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: 10271751
• 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 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10271751
• 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; Epithelial; 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.

在转移期间，癌细胞可以摆脱原发性肿瘤部位并迁移到细胞外 基质通过地下膜，以血液或淋巴管传播，然后定居在遥远的部位。肌动蛋白 重组，特别是在领先边缘的分支肌动蛋白网络中 这些转移过程的各个阶段。一个称为ARP2/3复合物的七蛋白络合物是 负责运动所需的新的肌动蛋白丝生长和分支 该复合物中的蛋白质通常在癌症中被上调。除了这种转录上调， MicroRNA还可以控制表达ARP2/3复合成分，从而影响迁移势。一种蛋白质 特别是，已知ARPC5受到转录的调节，在包括 头颈部鳞状细胞癌，非小细胞肺癌，膀胱癌，大肠癌，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 miRNA对表达的内源性抑制，也可能表达替代同工型的表达 ARPC5，ARPC5L，我们建议没有ARPC5，ARPC5或ARPC5L的ARP2/3复合物可以创建 为各种细胞功能调整的肌动蛋白结合复合物的版本。为了解决这个问题，在AIM 1中，我们将 从自然缺乏的模型系统中纯化和进行体外和体内表征复合物 ARPC5，一种在表面运动之外执行其他典型ARP2/3功能的绿色藻类。在AIM 2中，我们将 直接测试三分阴性乳腺癌中ARPC5和/或ARPC5L缺失的功能后果 细胞系分离出其迁移潜力各不相同。成功完成该项目将确定 ARPC5依赖性ARP2/3组成的分子和细胞后果以及 在转移性癌症的背景下调节ARPC5组成的潜在影响。生物群 Cobre将通过数十年的肌动蛋白生物化学和 特别是ARP2/3的经验以及肿瘤进展。毛绒还将提供核心设施 对这些目标的完成的生物化学和显微镜支持。最终，这将使PI能够 从她的实验室扩展基于微管的细胞骨架结构的核心项目的实验室。 与肌动蛋白调节有关的调查。