An epigenetic link from CXCL12-CXCR4 axis through nuclear LASP-1 in breast cancer

乳腺癌中 CXCL12-CXCR4 轴通过核 LASP-1 的表观遗传联系

• 批准号: 9185273
• 负责人: Dayanidhi Raman
• 金额: $ 16.48万
• 依托单位: UNIVERSITY OF TOLEDO HEALTH SCI CAMPUS
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-12-01 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9185273
• 关键词: AMD3100; Agar; Amino Acids; Anchorage-Independent Growth; Antibodies; Area; Binding; Biochemical; Biological; Biological Assay; Bone Resorption; Breast Cancer Cell; Breast Cancer cell line; Breast cancer metastasis; CXC Chemokines; CXCL12 gene; CXCR3 gene; CXCR4 Receptors; CXCR4 gene; Cancer Etiology; Cancer Patient; Cell Nucleus; Cell membrane; Cells; Cessation of life; Chemotaxis; Complex; Computer software; Confocal Microscopy; Cytoplasm; Dominant-Negative Mutation; E-Cadherin; Embryonic Development; Epigenetic Process; Epithelial; Estrogen Receptors; Event; Fructose-1,6-Bisphosphatase; Goals; Growth; Histone H3; Histones; IL8RA gene; IL8RB gene; In Situ; In Vitro; Individual; Knock-out; Label; Ligation; Light; Link; Lysine; Lytic Metastatic Lesion; MDA MB 231; Maps; Mediating; Mesenchymal; Metastatic breast cancer; Molecular; Mutagenesis; Mutation; Neoplasm Metastasis; Nuclear; Nucleosomes; Osteolytic; Parnate; Peptides; Phenotype; Play; Primary Neoplasm; Program Development; Proteins; Proteomics; Recombinants; Recruitment Activity; Research; Resistance; Role; SUM-159 Breast Cancer Cell Line; Shapes; Site; Site-Directed Mutagenesis; Snails; Survival Rate; Testing; Transforming Growth Factors; Tumor Initiators; Woman; bone; cancer cell; cell motility; chemokine; chemokine receptor; chemotherapeutic agent; chromatin immunoprecipitation; demethylation; epigenome; histone methylation; inhibitor/antagonist; knock-down; malignant breast neoplasm; matrigel; migration; mortality; mutant; new therapeutic target; novel; parathyroid hormone-related protein; promoter; public health relevance; slug; small molecule inhibitor; stemness; transcription factor; triple-negative invasive breast carcinoma; tumor microenvironment; tumor progression; AMD3100; Agar; Amino Acids; Anchorage-Independent Growth; Antibodies; Area; Binding; Biochemical; Biological; Biological Assay; Bone Resorption; Breast Cancer Cell; Breast Cancer cell line; Breast cancer metastasis; CXC Chemokines; CXCL12 gene; CXCR3 gene; CXCR4 Receptors; CXCR4 gene; Cancer Etiology; Cancer Patient; Cell Nucleus; Cell membrane; Cells; Cessation of life; Chemotaxis; Complex; Computer software; Confocal Microscopy; Cytoplasm; Dominant-Negative Mutation; E-Cadherin; Embryonic Development; Epigenetic Process; Epithelial; Estrogen Receptors; Event; Fructose-1,6-Bisphosphatase; Goals; Growth; Histone H3; Histones; IL8RA gene; IL8RB gene; In Situ; In Vitro; Individual; Knock-out; Label; Ligation; Light; Link; Lysine; Lytic Metastatic Lesion; MDA MB 231; Maps; Mediating; Mesenchymal; Metastatic breast cancer; Molecular; Mutagenesis; Mutation; Neoplasm Metastasis; Nuclear; Nucleosomes; Osteolytic; Parnate; Peptides; Phenotype; Play; Primary Neoplasm; Program Development; Proteins; Proteomics; Recombinants; Recruitment Activity; Research; Resistance; Role; SUM-159 Breast Cancer Cell Line; Shapes; Site; Site-Directed Mutagenesis; Snails; Survival Rate; Testing; Transforming Growth Factors; Tumor Initiators; Woman; bone; cancer cell; cell motility; chemokine; chemokine receptor; chemotherapeutic agent; chromatin immunoprecipitation; demethylation; epigenome; histone methylation; inhibitor/antagonist; knock-down; malignant breast neoplasm; matrigel; migration; mortality; mutant; new therapeutic target; novel; parathyroid hormone-related protein; promoter; public health relevance; slug; small molecule inhibitor; stemness; transcription factor; triple-negative invasive breast carcinoma; tumor microenvironment; tumor progression

