Fingerprinting Invasive Membrane Protrusions to Discover Metastatic Signatures

对侵入性膜突起进行指纹识别以发现转移特征

基本信息

批准号：
8913909
负责人：
Richard L. Klemke
金额：
$ 37.84万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8913909
关键词：
Adenocarcinoma Cell Adhesions Adhesives Animal Model Basement membrane Bioinformatics Biological Markers Blood Circulation Blood Vessels Cell Line Cell Surface Extensions Cells Complex Cues Development Diagnosis Diagnostic Disease Disseminated Malignant Neoplasm Distant Early Diagnosis Environment Enzymes Epigenetic Process Extracellular Matrix Extracellular Matrix Proteins Fibroblasts Fingerprint Genetically Engineered Mouse Goals Growth Factor Health Immune Invaded Malignant neoplasm of pancreas Mass Spectrum Analysis Mediating Membrane Metastatic to Methods Mining Neoplasm Metastasis Non-Malignant Nude Mice Pancreas Pancreatic Ductal Adenocarcinoma Patients Phosphoproteins Phosphorylation Phosphotransferases Primary Neoplasm Process Proteins Proteomics Pseudopodia RNA Interference Reaction Reporting Research Research Proposals Resources Serial Passage Shapes Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Protein Staging Structure Survival Rate Technology Testing Therapeutic Time Tissues Travel Variant Work base cancer cell capillary bed cell motility chemotherapy combat cytokine extracellular human tissue in vitro Model in vivo innovation new therapeutic target novel outcome forecast pancreatic cancer cells pre-clinical programs protein expression protein profiling stellate cell therapeutic development therapeutic target tumor microenvironment tumor progression

项目摘要

DESCRIPTION (provided by applicant): Because metastatic pancreatic cancer cells can disseminate from the primary tumor during the earliest stages of cancer progression, long before (10 years) the patient becomes symptomatic, the majority of patients have advanced stage metastatic disease at the time of diagnosis. This and the fact that patients resist all forms of conventional chemotherapy, pancreatic cancer has a dismal 5 year survival rate of less than 5%, and a median survival of 4-6 months. Clearly, to combat this devastating disease new biomarkers for early detection and new therapeutic targets directed at eliminating metastatic cells are urgently needed. Therefore, the objective of work in this proposal is to identify new proteins that mediate pancreatic ductal adenocarcinoma (PDAC), which can be used as biomarkers at and therapeutic targets to treat metastatic PDAC. To achieve this goal, in Aim 1 we will generate highly metastatic genetically engineered mouse and patient-derived PDAC cell lines by serial passaging of isolated metastatic cells in the pancreas of athymic nude mice. We will then biochemically purify their invadopodia, which drive PDAC cell invasion and metastasis, for proteomic and phosphoproteomic analyses. To identify metastatic signatures, the invasive invadopodia membranes will be interrogated for changes in total protein profile as well as kinase activation networks using established phosphosite identification methods, quantitative MudPIT mass spectrometry, and bioinformatics. In Aim 2, we will functionally test selected key signature proteins identified in Aim 1 for their ability to mediate PDAC cell metastasis using RNAi knockdown technology and preclinical animal models of PDAC cell metastasis. Our unique approach of using in vivo selected metastatic variants combined with invadopodia purification and comprehensive proteomics will reveal important new metastatic signature proteins that serve as 'fingerprints' to identify and treat metastatic PDAC cells. The identification of novel metastatic proteins is the first, and critical step, in the process that drives development of new PDAC therapeutics and diagnostics, which are sorely needed to treat this devastating disease.

描述（由申请人提供）：由于转移性胰腺癌细胞可以在癌症进展的最早阶段从原发肿瘤扩散，远在患者出现症状之前（10年），因此大多数患者在出现症状时已处于晚期转移性疾病诊断。这以及患者抵制所有形式的事实与传统化疗相比，胰腺癌的5年生存率不足5%，中位生存期为4-6个月。显然，为了对抗这种毁灭性的疾病，迫切需要用于早期检测的新生物标志物和旨在消除转移细胞的新治疗靶点。因此，本提案的工作目标是鉴定介导胰腺导管腺癌（PDAC）的新蛋白，其可用作治疗转移性PDAC的生物标志物和治疗靶点。为了实现这一目标，在目标 1 中，我们将通过连续传代无胸腺裸鼠胰腺中分离的转移细胞来产生高度转移的基因工程小鼠和患者来源的 PDAC 细胞系。然后，我们将通过生化方法纯化其驱动 PDAC 细胞侵袭和转移的侵袭伪足，用于蛋白质组和磷酸化蛋白质组分析。为了识别转移特征，将使用已建立的磷酸位点识别方法、定量 MudPIT 质谱法和生物信息学来询问侵入性侵袭伪足膜的总蛋白谱以及激酶激活网络的变化。在目标 2 中，我们将使用 RNAi 敲除技术和 PDAC 细胞转移的临床前动物模型对目标 1 中确定的选定关键特征蛋白进行功能测试，以确定其介导 PDAC 细胞转移的能力。我们使用体内选定的转移变体结合侵袭伪足纯化和全面蛋白质组学的独特方法将揭示重要的新转移特征蛋白，这些蛋白可作为识别和治疗转移性 PDAC 细胞的“指纹”。新型转移蛋白的鉴定是推动新 PDAC 疗法和诊断方法开发过程中的第一步，也是关键的一步，而新的 PDAC 疗法和诊断方法是治疗这种毁灭性疾病所迫切需要的。