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

使用 CE-RNA-SEQ 对单个胰腺腺癌细胞中的信号传导活性和基因表达进行分析

• 批准号: 10115487
• 负责人: Nancy L. Allbritton
• 金额: $ 59.84万
• 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/10115487
• 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; 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.

摘要 胰腺导管腺癌是一种毁灭性的疾病，迫切需要改进的治疗方法 针对特定的信号通路。对异常胰腺癌进行分子分析的策略 组织和告知靶向治疗决定将在患者中具有巨大价值 治疗然而，分子谱分析是极具挑战性的，因为活检的患者组织 是正常和恶性胰腺细胞的复杂混合物。此外，还有一个日益增长的 理解突变和基因表达本身并不与临床 反应在本申请中，一个多学科研究小组提出开发一种 最先进的单细胞平台技术来测量Sentinel的催化活性 KRAS通路中的激酶和通过RNA测序的基因表达。的 研究人员将优化和验证微取样和微电泳方法， 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报告基因 将设计和新的方法和仪器相结合的单细胞毛细管 将开创高效RNA捕获的电泳技术。人类肿瘤样本 将测定在小鼠异种移植物中维持的细胞，以获得对肿瘤性质的独特见解 目前无法访问。这项工作将直接将mRNA的产生与催化作用联系起来。 来自患者的单个肿瘤细胞中的激酶活性。这项技术将使 诸如哪种激酶信号模式驱动经典与基础表型的问题， 胰腺癌以及单个肿瘤是否具有经典和 基底型细胞从实施这项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，可能对以下方面产生深远影响： 精准医疗这一新兴领域的治疗策略。