Investigating the role of Eya3 in the regulation of innate immune signaling cascades in Triple Negative Breast Cancer

研究 Eya3 在三阴性乳腺癌先天免疫信号级联调节中的作用

• 批准号: 10415832
• 负责人: Connor J Hughes
• 金额: $ 3.57万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-13 至 2024-01-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10415832
• 关键词: Apoptosis; Binding; Biology; Breast Cancer Cell; Breast Cancer Patient; CD8-Positive T-Lymphocytes; CRISPR/Cas technology; Cell physiology; Cells; Complex; Cytokine Signaling; Data; Development; Disease; Distant; Distant Metastasis; ERBB2 gene; Embryonic Development; Estrogen Receptors; Eye; Family; Family member; Genetic; Genetic Transcription; Growth; Growth Factor Receptors; Homeodomain Proteins; Human; Immune; Immune Evasion; Immune signaling; Immunocompetent; Immunosuppression; Immunotherapy; In Vitro; Infiltration; Innate Immune Response; Laboratories; Lead; Malignant Neoplasms; Mediating; Mentors; Mind; Mus; NF-kappa B; Neoplasm Metastasis; Pathway interactions; Pharmacology; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Play; Point Mutation; Population; Prevention; Primary Neoplasm; Process; Prognosis; Protein Family; Protein Phosphatase 2A Regulatory Subunit PR53; Protein Serine/Threonine Phosphatase; Protein Tyrosine Phosphatase; Protein phosphatase; Proteins; Publishing; Regulation; Resistance; Role; Signal Pathway; Signal Transduction; Testing; Threonine; Tyrosine; Work; adaptive immune response; angiogenesis; breast cancer progression; chemokine; cofactor; cytokine; experimental study; immunoregulation; improved; in vivo; insight; knock-down; malignant breast neoplasm; migration; mouse model; neoplastic cell; new therapeutic target; novel; novel therapeutic intervention; overexpression; phosphoproteomics; prevent; programmed cell death ligand 1; small hairpin RNA; targeted treatment; therapeutic development; therapeutic target; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor microenvironment; tumor progression

PROJECT SUMMARY/ABSTRACT: Triple Negative Breast Cancer (TNBC) is a subtype of breast cancer characterized by low or absent expression of the estrogen receptor and the HER2 growth factor receptor. TNBC has high rates of metastasis and an overall poor prognosis in large part due to a lack of targeted therapeutics for treating both localized and disseminated disease. The Eyes absent (Eya) family of proteins are transcriptional cofactors that possess both intrinsic tyrosine phosphatase and associated serine/threonine phosphatase activity, and this family of proteins has been shown to play important roles in normal development in mice and humans. In addition to their developmental roles, Eya proteins have also been implicated in promoting nearly all known hallmarks of cancer. Recent published work from the laboratory of my mentor, Dr. Heide Ford, demonstrated that Eya3 plays an important role in suppressing CD8+ T-cell function within TNBC tumors by downregulating expression of PD-L1, primarily through its threonine phosphatase activity, and preliminary data suggests that it may also play a role in regulating the expression of various cytokines/chemokines in this context. Activation of innate immune signaling cascades in tumor cells, such as NF-kB, and downstream cytokine/chemokine signaling, has been shown to promote tumor progression, immune evasion, and distant metastasis in a variety of tumor types, including breast cancer. In preliminary experiments, we observed that shRNA-mediated Eya3 knockdown (KD) in murine TNBC cells reduced expression of multiple cytokines and chemokines and reduced NF-kB activation in vitro, suggesting a strong regulatory role of Eya3 in these signaling cascades. Additionally, in vivo experiments suggest that Eya3 KD in murine TNBC cells greatly reduces primary tumor growth and spontaneous metastasis in immune-competent mouse models, and this change correlates with significant alterations in innate immune populations within the Eya3 KD tumors compared to their control counterparts. With these preliminary findings in mind, we hypothesize that Eya3 expression regulates an innate immune signaling axis in TNBC cells which alters chemokine/cytokine signaling in the tumor microenvironment and facilitates enhanced primary tumor growth and metastasis. Our aims are as follows: 1. To determine the contribution of Eya3-mediated induction of innate immune signaling in TNBC cells to tumor-initiated immune suppression, tumor growth, and metastasis and 2. To identify the mechanism of action by which Eya3 regulates innate immune signaling pathways in TNBC. These studies present an opportunity not only to investigate the important biology governing tumor cell crosstalk and mechanisms of immune subversion, but additionally will identify potential novel therapeutic targets for prevention or treatment of disseminated TNBC.

项目摘要/摘要： 三重阴性乳腺癌（TNBC）是乳腺癌的亚型，其特征是低或不存在 雌激素受体和HER2生长因子受体的表达。 TNBC的转移率很高 而且总体预后不佳，这很大程度上是由于缺乏针对性的治疗局部和治疗局部和 传播疾病。 蛋白质不存在的眼睛（EYA）家族是具有固有性的转录辅助因子 酪氨酸磷酸酶和相关的丝氨酸/苏氨酸磷酸酶活性，这种蛋白质家族具有 被证明在小鼠和人类的正常发育中起着重要作用。除了他们 发育作用，EYA蛋白也与促进几乎所有已知标志有关 癌症。我的导师海德·福特（Heide Ford）实验室的最新发表的工作证明了Eya3 通过下调表达来抑制TNBC肿瘤中CD8+ T细胞功能的重要作用 PD-L1的主要是通过其苏氨酸磷酸酶活性，初步数据表明它也可以 在这种情况下，在调节各种细胞因子/趋化因子的表达方面发挥作用。先天的激活 肿瘤细胞中的免疫信号传导级联反应，例如NF-KB和下游细胞因子/趋化因子信号传导 已显示可促进多种肿瘤中的肿瘤进展，免疫逃避和远处转移 类型，包括乳腺癌。在初步实验中，我们观察到shRNA介导的eya3 鼠TNBC细胞中的敲低（KD）降低了多种细胞因子和趋化因子的表达，并降低 NF-KB在体外激活，表明EYA3在这些信号级联反应中的强烈调节作用。此外， 体内实验表明，鼠TNBC细胞中的EYA3 KD大大降低了原发性肿瘤的生长和 免疫能力小鼠模型中自发转移，这种变化与显着的 与对照组相比，EYA3 KD肿瘤内先天免疫种群的改变。 考虑到这些初步发现，我们假设EYA3表达可以调节先天性 TNBC细胞中的免疫信号传导轴改变了肿瘤中趋化因子/细胞因子信号传导 微环境和促进了原发性肿瘤的生长和转移。我们的目标是 以下内容：1。确定EYA3介导的TNBC细胞中先天免疫传导诱导的贡献 为了肿瘤引发的免疫抑制，肿瘤生长和转移以及2。 EYA3调节TNBC中先天免疫信号通路的作用。这些研究提出了 机会不仅要研究有关肿瘤细胞串扰和机制的重要生物学 免疫颠覆，但还将确定潜在的预防或治疗的新型治疗靶标 传播的TNBC。