Exploitation of RAS signaling to develop therapy and early detection strategies for PDA

利用 RAS 信号传导开发 PDA 的治疗和早期检测策略

基本信息

批准号：
9012030
负责人：
Rolf A Brekken
金额：
$ 43.59万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-02-10 至 2020-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9012030
关键词：
1-Phosphatidylinositol 3-Kinase Abraxane Acinar Cell Address Adult Apoptosis Biological Biological Assay Blood Cancer Biology Carcinoma Cell Proliferation Cells Chronic DDR1 gene DDR2 gene DNA Development Disease Drug Combinations Drug resistance Duct (organ) structure Ductal Ductal Epithelial Cell Early Diagnosis Early identification Endocrine Epithelial Event Feedback G Protein-Coupled Receptor Signaling G alpha q Protein Gene Expression Gene Fusion Genetic Goals Growth Health Intraepithelial Neoplasia Ligands Maintenance Malignant Neoplasms Malignant neoplasm of pancreas Mass Spectrum Analysis Metabolic Metaplasia Modeling Monitor Mouse Cell Line Mucinous Neoplasm Mus Neoplasm Metastasis Neoplasms Oncogenes Oncogenic Pancreas Pancreatic Ductal Adenocarcinoma Pancreatic Intraepithelial Neoplasia Pancreatic duct Pancreatitis Papillary Pathway interactions Patients Pharmaceutical Preparations Phenotype Physiology Plasma Point Mutation Process Proteinase 3 Proteomics Public Health RGS Family Gene RGS Proteins Ras Inhibitor Receptor Protein-Tyrosine Kinases Refractory Regulation Reporter Reporter Genes Research Signal Pathway Signal Transduction Staging Stem cells Stress System Techniques Testing Therapeutic Transcriptional Regulation Tumor Burden Tumor Suppressor Proteins Warfarin Xenograft procedure actionable mutation axl receptor tyrosine kinase base cell type chemotherapy connective tissue growth factor drug development effective therapy exome sequencing experience fusion gene gemcitabine gene induction improved in vivo in vivo Model inhibitor/antagonist innovation insight migration mimetics mouse model mutant neoplastic cell novel novel drug combination novel marker novel therapeutics pancreas development pancreatic neoplasm progenitor pup response screening small molecule standard of care therapy development tool transcription factor transcriptome sequencing tumor tumor growth tumor initiation tumor microenvironment tumor progression

项目摘要

DESCRIPTION (provided by applicant): The therapeutic problem we address is how to inhibit the development and progression of IPMN (intraductal papillary mucinous neoplasm), PanIN (Pancreatic Intraepithelial Neoplasia) and PDA (pancreatic ductal adenocarcinoma). The biological problem is to identify key signaling pathways that initiate and propagate IPMN, PanIN and PDA. A central debate in the field is whether PDA initiates in pancreatic progenitor cells or with dedifferentiation of parenchymal acinar cells to progenitor-like ductal cells, in a process termed acinar to ductal metaplasia (ADM). The function of the transcription factor Master Regulators (MRs) of acinar cell identity is critical to this process but post-translational regulaton of their activity through receptor tyrosine kinase (RTK) and G protein coupled receptor (GPCR) signaling is completely unexplored. Regulators of G protein signaling (RGS) proteins integrate multiple intracellular signals from RTKs and GPCRs to feedback regulate Gi and Gq protein signaling. We have developed a GFP reporter (Rgs16:GFP) that allows us to monitor neoplasia and tumor formation from early stages in in vivo mouse models of IPMN, PanIN and PDA. Combining the Rgs16:GFP reporter with mouse models for IPMN, PanIN and PDA is providing insights into development of these tumors and a novel and rapid in vivo screen for inhibitors of this process. We discovered RGS genes that suppress progression of IPMN to invasive carcinoma. Based on preliminary analyses using these models, we find that IPMN and PanIN initiate in discrete ductal progenitor cells in response to stress in the acinar or endocrine compartments of the pancreas. This interpretation challenges the currently favored paradigm that PDA initiates with cells that underwent ADM whereas we propose that chronic ADM stimulates proliferation of pre-existing progenitor cells in ducts. We find that the RTK Axl is expressed in pancreatic ducts and is highly expressed with Rgs16:GFP in PDA progenitor cells in primary culture. Our hypothesis is that the Axl-ligand Gas6 expressed in stroma evokes Axl signaling in epithelial progenitor cells and contributes to activation of KrasG12D, post-translational control of MR levels and/or activity, PDA initiation and progression. In our preliminary in vivo screens of new therapeutics to inhibit PDA progression, we determined that warfarin, which blocks Gas6 induced activation of Axl, combined with the standard of care Gemcitabine and Abraxane (GA), significantly reduced PDA progression. The Specific Aims of this proposal are 1) Identify optimal drug combinations, including inhibitors of Axl signaling, to inhibit PDA. 2) Identify early markers of IPMN, PanIN and PDA in plasma. 3) Determine Gas6-Axl function in PDA. The proposed project, if successful, will have a major impact on cancer by i) identifying early blood markers of PDA, and perhaps other epithelial cancers, ii) introducing a novel, general and rapid in vivo screening technique to find novel drug combinations that inhibit PDA progression, iii) providing novel genetic tools to determine if PDA initiates in progenitor cells in the pancreatic duct, Iv) identify previously unknown regulatory mechanisms of the MR transcription factors.

