The role of MIRO2 in tumor cell invasion and metastasis

MIRO2在肿瘤细胞侵袭和转移中的作用

• 批准号: 10704512
• 负责人: Dillon Boulton
• 金额: $ 3.72万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10704512
• 关键词: 3-Dimensional; 4T1; Ablation; Address; Binding; Binding Proteins; Biological Assay; Breast; Breast Cancer Model; Breast Cancer Patient; Breast cancer metastasis; Calcium; Calcium Binding; Cause of Death; Cell Death; Cell Survival; Cell physiology; Cells; Cessation of life; Co-Immunoprecipitations; Data; Detection; Development; Disease; Distal; Dominant-Negative Mutation; EF-Hand Domain; GTPase-Activating Proteins; Goals; Growth; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Immunohistochemistry; Immunoprecipitation; In Vitro; Injections; Invaded; Knockout Mice; Ligation; MDA MB 231; Malignant Neoplasms; Malignant neoplasm of prostate; Mass Spectrum Analysis; Matrix Metalloproteinases; Measurement; Mediating; Messenger RNA; Mitochondria; Mitochondrial Membrane Protein; Modeling; Molecular; Myosin ATPase; Myosin S-2; Neoplasm Metastasis; Neuroglia; Neurons; Normal tissue morphology; Organ; Organelles; Outer Mitochondrial Membrane; Pancreas; Patients; Phenocopy; Phenotype; Primary Neoplasm; Process; Production; Proliferation Marker; Prostate; Recombinant Proteins; Role; Sampling; Secondary to; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; System; Tertiary Protein Structure; Testing; Tissues; Transfection; Tumor Cell Invasion; Tumor Promotion; United States; Work; cell motility; cell type; hazard; in vitro Assay; in vivo; in vivo Model; in vivo imaging system; interest; knock-down; live cell imaging; mRNA Expression; melanoma; neoplastic cell; new therapeutic target; novel; novel therapeutics; pancreatic cancer cells; pharmacologic; prostate cancer cell line; protein function; rho GTP-Binding Proteins; rho GTPase-activating protein; small hairpin RNA; targeted treatment; therapeutic target; trafficking; tumor; tumor growth; tumorigenesis; tumorigenic

Project Abstract/Summary Cancer is the second leading cause of death in the United States. Overall survival for patients with tumors that remain localized to the tissue of origin remain relatively high, while it is estimated that 90% of all cancer related deaths are due to metastatic dissemination to secondary sites. This underscores the importance of identifying signaling pathways that promote metastasis. Mitochondria are multifunctional organelles with important roles in regulating many cellular processes outside of ATP production and are recognized for their roles in tumorigenesis and metastasis. This project seeks to define a novel mechanism that promotes tumor cell invasion and metastasis through a previously unidentified signaling pathway between Mitochondrial Rho GTPase 2 (MIRO2) and atypical myosin IXB (MYO9B). MIRO2 is an outer-mitochondrial membrane protein that is a part of the MIRO subfamily of Ras GTPases. MIROs were first characterized in neurons where they are involved in the anterograde and retrograde trafficking of mitochondria. MIRO2 has been shown by several groups to be dispensable for mitochondrial trafficking in many non-neuronal cell types. Our lab has previously shown that MIRO2 mRNA is enriched in tumorigenic tissues in many tumor types when compared to normal tissue. Additionally, we have shown in prostate cancer that MIRO2 is important in tumor cell viability, anchorage-dependent and -independent growth, and tumor growth in vivo. Despite this initial evidence, it remains unknown if MIRO2 exclusively impacts the primary tumor or if this is also important in metastasis. My initial results show that loss of MIRO2 reduces the invasive capacity of tumor cells from many tumor types including breast, melanoma, pancreas, and prostate. Furthermore, I have shown out of the top newly identified MIRO2 binding partners, loss of MYO9B reduces invasive capacity to the greatest extent. MYO9B is known to control cell motility by localizing to the leading edge of migrating cells and inactivating RhoA through MYO9B’s Rho GTPase activating protein (GAP) domain. Excitingly, I show that loss of both MIRO2 and MYO9B result in increased active RhoA. Given this evidence, I hypothesize MIRO2 promotes tumor cell invasion and metastasis through positively regulating MYO9B GAP activity. In this proposal I will 1) determine the role of MIRO2 in tumor cell invasion and metastasis and 2) determine the mechanism in which MIRO2 regulates tumor cell invasion. This proposal will utilize many models including, but not limited to: 2D/3D in vitro invasion systems, live cell imaging of migrating cells, in vivo breast cancer metastasis models, cell-free GTPase assays, co-immunoprecipitation, and proximity ligation assays. Overall, the goal of this proposal is to serve as the basis for the development of novel therapeutics to target metastatic disease.

项目摘要/摘要 癌症是美国第二大死因。肿瘤患者的总生存期 仍然局限于起源组织的比率仍然相对较高，而据估计，所有癌症中有90% 相关死亡是由于转移到继发部位造成的。这就强调了 确定促进转移的信号通路。线粒体是多功能细胞器，具有 在调节ATP生产之外的许多细胞过程中发挥重要作用，并因其 在肿瘤发生和转移中的作用。该项目试图定义一种促进肿瘤细胞生长的新机制 线粒体Rho GTP酶之间未知信号通路的侵袭和转移 2(MIRO2)和非典型肌球蛋白IXB(MYO9B)。 MIRO2是一种线粒体膜外膜蛋白，属于RAS GTP酶MIRO亚家族的一部分。 Miros最初的特征是神经元参与了顺行和逆行运输。 线粒体。几个研究小组已经证明，MIRO2对于线粒体的运输是必不可少的 许多非神经细胞类型。我们的实验室先前已经证明，MIRO2 mRNA在致癌过程中富含 与正常组织相比，许多肿瘤类型的组织。此外，我们已经在前列腺癌中显示了 MIRO2在肿瘤细胞活性、锚定依赖性和非依赖性生长以及肿瘤生长中起重要作用 在活体内。尽管有这些初步证据，但尚不清楚MIRO2是否只影响原发肿瘤，或者这是否 在转移过程中也很重要。我的初步结果显示，MIRO2的缺失降低了肿瘤的侵袭能力 来自多种肿瘤类型的细胞，包括乳腺癌、黑色素瘤、胰腺和前列腺癌。此外，我已经展示了 在新发现的顶级MIRO2结合伙伴中，MYO9B的缺失最大限度地降低了侵袭能力 范围。已知MYO9B通过定位于迁移细胞的前沿和失活来控制细胞的运动 RhoA通过MYO9B的Rho GTP酶激活蛋白(GAP)结构域。令人兴奋的是，我发现两个MIRO2的损失 和MYO9B导致活性RhoA增加。根据这一证据，我假设MIRO2促进肿瘤细胞 通过正向调节MYO9B GAP活性，促进肿瘤的侵袭转移。 在这项提案中，我将1)确定MIRO2在肿瘤细胞侵袭和转移中的作用；2) 确定MIRO2调节肿瘤细胞侵袭的机制。这项提议将利用许多模式 包括但不限于：2D/3D体外侵袭系统、迁移细胞的活细胞成像、活体乳腺 肿瘤转移模型、无细胞GTP酶分析、免疫共沉淀和邻近连接分析。 总体而言，这项提议的目标是作为开发新疗法的基础，以 转移性疾病。