Disrupting long noncoding RNA methylation to elicit antimorph behavior in breast cancer

破坏长链非编码 RNA 甲基化以引发乳腺癌的反形态行为

基本信息

批准号：
10646843
负责人：
Aaron M. Johnson
金额：
$ 18.17万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10646843
关键词：
Address Affect Antisense Oligonucleotides Atmosphere Behavior Breast Cancer Cell Breast Cancer Patient Breast Cancer cell line Cancer Biology Cell Line Cells Chemicals Clinic Code Diagnosis Future Gene Expression Gene Targeting Genes Genetic Transcription Goals Growth Intervention Lead Length MDA MB 231 Malignant Neoplasms Methylation Modification Molecular Mutate Nuclear Nucleotides Oncogenes Oncogenic Pathway interactions Patient-derived xenograft models of breast cancer Patients Pattern Phenotype Prevention Process Property Proteins Proteomics RNA RNA delivery RNA methylation RNA vaccine Role Site Suppressor Mutations Testing Therapeutic Therapeutic Intervention Transcript Tumor Suppressor Proteins Untranslated RNA Work Xenograft procedure anti-cancer breast cancer progression cancer cell cancer therapy cancer type carcinogenesis clinical application experimental study gene repression inhibitor malignant breast neoplasm mutant novel strategies novel therapeutic intervention novel therapeutics overexpression therapeutic RNA tumor vaccine delivery

项目摘要

PROJECT SUMMARY Specific long noncoding RNAs (LncRNAs) that are overexpressed in breast cancer aid in diagnosis and promote molecular mechanisms that drive breast cancer progression. Our recent work has identified a breast- cancer associated LncRNA that converts from an oncogene to a tumor suppressor by mutation of one single nucleotide. This “antimorphic” effect is caused by the prevention of RNA methylation at this one site, allowing overexpression of the converted LncRNA to decrease cancer phenotypes more so than depletion of the wild- type version. This exciting new finding prompts us to delve deeper into how this is occurring, with the ultimate goald of specifically disrupting this m6A pathway as a novel therapeutic approach. Proposed therapeutic intervention for breast cancer-associated LncRNAs has mainly been aimed at depleting the cancer-promoting RNA, with the assumption that this would be the most effective approach to reduce cancer phenotypes. Our results suggest a novel therapeutic approach to oncogenic LncRNAs, where disruption of RNA methylation alone has a greater impact than simple elimination of the RNA. The current proposal provides a roadmap to understand how to elicit antimorphic effects in an oncogenic LncRNA. We aim to use this proof-of-principle project as a springboard for future work that would lead this therapeutic approach to the clinic. We hypothesize that m6A modification sites on a specific breast cancer-associated LncRNAs can be targeted and disrupted to reverse oncogenic behavior below baseline, providing a roadmap for a potential new therapeutic approach. In our first aim, a mutant HOTAIR LncRNA that behaves as an antimorph will be studied for physical interactions in triple negative MDA-MB-231 breast cancer cells using proteomic approaches. Physical and functional interactors will then be depleted in a breast cancer cell line that overexpresses the antimorph HOTAIR to determine their role in the antimorph mechanism. Recently-derived primary cell lines from breast cancer patient-derived xenografts will be used to determine the generalizability of this mechanism. In the second aim, anti-sense oligonucleotides (ASOs) will be used to disrupt the m6A site on HOTAIR to elicit antimorph behavior. In a parallel approach, we will developed an optimized strategy to deliver the antimorph HOTAIR RNA directly to suppress cancer phenotypes. These two approaches are complementary in cases of breast cancer where HOTAIR is overexpressed and those where it is not. Our work would be a foundational test of a novel therapeutic approach in breast cancer by inducing antimorphic behavior of an m6A-modified LncRNA. In general, the ASO-based strategy is a novel approach to breast cancer therapeutics. RNA therapeutics are now a reality and are gaining wide acceptance. Our project capitalizes on this exciting atmosphere with an ambitious, novel approach to noncoding RNA interventions in breast cancer. By taking advantage of a molecular switch provided by m6A, we can reverse the activity of an oncogenic LncRNA to reverse breast cancer properties.

项目摘要在乳腺癌中过表达诊断和促进驱动乳腺癌进展的分子机制。我们最近的工作已经确定了乳房癌症相关的lncRNA，通过一个单一的突变将癌基因转化为肿瘤抑制剂核苷酸。这种“抗态”效应是由于预防RNA甲基化在一个部位引起的，允许转化后的lncRNA的过表达比野生的深度更降低癌症表型类型版本。这一令人兴奋的新发现促使我们更深入地研究了这种情况的发生方式，最终将这种M6A途径特别破坏作为一种新型治疗方法的目标。建议用于乳腺癌相关的LNCRNA的治疗干预措施主要针对耗尽癌症的RNA，假设这将是最有效的方法减少癌症表型。我们的结果表明一种新型的热方法来致癌lncrnas，其中与简单消除RNA相比，仅RNA甲基化的破坏具有更大的影响。电流提案提供了一个路线图，以了解如何在致癌性LNCRNA中引起抗鲜相作用。我们的目标使用此原理证明项目作为未来工作的跳板，以领导这种治疗方法去诊所。我们假设在特定的乳腺癌相关LNCRNA上的M6A修饰位点可以是有针对性和破坏基线以下的致癌行为，为潜在的新的路线图提供了路线图治疗方法。在我们的第一个目标中，表现为抗微孔的突变热热lncRNA将被研究用于使用蛋白质组学方法的三个阴性MDA-MB-231乳腺癌细胞中的物理相互作用。然后，身体和功能相互作用的人将在乳腺癌细胞系中耗尽，该细胞过表达抗微孔热水以确定它们在抗肌电机理中的作用。最近衍生的主要细胞系乳腺癌患者衍生的Xenographictic将用于确定该机制的普遍性。在第二个目的，反义寡核苷酸（ASO）将用于破坏Hotair上的M6A站点以激发抗肌行为。在一种平行的方法中，我们将制定一种优化的策略来提供抗肌层 Hotair RNA直接抑制癌症表型。这两种方法是完整的热带过表达的乳腺癌，而不是那些没有表达的乳腺癌。我们的工作将是通过诱导的乳腺癌治疗方法的基础测试 M6a修饰的lncRNA的抗鲜齿行为。通常，基于ASO的策略是一种新颖的方法乳腺癌疗法。 RNA疗法现在已成为现实，并且正在广泛接受。我们的项目通过一种雄心勃勃的新型方法来利用这种令人兴奋的氛围乳腺癌。通过利用M6A提供的分子开关，我们可以逆转致癌性lncRNA逆转乳腺癌特性。