SAM-Competitive PRMT5 Inhibitor PF-06939999 Demonstrates Antitumor Activity in Splicing Dysregulated NSCLC with Decreased Liability of Drug Resistance.

Abstract

Protein arginine methyltransferase 5 (PRMT5) overexpression in hematologic and solid tumors methylates arginine residues on cellular proteins involved in important cancer functions including cell-cycle regulation, mRNA splicing, cell differentiation, cell signaling, and apoptosis. PRMT5 methyltransferase function has been linked with high rates of tumor cell proliferation and decreased overall survival, and PRMT5 inhibitors are currently being explored as an approach for targeting cancer-specific dependencies due to PRMT5 catalytic function. Here, we describe the discovery of potent and selective S-adenosylmethionine (SAM) competitive PRMT5 inhibitors, with in vitro and in vivo characterization of clinical candidate PF-06939999. Acquired resistance mechanisms were explored through the development of drug resistant cell lines. Our data highlight compound-specific resistance mutations in the PRMT5 enzyme that demonstrate structural constraints in the cofactor binding site that prevent emergence of complete resistance to SAM site inhibitors. PRMT5 inhibition by PF-06939999 treatment reduced proliferation of non-small cell lung cancer (NSCLC) cells, with dose-dependent decreases in symmetric dimethyl arginine (SDMA) levels and changes in alternative splicing of numerous pre-mRNAs. Drug sensitivity to PF-06939999 in NSCLC cells associates with cancer pathways including MYC, cell cycle and spliceosome, and with mutations in splicing factors such as RBM10. Translation of efficacy in mouse tumor xenograft models with splicing mutations provides rationale for therapeutic use of PF-06939999 in the treatment of splicing dysregulated NSCLC.

• Authors: Kristen Jensen-Pergakes, John Tatlock, Karen A Maegley, Indrawan J McAlpine, Michele McTigue, Tao Xie, Christopher P Dillon, Yuli Wang, Shinji Yamazaki, Noah Spiegel, Manli Shi, Amy Nemeth, Natalie Miller, Eleanore Hendrickson, Hieu Lam, John Sherrill, Chi-Yeh Chung, Elizabeth A McMillan, Shannon Karlicek Bryant, Prakash Palde, John Braganza, Alexei Brooun, Ya-Li Deng, Vesta Goshtasbi, Susan E Kephart, Robert A Kumpf, Wei Liu, Ryan L Patman, Eugene Rui, Stephanie Scales, Michelle Tran-Dube, Fen Wang, Martin Wythes, Thomas A Paul
• Journal: Molecular cancer therapeutics (Mol Cancer Ther) Vol. 21 Issue 1 Pg. 3-15 (01 2022) ISSN: 1538-8514 [Electronic] United States
• PMID: 34737197 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: ©2021 American Association for Cancer Research.

Chemical References

• S-Adenosylmethionine
• Prmt5 protein, mouse
• Protein-Arginine N-Methyltransferases

Topics

• Animals
• Apoptosis
• Carcinoma, Non-Small-Cell Lung (drug therapy, genetics, pathology)
• Cell Line, Tumor
• Cell Proliferation
• Drug Resistance
• Female
• Humans
• Lung Neoplasms (drug therapy, genetics, pathology)
• Mice
• Protein-Arginine N-Methyltransferases (antagonists & inhibitors)
• S-Adenosylmethionine (metabolism)