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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.

项目摘要在乳腺癌中过表达的特异性长非编码RNA（LncRNA）有助于诊断和治疗。促进推动乳腺癌进展的分子机制。我们最近的研究发现了一个乳房- 一种癌症相关的LncRNA，通过一个单一的核苷酸这种"反形态"效应是由防止RNA甲基化引起的，过度表达转化的LncRNA以减少癌症表型比消耗野生型LncRNA更明显。类型版本。这一令人兴奋的新发现促使我们深入研究这是如何发生的，最终目标是特异性破坏这种m6A通路作为一种新的治疗方法。针对乳腺癌相关LncRNA提出的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。我们的研究结果提示了一种新的致癌LncRNA治疗方法，单独破坏RNA甲基化比简单消除RNA具有更大的影响。当前该提案提供了一个路线图，以了解如何在致癌LncRNA中引发抗形态效应。我们的目标将这个原理验证项目作为未来工作的跳板，去诊所我们假设特定乳腺癌相关LncRNA上的m6A修饰位点可能是靶向和破坏，以逆转低于基线的致癌行为，为潜在的新的治疗方法在我们的第一个目标中，将研究表现为反形的突变HOTAIR LncRNA 使用蛋白质组学方法在三阴性MDA-MB-231乳腺癌细胞中进行物理相互作用。物理和功能相互作用物然后将在过表达该相互作用物的乳腺癌细胞系中耗尽。 HOTAIR，以确定它们在抗变形机制中的作用。最近衍生的原代细胞系来自将使用乳腺癌患者来源的异种移植物来确定该机制的普遍性。在第二个目标是使用反义寡核苷酸（ASO）破坏HOTAIR上的m6A位点，反形行为在一个平行的方法中，我们将开发一个优化的策略来提供反形态 HOTAIR RNA直接抑制癌症表型。在以下情况下，这两种方法是互补的： HOTAIR过表达的乳腺癌和HOTAIR未过表达的乳腺癌。我们的工作将是一个基础性的测试，一种新的治疗方法在乳腺癌中诱导 m6A修饰的LncRNA的反形态行为。总的来说，基于ASO的策略是一种新颖的方法，乳腺癌治疗学。RNA疗法现在已经成为现实，并得到广泛接受。我们的项目利用这一令人兴奋的气氛，提出了一种雄心勃勃的新方法来干预非编码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逆转乳腺癌特性。