Characterization of ILEI/LIFR Axis-induced Intracellular Signaling

ILEI/LIFR 轴诱导的细胞内信号传导的表征

基本信息

批准号：
10547814
负责人：
William Scott Streitfeld
金额：
$ 4.26万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-01 至 2023-11-22
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10547814
关键词：
AKT2 gene Affect Attenuated Binding Breast Carcinoma Breast Epithelial Cells CRISPR/Cas technology Cell Culture Techniques Cell Maintenance Cells Characteristics Clinical Clinical Treatment Consensus Data Disease Progression Elements Epithelial Cells Event Exposure to Fatty acid glycerol esters Gene Expression Gene Expression Profile Genes Genetic Transcription Goals Heterogeneous-Nuclear Ribonucleoproteins Human Human papillomavirus 16 E1 protein Immunodeficient Mouse In Vitro Injections Interleukins JAK2 gene Knock-out Laboratories Literature Luciferases Lung Maintenance Mammary Neoplasms Mammary gland Measures Mediating Metastatic Neoplasm to the Lung Metastatic breast cancer Modeling Mus Neoplasm Metastasis Nuclear Participant Phenotype Phosphorylation Promoter Regions Property Proteins Publishing RNA analysis Receptor Signaling Receptor Up-Regulation Regulation Relapse Research Residencies Resistance Ribonucleoproteins Role STAT3 gene Signal Pathway Signal Transduction Site-Directed Mutagenesis Tail Therapeutic Transforming Growth Factor beta Translating Translations Up-Regulation Veins cancer stem cell chromatin immunoprecipitation epithelial to mesenchymal transition experimental study in vivo knock-down leukemia inhibitory factor receptor malignant breast neoplasm mammary mammary epithelium neoplastic cell new therapeutic target novel progenitor promoter protein expression receptor receptor expression receptor upregulation response self-renewal stem cell self renewal stem cells stemness transcription factor transcriptome sequencing transcriptomic profiling transcriptomics tumor tumor growth tumor initiation tumor progression

项目摘要

Project Summary Our lab has previously identified a mechanism of transforming growth factor beta (TGFβ)-mediated epithelial-to-mesenchymal transition (EMT) in murine mammary epithelium. We have shown that expression of the interleukin-like EMT inducer (ILEI) protein is necessary to induce EMT in murine mammary gland cells following exposure to TGFβ. ILEI has also been shown to increase the self-renewal capacity of epithelial cells following EMT, through its interaction with leukemia inhibitory factor receptor (LIFR), suggesting that the ILEI/LIFR signaling axis promotes breast cancer stem-cell (BCSC) phenotype. Further, our lab has shown that TGFβ-induced upregulation of LIFR is ILEI-dependent. In cells derived from a mouse tumor progression model created by our lab, spheroid formation capacity in non-adherent cell culture conditions is attenuated following either ILEI or LIFR knockdown. Additionally, orthotopic grafts of cells with ILEI or LIFR knockdown display a decrease in tumor growth and metastasis relative to control cells. We hypothesize that TGFβ-induced LIFR regulation contributes to metastases. The precise mechanisms of ILEI-mediated EMT and BCSC induction are unknown. Herein we aim to interrogate ILEI/LIFR axis-mediated mechanisms downstream of TGFβ exposure that influence (1) the regulation of LIFR protein expression and (2) the ensuing maintenance of self-renewal capacity and disease progression in our model. In Specific Aim 1, the LIFR promoter sequence will be examined to identify key factors regulating LIFR expression. In Specific Aim 2, transcriptomic data will be examined following ILEI/LIFR knockout to identify a signature of ILEI/LIFR-regulated gene expression and its association with self-renewal capacity. In Specific Aim 3, the role of LIFR will be examined in vivo to determine the impact of its expression upon outgrowth of pulmonary tumors following tail vein injection of cells into immunodeficient mice. Data from our experiments will characterize a signaling pathway associated with BCSC maintenance and will potentially identify novel therapeutic targets. Our findings may translate to novel treatments for dormancy and relapse in human metastatic breast cancer.

项目摘要我们的实验室以前已经确定了转化生长因子β（TGFβ）介导的机制鼠乳腺上皮中的上皮到间质转变（EMT）。我们已经表明了白介素样EMT诱导蛋白（ILEI）蛋白对于诱导鼠乳腺细胞诱导EMT是必要的暴露于TGFβ之后。 ILEI还显示出增加上皮细胞的自我更新能力在EMT之后，通过与白血病抑制因子受体（LIFR）的相互作用，表明 ILEI/LIFR信号轴促进乳腺癌干细胞（BCSC）表型。此外，我们的实验室表明 TGFβ诱导的LIFR上调ILEI依赖性。在源自小鼠肿瘤进展模型的细胞中由我们的实验室创建的，在非粘附细胞培养条件下的球状形成能力受到衰减 Ilei或LIFR敲低。此外，具有ILEI或LIFR敲低的细胞的原位移植物显示相对于对照细胞的肿瘤生长和转移减少。我们假设TGFβ诱导的LIFR 调节有助于转移。 ILEI介导的EMT和BCSC诱导的精确机制尚不清楚。我们的目标是质疑ILEI/LIFR轴介导的TGFβ暴露的机制，影响（1）调节 LIFR蛋白表达和（2）确保维持自我更新能力和疾病进展在我们的模型中。在特定目标1中，将检查LIFR启动子序列以识别调节的关键因素 LIFR表达。在特定的目标2中，将在ILEI/LIFR敲除之后检查转录组数据以识别 ILEI/LIFR调节的基因表达的签名及其与自我更新能力的关联。具体 AIM 3，将在体内检查LIFR的作用，以确定其表达对生长的影响尾静脉注射细胞后，肺部肿瘤进入免疫缺陷小鼠。我们实验的数据将表征与BCSC维护相关的信号通路和将有可能识别新的治疗靶标。我们的发现可能转化为休眠的新治疗方法并缓解人类转移性乳腺癌。