Signaling of Integrin Alpha 7

整合素 Alpha 7 的信号转导

• 批准号: 8676444
• 负责人: JIANHUA LUO
• 金额: $ 24.78万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8676444
• 关键词: Amino Acid Motifs; Amino Acids; Anchorage-Independent Growth; Apoptosis; Binding; Biological; Biological Assay; Breast; Cell Death; Cell Death Induction; Cell Death Inhibition; Cell Differentiation process; Cell Line; Cells; Clinical; Cyclin-Dependent Kinase Inhibitor 3; Cytoskeleton; DU145; Data; Deceleration; Detection; Development; Diagnosis; Disease; Down-Regulation; Epithelial Cells; Event; Extracellular Matrix; Frequencies; Genes; Genome; Glioblastoma; Growth; Heterogeneity; ITGA7 gene; In Situ Nick-End Labeling; In Vitro; Indolent; Induction of Apoptosis; Integrins; Knock-out; Knockout Mice; Lead; Life; Link; Liver; Malignant Neoplasms; Malignant neoplasm of prostate; Mass Spectrum Analysis; Mediating; Meta-Analysis; Micro Array Data; Mouse Strains; Mus; Mutate; Mutation; Mutation Analysis; Neoplasm Metastasis; Organogenesis; PC3 cell line; Patients; Peptide Hydrolases; Phosphorylation; Phosphorylation Site; Phosphotransferases; Play; Point Mutation; Primary carcinoma of the liver cells; Prostate; Prostate Adenocarcinoma; Prostate Leiomyosarcoma; Prostate-Specific Antigen; Prostatic Neoplasms; Protein-Serine-Threonine Kinases; Proteins; Publishing; Relapse; Respiratory Diaphragm; Role; Serum; Signal Pathway; Signal Transduction; Small Interfering RNA; Staining method; Stains; Structural Protein; System; TdT-Mediated dUTP Nick End Labeling Assay; Testing; Tissues; Tumor Suppression; Tumor Suppressor Genes; Tumor Volume; Wound Healing; Xenograft procedure; annexin A5; base; cancer cell; caspase-3; cell growth; cell motility; clinically relevant; expression vector; human ITGA7 protein; integrin-linked kinase; knock-down; leiomyosarcoma; matrigel; migration; mortality; mouse model; mutant; myopodin; neoplastic cell; overexpression; programs; restoration; screening; soft tissue; tumor growth; tumorigenesis; vector; yeast two hybrid system

DESCRIPTION (provided by applicant): Studies in liver and breast have indicated that extracellular matrix plays a critical role in limiting the growth and promoting the differentiation of epithelial cells. However, the signaling mechanism that leads to inhibition of over-growth of epithelial cells is not well understood. Recently, we performed a mutational analysis on ITGA7 gene (ITGA7). We found mutations on ITGA7 in prostate cancer, hepatocellular carcinoma, leiomyosarcoma and glioblastoma multiforms with frequencies ranging from 25% to 83%. Many of these mutations caused truncation, micro-deletion or frameshift of the protein. Interestingly, patients with ITGA7 mutations had higher rate of clinical relapse in both prostate cancer and hepatocellular carcinoma. Mouse model of PC3 and Du145 xenografted prostate tumors showed a dramatic reduction in tumor volume, rate of metastasis and rate of mortality when expression of ITGA7 was restored in these cell lines. ITGA7 induced the expression of cyclin dependent kinase inhibitor 3 (CDKN3) and RACGAP1. siRNA knocking down of CDKN3 and RACGAP1 completely reversed the growth inhibition effect of IGGA7, while only partially reversed the migration inhibition activity. Recently, Annexin V staining and TUNEL assays indicated that over-expression of ITGA7 induced apoptosis in PC3 cells. Through a Yeast two-hybrid system, we identified that the C-terminus of ITGA7 interacted with HtrA2, a cell death protein. A 30 amino acid motif located in the C-terminus of ITGA7 was identified as critical for its interaction with HtrA2. Expression of ITGA7 induced protease activity of HtrA2. siRNA knocking down of HtrA2 abolished the cell death induction by ITGA7. Separately, expression of ITGA7 induced the kinase activity of integrin link kinease (ILK). Interestingly, we also found that ILK, a serine/threonine kinase and structural protein associated with integrins and the cytoskeleton, bound and phosphorlyated a tumor suppressor gene called myopodin, which regulates cell growth and cell motility. Based on these preliminary data, we hypothesize that ITGA7 activates HtrA2 and ILK signaling pathways to achieve induction of apoptosis, cell growth inhibition and migration deceleration. In this study, we propose: 1) To elucidate the role of ITGA7/HtrA2 interaction in mediating cell death and inhibition of cancer invasion by analyzing the mutant ITGA7 molecule defective of binding with HtrA2 in PC3 and DU145 cells; 2) To investigate the role of ILK/myopodin interaction in ITGA7 regulated cell motility, cell growth and inhibition of invasiveness by analyzing myopodin defective cell system; 3) To study the impact of ITGA7 knock-out on the carciongenesis of TRAMP mice, and to investigate the biological role of ITGA7 in prostate gland tissue development, cell differentiation and tumorigenesis in pure ITGA7 knockout mice.

