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

• 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
• 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

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.

项目总结/摘要 胰腺导管腺癌（PDAC）的五年生存率仍然保持在10%。这个可怜 结果归因于PDAC复杂的肿瘤微环境（TME）， 其由癌症相关的成纤维细胞（CAF）和致密的细胞外基质（ECM）组成。该基质 影响胰腺癌细胞（PCCs）的增殖、侵袭和耐药性。出乎意料的是， CAFs或基质中某些ECM蛋白的靶向作用导致了实际上加速肿瘤生长的变化 和受损的治疗结果。为了应对这一挑战，我们正在寻求有选择地杀死PCC的目标 而不是咖啡馆。氧化还原效应因子-1（Ref-1）作为一种调节因子，在几个关键信号通路的上游 转录因子（TF）。使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，包括HIF-1α、NFkB和AP-1，所有这些都与生长和转移有关 肿瘤中的信号。然而，更有效的第二代化合物是必要的， PDAC。我们最近合成了新的类似物，并获得了数据显示，这些类似物在与 肿瘤细胞存活率显著降低，CAF存活率变化最小。这种选择性的癌细胞杀伤 是我们的新类似物的一个新特征，因为吉西他滨可以杀死PCC和CAF。底层 这些化合物的作用机制和治疗功效是本提案的重点。我们的总目标 通过比较Ref-1的作用机制，评价第二代Ref-1抑制剂治疗PDAC的效果 抑制PCC和CAF。我们的假设是，通过抑制Ref-1，多种癌症相关的肿瘤细胞可以被抑制。 同时有效地靶向通路，从而抑制PCC中的存活信号传导， 避免咖啡因。PDAC的异质性和复杂性对PDAC的筛选和治疗提出了重大挑战。 评价新化合物对肿瘤和基质成分的功效。为了克服这些技术 挑战，并最终导致新的治疗方法，我们将进行机械筛选的Ref-1抑制剂使用 新开发的基于患者来源的PCC和CAF的体外肿瘤模型- 1）3D共培养球体 用于筛选和肿瘤-基质相互作用;和2）微流体芯片上肿瘤微环境（T-MOC） 力学评价模型。使用这些新的体外系统的发现将通过原位 PDAC模型。具体目标是：目标1。研究Ref-1信号传导的选择性及其对细胞凋亡的影响。 使用T-MOC模型在PCC和CAF中的抑制;目的2.表征Ref-1抑制 第二代化合物;和目标3.验证Ref-1类似物的功效和药效学特征 在原位PDAC模型中。本提案的目的将提供关于以下两方面的宝贵信息： 新Ref-1类似物在复杂的体内和体外PDAC模型中的有效性，并验证我们的体外 共培养模型，用于在需要治疗选择的癌症中进行新靶点和化合物筛选。