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Principal Investigator: Aaron M. Johnson
Organization: UNIVERSITY OF COLORADO DENVER
Fiscal Year: 2023
Award: $181,741
Funding agency: National Cancer Institute
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.
Terms: <Address><Affect><Anti-Sense Oligonucleotides><Antisense Agent><Antisense Oligonucleotides><Atmosphere><Behavior><Breast Cancer><Breast Cancer Cell><Breast Cancer Patient><Breast Cancer cell line><Breast PDX models><Breast Tumor Patient><Breast tumor cell line><Cancer Biology><Cancer Genes><Cancer Induction><Cancer Treatment><Cancer-Promoting Gene><Cancers><Cell Body><Cell Line><CellLine><Cells><Chemicals><Clinic><Code><Coding System><Diagnosis><Functional RNA><Future><Gene Down-Regulation><Gene Expression><Gene Targeting><Gene Transcription><Generalized Growth><Genes><Genetic Transcription><Goals><Growth><Heterograft><Heterologous Transplantation><Intervention><Intervention Strategies><Lead><Length><MDA MB 231><MDA-231><MDA-MB231><Malignant Cell><Malignant Neoplasm Therapy><Malignant Neoplasm Treatment><Malignant Neoplasms><Malignant Tumor><Methylation><Modification><Molecular><Mutate><Non-Coding><Non-Coding RNA><Non-Polyadenylated RNA><Non-translated RNA><Noncoding RNA><Nontranslated RNA><Nuclear><Nucleotides><Oncogenes><Oncogenic><Pathway interactions><Patient-derived xenograft models of breast cancer><Patients><Pattern><Pb element><Phenotype><Prevention><Process><Property><Proteins><Proteomics><RNA><RNA Expression><RNA Gene Products><RNA based therapeutics><RNA based therapy><RNA delivery><RNA methylation><RNA therapy><RNA vaccine><RNA-based vaccine><Ribonucleic Acid><Role><Site><Strains Cell Lines><Suppressor Mutations><Testing><Therapeutic><Therapeutic Intervention><Tissue Growth><Transcript><Transcription><Transcription Repression><Transcriptional Repression><Transforming Genes><Tumor Suppressor Proteins><Untranslated RNA><Work><Xenograft><Xenograft procedure><Xenotransplantation><anti-cancer><anti-cancer therapy><anti-sense agent><anti-sense oligo><anticancer><anticancer therapy><antisense oligo><breast cancer PDX><breast cancer patient-derived xenograft><breast cancer progression><breast tumor cell><cancer cell><cancer therapy><cancer type><cancer-directed therapy><carcinogenesis><clinical applicability><clinical application><cultured cell line><deliver vaccines><experiment><experimental research><experimental study><experiments><gene repression><heavy metal Pb><heavy metal lead><inhibitor><intervention therapy><interventional strategy><mRNA vaccine><mRNA-based vaccine><malignancy><malignant breast neoplasm><malignant breast tumor><mutant><neoplasm/cancer><new approaches><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutic approach><new therapeutic intervention><new therapeutic strategies><new therapeutics><new therapy><new therapy approaches><new treatment approach><new treatment strategy><next generation therapeutics><noncoding><novel approaches><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel strategies><novel strategy><novel therapeutic approach><novel therapeutic intervention><novel therapeutic strategies><novel therapeutics><novel therapy><novel therapy approach><ontogeny><overexpress><overexpression><pathway><social role><therapeutic RNA><tumor><tumor suppressor><vaccine delivery><xeno-transplant><xeno-transplantation>