Effects of the PI3K Pathway on p27 Function and Cancer Progression

PI3K 通路对 p27 功能和癌症进展的影响

基本信息

批准号：
8608487
负责人：
JOYCE MARIE SLINGERLAND
金额：
$ 23.06万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-03-23 至 2016-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8608487
关键词：
3-Phosphoinositide Dependent Protein Kinase-1 Actins Actomyosin Binding Binding Sites Cancer Detection Cell Cycle Cell Cycle Progression Cell Proliferation Cells Cyclin E Cyclins Cytoplasm Cytoskeleton Development Family Gene Deletion Goals Grant Growth Guanosine Triphosphate Human In Vitro Injection of therapeutic agent Knock-in Mouse Knockout Mice Knowledge Link Lung Lung Neoplasms MDA MB 231 Malignant Neoplasms Mediating Mediator of activation protein Metastatic Neoplasm to the Lung Modeling Neoplasm Metastasis Nuclear Nuclear Import Oncogene Activation Oncogenic PTEN gene Pathway interactions Patients Pharmaceutical Preparations Phosphorylation Phosphotransferases Play Post-Translational Protein Processing Precipitation Predisposition ROCK1 gene Raptors Reporting Role Signal Pathway Signal Transduction Site Stress Fibers Tail Testing Tissues Transfection Tumor Cell Invasion Tumorigenicity Variant Veins Work Xenograft procedure bone cancer cell cell growth cell motility cofilin cyclin-dependent kinase inhibitor 1B design gain of function human FRAP1 protein in vivo inhibitor/antagonist knock-down mTOR Inhibitor mouse model mutant neoplastic cell new therapeutic target outcome forecast overexpression prevent progenitor public health relevance response rho small hairpin RNA stem cell population therapeutic target tool tumor tumor progression tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): While nuclear p27 is often reduced, p27 gene deletion and complete p27 loss is rare in human cancers. Cytoplasmic p27, seen in many cancers, is associated with a poor prognosis. Cytoplasmic p27 acquires an oncogenic gain of function to promote cell motility by binding RhoA to inhibit RhoA-ROCK activation needed for cytoskeletal stability. p27CK- knock- in mice show cytoplasmic p27, and increased cell motility, progenitor/stem cell populations and lung tumor formation. Thus, p27 regulates both cell proliferation and migration, and has a pro-oncogenic action to promote cell motility independent of its cell cycle effect, which may explain why p27 is rarely entirely lost in human cancers. Here we investigate how p27 phosphorylation regulates its motility function. We showed that p27 is phosphorylated by AGC family kinases downstream of PI3K at T157, T198, or both, which impairs nuclear p27 import, stabilizes p27 in the cytoplasm and stimulates cell motility. Cells overexpressing AGC kinases show a p27-dependent increased cell motility which is reversed by p27 knock- down via shRNAp27. T198 phosphorylation increased p27:RhoA binding in vitro and p27T157AT198A bound RhoA poorly, suggesting that T198 phosphorylation may increase cell motility via Rho-ROCK1 inhibition. p27 also appears to mediate PI3K driven metastasis. High metastatic MDA-MB-231-derivatives (4175 and 1833) showed PI3K activation, increased cytoplasmic p27 and increased cell motility that were reversed by p27 knockdown. p27 knock-down reversed the high lung metastasis of MDA-MB-231-4715 to levels similar to those in parental MDA-MB-231. Thus, PI3K may stimulate invasion and metastasis via oncogenic effects of cytoplasmic p27. Our hypothesis is that p27 phosphorylation at T198 promotes its association with RhoA to increase tumor cell motility and potentiate tumor invasion and metastasis. This grant investigates further how p27 phosphorylation by PI3K effector kinases increases cell motility and tests if p27pT157pT198 promotes tumorigenesis and metastasis in vivo. AIM1 will identify the p27:RhoA binding site and test the importance of phosphorylation to RhoA binding and increased cell motility. AIM 2 will test effects of pT198 and cytoplasmic p27 on local tumor invasion and metastasis in xenografts. AIM 3 will create p27CK-T198D and p27CK-T198A knock-in mice and test their susceptibility to tumorigenesis. Cytoplasmic p27pT198 and its cytoskeleton effects may be major drivers of tumorigenicity and metastasis activated by RTK/PI3K in human cancers. Therapeutic targeting agents that disrupt p27:RhoA interaction may prevent tumor invasion and metastasis. In cancers, cytoplasmic p27 may predict potential for response to PI3K/mTOR inhibitor drugs.

