Nucleoporin NUP205-driven Novel Pathway in Melanoma Tumor Growth and Metastasis

核孔蛋白 NUP205 驱动的黑色素瘤生长和转移新途径

• 批准号: 9104466
• 负责人: Narendra Wajapeyee
• 金额: $ 38.3万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9104466
• 关键词: Accounting; Anoikis; BRAF gene; Biological Process; Cell Culture Techniques; Cell Line; Cessation of life; Chromosomal translocation; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Data Set; Doxycycline; Drug Targeting; Drug resistance; Enzymes; Genetic Predisposition to Disease; Genomic approach; Growth; Guide RNA; Hematologic Neoplasms; Heparitin Sulfate; Individual; Malignant Neoplasms; Measures; Melanoma Cell; Mitogen-Activated Protein Kinase Kinases; Mitogen-Activated Protein Kinases; Mutation; Neoplasm Metastasis; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Oncogenic; Organ; Pathway interactions; Patients; Phenotype; Phosphotransferases; Play; Process; Proteins; RNA Interference; RNA interference screen; Resistance; Role; Sampling; Skin Cancer; Solid Neoplasm; Testing; The Cancer Genome Atlas; TimeLine; United States; WNT Signaling Pathway; base; clinically significant; follow-up; functional genomics; genetic approach; human disease; in vivo; inhibitor/antagonist; macromolecule; melanoma; mouse model; mutant; novel; nucleocytoplasmic transport; prevent; prototype; public health relevance; raf Kinases; research study; small molecule inhibitor; sulfotransferase; trafficking; transcriptome; transcriptome sequencing; tumor; tumor growth

 DESCRIPTION (provided by applicant): Nucleocytoplasmic trafficking of macromolecules is a highly specific and tightly regulated process that occurs exclusively through the nuclear pore complex (NPC). NPC is composed of approximately 30 proteins, termed nucleoporins, which play crucial roles in several biological processes. Although, chromosomal translocation involving some nucleoporins are observed in hematological malignancies, however, their role in solid tumors is not known. Therefore, to study the role of nucleoporins in melanoma tumor growth and metastasis, we developed an integrative genomics approach by combining The Cancer Genome Atlas (TCGA) melanoma sample dataset analyses with functional genomics approach of an in vivo RNA interference (RNAi) screening. This approach identified NUP205 as the only nucleoporin that is essential for tumor growth and metastasis of BRAF- and NRAS-mutant melanoma. Additional experiments revealed that NUP205 confers anoikis resistance and stimulates MAK kinase and WNT signaling pathways. Based on these results, we hypothesize that NUP205 by regulating MAP kinase and WNT signaling pathways drives melanoma tumor growth and metastasis. The overall objective is to determine the role and mechanism of NUP205 in tumor growth and metastasis and test if NUP205-driven pathway can be targeted to treat melanoma. In Aim 1, we will determine the in vivo role of NUP205 in melanoma tumor growth and metastasis. To this end, using in vivo mouse models of melanoma tumor growth and organ-specific and spontaneous mouse models of melanoma metastasis, we will determine the role of NUP205 in facilitating melanoma tumor growth and metastasis. In Aim 2, we will determine the mechanism of NUP205 action. Based on our results, we will test whether the ability of NUP205 to stimulate MAP kinase via Raf kinase inhibitory protein (RKIP) and/or WNT signaling pathway via heparan sulfate 2-O-sulfotransferase 1 (HS2ST1) is required for it to confer anoikis resistance and promote melanoma tumor growth and metastasis. In Aim 3, we will determine the utility of targeting NUP205 regulated pathway for melanoma therapy. First, we will test if pharmacological inhibition of NUP205 downstream effector HS2ST1 enzyme by small molecule inhibitors can block metastatic tumor growth in vivo. Additionally, because NUP205 protein stabilization can confer resistance to BRAFV600E inhibitor vemurafenib, therefore, we will determine if inhibition of its downstream effector HS2ST1 can restore sensitivity to vemurafenib and prevent emergence of drug resistance. Clinical significance of these results will be established by analyzing pre- and post- vemurafenib treatment melanoma patient samples for the expression of NUP205 and its downstream effectors. Collectively, we expect to uncover a novel NUP205-driven druggable genetic vulnerability pathway that can be targeted for treating metastatic and drug resistant melanoma.

 描述（由申请人提供）：大分子的核细胞质运输是一个高度特异性且严格调控的过程，仅通过核孔复合体（NPC）发生。 NPC 由大约 30 种蛋白质组成，称为核孔蛋白，在多种生物过程中发挥着至关重要的作用。尽管在血液恶性肿瘤中观察到涉及某些核孔蛋白的染色体易位，但它们在实体瘤中的作用尚不清楚。因此，为了研究核孔蛋白在黑色素瘤肿瘤生长和转移中的作用，我们通过将癌症基因组图谱 (TCGA) 黑色素瘤样本数据集分析与体内 RNA 干扰 (RNAi) 筛选的功能基因组学方法相结合，开发了一种综合基因组学方法。该方法确定 NUP205 是唯一对 BRAF 和 NRAS 突变黑色素瘤的肿瘤生长和转移至关重要的核孔蛋白。其他实验表明，NUP205 赋予失巢凋亡抗性并刺激 MAK 激酶和 WNT 信号通路。基于这些结果，我们假设NUP205通过调节MAP激酶和WNT信号通路驱动黑色素瘤肿瘤的生长和转移。总体目标是确定NUP205在肿瘤生长和转移中的作用和机制，并测试NUP205驱动的通路是否可以靶向治疗黑色素瘤。在目标 1 中，我们将确定 NUP205 在黑色素瘤肿瘤生长和转移中的体内作用。为此，我们将使用黑色素瘤生长的体内小鼠模型和黑色素瘤转移的器官特异性和自发性小鼠模型，确定NUP205在促进黑色素瘤生长和转移中的作用。在目标2中，我们将确定NUP205的作用机制。根据我们的结果，我们将测试NUP205是否能够通过Raf激酶抑制蛋白（RKIP）和/或通过硫酸乙酰肝素2-O-磺基转移酶1（HS2ST1）刺激WNT信号通路刺激MAP激酶，从而赋予失巢凋亡抗性并促进黑色素瘤肿瘤生长和转移。在目标 3 中，我们将确定针对 NUP205 调节途径的黑色素瘤治疗的效用。首先，我们将测试小分子抑制剂对 NUP205 下游效应子 HS2ST1 酶的药理学抑制是否可以阻止体内转移性肿瘤的生长。此外，由于NUP205蛋白稳定可以赋予对BRAFV600E抑制剂vemurafenib的耐药性，因此，我们将确定抑制其下游效应器HS2ST1是否可以恢复对vemurafenib的敏感性并防止耐药性的出现。这些结果的临床意义将通过分析维莫非尼治疗前和治疗后黑色素瘤患者样本中 NUP205 及其下游效应子的表达来确定。总的来说，我们期望发现一种新的 NUP205 驱动的可药物遗传脆弱性途径，可用于治疗转移性和耐药性黑色素瘤。