Malignant melanoma: Regulation by AP-2 and PAR-1

恶性黑色素瘤：AP-2 和 PAR-1 的调节

基本信息

批准号：
8065375
负责人：
MENASHE BARELI
金额：
$ 26.29万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-04-01 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8065375
关键词：
Angiopoietin-2 Applications Grants Cell Nucleus Coagulation Process Complementary DNA Connexin 43 Coupled Cutaneous Melanoma Data Development Dominant-Negative Mutation Event Funding Gelatinase A Gene Targeting Genes Health Human IL8 gene In Vitro Laboratories Laser Scanning Cytometry Lead Life Ligands Link Liposomes MCAM gene Melanoma Cell Membrane Metastatic Melanoma Metastatic to Modality Molecular Neoplasm Metastasis Nuclear Phenotype Progress Reports Radial Growth Phase Regulation Research Role Small Interfering RNA System TFAP2A gene TFAP2B gene Technology Testing Thrombin Thrombin Receptor Tissue Microarray Tumor Suppressor Genes Tumorigenicity Up-Regulation Vascular Endothelial Growth Factors Vertical Growth Phase Work base cytokine improved in vivo malignant phenotype maspin melanoma nanoparticle novel therapeutics small hairpin RNA success transcription factor tumor growth

项目摘要

DESCRIPTION (provided by applicant): The molecular changes associated with the transition of melanoma cells from Radial Growth Phase (RGP) to Vertical Growth Phase (VGP) and the acquisition of the metastatic phenotype is not very well-defined. Recent work from this laboratory demonstrated that this transition is associated with loss of nuclear expression of AP-2a in metastatic melanoma cells which resulted in deregulation of c-KIT, MCAM/MUC18, MMP-2, and VEGF, all of which are involved in the progression of human melanoma. However, the mechanisms for the lack of AP-2a expression in metastatic melanoma cells are not known. Using melanoma tissue microarrays combined with LSC and AQUA technologies, here, we further validated the lack of nuclear AP-2a expression in metastatic melanoma cells. In an effort to search for other target genes regulated by AP-2a, we have identified the thrombin receptor (PAR-1) and provided evidence for inverse correlation between AP-2a and PAR-1 expression in metastatic melanoma cells. Regulation of PAR-1 by AP-2a was demonstrated in vitro and in vivo, thus providing a unique link between the coagulation system and the progression of human melanoma. The role of PAR-1 in the progression of human melanoma is now established through the efforts of this continued research. We have used stable silencing of PAR-1 by lentiviral shRNA combined with cDNA chip arrays and cytokine membrane arrays to identify possible downstream genes regulated by the axis of thrombin/PAR-1. These genes include MCAM/MUC18, Connexin 43, Maspin, RUNX3, IL-8, Ang-2, GRO, ACRP30, and GITR-ligand. The hypothesis to be tested in this proposal is that loss of nuclear AP-2a results in upregulation of PAR-1 and contributes to the acquisition of the malignant phenotype in human melanoma. To test this hypothesis, we now propose: 1) To determine the mechanism(s) for the loss of nuclear AP-2a expression during melanoma progression; 2) To investigate how the axis of thrombin/PAR-1 contributes to the metastatic phenotype; and 3) To inhibit PAR-1 expression in vivo via delivery of siRNA packaged in neutral liposome nanoparticles as a possible new therapeutic modality. It is expected that the results obtained from this study will provide a better understanding of the role of AP-2a and PAR-1 in the progression of human melanoma that could thereby lead to new modalities to inhibit melanoma metastasis. PUBLIC HEALTH RELEVANCE: The molecular changes that are associated with the progression of human melanoma from radial growth phase into a vertical growth phase (metastatic phenotype) are largely unknown. Previously, we identified that the loss of the transcription factor AP-2a is associated with this transition. In this grant application, we will continue to investigate the role of AP-2a and its downstream target gene, the thrombin receptor (PAR-1), in the progression of human melanoma.

描述（由申请人提供）：与黑色素瘤细胞从径向生长阶段（RGP）转变为垂直生长阶段（VGP）的分子变化以及转移表型的获取不是很好。该实验室的最新工作表明，这种过渡与转移性黑色素瘤细胞中AP-2A的核表达丧失有关，从而导致C-KIT，MCAM/MUC18，MMP-2和VEGF的管制管制，所有这些都参与了人类黑色素瘤的进展。但是，尚不清楚转移性黑色素瘤细胞中缺乏AP-2A表达的机制。使用黑色素瘤组织微阵列结合LSC和Aqua技术，我们进一步验证了转移性黑色素瘤细胞中缺乏核AP-2A表达。为了搜索由AP-2A调控的其他靶基因，我们已经鉴定了凝血酶受体（PAR-1），并提供了转移性黑素瘤细胞中AP-2A与PAR-1表达之间相关性相关性的证据。在体外和体内证明了通过AP-2A对PAR-1的调节，因此在凝结系统与人类黑色素瘤的进展之间提供了独特的联系。现在，通过这项持续研究的努力确定了PAR-1在人黑色素瘤进展中的作用。我们已经使用慢病毒shRNA和cDNA芯片阵列和细胞因子膜阵列对PAR-1的稳定沉默来鉴定由凝血酶/PAR-1轴调节的可能的下游基因。这些基因包括MCAM/MUC18，Connexin 43，Maspin，Runx3，IL-8，Ang-2，Gro，GRO，ACRP30和GITR-配合。该提议中要检验的假设是核AP-2A的损失导致PAR-1上调，并有助于人类黑色素瘤中恶性表型的获取。为了检验该假设，我们现在建议：1）确定黑色素瘤进展过程中核AP-2a表达丧失的机制； 2）研究凝血酶/PAR-1的轴如何促进转移表型； 3）通过在中性脂质体纳米颗粒中包装的siRNA作为可能的新治疗方式来抑制体内PAR-1表达。可以预期，从这项研究中获得的结果将更好地理解AP-2A和PAR-1在人黑色素瘤进展中的作用，从而导致新的方式抑制黑色素瘤转移。公共卫生相关性：与人类黑色素瘤从径向生长阶段发展为垂直生长阶段（转移表型）相关的分子变化在很大程度上是未知的。以前，我们确定转录因子AP-2A的丢失与该过渡有关。在此赠款应用中，我们将继续研究AP-2A及其下游靶基因凝血酶受体（PAR-1）在人类黑色素瘤进展中的作用。