Role of DHHC Protein Palmitoyltransferases in Non-Small Cell Lung Cancer

DHHC 蛋白棕榈酰转移酶在非小细胞肺癌中的作用

• 批准号: 8691304
• 负责人: SANDRA L HOFMANN
• 金额: $ 17.29万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8691304
• 关键词: Biological Assay; Cancer Center; Cancer cell line; Catalysis; Cell Line; Cell physiology; Cells; Cysteine; Databases; Development; Drug Targeting; Enzymes; Erlotinib; Family; Family member; Fatty Acids; Genes; Genetic Screening; Goals; Growth; Human; Individual; Laboratories; Learning; Lipids; Maintenance; Malignant Neoplasms; Malignant neoplasm of lung; Membrane Proteins; Methods; Modeling; Modification; Names; Neurodegenerative Disorders; Neurons; Non-Small-Cell Lung Carcinoma; Palmitates; Palmitic Acylation Site; Pathway interactions; Pharmaceutical Preparations; Phosphorylation; Physiological; Post-Translational Protein Processing; Process; Protein Family; Protein Kinase; Protein S; Proteins; Publishing; Research; Resistance; Role; Small Interfering RNA; Technology; Testing; Therapeutic; Traction; Transferase; Ursidae Family; Xenograft procedure; addiction; base; cancer cell; cancer type; cell growth; design; enzyme substrate; fatty acylation; flotillin; genome wide association study; inhibitor/antagonist; interest; killings; member; neoplastic; new therapeutic target; novel therapeutics; oncology; overexpression; palmitoyl-protein hydrolase; palmitoylation; protein protein interaction; protein transport; public health relevance; research study; screening; therapeutic development; therapeutic target; tumor; tumor growth; tumor xenograft; yeast two hybrid interaction screening

DESCRIPTION (provided by applicant): Palmitoylation is a well-recognized post-translational lipid modification important for protein-membrane association, protein-protein interaction, and protein stability. Intracellular protein palmitoylation is carried out by a newly described family f protein S-acyl transferases (PATs), referred to as the DHHC family, named for a common catalytic motif. The family is comprised of 23 members in humans and the role of these in cancer is just beginning to be explored. In a recent genome-wide screen conducted at UT Southwestern, and a more targeted screen performed in the Hamon Center for Therapeutic Oncology research, the DHHC5 gene was found to contribute to the growth of non-small lung cancer cell lines and tumor xenografts. The goal of this proposal is to gain a better understanding of the mechanism(s) whereby DHHC5 contributes to tumor growth and to develop assays that will serve as the basis for development of inhibitors. Two specific aims are proposed. In Specific Aim 1 we will determine the basis for selective vulnerability of NSCLCs to DHHC5 knockdown. We will test the hypothesis that DHHC knockdown inhibits cell growth and tumor xenograft formation though inhibition of palmitoylation of substrates important for cancer maintenance. To test this hypothesis, we will identify further cell lines sensitive and resistant t DHHC5 knockdown and identify potential pathways for DHHC5 addiction, drawing upon the extensive database available at UT Southwestern. We will identify candidate substrates by profiling palmitoylation of proteins in control and DHHC5 knockdown cell lines, using methods we have previously published. Candidate DHHC5-substrate pairs will be validated through expression assays involving heterologous cells, and then evaluated as targets in lung cancer cell lines using appropriately designed siRNA and overexpression experiments. In Specific Aim 2, we will determine structural features of DHHC5 needed to support growth of NSCLC lines and tumors. We have identified flotillin-2 as a physiological substrate of DHHC5 in neuronal cells, and it is a candidate substrate in NSCLC. Using flotillin-2 as a model substrate for DHHC5, we aim to learn about residues involved in catalysis and substrate recognition that would support the eventual development of inhibitors. These studies may be of significant value to the 160,000 individuals in the US who will succumb to lung cancer annually.

描述（由申请人提供）：棕榈酰化是一种公认​​的翻译后脂质修饰，对蛋白质 - 膜缔合，蛋白质 - 蛋白质相互作用和蛋白质稳定性很重要。细胞内蛋白质棕榈酰化是由新描述的Family F蛋白S-酰基转移酶（PATS）进行的，称为DHHC家族，以常见的催化基序命名。该家庭由人类的23名成员组成，这些成员在癌症中的作用才刚刚开始探索。在最近在UT西南部进行的全基因组筛查中，在哈蒙治疗肿瘤学研究中心进行了更具针对性的筛查，发现DHHC5基因有助于非小肺癌细胞系和肿瘤异种移植物的生长。该提案的目的是更好地了解DHHC5有助于肿瘤生长的机制，并开发将作为抑制剂开发的基础的测定。提出了两个具体目标。在特定目标1中，我们将确定NSCLC对DHHC5敲低的选择性脆弱性的基础。我们将测试以下假设：DHHC敲低抑制细胞生长和肿瘤异种移植的形成，尽管抑制了对癌症维持至关重要的底物的底层化。为了检验该假设，我们将确定进一步的细胞系敏感和抗性T DHHC5敲低，并确定DHHC5成瘾的潜在途径，利用UT西南地区可用的广泛数据库。我们将使用我们先前发表的方法来鉴定蛋白质和DHHC5敲低细胞系中蛋白质的棕榈酰化来鉴定候选底物。候选DHHC5 - 基底对将通过涉及异源细胞的表达测定进行验证，然后使用适当设计的siRNA和过表达实验对肺癌细胞系中的靶标进行评估。在特定的目标2中，我们将确定支持NSCLC系和肿瘤生长所需的DHHC5的结构特征。我们已经确定了Flotillin-2是神经元细胞中DHHC5的生理底物，并且它是NSCLC中的候选底物。使用Flotillin-2作为DHHC5的模型底物，我们旨在了解涉及催化和底物识别的残基，这些残基将支持最终发展抑制剂的发展。这些研究对于每年将屈服于肺癌的160,000个人可能具有重要价值。