Disrupting two cancer hallmarks with one target: DHX36, the major G-quadruplex helicase

用一个靶点破坏两种癌症标志：DHX36，主要的 G 四链体解旋酶

• 批准号: 10045972
• 负责人: ROBERT HANEY
• 金额: $ 42.8万
• 依托单位: BALL STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10045972
• 关键词: Address; Animal Model; B-Lymphocytes; BCL2 gene; Binding; Bioinformatics; Biology; Biopsy; Biotechnology; Cell Proliferation; Cell Survival; Cells; Collaborations; DNA; Data; Development; Disease remission; Emu species; Enhancers; Enrollment; Enterobacteria phage P1 Cre recombinase; Epigenetic Process; G-Quartets; Genes; Genetic; Genetic Transcription; Guanine; High School Student; Holoenzymes; Human; KRAS2 gene; Knock-out; Knockout Mice; Laboratories; Lead; Length; Ligands; LoxP-flanked allele; Lymphoma; Malignant - descriptor; Malignant Neoplasms; Mass Spectrum Analysis; Mentors; Mentorship; Metabolism; Modality; Molecular; Monitor; Morbidity - disease rate; Mus; Non-Hodgkin' s Lymphoma; Oncogenes; Oncogenic; Pathway interactions; Patients; Pharmaceutical Preparations; Prevention; Promoter Regions; Proteins; Proteomics; Proto-Oncogene Protein c-kit; Proto-Oncogene Proteins c-akt; RNA; RNA-Binding Proteins; Research; Role; Small Interfering RNA; Structure; Students; Technology; Telomerase; Telomere Shortening; Testing; Therapeutic; Tissues; Transgenic Mice; Transgenic Organisms; Translations; Tumor Suppressor Genes; Underrepresented Students; Universities; Up-Regulation; Vascular Endothelial Growth Factors; Work; c-myc Genes; cancer cell; cell killing; cohort; differential expression; experience; graduate student; helicase; human disease; innovation; knock-down; mortality; mouse model; multidisciplinary; neoplastic cell; new therapeutic target; novel; nucleic acid structure; overexpression; prevent; promoter; response; telomere; therapeutic target; transcriptome sequencing; treatment strategy; tumor; tumor heterogeneity; tumor initiation; tumor initiators; tumor progression; tumorigenesis; undergraduate student

PROJECT SUMMARY This proposal uses innovative approaches and novel mouse models to define an underlying mechanism of tumorigenesis. This proposal further seeks to identify the major G-quadruplex helicase, DHX36 (aliases: G4R1 and RHAU), as a novel therapeutic target. G-quadruplexes (G4s) are dynamic, “knot-like” DNA or RNA structures that shut down transcription and translation, respectively. Of relevance to cancer, genes that promote cell proliferation and survival (i.e. oncogenes) are more likely to contain G4 sequences, while genes that suppress cell proliferation and survival (i.e. tumor suppressor genes) are depleted of G4s. Therefore, G4s represent an epigenetic feature that generally distinguishes oncogenes from tumor suppresser genes. As such, G4s are attractive cancer therapeutic targets. Moreover, >80% of tumors rely on telomerase to prevent telomere shortening, which confers cellular immortality, a classic hallmark of cancer. Extensive G4 structures form in telomere DNA that inhibit telomerase. Taken together, G4 structures reduce oncogene expression and cellular immortality. Conversely, G4 helicases unwind G4 structures increasing oncogene expression and limiting telomere elongation. DHX36 accounts for the majority of G4 helicase activity in human cells and is commonly overexpressed in cancer. DHX36-overexpression is correlated with a significantly reduction in patient survival. Thus, DHX36 is a prime candidate to explore as a therapeutic target. In this proposal, we will pursue two aims to determine the potential of DHX36 as a therapeutic target. In the first aim, we hypothesize that DHX36- overexpression initiates tumors and exacerbates tumor progression. In the second aim, we hypothesize that decreased DHX36 expression alone or in combination with G4 ligands will reduce oncogene expression, telomere elongation, and result in tumor remission. We will test these hypotheses using a novel Dhx36- overexpression mouse as well as a Dhx36-knockout mouse crossed to a transgenic mouse tumor line. The proposed studies will be the first to determine the role of DHX36 in tumorigenesis using mouse models. This work is poised to identify DHX36 as a novel therapeutic target that inhibition of will disrupt two fundamental cancer pathways: oncogene expression and telomere elongation. Undergraduate and graduate students will be integrated at every stage of the project allowing them to gain authentic experience with innovative mouse and bioinformatics technologies applied to a deadly human disease.

项目概要 该提案使用创新方法和新颖的小鼠模型来定义潜在的机制 肿瘤发生。该提案进一步寻求鉴定主要的 G-四链体解旋酶 DHX36（别名：G4R1） 和 RHAU），作为一个新的治疗靶点。 G-四链体 (G4) 是动态的“结状”DNA 或 RNA 结构 分别关闭转录和翻译。与癌症相关，促进细胞生长的基因 增殖和存活（即癌基因）更有可能包含 G4 序列，而抑制基因 细胞增殖和存活（即肿瘤抑制基因）的 G4 被耗尽。因此，G4代表 通常区分癌基因和抑癌基因的表观遗传特征。因此，G4 是 有吸引力的癌症治疗靶点。此外，>80%的肿瘤依赖端粒酶来预防端粒 缩短，赋予细胞永生性，这是癌症的典型标志。广泛的 G4 结构形成于 抑制端粒酶的端粒DNA。总而言之，G4 结构可减少癌基因表达和细胞 不朽。相反，G4 解旋酶解旋 G4 结构，增加癌基因表达并限制 端粒伸长。 DHX36 负责人类细胞中 G4 解旋酶活性的大部分，并且通常被认为是 在癌症中过度表达。 DHX36 过度表达与患者生存率显着降低相关。 因此，DHX36 是探索治疗靶点的主要候选者。在这个提案中，我们将追求两个目标 确定 DHX36 作为治疗靶点的潜力。在第一个目标中，我们假设 DHX36- 过度表达会引发肿瘤并加剧肿瘤进展。在第二个目标中，我们假设 单独或与 G4 配体组合降低 DHX36 表达将减少癌基因表达， 端粒延长，从而导致肿瘤缓解。我们将使用一种新颖的 Dhx36- 来测试这些假设 过表达小鼠以及 Dhx36 敲除小鼠与转基因小鼠肿瘤系杂交。这 拟议的研究将是第一个使用小鼠模型确定 DHX36 在肿瘤发生中的作用的研究。这 工作准备将 DHX36 确定为一种新的治疗靶点，抑制该靶点将破坏两个基本的治疗靶点 癌症途径：癌基因表达和端粒延长。本科生和研究生将 集成在项目的每个阶段，使他们能够通过创新的鼠标获得真实的体验 生物信息学技术应用于致命的人类疾病。

项目成果

"Important enough to show the world": Using Authentic Research Opportunities and Micropublications to Build Students' Science Identities.

“重要到足以向世界展示”：利用真实的研究机会和微型出版物来建立学生的科学身份。

• DOI: 10.1101/2023.08.17.553701
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Rubenstein,LisaDaVia;Woodruff,KelseyA;Taylor,AprilM;Olesen,JamesB;Smaldino,PhilipJ;Rubenstein,EricM
• 通讯作者: Rubenstein,EricM