描述（由申请人提供）：肝脏和乳腺的研究表明，细胞外基质在限制上皮细胞生长和促进上皮细胞分化方面起着关键作用。然而，导致抑制上皮细胞过度生长的信号传导机制还不清楚。最近，我们对ITGA 7基因（ITGA 7）进行了突变分析。我们在前列腺癌、肝细胞癌、平滑肌肉瘤和多形性胶质母细胞瘤中发现ITGA 7突变，频率范围为25%至83%。这些突变中的许多导致蛋白质的截短、微缺失或移码。有趣的是，具有ITGA 7突变的患者在前列腺癌和肝细胞癌中具有更高的临床复发率。PC 3和Du 145异种移植前列腺肿瘤的小鼠模型显示，当ITGA 7的表达在这些细胞系中恢复时，肿瘤体积、转移率和死亡率显著降低。ITGA 7诱导细胞周期蛋白依赖性激酶抑制剂3（CDKN 3）和RACGAP 1的表达。siRNA敲低CDKN 3和RACGAP 1可完全逆转IGGA 7的生长抑制作用，但仅部分逆转其迁移抑制作用。最近，Annexin V染色和TUNEL检测表明ITGA 7的过表达诱导PC 3细胞凋亡。通过酵母双杂交系统，我们鉴定了ITGA 7的C-末端与细胞死亡蛋白HtrA 2相互作用。位于ITGA 7的C-末端的30个氨基酸基序被鉴定为其与HtrA 2相互作用的关键。ITGA 7的表达诱导HtrA 2的蛋白酶活性。siRNA敲低HtrA 2消除了ITGA 7诱导的细胞死亡。另外，ITGA 7的表达诱导了整合素连接激酶（ILK）的激酶活性。有趣的是，我们还发现，ILK，丝氨酸/苏氨酸激酶和结构蛋白与整合素和细胞骨架，绑定和磷酸化的肿瘤抑制基因称为肌足蛋白，调节细胞生长和细胞运动。基于这些初步数据，我们假设ITGA 7激活HtrA 2和ILK信号通路，以实现诱导细胞凋亡、细胞生长抑制和迁移减速。在这项研究中，我们建议：1）通过分析PC 3和DU 145细胞中ITGA 7与HtrA 2结合缺陷的突变体，阐明ITGA 7/HtrA 2相互作用在介导细胞死亡和抑制肿瘤侵袭中的作用：2）通过分析肌足蛋白缺陷细胞系统，研究ILK/肌足蛋白相互作用在ITGA 7调节细胞运动、细胞生长和抑制侵袭中的作用; 3）研究ITGA 7基因敲除对TRAMP小鼠前列腺癌发生的影响，并探讨ITGA 7基因在前列腺组织发育、细胞分化和肿瘤发生中的生物学作用。

项目成果

Investigating Multi-cancer Biomarkers and Their Cross-predictability in the Expression Profiles of Multiple Cancer Types.

• DOI: 10.4137/bmi.s930
• 发表时间: 2009-05-01
• 期刊: Biomarker insights
• 影响因子: 3.8
• 作者: Tseng GC;Cheng C;Yu YP;Nelson J;Michalopoulos G;Luo JH
• 通讯作者: Luo JH

Targeting genomic rearrangements in tumor cells through Cas9-mediated insertion of a suicide gene.

• DOI: 10.1038/nbt.3843
• 发表时间: 2017-06
• 期刊: Nature biotechnology
• 影响因子: 46.9
• 作者: Chen ZH;Yu YP;Zuo ZH;Nelson JB;Michalopoulos GK;Monga S;Liu S;Tseng G;Luo JH
• 通讯作者: Luo JH

Detection of fusion gene transcripts in the blood samples of prostate cancer patients.

• DOI: 10.1038/s41598-021-96528-9
• 发表时间: 2021-08-20
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Yu YP;Liu S;Nelson J;Luo JH
• 通讯作者: Luo JH

Phosphorylation and interaction of myopodin by integrin-link kinase lead to suppression of cell growth and motility in prostate cancer cells.

• DOI: 10.1038/onc.2011.200
• 发表时间: 2011-12-08
• 期刊: ONCOGENE
• 影响因子: 8
• 作者: Yu, Y-P;Luo, J-H
• 通讯作者: Luo, J-H

Metallothionein 1 h tumour suppressor activity in prostate cancer is mediated by euchromatin methyltransferase 1.

• DOI: 10.1002/path.4169
• 发表时间: 2013-06
• 期刊: JOURNAL OF PATHOLOGY
• 影响因子: 7.3
• 作者: Han, Yu-Chen;Zheng, Zhong-Liang;Zuo, Ze-Hua;Yu, Yan P.;Chen, Rui;Tseng, George C.;Nelson, Joel B.;Luo, Jian-Hua
• 通讯作者: Luo, Jian-Hua