Targeting PPT1 in the Tumor Microenvironment

靶向肿瘤微环境中的 PPT1

基本信息

批准号：
10471234
负责人：
RAVI K AMARAVADI
金额：
$ 48.1万
依托单位：
WISTAR INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10471234
关键词：
3-Dimensional Acidity Acids Address Affect Animals Antimalarials Antineoplastic Agents Autologous Autophagocytosis BRAF gene Binding Biological Assay Cell Communication Cell Death Cell-Mediated Cytolysis Cells Cellular Assay Chemicals Chloroquine Clinical Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Critiques Data Enzymes Family Fibroblasts Future Genetic study Goals Hydroxychloroquine Immunocompetent Immunotherapy In Vitro Knock-out Knowledge Lead Length Lipids Literature Lysosomes MEKs Malignant Neoplasms Mediating Melanoma Cell Metabolic Modeling Molecular Target Neoplasm Metastasis Normal Cell Nude Mice Nutrient Pathway interactions Patient-Focused Outcomes Patients Penetration Pharmaceutical Preparations Phenotype Post-Translational Protein Processing Primary Neoplasm Proteins Quinacrine Refractory Reporting Resistance Role Signal Transduction Solubility Stress T-Cell Activation T-Lymphocyte Testing The Cancer Genome Atlas Tumor Immunity Tumor-associated macrophages Water Xenograft procedure aged analog anti-cancer aqueous base cancer cell cancer therapy cell type clinical development conditional knockout cytotoxicity dimer immunosuppressive macrophages improved in vivo in vivo evaluation inhibition of autophagy inhibitor innovation lipid transport macrophage melanoma mouse model neoplastic cell new therapeutic target next generation novel novel therapeutics overexpression palmitoylation prevent programmed cell death protein 1 resistance mechanism response targeted treatment therapy resistant thioesterase PPT1 gene product three dimensional cell culture tool translational potential tumor tumor growth tumor microenvironment tumor-immune system interactions tumorigenic

项目摘要

Project Summary – Project 2 Therapy resistance in melanoma is a major hurdle to improved survival. This project will determine if therapy resistance can be reversed by targeting the lysosomal enzyme protein palmitoyl thioesterase 1 (PPT1). Autophagy is a lysosome-dependent pathway that promotes tumor growth and resistance to therapy in melanoma. Autophagy inhibition with chloroquine (CQ) derivatives augments the efficacy of many anticancer therapies, but has limited activity as a single agent. Clinical trials involving HCQ in melanoma show promising activity but concerns have been raised about the potency of HCQ, and its poorly understood mechanism of action. We have prepared dimeric antimalarial compounds that are 10-1000 fold more potent in vitro and in vivo than CQ or HCQ. Dimeric quinacrines (DQs) (Rebecca Cancer Discovery 2017) and dimeric chloroquines (DCs) (Rebecca Cancer Discovery in revision) look especially promising as both tool compounds and potential clinical drugs. In the current cycle, we have found that extending the linker length of these dimeric compounds increases lysosomal localization and anti-melanoma activity. These DQs and DCs with longer linkers as well as CQ were used to pull a new lysosomal target, PPT1, which is overexpressed in cancer, especially in metastatic lesions. Efforts to target the lysosome and autophagy in cancer cells have focused on the effects within cancer cells but recent literature suggests targeting this pathway in immunosuppressive cells within the TME also contributes to antitumor activity. We will leverage innovative collaborations with Projects 1, 3 and 4 and heavy support from the P01 cores to understand the effects of PPT1 inhibition in both tumor cells, the interaction between tumor cells and fibroblasts, tumor associated macrophages and T cells. The proposal is based on extensive new preliminary data in response to the reviewers’ critiques from the September 2017 submission. We will test the hypothesis that targeting PPT1 in tumor cells and macrophages overcomes therapy resistance in melanoma through completion of 3 aims: Aim 1 will develop innovative new compounds by introducing heteroatom substitutions into the linker, and developing the first ever dimeric ferroquine derivatives that could have better penetration in the acidic TME. We will also develop a novel assay for PPT1 that is compatible with live cells and animal studies. Aim 2 will leverage collaborations within the P01 to study the role of PPT1 in blocking lipid trafficking from aged fibroblasts to melanoma cells and reversing resistance to targeted therapy. Aim 3 will study the effects of PPT1 inhibition on tumor cell interactions with T cells and macrophages in 3D culture, immunocompetent mouse models, and a new conditional KO model of Ppt1, with the goal of reversing resistance to immunotherapy. The impact of these studies will be to unravel a deeper mechanistic understanding of the consequences of lysosomal inhibition within the TME, which will support clinical development of these agents in the future. Knowledge gained will ultimately lead to improved patient outcomes.

项目总结 – 项目 2 黑色素瘤的治疗耐药性是提高生存率的主要障碍，该项目将确定治疗是否有效。通过靶向溶酶体酶蛋白棕榈酰硫酯酶 1 (PPT1) 可以逆转耐药性。自噬是一种溶酶体依赖性途径，可促进肿瘤生长和对治疗的抵抗力氯喹 (CQ) 衍生物抑制自噬可增强许多抗癌药物的功效。治疗，但涉及 HCQ 治疗黑色素瘤的临床试验显示其活性有限。有希望的活动，但人们对 HCQ 的效力及其了解甚少表示担忧我们制备了二聚体抗疟化合物，其效力提高了 10-1000 倍。在体外和体内均优于 CQ 或 HCQ (DQ) (Rebecca Cancer Discovery 2017) 和二聚氯喹 (DC)（丽贝卡癌症发现修订版）作为这两种工具看起来特别有前途在当前周期中，我们发现延长接头长度。这些二聚体化合物增加了溶酶体定位和抗黑色素瘤活性。具有较长接头的 DC 以及 CQ 用于拉出新的溶酶体靶点 PPT1，该靶点在癌症中过度表达，特别是在转移性病变中，努力靶向溶酶体和自噬。癌细胞关注的是癌细胞内部的影响，但最近的文献表明针对这一点 TME 内免疫抑制细胞的通路也有助于抗肿瘤活性。与项目 1、3 和 4 的创新合作以及 P01 核心的大力支持，以了解 PPT1抑制对两种肿瘤细胞的影响，肿瘤细胞与成纤维细胞之间的相互作用，肿瘤该提案基于大量新的初步数据作为回应。根据 2017 年 9 月提交的审稿人的评论，我们将测试目标假设。肿瘤细胞和巨噬细胞中的 PPT1 通过完成 3 个步骤克服了黑色素瘤的治疗耐药性目标：目标 1 将通过在连接基中引入杂原子取代来开发创新的新化合物，并开发了第一个二聚二茂铁喹衍生物，该衍生物在酸性环境中具有更好的渗透性 TME。我们还将开发一种与活细胞和动物研究兼容的新型 PPT1 检测方法。 2 将利用 P01 内部的合作来研究 PPT1 在阻止老年人脂质贩运中的作用目标 3 将研究成纤维细胞对黑色素瘤细胞的影响以及逆转对靶向治疗的耐药性。 PPT1 对 3D 培养中肿瘤细胞与 T 细胞和巨噬细胞相互作用的抑制，免疫活性小鼠模型，以及 Ppt1 的新条件 KO 模型，目标是逆转耐药性这些研究的影响将是加深对免疫疗法的机制理解。 TME 内溶酶体抑制的后果，这将支持这些药物的临床开发未来，获得的知识最终将改善患者的治疗效果。