Investigation of novel signaling protein in 3D and in vivo PDAC models using second generation Ref-1 inhibitors

使用第二代 Ref-1 抑制剂研究 3D 和体内 PDAC 模型中的新型信号蛋白

基本信息

批准号：
10297976
负责人：
Melissa L Fishel
金额：
$ 44.81万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10297976
关键词：
3-Dimensional Ablation Address Affect Antineoplastic Agents Biological Models Cancer Cell Growth Cancer Etiology Cell Survival Cells Cessation of life Characteristics Chemicals Clinical Coculture Techniques Complex Critical Pathways DNA Binding Data Desmoplastic Drug Kinetics Drug Targeting Drug resistance Effectiveness Evaluation Extracellular Matrix Extracellular Matrix Proteins Fibroblasts Foundations Generations Goals Growth Healthcare Systems Heterogeneity Human Impairment In Vitro Investigation Lead Liver Malignant Neoplasms Malignant neoplasm of pancreas Metabolism Microfluidics Microsomes Modeling Molecular Target Mus Neoplasm Metastasis Outcome Oxidation-Reduction Pancreatic Ductal Adenocarcinoma Pathway interactions Patients Pharmacodynamics Pharmacology Phase Role STAT3 gene Signal Pathway Signal Transduction Signaling Protein Survival Rate System Therapeutic Tissues Toxic effect Transcription Factor AP-1 Translating Translations Treatment Efficacy Treatment outcome Triage analog base cancer cell cancer clinical trial cell killing effective therapy efficacy evaluation efficacy validation gemcitabine improved in vivo in vivo Model inhibitor/antagonist insight neoplastic cell novel pancreatic cancer cells pancreatic cancer patients pancreatic ductal adenocarcinoma model pancreatic neoplasm phase I trial screening targeted agent three-dimensional modeling transcription factor treatment response tumor tumor growth tumor microenvironment

项目摘要

PROJECT SUMMARY/ABSTRACT Five-year survival rate of pancreatic ductal adenocarcinoma (PDAC) still remains at a dismal 10%. This poor outcome is attributed to PDAC’s complex tumor microenvironment (TME) containing a desmoplastic stroma, which consists of cancer-associated fibroblasts (CAFs) and a dense extracellular matrix (ECM). This stroma affects proliferation, invasion and drug resistance of pancreatic cancer cells (PCCs). Unexpectedly, the ablation of CAFs or targeting of certain ECM proteins in the stroma led to changes that actually accelerated tumor growth and impaired treatment outcome. To address this challenge, we are pursuing targets that selectively kill PCCs rather than CAFs. Redox effector factor-1 (Ref-1) is upstream of several key signaling pathways as a regulator of transcription factors (TFs). Blockade of Ref-1 redox function with our first-generation inhibitor blocks the activation of key TFs including HIF-1α, NFkB, and AP-1, all of which are implicated in growth and metastasis signaling in the tumor. However more potent second-generation compounds are necessary to effectively treat PDAC. We recently synthesized new analogs and obtained data showing that these are more potent with a significant decrease in tumor cell survival with minimal changes in CAF survival. This selective cancer cell killing is a novel characteristic of our new analogs, as gemcitabine kills both PCCs and CAFs. The underlying mechanism and therapeutic efficacy of these compounds are the focus of this proposal. Our overarching goal is to evaluate second-generation Ref-1 inhibitors for treatment of PDAC by comparing the mechanisms of Ref-1 inhibition in PCCs and CAFs. Our hypothesis is that through inhibition of Ref-1, multiple cancer-associated pathways are simultaneously and effectively targeted leading to suppression of survival signaling in PCCs while sparing the CAFs. The heterogeneity and complexity of PDAC poses a significant challenge in screening and evaluating the efficacy of new compounds on tumors and on stromal components. To overcome these technical challenges and ultimately lead to new treatments, we will perform mechanistic screening of Ref-1 inhibitors using newly developed in vitro tumor models based on patient-derived PCCs and CAFs - 1) 3D co-culture spheroids for screening and tumor-stroma interaction; and 2) a microfluidic tumor-microenvironment-on-chip (T-MOC) model for mechanistic evaluation. Findings using these novel in vitro systems will be validated with orthotopic PDAC models. Specific aims are: Aim 1. Investigate the selectivity of Ref-1 signaling and the effects of its inhibition in PCCs and CAFs using T-MOC models; Aim 2. Characterize the mechanism of Ref-1 inhibition for second-generation compounds; and Aim 3. Validate the efficacy and pharmacodynamic profiles of Ref-1 analogs in orthotopic PDAC models. The aims of this proposal will provide invaluable information regarding both the effectiveness of new Ref-1 analogs in sophisticated in vivo and in vitro PDAC models and validate our in vitro co-culture models for novel target and compound screening in a cancer that is in need of therapeutic options.

项目摘要/摘要胰腺导管腺癌（PDAC）的五年生存率仍然保持10％。这个穷人结果归因于PDAC的复杂肿瘤微环境（TME），其中含有脱肿瘤基质，由癌症相关的成纤维细胞（CAF）和密集的细胞外基质（ECM）组成。这个基质影响胰腺癌细胞（PCC）的增殖，侵袭和耐药性。出乎意料的是，消融咖啡馆或靶向基质中某些ECM蛋白的靶向导致实际加速肿瘤生长的变化和治疗结果受损。为了应对这一挑战，我们正在追求选择性杀死PCC的目标而不是CAF。氧化还原效应因子1（REF-1）是几个关键信号通路的上游作为调节器转录因子（TFS）。用我们的第一代抑制剂阻断Ref-1氧化还原功能关键TF的激活在内肿瘤中的信号传导。但是，必须有效治疗更多潜在的第二代化合物 PDAC。我们最近合成了新的类似物，并获得了数据，表明这些数据具有更大的潜力肿瘤细胞存活的显着降低，CAF存活的最小变化。这种选择性癌细胞杀死是我们新类似物的新颖特征，因为吉西他滨杀死了PCC和CAFS。基础这些化合物的机制和治疗功效是该提议的重点。我们的总体目标是通过比较Ref-1的机制来评估第二代REF-1抑制剂以治疗PDAC PCC和CAFS的抑制作用。我们的假设是，通过抑制Ref-1，多个癌症相关途径是简单有效地靶向的，导致抑制PCC中的生存信号传导保留咖啡馆。 PDAC的异质性和复杂性在筛查和评估新化合物在肿瘤和基质成分上的效率。克服这些技术挑战并最终导致新治疗，我们将使用Ref-1抑制剂进行机械筛查基于患者衍生的PCC和CAF的新开发的体外肿瘤模型-1）3D共培养球体用于筛查和肿瘤的相互作用； 2）微流体肿瘤 - 微环境片（T-MOC）机械评估模型。使用这些新型体外系统的发现将通过原位验证 PDAC模型。具体目的是：目标1。研究ref-1信号的选择性及其效果使用T-MOC模型在PCC和CAF中抑制； AIM 2。表征Ref-1抑制的机制第二代化合物；和目标3。验证Ref-1类似物的效率和药效学概况在原位PDAC模型中。该提案的目的将提供有关同时的宝贵信息新的Ref-1类似物在体内和体外PDAC模型中的有效性，并验证我们的体外需要热选择的癌症中新型目标和化合物筛查的共同培养模型。