Defining oncogenic capacities of PAK7 mutations in human cancer

定义人类癌症中 PAK7 突变的致癌能力

• 批准号: 9052124
• 负责人: STEEN HENNING HANSEN
• 金额: $ 19.25万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-09 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9052124
• 关键词: Address; Advanced Malignant Neoplasm; Affect; Apical; Apoptosis; Architecture; Attention; Automobile Driving; BRAF gene; Biological Assay; Caco-2 Cells; Cadherins; Cancer Patient; Cancer cell line; Carcinoma; Cell Line; Cell Polarity; Cell Proliferation; Cell-Cell Adhesion; Cells; Colon Carcinoma; Colorectal Cancer; Controlled Study; Data; Epithelial; Epithelial Cells; Family; Gastric Adenocarcinoma; Genes; Genome; Health; Human; MEL Gene; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Measures; Mediating; Missense Mutation; Mus; Mutate; Mutation; Non-Small-Cell Lung Carcinoma; Nude Mice; Oncogenes; Oncogenic; Phosphotransferases; Probability; Property; Protein Kinase; Resistance; Role; STK11 gene; Sequence Analysis; Staging; Technology; Testing; Therapeutic; Tumor Suppressor Proteins; Tumorigenicity; Work; actionable mutation; anaplastic lymphoma kinase; cancer cell; cancer genome; gastrointestinal carcinoma; genome editing; genome sequencing; improved; inhibitor/antagonist; insertion/deletion mutation; knock-down; lung Carcinoma; matrigel; melanoma; oncology; p21 activated kinase; preclinical trial; research study; targeted treatment; three dimensional cell culture; trait; tumorigenesis; zinc finger nuclease

 DESCRIPTION (provided by applicant): Genome sequencing efforts have identified PAK7 (p21-activated kinase 7) as a potentially important cancer kinase. PAK7 ranked 17 among all 518 kinases in probability of harboring driver mutations for human cancer. The vast majority of PAK7 mutations in human cancer are missense and thus suggestive of a role as an oncogene. PAK7 mutations occur in a significant proportion of lung and gastrointestinal carcinomas, as well as in malignant melanomas. Our preliminary data suggest that cancer mutations in PAK7 potently stimulate kinase activity. However, PAK7 is a minimally characterized kinase, thus providing merit to address the following aims: (1) Are putative PAK7 driver mutations required for oncogenic capacities in cancer cells? (2) Are putative PAK7 driver mutations sufficient to promote epithelial oncogenesis? We will rigorously address these aims by genome editing to avoid confounding effects associated with exogenous expression or knockdown of genes. First, we will correct PAK7 missense mutations in human cancer cell lines. We will then determine whether correction of these mutations reverts any transformed capacities that these cell lines possess. Specifically, we will assay cell proliferation, apoptosis, invasion, and tumorigenicity in athymic mice. Second, we will introduce putative PAK7 driver mutations into the relatively well-differentiated colonic Caco-2 cells and bronchial 16HBE14o- cells. In addition to the capacities listed above, we will determine effects of PAK7 cancer mutations on parameters of normal epithelial architecture; a hallmark of differentiated epithelial cells with established tumor suppressor capacities. Specifically, we will test whether naturally occurring PAK7 mutations in cancer affect epithelial cell polarity, cell-cell adhesion, and lumen formation in 3D culture. Collectively, the proposed studies will provide a stringent initial approach to define the oncogenic potential of PAK7 mutations in human cancer and elucidate whether PAK7 represents a promising candidate in expanding targets for cancer therapeutics.

 描述(由申请人提供)：基因组测序工作已确定PAK7(p21激活的激酶7)是一种潜在的重要的癌症激酶。在所有518个激酶中，PAK7在人类癌症驱动程序突变的概率中排名第17。人类癌症中绝大多数的PAK7突变是错误的，因此暗示了作为癌基因的作用。PAK7突变发生在相当大比例的肺癌和胃肠道癌以及恶性黑色素瘤中。我们的初步数据表明，PAK7的癌症突变可以有效地刺激激酶活性。然而，PAK7是一个最小特征的激酶，因此提供了解决以下目的的优点：(1)假定的PAK7驱动突变是癌细胞致癌能力所必需的吗？(2)假设的PAK7驱动突变是否足以促进上皮肿瘤的发生？我们将通过基因组编辑严格解决这些目标，以避免与外源表达或基因敲除相关的混淆效应。首先，我们将纠正人类癌细胞系中的PAK7错义突变。然后，我们将确定这些突变的纠正是否会逆转这些细胞系拥有的任何转化能力。具体地说，我们将分析细胞的增殖、凋亡、侵袭和致瘤性 无性系小鼠。其次，我们将把可能的PAK7驱动突变引入相对分化较好的结肠癌Caco-2细胞和支气管16HBE14o-细胞。除了上面列出的能力，我们还将确定PAK7癌症突变对正常上皮结构参数的影响；正常上皮结构是分化的上皮细胞的标志，具有既定的肿瘤抑制能力。具体地说，我们将测试癌症中自然发生的PAK7突变是否会影响3D培养中的上皮细胞极性、细胞-细胞黏附和管腔形成。总之，拟议的研究将提供一种严格的初步方法来定义人类癌症中PAK7突变的致癌潜力，并阐明PAK7是否代表着扩大癌症治疗靶点的有前途的候选者。