PROFILING SIGNALING ACTIVITY AND GENE EXPRESSION IN SINGLE, PANCREATIC ADENOCARCINOMA CELLS USING CE-RNA-SEQ

使用 CE-RNA-SEQ 分析单个胰腺腺癌细胞中的信号传导活性和基因表达

基本信息

批准号：
10200700
负责人：
Nancy L. Allbritton
金额：
$ 58.45万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-04-01 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10200700
关键词：
Adenocarcinoma Cell Adjuvant Therapy Adult American Cancer Society Biological Assay Biological Models Biopsy Cancer Etiology Capillary Electrophoresis Cell Count Cell Line Cells Cessation of life Characteristics Chemotherapy and/or radiation Classification Clinical Collection Complex Mixtures Computing Methodologies Consensus Data Data Set Development Diagnosis Disease Early Diagnosis Excision Future Gene Expression Gene Expression Profile Gene Expression Profiling Genes Goals Heterogeneity Human Individual Interdisciplinary Study International Invaded KRAS2 gene Lead Link Malignant - descriptor Malignant Epithelial Cell Malignant Neoplasms Malignant neoplasm of pancreas Measures Messenger RNA Methods Minority Molecular Molecular Profiling Mus Mutation Neoplasm Metastasis Nomenclature Normal tissue morphology Nude Mice Operative Surgical Procedures Organ PI3 gene Pancreas Pancreatic Adenocarcinoma Pancreatic Ductal Adenocarcinoma Pathway interactions Patients Pattern Phenotype Phosphotransferases Process Production Property Protein Kinase C Proto-Oncogene Proteins c-akt RNA Regimen Reporter Reporting Research Research Personnel Ribosomal Protein S6 Kinase Sample Size Sampling Sentinel Signal Pathway Signal Transduction Signal Transduction Pathway Stromal Cells System Technology Testing The Cancer Genome Atlas Therapeutic Time Tissues Tumor Subtype Tumor Tissue United States Work Xenograft procedure cancer genome cancer subtypes cell type cohort design improved innovation insight instrumentation molecular targeted therapies multidisciplinary neoplastic cell novel novel strategies p21 activated kinase pancreatic cancer cells pancreatic ductal adenocarcinoma cell patient derived xenograft model precision medicine progenitor prognostic significance prognostic value rare cancer reduce symptoms response targeted treatment technology validation tool transcriptome sequencing tumor tumor heterogeneity

项目摘要

Abstract Pancreatic ductal adenocarcinoma is a devastating disease in dire need of improved therapies targeted at specific signaling pathways. Strategies to molecularly profile aberrant pancreatic tissue and inform targeted therapeutic decisions would be of immense value in patient treatment. However, molecular profiling is extremely challenging since biopsied patient tissue is a complex mixture of normal and malignant pancreatic cells. Furthermore, there is a growing understanding that mutations and gene expression alone do not tightly correlate with clinical response. In the current application, a multidisciplinary research team proposes to develop a state-of-the-art, single-cell, platform technology to measure the catalytic activity of sentinel kinases within the KRAS pathway and gene expression through RNA sequencing. The investigators will optimize and validate microsampling and microelectrophoresis methods to assay single cells, simultaneously overcoming the challenges of cellular heterogeneity and sample-size limits. Novel reporters of kinase activity within KRAS-outflow signaling pathways will be designed and new methods and instrumentation combining single-cell capillary electrophoresis with efficient RNA capture will be pioneered. Human tumor samples maintained in murine xenografts will be assayed to gain unique insights into tumor properties not currently addressable. The work will directly link mRNA production with the catalytic activity of kinases in individual tumor cells derived from patients. The technology will enable questions such as which kinase signaling patterns drive the classical vs the basal phenotypes of pancreatic adenocarcinoma and whether a single tumor possesses a mixture of classical and basal-type cells. The data and the insights gained from implementation of this technology will provide a new approach for clinical assays with the potential for a profound impact on therapeutic strategies in the emerging field of precision medicine.

抽象的胰腺导管腺癌是一种毁灭性疾病，迫切需要改进的治疗方法针对特定的信号通路。异常胰腺的分子分析策略组织并告知有针对性的治疗决策对患者具有巨大价值治疗。然而，分子分析极具挑战性，因为活检患者组织是正常和恶性胰腺细胞的复杂混合物。此外，还有越来越多的了解突变和基因表达本身与临床并不紧密相关回复。在当前的应用中，一个多学科研究团队建议开发一种最先进的单细胞平台技术，用于测量哨兵的催化活性通过 RNA 测序了解 KRAS 通路中的激酶和基因表达。这研究人员将优化和验证微采样和微电泳方法分析单细胞，同时克服细胞异质性的挑战样本量限制。 KRAS 流出信号通路中激酶活性的新型报告基因将设计结合单细胞毛细管的新方法和仪器具有高效 RNA 捕获功能的电泳将成为首创。人类肿瘤样本将对小鼠异种移植物中保存的细胞进行分析，以获得对肿瘤特性的独特见解目前无法寻址。这项工作将直接将 mRNA 的产生与催化联系起来来自患者的个体肿瘤细胞中激酶的活性。该技术将使诸如哪些激酶信号传导模式驱动经典表型与基础表型等问题胰腺癌以及单个肿瘤是否具有经典和经典的混合基底型细胞。从实施该技术中获得的数据和见解将为临床检测提供了一种新方法，有可能对临床检测产生深远影响精准医学这一新兴领域的治疗策略。