描述（由申请人提供）：虽然核P27通常会降低，但P27基因缺失和完全的P27损失在人类癌症中很少。在许多癌症中看到的细胞质p27与预后不良有关。细胞质p27通过结合RhoA抑制细胞骨架稳定性所需的Rhoa-Rock激活，从而获得了功能的致癌性增益，以促进细胞运动。小鼠中的p27ck-敲击显示细胞质P27，细胞运动，祖/干细胞群体和肺部肿瘤的形成增加。因此，p27调节细胞增殖和迁移，并具有促疾病，可以促进细胞运动与其细胞周期效应无关，这可以解释为什么p27在人类癌症中很少完全丢失。在这里，我们研究了P27磷酸化如何调节其运动功能。我们表明，p27在T157，T198或两者兼而有之的PI3K下游AGC家族激酶磷酸化，这损害了核P27的进口，使P27稳定在细胞质中并刺激细胞运动。过表达AGC激酶的细胞显示P27依赖性增加的细胞运动性，这被p27敲击通过SHRNAP27逆转。 T198磷酸化增加了P27：RHOA在体外结合，P27T157AT198A结合了RhoA的较差，这表明T198磷酸化可能通过Rho-Rock1抑制增加细胞运动性。 p27似乎还介导了PI3K驱动的转移。高转移性MDA-MB-231衍生物（4175和1833）显示PI3K激活，细胞质P27增加，并增加了细胞运动性，而细胞运动性则被P27敲低反转。 p27敲除将MDA-MB-231-4715的高肺转移扭转到与父母MDA-MB-231相似的水平。因此，PI3K可以通过细胞质P27的致癌作用刺激侵袭和转移。我们的假设是，T198处的p27磷酸化促进了其与RhoA的关联，以增加肿瘤细胞运动性并增强肿瘤侵袭和转移。该赠款进一步研究了PI3K效应子激酶的P27磷酸化如何增加细胞运动性并测试P27PT157PT198在体内促进肿瘤发生和转移。 AIM1将识别p27：RhoA结合位点，并测试磷酸化对RhoA结合和增加细胞运动的重要性。 AIM 2将测试PT198和细胞质P27对异种移植物中局部肿瘤侵袭和转移的影响。 AIM 3将创建P27CK-T198D和P27CK-T198A敲入小鼠，并测试其对肿瘤发生的敏感性。细胞质P27PT198及其细胞骨架作用可能是RTK/PI3K在人类癌症中激活的肿瘤性和转移的主要驱动因素。破坏p27的治疗靶向剂：RhoA相互作用可能预防肿瘤侵袭和转移。在癌症中，细胞质P27可能预测对PI3K/MTOR抑制剂药物的反应潜力。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

p27: a barometer of signaling deregulation and potential predictor of response to targeted therapies.

p27：信号放松管制的晴雨表和靶向治疗反应的潜在预测因子。

DOI：
10.1158/1078-0432.ccr-10-0752
发表时间：
2011-01-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
0
作者：
Wander SA;Zhao D;Slingerland JM
通讯作者：
Slingerland JM

PI3K/mTOR inhibition can impair tumor invasion and metastasis in vivo despite a lack of antiproliferative action in vitro: implications for targeted therapy.

DOI：
10.1007/s10549-012-2389-6
发表时间：
2013-04
期刊：
BREAST CANCER RESEARCH AND TREATMENT
影响因子：
3.8
作者：
Wander, Seth A.;Zhao, Dekuang;Besser, Alexandra H.;Hong, Feng;Wei, Jianqin;Ince, Tan A.;Milikowski, Clara;Bishopric, Nanette H.;Minn, Andy J.;Creighton, Chad J.;Slingerland, Joyce M.
通讯作者：
Slingerland, Joyce M.

Erratum to: PI3K/mTOR inhibition can impair tumor invasion and metastasis in vivo despite a lack of antiproliferative action in vitro: implications for targeted therapy.

勘误表：尽管体外缺乏抗增殖作用，但 PI3K/mTOR 抑制可损害体内肿瘤侵袭和转移：对靶向治疗的影响。

DOI：
10.1007/s10549-016-3752-9
发表时间：
2016
期刊：
Breast cancer research and treatment
影响因子：
3.8
作者：
Wander,SethA;Zhao,Dekuang;Besser,AlexandraH;Hong,Feng;Wei,Jianqin;Ince,TanA;Milikowski,Clara;Bishopric,NanetteH;Minn,AndyJ;Creighton,ChadJ;Slingerland,JoyceM
通讯作者：
Slingerland,JoyceM