Role of Autophagy in Bortezomib-induced HNSCC Apoptosis

自噬在硼替佐米诱导的 HNSCC 细胞凋亡中的作用

基本信息

批准号：
8909536
负责人：
Insoon Chang
金额：
$ 4.05万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8909536
关键词：
26S proteasome Acetylation Address Americas Antibodies Apoptosis Autophagocytosis Basic Science Biological Assay Bortezomib Cell Line Cell Survival Confocal Microscopy Cytoplasm Data Development Flow Cytometry Genes Genetic Green Fluorescent Proteins HDAC6 gene Head and Neck Cancer Head and Neck Squamous Cell Carcinoma Head and neck structure Heat shock proteins Heat-Shock Response Histone Deacetylase Inhibitor Human Immunofluorescence Immunologic Immunohistochemistry Immunoprecipitation In Vitro Investigation Light Link Lysine Malignant Epithelial Cell Malignant Neoplasms Mediating Messenger RNA Microtubule-Associated Proteins Modality Molecular Molecular and Cellular Biology Monitor Morbidity - disease rate Noxae Nuclear Translocation Nude Mice Pathway interactions Patients Phosphorylation Phosphotransferases Polymerase Chain Reaction Proteasome Inhibitor Proteins RNA Interference Recurrence Regimen Research Research Personnel Research Training Resistance Reverse Transcription Role Specificity Staining method Stains Therapeutic Time Trichostatin A Untranslated RNA Vesicle Western Blotting Xenograft procedure annexin A5 antitumor effect base carcinogenesis career cellular targeting chemotherapeutic agent chemotherapy chromatin immunoprecipitation conventional therapy cost cytotoxic cytotoxicity design endoplasmic reticulum stress heat-shock factor 1 in vivo inhibition of autophagy mortality neoplastic cell novel novel therapeutics outcome forecast prevent protein activation protein aggregate protein expression public health relevance response screening small hairpin RNA transcription factor translational medicine tumor

项目摘要

DESCRIPTION (provided by applicant): Head and Neck Squamous Cell Carcinoma (HNSCC) comprise 90% of head and neck cancer and typically presents poor prognosis and resistant to chemotherapy. Hence, the development of more effective therapeutic strategy is critical. Previously, Bortezomib has been proposed as an alternate chemotherapeutic agent to conventional chemotherapy in HNSCC, but infers chemoresistance through a largely unknown pathway. The main objectives of this application are to investigate the potential utilization of Trichostatin A (TSA) and Bortezomib (aka PS-341) combinatory regimen to treat HNSCC and to understand the molecular mechanisms by which TSA enhances Bortezomib-induced apoptosis to effectively overcome chemoresistance. Our preliminary studies suggest that histone deacetylase 6 (HDAC6) promotes autophagy to reduce the Bortezomib cytotoxicity. When we abolished expression of HDAC6 in HNSCC cells, autophagy induction was inhibited and the Bortezomib cytotoxicity was significantly enhanced. Based on these novel findings, we hypothesize that TSA may enhance antitumor effects of Bortezomib through inhibition of HDAC6 activity, which may serve as a critical causal link between autophagy, apoptosis, and cellular survival response in HNSCC. Our hypotheses will be addressed in the following specific aims: (1) Determine if TSA enhances Bortezomib- induced apoptosis while inhibiting HDAC6-mediated autophagy; and (2) Investigate the molecular mechanisms by which HDAC6 initiates autophagy induced by Bortezomib in HNSCC. This plan will utilize several features from both basic science and translational medicine aspects: Green fluorescent protein (GFP) - LC3 puncta formation assay, flow cytometry, western blotting, real-time qRT-PCR, chromatin immunoprecipitation, and immunofluorescence staining will be used to examine changes in expression of molecules associated with HDAC6-induced autophagy and accumulation of cytotoxic protein aggregates. Further, we plan to utilize xenotransplantation of HDAC6-depleted human HNSCC cells in nude mice to investigate molecular and cellular changes associated with autophagy, endoplasmic reticulum stress, and apoptosis. This research strategy is an integral component of a research training plan designed to develop the trainee's expertise in molecular/cellular biology and genetics of head and neck carcinogenesis, chemoresistance, and apoptosis and will prepare the trainee for an academic career as an independent researcher.

描述（由适用提供）：头部和颈部方形细胞癌（HNSCC）完成了90％的头颈癌，通常表现出较差的预后和对化学疗法的耐药性。因此，更有效的治疗策略的制定至关重要。以前，已提出硼替佐米作为HNSCC常规化学疗法的替代化学治疗剂，但通过在很大程度上未知的途径中渗透化学抗性。本应用的主要目标是研究毛乳杆菌A（TSA）和硼替佐米（又名PS-341）组合方案的潜在利用，以治疗HNSCC，并了解TSA可以增强bortezomib诱导的凋亡，从而有效地胜任化学化的分子机制。我们的初步研究表明，组蛋白脱乙酰基酶6（HDAC6）促进自噬以降低硼替佐米的细胞毒性。当我们废除HNSCC细胞中HDAC6的表达时，抑制了自噬诱导，并且硼替佐米的细胞毒性显着增强。基于这些新的发现，我们假设TSA可以通过抑制HDAC6活性来增强硼替佐米的抗肿瘤作用，这可能是HNSCC中自噬，凋亡和细胞生存反应之间的关键因果关系。我们的假设将在以下特定目的中解决：（1）确定TSA是否增强了硼替唑诱导的凋亡，同时抑制HDAC6介导的自噬；（2）研究HDAC6在HNSCC中启动HDAC6启动自噬的分子机制。该计划将利用基础科学和翻译医学方面的几个功能：绿色荧光蛋白（GFP） - LC3点状形成测定法，流式细胞术，蛋白质印迹，实时QRT-PCR，染色质质免疫沉淀，染色质蛋白免疫沉淀，以及与Hydac6-相关的HDAC6-的累积变化，用于研究聚合。此外，我们计划利用裸鼠中HDAC6耗尽的人类HNSCC细胞的异种移植来研究与自噬，内质网应激和凋亡相关的分子和细胞变化。研究策略是研究培训计划的不可或缺的组成部分，旨在发展学员在分子/细胞生物学以及头颈癌变，化学耐药性和凋亡的遗传学方面的专业知识，并将为学员作为独立研究人员的学术生涯做好准备。