描述（由申请人提供）：我们解决的治疗问题是如何抑制IPMN的发育和进展（导管乳头状粘膜肿瘤），Panin（胰腺上皮内肿瘤）和PDA（胰腺内肿瘤）（胰腺导管腺癌）。生物学问题是确定启动和传播IPMN，Panin和PDA的关键信号通路。该领域的一个中心辩论是，PDA是在胰腺腺泡细胞中启动的，还是将实质细胞推导到祖细胞样性导管细胞中，这是在被称为导管性化合物的腺泡的过程中（ADM）。腺泡细胞身份的转录因子主调节剂（MRS）的功能对于此过程至关重要，但是通过受体酪氨酸激酶（RTK）和G蛋白偶联受体（GPCR）信号传导的翻译后调节量是至关重要的。 G蛋白信号传导（RGS）蛋白的调节剂会整合来自RTK和GPCR的多个细胞内信号，以调节GI和GQ蛋白信号传导。我们已经开发了一个GFP报告基因（RGS16：GFP），使我们能够从IPMN，Panin和PDA的体内小鼠模型中的早期阶段监测肿瘤和肿瘤形成。 Panin和PDA将RGS16：GFP报告基因与IPMN的小鼠模型相结合，为这些肿瘤的发展提供了见解，以及用于此过程抑制剂的新型且快速的体内筛选。我们发现RGS基因抑制了IPMN向入侵癌的进展。基于使用这些模型的初步分析，我们发现IPMN和Panin在离散的导管祖细胞中启动，以响应胰腺的腺泡或内分泌室中的胁迫。这种解释挑战了PDA使用ADM的细胞启动的当前偏爱的范式，而我们提出慢性ADM刺激管道中现有的祖细胞的增殖。我们发现RTK AXL在胰管中表达，在原代培养物中PDA祖细胞中用RGS16：GFP高度表达。我们的假设是，在基质中表达的AXL-配体GAS6唤起上皮祖细胞中的AXL信号传导，并有助于激活KRASG12D，MR水平的翻译后控制和/或活性，PDA启动和进展。在我们的初步体内体内筛查新疗法以抑制PDA进展的过程中，我们确定阻止GAS6诱导AXL激活的华法蛋白结合了护理标准的吉西他滨和Abraxane（GA），可显着降低PDA进展。该提案的具体目的是1）确定最佳药物组合，包括AXL信号传导的抑制剂，以抑制PDA。 2）在血浆中确定IPMN，Panin和PDA的早期标记。 3）确定PDA中的GAS6-AXL功能。 The proposed project, if successful, will have a major impact on cancer by i) identifying early blood markers of PDA, and perhaps other epithelial cancers, ii) introducing a novel, general and rapid in vivo screening technique to find novel drug combinations that inhibit PDA progression, iii) providing novel genetic tools to determine if PDA initiates in progenitor cells in the pancreatic duct, Iv) identify previously unknown MR转录因子的调节机制。