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：
9208125
负责人：
Insoon Chang
金额：
$ 4.09万
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/9208125
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 Health 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 screen 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 gene product heat-shock factor 1 in vivo inhibition of autophagy knock-down mortality neoplastic cell novel novel therapeutics outcome forecast prevent protein activation protein aggregate protein expression response 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（申请人提供）：头颈鳞状细胞癌（HNSCC）占头颈癌的90%，通常预后不良且对化疗产生耐药性，因此，开发更有效的治疗策略至关重要。已被提议作为 HNSCC 常规化疗的替代化疗药物，但通过很大程度上未知的途径推断化疗耐药性。该应用的主要目的是研究潜在的潜力。利用曲古抑菌素 A (TSA) 和硼替佐米 (又名 PS-341) 联合方案治疗 HNSCC，并了解 TSA 增强硼替佐米诱导的细胞凋亡以有效克服化疗耐药性的分子机制。促进自噬以降低硼替佐米的细胞毒性当我们消除 HNSCC 细胞中 HDAC6 的表达时，会诱导自噬。基于这些新发现，我们发现 TSA 可能通过抑制 HDAC6 活性来增强硼替佐米的抗肿瘤作用，这可能是自噬、细胞凋亡和细胞存活反应之间的关键因果关系。 HNSCC。我们的假设将在以下具体目标中得到解决：（1）确定 TSA 是否增强硼替佐米诱导的细胞凋亡，同时抑制 HDAC6 介导的自噬；以及（2）研究 HDAC6 在 HNSCC 中启动硼替佐米诱导自噬的分子机制。该计划将利用基础科学和转化医学方面的几个特征：绿色荧光蛋白 (GFP) - LC3 斑点形成测定、流式细胞术、蛋白质印迹、real- qRT-PCR、染色质免疫沉淀和免疫荧光染色将用于检查与 HDAC6 诱导的自噬和细胞毒性积累相关的分子表达变化此外，我们计划利用 HDAC6 耗尽的人 HNSCC 细胞在裸鼠中进行异种移植来研究与自噬、内质网应激和细胞凋亡相关的分子和细胞变化。培养学员在头颈癌、化疗耐药和细胞凋亡的分子/细胞生物学和遗传学方面的专业知识，并为学员作为独立研究员的学术生涯做好准备。