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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.

项目概要在乳腺癌中过度表达的特定长非编码 RNA (LncRNA) 有助于诊断和治疗促进推动乳腺癌进展的分子机制。我们最近的工作发现了一个乳房癌症相关的 LncRNA，通过一个单一突变从癌基因转变为肿瘤抑制基因核苷酸。这种“反形态”效应是由阻止该位点的 RNA 甲基化引起的，从而允许转化后的 LncRNA 的过度表达比野生型 LncRNA 的耗竭更能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这是如何发生的，并提供最终的结果目标是专门破坏这种 m6A 途径作为一种新的治疗方法。拟议的乳腺癌相关 LncRNA 治疗干预主要针对耗尽促癌RNA，假设这将是最有效的方法减少癌症表型。我们的结果提出了一种致癌 LncRNA 的新治疗方法，其中单独破坏 RNA 甲基化比简单消除 RNA 具有更大的影响。目前的该提案提供了了解如何在致癌 LncRNA 中引发反态效应的路线图。我们的目标使用这个原理验证项目作为未来工作的跳板，从而引领这种治疗方法到诊所。我们假设特定乳腺癌相关 LncRNA 上的 m6A 修饰位点可能是有针对性和破坏性地扭转低于基线的致癌行为，为潜在的新方法提供了路线图治疗方法。在我们的第一个目标中，将研究一种充当反形态的突变型 HOTAIR LncRNA 使用蛋白质组学方法研究三阴性 MDA-MB-231 乳腺癌细胞中的物理相互作用。然后，过度表达该相互作用的乳腺癌细胞系中的物理和功能相互作用因子将被耗尽。 antimorph HOTAIR 以确定其在 antimorph 机制中的作用。最近衍生的原代细胞系乳腺癌患者来源的异种移植物将用于确定该机制的普遍性。在第二个目标，反义寡核苷酸（ASO）将用于破坏 HOTAIR 上的 m6A 位点以引发反形态行为。在并行方法中，我们将开发一种优化策略来提供反形态 HOTAIR RNA 直接抑制癌症表型。这两种方法在以下情况下是互补的： HOTAIR 过度表达和非过度表达的乳腺癌。我们的工作将是对乳腺癌新治疗方法的基础测试，通过诱导 m6A 修饰的 LncRNA 的反态行为。总的来说，基于 ASO 的策略是一种新颖的方法乳腺癌治疗。 RNA 疗法现已成为现实，并获得广泛接受。我们的项目利用这种令人兴奋的氛围，采用雄心勃勃的新颖方法来干预非编码 RNA 乳腺癌。通过利用 m6A 提供的分子开关，我们可以逆转致癌 LncRNA 可逆转乳腺癌特性。