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.

项目概要/摘要： 三阴性乳腺癌 (TNBC) 是乳腺癌的一种亚型，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 转移率高 总体预后不良，很大程度上是由于缺乏治疗局部和局部的靶向治疗方法 传播疾病。 眼睛缺失 (Eya) 蛋白家族是转录辅助因子，具有内在的 酪氨酸磷酸酶和相关的丝氨酸/苏氨酸磷酸酶活性，并且该蛋白质家族具有 已被证明在小鼠和人类的正常发育中发挥重要作用。除了他们的 除了发育作用外，Eya 蛋白还与促进几乎所有已知的特征有关 癌症。我的导师 Heide Ford 博士的实验室最近发表的工作表明，Eya3 通过下调表达，在 TNBC 肿瘤内抑制 CD8+ T 细胞功能中发挥重要作用 PD-L1 的作用，主要是通过其苏氨酸磷酸酶活性，初步数据表明，它也可能 在这种情况下，在调节各种细胞因子/趋化因子的表达中发挥作用。先天激活 肿瘤细胞中的免疫信号级联，例如 NF-kB 和下游细胞因子/趋化因子信号传导，已 已被证明可促进多种肿瘤的肿瘤进展、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 小鼠 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，并减少了 NF-kB 体外激活表明 Eya3 在这些信号级联中具有强大的调节作用。此外， 体内实验表明，小鼠 TNBC 细胞中的 Eya3 KD 极大地减少了原发性肿瘤的生长，并且 免疫活性小鼠模型中的自发转移，这种变化与显着相关 与对照肿瘤相比，Eya3 KD 肿瘤内先天免疫群体的变化。 考虑到这些初步发现，我们假设 Eya3 表达调节先天性 TNBC 细胞中的免疫信号轴改变肿瘤中的趋化因子/细胞因子信号 微环境并促进原发肿瘤生长和转移。我们的目标是 如下： 1. 确定 Eya3 介导的 TNBC 细胞中先天免疫信号传导诱导的贡献 2. 确定肿瘤引发的免疫抑制、肿瘤生长和转移的机制 Eya3 在 TNBC 中调节先天免疫信号通路的作用。这些研究提出了 不仅有机会研究控制肿瘤细胞串扰的重要生物学及其机制 免疫颠覆，而且还将确定潜在的新治疗靶点用于预防或治疗 传播的 TNBC。