KLF8 Regulation of EMT Tumor Metastasis

KLF8对EMT肿瘤转移的调节

基本信息

批准号：
8517026
负责人：
JIHE ZHAO
金额：
$ 24.54万
依托单位：
UNIVERSITY OF CENTRAL FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-01 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8517026
关键词：
Acinus organ component Adhesions Biochemical Breast Cancer Cell Cause of Death Cells E-Cadherin Epithelial Cells Event Fatty acid glycerol esters Focal Adhesion Kinase 1 Genetic Genetic Transcription Goals Human Injection of therapeutic agent Integrins Kruppel-like transcription factors Lung MMP2 gene MMP9 gene Mammary gland Matrix Metalloproteinases Mediating Modeling Molecular N-Cadherin Neoplasm Circulating Cells Neoplasm Metastasis Nude Mice Oncogenic Pathway interactions Patients Phenotype Pilot Projects Play Proteins RNA Regulation Repression Role Signal Transduction Snails Sorting - Cell Movement Specimen Tail Testing Tumor Cell Invasion Variant Veins Woman Xenograft procedure bioluminescence imaging cancer cell cancer imaging catenin p120ctn protein design epithelial to mesenchymal transition human MMP14 protein in vivo in vivo Model knock-down malignant breast neoplasm mouse model novel novel therapeutic intervention novel therapeutics overexpression prevent slug tumor tumor progression

项目摘要

Project summary Breast cancer remains one of the leading causes of death in women and most patients die from the tumor metastasis. Our long-term goal is to understand the molecular mechanisms of metastasis so that novel therapeutic strategy can be designed. EMT (epithelial to mesenchymal transition) is a determining step for a cancer cell to progress from a non-invasive to invasive state. Disruption of intercellular adhesion via loss of E- cadherin, the defining event for EMT, is often correlated with gain of the matrix metalloproteinases (MMP) that catalyze proteolytic matrix remodeling important for tumor cell invasion. Such a correlation has been found in invasive breast cancer. However how EMT is regulated during and contributes to breast cancer metastasis is largely unknown. We previously identified Kr¿ppel-like transcription factor 8 (KLF8) as a downstream effector of FAK that plays a crucial role in breast cancer metastasis. Recently, we found that KLF8 is important for oncogenic transformation and overexpressed in invasive human breast cancer, suggesting an uninvestigated role for KLF8 in breast cancer metastasis. Our preliminary studies, using inducible and constitutive expression of KLF8 in MCF-10A cells, suggest that KLF8 is a potent inducer of EMT and invasion by repressing E- cadherin and activating MMP9, MMP2 and MT1-MMP. We also show that KLF8 plays a key role in the loss of E-cadherin and gain of the MMPs in invasive human breast cancer cells and that aberrant KLF8 elevation is tightly correlated with the loss of E-cadherin in patient specimens. Finally, our pilot studies suggest a role for KLF8 in supporting MDA-MB-231 lung metastasis. These results strongly support our hypothesis that KLF8 promotes breast cancer metastasis by initiating EMT and invasion through repression of E-cadherin and activation of the MMPs. We propose three specific aims to test this hypothesis using in semi-in vivo 3D acinus culture and in vivo mouse models and our MCF-10A cells expressing inducible KLF8 and MDA-MB-231 cells expressing inducible KLF8 sh RNA. In Aim 1, we will determine the role of E-cadherin and the MMPs in KLF8- induced EMT and invasion by genetically modifying these proteins. In Aim 2, we will identify the molecular mechanisms by which E-cadherin, the MMPs and intergrin-FAK signaling interplay to regulate KLF8-induced EMT and invasion using genetic, biochemical or pharmacological approaches. In Aim 3, we will determine the role of KLF8 in tumor metastasis using a mouse model of human breast cancer and bioluminescence imaging. We will also evaluate role for other EMT regulators such as Snails, ZEBs and Twist in the regulation of metastasis by KLF8. Completion of this project will shed new lights on mechanisms responsible for breast cancer metastasis which may leads to new therapeutic intervention.

项目摘要乳腺癌仍然是女性死亡的主要原因之一，大多数患者死于肿瘤转移。我们的长期目标是了解转移的分子机制，以便新颖可以设计治疗策略。 EMT（上皮到间充质转变）是A的确定步骤癌细胞从非侵入性到侵入性状态发展。通过损失E-的细胞间粘附破坏 Cadherin是EMT的定义事件，通常与基质金属蛋白酶（MMP）的增益相关。催化蛋白水解基质重塑对于肿瘤细胞浸润很重要。在侵入性乳腺癌。但是，如何调节EMT并在乳腺癌转移过程中有效在很大程度上未知。我们以前以前将kr¿ppel样转录因子8（KLF8）识别为下游效应器 FAK在乳腺癌转移中起着至关重要的作用。最近，我们发现KLF8对于致癌性转化并在侵入性的人类乳腺癌中过表达，表明未投资 KLF8在乳腺癌转移中的作用。我们的初步研究，使用诱导和本构表达 MCF-10A细胞中KLF8的作用，表明KLF8是EMT和侵袭的潜在诱导剂，通过抑制E- 钙粘蛋白和激活MMP9，MMP2和MT1-MMP。我们还表明，KLF8在损失中起关键作用 E-钙黏着蛋白和MMP在侵入性人类乳腺癌细胞中的增益，而异常KLF8升高为与患者标本中电子钙粘蛋白的损失密切相关。最后，我们的试点研究提出了 KLF8支持MDA-MB-231肺转移。这些结果强烈支持了我们KLF8的假设通过反射电子钙粘蛋白和 MMP的激活。我们提出了三个特定的目的，以测试使用半体内3D acinus中使用该假设培养和体内小鼠模型以及我们表达诱导KLF8和MDA-MB-231细胞的MCF-10A细胞表达可诱导的KLF8 SH RNA。在AIM 1中，我们将确定E-钙粘蛋白和MMP在KLF8-中的作用。通过遗传修饰这些蛋白质引起的EMT和侵袭。在AIM 2中，我们将确定分子 E-钙粘蛋白，MMP和Grin-Fak信号传导相互作用以调节KLF8诱导的机制使用遗传，生化或药物方法的EMT和入侵。在AIM 3中，我们将确定 KLF8使用人类乳腺癌和生物发光成像的小鼠模型在肿瘤转移中的作用。我们还将评估其他EMT调节剂的作用，例如蜗牛，Zebs和Twist KLF8的转移。该项目的完成将为负责乳房的机制开发新的灯光癌症转移可能导致新的治疗干预。