AbstractDysregulated transcription factors (TFs) represent a unique class of drug targets that drive aberrant gene expression programs involved in the major hallmarks of cancers. However, developing drug-like small molecules targeting TF proteins for clinical purposes has proven to be extremely challenging. Here, we present a novel and innovative approach to degrade the RNA transcript of the master oncogenic transcription factor C-MYB, thereby preventing its undesirable expression, using potent and orally available small molecules.C-MYB is a transcription factor that regulates differentiation and proliferation programs in normal cells, including hematopoietic, stem and epithelial cells. In numerous cancer types, such as adenoid cystic carcinoma (ACC), leukemia, colorectal cancer, and breast cancer, C-MYB is a well-established oncogenic transcription factor, overactivated via different mechanisms, including chromosomal translocations and gene amplification.Here, we leveraged our integrative RNA-targeting platform to identify and validate actionable cryptic exon target sites in the human C-MYB gene. We identified RGT-M001, a potent small molecule that can selectively induce the inclusion of the cryptic exon into the final C-MYB transcripts, resulting in a robust decrease of C-MYB canonical transcripts and C-MYB protein in cells. In a functional assay, we demonstrated that RGT-M001 has potent cell-killing activity against a large panel of cancer cell lines overexpressing C-MYB (EC50 ~ 20 - 300nM), while sparing normal cells. To confirm RGT-M001's on-target effect, we demonstrated a robust correlation between RGT-M001 cell-killing activity and the knockdown of C-MYB RNA and protein.We further investigated the anti-tumor activity of RGT-M001 in an ACC patient-derived xenograft (PDx) mouse model harboring C-MYB-NFBI fusion. Recurrent or metastatic ACC is a malignant neoplasm of predominantly salivary gland origin for which effective approved therapies are lacking; the best reported ORR being 15.6% for lenvatinib. As a single agent, RGT-M001 reduced in vivo C-MYB transcript levels by >80% at peak drug exposure and induced a remarkable tumor growth inhibition response (~70% TGI) in the ACCX11 PDx mouse model, surpassing the therapeutic benchmark Lenvatinib (40% TGI). Importantly, the RGT-M001 regimen was well tolerated. Finally, we showed that the combination of RGT-M001 with the Notch Inhibitor AL-101 resulted in complete inhibition of tumor growth.In conclusion, these data demonstrate that small molecules targeting RNA are a safe and effective approach to address previously undruggable protein targets. Down-regulation of C-MYB by our RNA-targeting small molecules is an attractive therapeutic strategy to treat ACC and other cancers driven by C-MYB dysregulations.Citation Format: Norman Lu, Patricia Soulard, Kay Li, Heather Sadlish, Chris Yates, Xiubin Gu, Ibrahim Kays, Jae Lee, Sam Hasson, Zhiping Weng, Simon Xi, Travis Wager. RGT-M001, a first-in-class small molecule mRNA degrader of the oncogenic transcription factor c-Myb, demonstrated remarkable single agent anti-tumor efficacy in cancer patient-derived xenograft model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3306.
