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

• 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
• 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

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迫切需要改善疗法的毁灭性疾病 针对特定的信号通路。分子剖面异常胰腺的策略 组织并告知有针对性的治疗决定在患者中具有巨大的价值 治疗。但是，由于活检的患者组织，分子分析非常具有挑战性 是正常和恶性胰腺细胞的复杂混合物。此外，还有一个增长 了解仅突变和基因表达不会与临床密切相关 回复。在当前的应用程序中，一个多学科研究小组建议开发 最先进的单细胞平台技术，用于测量哨兵的催化活动 KRAS途径内的激酶和基因表达通过RNA测序。这 研究人员将优化和验证微采样和微电泳方法 测定单细胞，同时克服细胞异质性和 样品尺寸限制。 KRAS流出信号通路中激酶活性的新记者 将设计和结合单细胞毛细管的新方法和仪器 具有有效RNA捕获的电泳将开创。人类肿瘤样品 将测定在鼠异种移植物中维持的，以获得对肿瘤特性的独特见解 目前不可解决。这项工作将直接将mRNA产生与催化 激酶在来自患者的单个肿瘤细胞中的活性。该技术将启用 诸如哪些激酶信号传导模式驱动经典与基础表型的问题 胰腺腺癌以及单个肿瘤是否具有经典和 基础型细胞。该技术实施中获得的数据和见解将会 提供一种新的临床测定方法，有可能对 精密医学新兴领域的治疗策略。