 DESCRIPTION (provided by applicant): The goal of the proposed research is to investigate the role of CXCL12-driven nuclear LASP1 in modulation of epigenetic events in breast cancer. LASP-1 mediates cell migration, proliferation and survival in several breast cancer cell lines. Silencing of LASP-1 inhibits proliferation and migration by 45%. Previously, LASP-1 was demonstrated to directly interact with CXC chemokine receptors, CXCR1, CXCR2, CXCR3 and CXCR4 that play a key role in the tumor microenvironment facilitating breast cancer progression and metastasis. In particular, LASP-1 augmented CXCR2-mediated cell migration. Epigenetic alterations in breast cancer cells convert them into aggressive and metastatic phenotype. In this study, through proteomics, UHRF1 was discovered as a novel LASP-1 associating protein. Subsequently, DNMT1, G9a and Snail1 were also observed to associate with LASP-1. In particular, Snail1 was found to directly bind to LASP-1. The biological implication of this direct interaction is unclear. First, I propose to map the interaction sites between LASP-1 and Snail1 by mutational and biochemical approaches. I further propose to study the LASP1-Snail1 complex for its ability to modulate the E-cadherin promoter. Additionally, as Snail1 directly binds to lysine demethyalse1 (LSD1), LSD1 will be analyzed whether it associates with LASP1-Snail1 complex by biochemical studies. Functionally, I propose to perform a chromatin immunoprecipitation (ChIP) analysis for LASP-1, LSD1, and K4/9 methylation of histone H3 to see if LASP-1/LSD1 co-localize to regions with demethylated histones upon stimulation with CXCL12. Alternatively, recombinant LASP-1 will be mixed with purified LSD-1 and see if LASP-1 enhances in vitro demethylation activity on labeled histone H3 or nucleosome substrates. Additionally, non-silenced and LASP1-knock down basal- like breast cancer cells will be tested for their ability to facilitate bone resorption in a dentine disk assay. LASP1-Snail1 axis would become a novel drug target for small molecule inhibitors with the aim of prolonging the survival rate especially in triple-negative breast cancer patients. Novel inhibitors might be even more useful in cases of breast cancer that are resistant to current chemotherapeutic agents.

 描述（由适用提供）：拟议的研究的目的是研究CXCL12驱动的核LASP1在调节乳腺癌表观遗传事件中的作用。 LASP-1介导了几种乳腺癌细胞系中的细胞迁移，增殖和存活。 LASP-1的沉默抑制了45％的增殖和迁移。以前，LASP-1已证明与CXC趋化因子受体CXCR1，CXCR2，CXCR3和CXCR4直接相互作用，这些受体在支持乳腺癌进展和转移的肿瘤微环境中起关键作用。特别是，LASP-1增强了CXCR2介导的细胞迁移。乳腺癌细胞的表观遗传学改变将其转化为侵略性和转移性表型。在这项研究中，通过蛋白质组学，UHRF1被发现是一种新型LASP-1关联蛋白。随后，还观察到DNMT1，G9A和SNAIL1与LASP-1相关。特别是，发现Snail1直接与LASP-1结合。这种直接相互作用的生物学意义尚不清楚。首先，我建议通过突变和生化方法绘制LASP-1和SNAIL1之间的相互作用位点。我进一步建议研究LASP1-SNAIL1复合物的调节E-钙粘蛋白启动子的能力。另外，如Snail1直接绑定 对于赖氨酸甘甲基1（LSD1），将通过生化研究分析LSD1是否与LASP1-SNAIL1复合物相关。在功能上，我建议对组蛋白H3的LASP-1，LSD1和K4/9甲基化进行染色质免疫沉淀（CHIP）分析，以查看LASP-1/LSD1在用CXCL12刺激后与脱甲基化组蛋白的区域是否共定位。或者，重组LASP-1将与纯化的LSD-1混合，看看LASP-1在标记的组蛋白H3或核小体底物上是否增强了体外脱甲基化活性。此外，将测试非脱毛和LASP1敲击基本的乳腺癌细胞，以便在牙本质磁盘测定中促进骨骼修复的能力。 LASP1-SNAIL1轴将成为小分子抑制剂的新型药物靶标，目的是延长生存率，尤其是在三阴性乳腺癌患者中。在抗当前化学治疗剂的乳腺癌病例中，新型抑制剂可能更有用。