，可以轻松地在血液中检测到 CEC。此外，我们将利用优化的设备以及其他单细胞分离技术（例如 DEPArray(tm) 技术或流式细胞术）对 CEC 进行新一代测序，以便 (i) 识别新的生物标志物以进行更有效的筛查和诊断，以及 (ii) 研究胰腺癌进展不同阶段 CEC 之间的功能差异。最后，我们将在一组患有已知 PDA 的患者中实施一项试点研究，以使用先前目标中开发的检测方法富集和检测血液中的 CEC。我们设想该策略可以适用于许多其他应用，包括评估治疗反应、疾病分期和高危人群筛查，从而提高胰腺癌的生存率。