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.

肿瘤细胞通过细胞外基质（ECM）侵袭促进局部和远处的癌症扩散， 这是癌症最致命的方面。细胞在不同侵入模式之间切换的能力， 当面临不同的物理或化学挑战时，称为可塑性或适应性 无法开发抗侵入疗法。确定有针对性的适应性反应以阻止入侵 由于缺乏识别、表征和测试密钥丢失的实验模型而受到阻碍 促进可塑性的分子。为了满足这一关键需求，我们重点关注矩阵 金属蛋白酶（MMP），因其强大的作用而成为广泛的临床试验的目标 与癌症的关联以及在降解 ECM 中的作用。然而，抗 MMP 疗法无效， 可能是因为侵入可塑性。为了识别和了解侵袭细胞如何适应 MMP 损失， 我们正在使用秀丽隐杆线虫锚细胞（AC）入侵的体内模型。我们发现遗传 MMP 的去除会导致适应性入侵反应，其中 AC 不是 ECM 降解，而是 增加 F-肌动蛋白聚合以强力渗透 ECM。使用 MMP 缺失的动物，我们进行了 首次协同入侵筛选，以查明促进适应性 AC 入侵的基因，并确定 线粒体 ATP/ADP 转位酶 ant-1.1 是最强的候选者。 ANT 有多个 线粒体功能（ATP/ADP 交换、线粒体自噬、线粒体动力学）和 ANT-1.1 蛋白质在 AC 线粒体中高度富集，极化至 ECM 入侵部位。 ANT-1.1 MMP 缺失动物中的敲低可阻止适应性 F-肌动蛋白形成并抑制 AC 入侵。整体 本申请的目的是（目标 1）阐明 ant-1.1 如何促进 MMP 后的适应性入侵 线虫中的损失，以及（目标 2）确定 4-D 中 MMP 和 ANT 活性是否同时损失 胶质母细胞瘤（GBM）的器官型脑切片模型可阻止侵袭性活动。我们的中心假设是 了解 ANT-1.1 在线粒体中如何发挥适应性入侵功能将有助于靶向 临床相关的 GBM 侵袭脑切片模型中的 ANT 和 MMP。要了解如何 ANT-1.1促进适应性入侵，将利用遗传分析、荧光报告基因、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 ANT 和 MMP 联合疗法对 GBM 的疗效 细胞侵袭。我们期望确定 ANT-1.1 在线粒体内如何发挥作用以促进适应性 入侵（可能通过多种功能）并开发联合治疗方法 有效阻断GBM侵袭。这些贡献将意义重大，因为它们将揭示侵入性的方式 细胞在没有 MMP 的情况下适应性地侵入并建立可用于识别和 表征协同侵入靶标，从而产生更有效的癌症治疗。