Targeting invasive plasticity by inhibiting mitochondrial adaptations to matrix metalloproteinase loss

通过抑制线粒体对基质金属蛋白酶损失的适应来靶向侵入可塑性

• 批准号: 10684722
• 负责人: Laura Catherine Kelley
• 金额: $ 18.44万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-16 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10684722
• 关键词: 4D Imaging; Actins; Address; Adenine Nucleotide Translocase; Adopted; Animals; Ants; Basic Science; Behavior; Biosensor; Brain; Caenorhabditis elegans; Cell Line; Cell Survival; Cells; Chemicals; Clinical Research; Clinical Trials; Compensation; Data; Developmental Process; Distant Metastasis; Event; Excision; Experimental Models; Extracellular Matrix; Extracellular Matrix Degradation; F-Actin; Fluorescence; Genes; Genetic; Glioblastoma; Goals; Human; Immunologic Surveillance; In Vitro; Invaded; Localized Malignant Neoplasm; Malignant Neoplasms; Matrix Metalloproteinases; Metabolic; Mission; Mitochondria; Modeling; Neoplasm Metastasis; Output; Oxidative Stress; Pathway interactions; Penetration; Pharmacologic Substance; Polymers; Porosity; Production; Proteins; Proteolysis; Public Health; RNA Interference; Reporter; Research; Role; Site; Slice; Testing; Therapeutic; Time; Tissues; Tumor Cell Invasion; Tumor-Derived; United States National Institutes of Health; Work; brain tissue; cancer cell; cancer invasiveness; cancer therapy; clinically relevant; combinatorial; confocal imaging; efficacy testing; genetic analysis; improved; in vivo; in vivo Model; inhibitor; knock-down; live cell imaging; mutant; neoplastic cell; overexpression; patient prognosis; polymerization; prevent; programs; response; tissue culture; tumor; tumor progression

Tumor cell invasion through extracellular matrix (ECM) facilitates localized and distant cancer spread, which is the most lethal aspect of cancer. The ability of cells to switch between distinct invasive modes, termed plasticity or adaptation, when faced with varying physical or chemical challenges underlies the inability to develop anti-invasive therapies. Identifying targetable adaptive responses to halt invasion has been hindered by the lack of experimental models to identify, characterize, and test the loss of key molecules that facilitate plasticity. To address this critical need we have focused on matrix metalloproteinases (MMPs), which have been targeted in extensive clinical trials because of their strong association with cancer and role in degrading ECM. Anti-MMP therapies, however, have been ineffective, likely because of invasive plasticity. To identify and understand how invasive cells adapt to MMP loss, we are using the in vivo model of anchor cell (AC) invasion in C. elegans. We found that the genetic removal of MMPs results in an adaptive invasion response where instead of ECM degradation, the AC increases F-actin polymerization to forcefully penetrate ECM. Using MMP-null animals, we performed the first synergistic invasion screen to pinpoint genes that promote adaptive AC invasion and identified the mitochondrial ATP/ADP translocase, ant-1.1, as the strongest candidate. ANTs have multiple mitochondrial functions (ATP/ADP exchange, mitophagy, mitochondrial dynamics) and the ANT-1.1 protein is highly enriched in AC mitochondria that polarize to the site of ECM invasion. ANT-1.1 knockdown in MMP-null animals prevents adaptive F-actin formation and inhibits AC invasion. The overall objective of this application is to (Aim 1) elucidate how ant-1.1 promotes adaptive invasion after MMP loss in C. elegans, and (Aim 2) determine if the concurrent loss of MMP and ANT activity in a 4-D organotypic brain slice model of glioblastoma (GBM) blocks invasive activity. Our central hypothesis is that understanding how ANT-1.1 functions in mitochondrial for adaptive invasion will facilitate targeting ANTs along with MMPs in a clinically relevant brain slice model of GBM invasion. To understand how ANT-1.1 promotes adaptive invasion, will use genetic analysis, fluorescence reporters, metabolic biosensors, cell-specific metabolic analysis, and quantitative live-cell imaging. We will then use quantitative confocal imaging to directly test the efficacy of combined ANT and MMP therapies on GBM cell invasion. We expect to establish how ANT-1.1 functions within mitochondria to facilitate adaptive invasion (possibly through multiple functions) and to develop combined therapeutic approaches to effectively block GBM invasion. These contributions will be significant as they will reveal how invasive cells adaptively invade in the absence of MMPs and establish a pipeline that can be used to identify and characterize synergistic invasive targets resulting in more effective cancer therapies.

通过细胞外基质（ECM）侵袭肿瘤细胞，促进了局部和遥远的癌症扩散， 这是癌症最致命的方面。细胞在不同的侵入性模式之间切换的能力， 面对不同的物理或化学挑战时，称为可塑性或适应 无法开发抗侵入性疗法。确定对停止入侵的适应性自适应反应 缺乏实验模型来识别，表征和测试钥匙丢失的阻碍 促进可塑性的分子。为了满足这种关键需求，我们专注于矩阵 金属蛋白酶（MMP），由于其强大而在广泛的临床试验中。 与癌症和降解ECM的作用相关。然而，抗MMP疗法无效， 可能是由于侵入性可塑性。识别和了解侵入性细胞如何适应MMP损失， 我们正在使用秀丽隐杆线虫中的锚固细胞（AC）入侵的体内模型。我们发现遗传 去除MMP会导致自适应侵袭反应，其中AC而不是ECM降解而不是ECM降解 增加F-肌动蛋白聚合以有效穿透ECM。使用MMP无动物，我们进行了 第一个协同入侵屏幕以查明促进自适应AC侵袭并确定的基因 线粒体ATP/ADP易位酶，ANT-1.1，是最强的候选者。蚂蚁有多个 线粒体功能（ATP/ADP交换，线粒体，线粒体动力学）和ANT-1.1 蛋白质高度富集在AC线粒体上，该线粒体偏向于ECM侵袭部位。 ANT-1.1 MMP无效动物的敲低可防止自适应F-肌动蛋白形成并抑制AC侵袭。总体 该应用的目的是（AIM 1）阐明ANT-1.1如何促进MMP后的适应性侵袭 秀丽隐杆线虫的损失，以及（目标2）确定4-D中MMP和ANT活性的并发损失是否 胶质母细胞瘤（GBM）的器官脑切片模型阻断了侵入性活性。我们的中心假设是 理解ANT-1.1在线粒体中如何进行适应性侵袭的功能将有助于靶向 在GBM侵袭的临床相关脑切片模型中，蚂蚁与MMP一起。了解如何 ANT-1.1促进适应性侵袭，将使用遗传分析，荧光报告，代谢 生物传感器，细胞特异性代谢分析和定量活细胞成像。然后我们将使用 定量共聚焦成像直接测试GBM中联合ANT和MMP疗法的功效 细胞入侵。我们希望建立ANT-1.1在线粒体内的功能如何促进适应性 入侵（可能是通过多个功能），并开发合并的治疗方法 有效阻止GBM入侵。这些贡献将非常重要，因为它们将揭示出侵入性的方式 细胞在没有MMP的情况下适应侵入并建立可用于识别和 表征导致更有效癌症疗法的协同侵入性靶标。