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: 10548875
• 负责人: Connor J Hughes
• 金额: $ 3.68万
• 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/10548875
• 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; Innate Immune Response; Invaded; Laboratories; Malignant Neoplasms; Mediating; Mentors; Mind; Mus; NF-kappa B; Neoplasm Metastasis; Pathway interactions; Phenotype; Phosphoric Monoester Hydrolases; Play; Point Mutation; Population; Prevention; Primary Neoplasm; Process; Prognosis; Proliferating; Protein Dephosphorylation; Protein Family; Protein Phosphatase 2A Regulatory Subunit PR53; Protein Tyrosine Phosphatase; Protein phosphatase; Proteins; Publishing; Regulation; Resistance; Role; Serine; Signal Pathway; Signal Transduction; Testing; Threonine; Tumor Promotion; Tyrosine; Viral; Work; adaptive immune response; angiogenesis; breast cancer progression; chemokine; cofactor; cytokine; experimental study; immune cell infiltrate; immunoregulation; improved; in vivo; insight; knock-down; malignant breast neoplasm; migration; mouse model; neoplastic cell; new therapeutic target; novel; novel therapeutic intervention; overexpression; pharmacologic; 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 initiation; 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.

项目总结/摘要： 三阴性乳腺癌（TNBC）是乳腺癌的一种亚型，其特征在于低或无肿瘤标志物。 雌激素受体和HER 2生长因子受体的表达。TNBC转移率高 并且总体预后不良，这在很大程度上是由于缺乏用于治疗局部和局部性的靶向治疗剂， 传播性疾病 Eyes absent（Eya）家族蛋白是一种转录辅因子， 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 在小鼠和人类的正常发育中发挥重要作用。除了它们 Eya蛋白还参与促进几乎所有已知的发育特征， 癌我的导师Heide福特博士的实验室最近发表的工作表明，Eya 3 在通过下调表达来抑制TNBC肿瘤内的CD 8 + T细胞功能中起重要作用 PD-L1，主要是通过其苏氨酸磷酸酶活性，初步数据表明，它也可能 在此背景下在调节各种细胞因子/趋化因子的表达中起作用。先天激活 肿瘤细胞中的免疫信号级联，如NF-κ B和下游细胞因子/趋化因子信号， 在多种肿瘤中显示促进肿瘤进展、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Eya 3 在鼠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，并且降低了细胞因子和趋化因子的表达。 NF-kB在体外激活，表明Eya 3在这些信号级联中的强调节作用。此外，本发明还 体内实验表明，鼠TNBC细胞中的Eya 3 KD大大降低了原发性肿瘤生长， 免疫活性小鼠模型中的自发转移，并且这种变化与显著的 Eya 3 KD肿瘤内先天免疫群体与其对照对应物相比的改变。 考虑到这些初步的发现，我们假设Eya 3表达调节先天性免疫缺陷。 TNBC细胞中改变肿瘤中趋化因子/细胞因子信号传导的免疫信号传导轴 微环境和促进增强的原发性肿瘤生长和转移。我们的目标是 如下：1.确定Eya 3介导的TNBC细胞中先天免疫信号传导诱导的贡献 肿瘤引发的免疫抑制、肿瘤生长和转移，以及2.为了确定 Eya 3通过其调节TNBC中的先天免疫信号传导途径的作用。这些研究提出了一个 这不仅是研究控制肿瘤细胞串扰的重要生物学和肿瘤细胞相互作用的机制的机会， 免疫颠覆，但另外将确定潜在的新的治疗靶点，用于预防或治疗 传播性TNBC。