Regulation of EGFR by SC4MOL- and NSDHL-Dependent Trafficking

SC4MOL 和 NSDHL 依赖性贩运对 EGFR 的调节

基本信息

批准号：
8537870
负责人：
Igor Astsaturov
金额：
$ 21.9万
依托单位：
RESEARCH INST OF FOX CHASE CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-01 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8537870
关键词：
Antineoplastic Agents Bioinformatics Biological Biological Markers Biological Testing Candidate Disease Gene Cell Survival Cell membrane Cetuximab Development Dimerization Drug Targeting Drug resistance Engineering Enzymes Epidermal Growth Factor Receptor Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor Epithelial Cells Erlotinib Eukaryota Excision Future Genes Goals Growth Ligands Link Literature Lysosomes Malignant Neoplasms Metabolic Metabolism Methods Modeling NADP Oncogenes Orthologous Gene Oxidases Oxidoreductase Pharmaceutical Preparations Phenotype Positioning Attribute Process Proteins RNA Interference Receptor Signaling Regulation Resistance Role Science Selection for Treatments Signal Transduction Small Interfering RNA Steroids Sterol Biosynthesis Pathway Sterols System Systems Biology Testing Therapeutic Therapeutic Agents Ubiquitination Work base cancer cell cancer prevention cancer therapy cell growth desensitization drug development enzyme pathway genetic analysis in vivo insight neoplastic cell network models promoter protein degradation protein transport public health relevance receptor expression resistance mechanism response screening small hairpin RNA therapeutic target tool trafficking

项目摘要

DESCRIPTION (provided by applicant): Our goal is to integrate systems biology approaches into the analysis of cancer drug resistance mechanisms, and thus to develop and evaluate new strategies for the efficient selection of optimal therapeutic combinations. In our recent work, we used siRNA screening to interrogate over 600 candidate genes from a set we nominated by network modeling to be enriched for regulators of EGFR signaling. This and subsequent analysis identified SC4MOL (sterol C4-methyl oxidase-like), a little-studied intermediate in the sterol biosynthesis pathway, as a potent regulator of cell viability following treatment with the EGFR inhibitors erlotinib and cetuximab. To understand the mechanism of SC4MOL action, we again used a bioinformatics strategy to model an interaction network for evolutionarily conserved orthologs of SC4MOL and its partner protein, NSDHL (NADP-dependent steroid dehydrogenase-like). This predicted multiple connections to regulators of exocytic trafficking and protein degradation. In direct preliminary test of the network predictions, we demonstrated that RNAi depletion of SC4MOL or NSDHL resulted in loss of EGFR from the plasma membrane, and enhanced its ubiquitination and degradation. Our hypothesis is that loss of interactions between SC4MOL and NSDHL with partner proteins influences trafficking so as to accelerate EGFR degradation, thereby potentiating EGFR-targeting drugs. To further validate this approach to identifying modifiers of targeted response, and potentially validate SC4MOL and NSDHL as biomarkers of drug response and/or targets for drug development, we will perform the following two Aims: Aim 1. Investigate the mechanism by which SC4MOL and NSDHL regulate EGFR trafficking. We will assess how SC4MOL and NSDHL influence EGFR trafficking and degradation, and whether the catalytic activity of SC4MOL and NSDHL is required for their actions. We will analyze interactions between SC4MOL and a small set of predicted conserved interaction partners with known roles in regulating vesicular trafficking predicted by bioinformatics analysis of the orthologs in lower eukaryotes. Aim 2. Investigate the interaction of SC4MOL and NSDHL with ERAD proteins. We have found silencing of SC4MOL or NSDHL enhances EGFR ubiquitination and lysosomal degradation. Our analysis identified a number of candidates within the ER-associated protein degradation (ERAD) system, which will be tested for physical association with SC4MOL and NSDHL, and for a role in SC4MOL- and NSDHL-dependent regulation of EGFR expression, trafficking, and degradation. Given the prominence of EGFR as a target in cancer therapy, this work has potentially high impact to reduce drug resistance relevant to many clinically valuable therapeutic agents.

描述（由申请人提供）：我们的目标是将系统生物学方法整合到癌症耐药机制的分析中，从而制定和评估新的策略，以有效选择最佳的最佳治疗组合。在最近的工作中，我们使用siRNA筛选从我们通过网络建模提名的集合中询问了600多个候选基因，以丰富EGFR信号的调节剂。此和随后的分析确定了SC4MOL（类固醇C4-甲基氧化酶样），在固醇生物合成途径中有点研究中间体，是用EGFR抑制剂Erlotinib和Cetuximab处理后的有效调节细胞活力的调节剂。为了了解SC4mol作用的机制，我们再次使用生物信息学策略来对SC4MOL及其伴侣蛋白NSDHL（NADP依赖性类固醇脱氢酶样）的进化保守直系同源物进行相互作用网络进行建模。这预测了与外囊肿运输和蛋白质降解的调节因子的多次联系。在对网络预测的直接初步测试中，我们证明了SC4mol或NSDHL的RNAi耗竭导致质膜中的EGFR损失，并增强了其泛素化和降解。我们的假设是，SC4mol和NSDHL与伴侣蛋白之间的相互作用丧失会影响运输以加速EGFR降解，从而增强EGFR靶向药物。为了进一步验证这种方法来识别有针对性反应的修饰符，并可能验证SC4mol和NSDHL为药物反应和/或药物开发目标的生物标志物，我们将执行以下两个目标：目标1。研究SC4mol和NSDHL调节EGFR贩运的机制。我们将评估SC4MOL和NSDHL如何影响EGFR的运输和退化，以及SC4mol和NSDHL的催化活性是否需要其行为。我们将分析SC4MOL与一组预测的保守相互作用伙伴之间的相互作用，在调节较低真核生物中直系同源物的生物信息学分析预测的囊泡贩运方面具有已知作用。 AIM 2。研究SC4MOL和NSDHL与ERAD蛋白的相互作用。我们发现SC4mol或NSDHL的沉默增强了EGFR泛素化和溶酶体降解。我们的分析确定了与ER相关蛋白质降解（ERAD）系统中的许多候选物，该系统将与SC4mol和NSDHL进行物理关联，并在SC4MOL-和NSDHL依赖性EGFR表达，运输和脱粒的调控中发挥作用。鉴于EGFR作为癌症治疗的目标，这项工作具有很大的影响，可降低与许多临床上有价值的治疗剂相关的耐药性。