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，癌细胞可以从原发肿瘤部位脱离并迁移到细胞外基质中。 基质通过基底膜，在血液或淋巴液中移动，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，负责细胞运动， 这些转移过程的不同阶段。一种称为Arp 2/3复合物的七蛋白复合物， 负责新的肌动蛋白丝的生长和运动所需的前沿分支， 这种复合物中的蛋白质通常在癌症中上调。除了这种转录上调， microRNA还可以控制Arp 2/3复合物组分的表达，影响迁移潜力。一种蛋白质 特别是ARPC 5，已知在许多癌症中受转录调节和miRNA调节，包括 头颈部鳞状细胞癌、非小细胞肺癌、膀胱癌、结肠直肠癌、乳腺癌 癌症、肾细胞癌和前列腺癌，表明其在癌细胞迁移中的关键作用。鉴于 内源性抑制表达的miRNA，也可能表达的替代亚型， ARPC 5，ARPC 5L，我们认为没有ARPC 5，ARPC 5或ARPC 5L的Arp 2/3复合物可以产生 肌动蛋白结合复合物的各种版本，这些版本针对各种细胞功能进行了调整。为了解决这个问题，在目标1中，我们将 从天然缺乏的模型系统中纯化复合物并进行体外和体内表征 ARPC 5，一种绿色蛋白，在表面运动性之外执行其他典型的Arp 2/3功能。在目标2中，我们将 直接测试三阴性乳腺癌中ARPC 5和/或ARPC 5L缺失的功能后果 细胞系分离物，其迁移潜力不同。这个项目的成功完成将决定 ARPC 5依赖性Arp 2/3组成的分子和细胞后果以及 在转移性癌症的情况下调节ARPC 5组合物的潜在影响。关于BioMT COBRE将提供必要的指导和合作，从专家与几十年的肌动蛋白生物化学， 特别是Arp 2/3经历以及在肿瘤进展中。COBRE还将提供核心设施 在生物化学和显微镜方面提供支持，以完成这些目标。最终，这将允许PI 将她的实验室核心项目以外的工作扩展到基于微管的细胞骨架结构的新领域， 与肌动蛋白调节相关的研究。