Arp2/3 complex heterogeneity as a target for metastatic cancer

Arp2/3 复合体异质性作为转移性癌症的靶标

• 批准号: 10460277
• 负责人: PRACHEE AVASTHI
• 金额: $ 26.24万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-15 至 2021-08-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10460277
• 关键词: ANGPTL2 gene; Actins; Address; Affect; Area; Basement membrane; Binding; Biochemical; Biochemistry; Biological Assay; Biological Models; Blood; Breast Cancer Cell; Breast Cancer cell line; Cancer cell line; Cell Nucleus; Cell physiology; Cells; Centers of Research Excellence; Chlamydomonas; Chlamydomonas reinhardtii; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Colorectal Cancer; Complex; Core Facility; Cytoplasmic Vesicles; Cytoskeleton; Disseminated Malignant Neoplasm; Distant; Double Strand Break Repair; Endocytosis; Epithelium; Eukaryota; Extracellular Matrix; Extravasation; Fluorescence Microscopy; Future; Genes; Genetic Transcription; Goals; Green Algae; Growth; Head and Neck Squamous Cell Carcinoma; Heterogeneity; Human; Immunocompromised Host; In Vitro; Institutes; Investigation; Knowledge; Laboratories; Lymph; Malignant Neoplasms; Malignant neoplasm of prostate; Malignant neoplasm of urinary bladder; Mediating; Mentors; Mesenchymal; Messenger RNA; MicroRNAs; Microfilaments; Microscopy; Microtubules; Molecular; Mus; Nature; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Normal Cell; Nuclear; Pathologic; Phenotype; Play; Primary Neoplasm; Property; Protein Isoforms; Proteins; Regulation; Renal Cell Carcinoma; Role; Site; Structure; Surface; System; Testing; Tissues; Travel; Tumor Cell Invasion; Up-Regulation; Work; base; cancer cell; cell motility; circulating cancer cell; experience; experimental study; gene product; in vivo; knock-down; malignant breast neoplasm; metastatic process; migration; neoplastic cell; polymerization; protein complex; triple-negative invasive breast carcinoma; tumor progression

During metastasis, cancer cells can break free from primary tumor sites and migrate into the extracellular matrix, through the basement membrane, travel in the blood or lymph, and settle in distant sites. Actin reorganization, specifically in branched actin networks at the leading edge, is responsible for cell motility at various stages of these metastatic processes. A seven-protein complex called the Arp2/3 complex is responsible for the new actin filament growth and branching at the leading edge required for motility and proteins in this complex are often upregulated in cancer. In addition to this transcriptional upregulation, microRNAs can also control expression Arp2/3 complex components, affecting migratory potential. One protein in particular, ARPC5, is known to be regulated transcriptionally and by miRNAs in many cancers including head and neck squamous cell carcinoma, non-small cell lung cancer, bladder cancer, colorectal cancer, breast cancer, renal cell carcinoma, and prostate cancer, suggesting its key role in cancer cell migration. Given the endogenous suppression of expression by miRNAs and also the possible expression of an alternate isoform of ARPC5, ARPC5L, we propose that Arp2/3 complexes with no ARPC5, ARPC5, or ARPC5L can create versions of the actin-binding complex that are tuned for various cell functions. To address this, in Aim 1, we will purify and perform in vitro and in vivo characterization of complexes from a model system that naturally lacks ARPC5, a green alga that performs other typical Arp2/3 functions outside of surface motility. In Aim 2, we will directly test the functional consequences of ARPC5 and/or ARPC5L deletion in triple negative breast cancer cell line isolates that vary in their migration potential. Successful completion of this project will determine the molecular and cellular consequences of ARPC5-dependent Arp2/3 composition as well as the potential impact of modulating ARPC5 composition in the context of metastatic cancer. The BioMT COBRE will provide essential mentoring and collaboration from experts with decades of actin biochemistry and specifically Arp2/3 experience as well as in tumor progression. The COBRE will also provide core facility support in biochemistry and microscopy for the completion of these aims. Ultimately, this will allow the PI to expand work from her lab outside core projects in microtubule-based cytoskeletal structures into a new area of investigation related to actin regulation.

在转移过程中，癌细胞可以脱离原发肿瘤部位，迁移到细胞外。 基质，通过基底膜，在血液或淋巴中移动，并在远处定居。肌动蛋白 重组，特别是在前沿的分支肌动蛋白网络中，负责细胞的运动 这些转移过程的不同阶段。一种称为Arp2/3复合体的七种蛋白质复合体 负责运动和运动所需前沿的新肌动蛋白细丝的生长和分支 这种复合体中的蛋白质通常在癌症中上调。除了这种转录上调外， MicroRNAs还可以调控Arp2/3复合体的表达，影响细胞的迁移潜能。一种蛋白质 尤其是ARPC5，已知在许多癌症中都受到转录和miRNAs的调控，包括 头颈部鳞状细胞癌、非小细胞肺癌、膀胱癌、结直肠癌、乳腺癌 癌症、肾细胞癌和前列腺癌，表明它在癌细胞迁移中起关键作用。给定 MiRNAs对内源性表达的抑制以及可能的另一种异构体的表达 ARPC5、ARPC5L，我们提出不含ARPC5、ARPC5或ARPC5L的Arp2/3复合体可以创建 为各种细胞功能调整的肌动蛋白结合复合体的版本。为了解决这个问题，在目标1中，我们将 从天然缺乏的模型体系中提纯并进行体外和体内表征 ARPC5，一种绿藻，在表面运动之外执行其他典型的Arp2/3功能。在目标2中，我们将 直接检测ARPC5和/或ARPC5L缺失在三阴性乳腺癌中的功能后果 迁移能力不同的细胞系分离株。该项目的成功完成将决定 依赖ARPC5的Arp2/3组成的分子和细胞后果以及 调节ARPC5成分在转移性癌症中的潜在影响。BIOMT Cobre将提供必要的指导和专家的合作，拥有数十年的肌动蛋白生物化学和 尤其是Arp2/3的经验以及肿瘤的进展。科布雷还将提供核心设施 在生物化学和显微镜方面的支持，以实现这些目标。最终，这将使PI能够 将她的实验室工作从基于微管的细胞骨架结构的核心项目扩展到一个新的领域 与肌动蛋白调控相关的调查。