Low-Dose Vertical Inhibition of the RAF-MEK-ERK Cascade Causes Apoptotic Death of KRAS Mutant Cancers

Irem Ozkan-Dagliyan; J Nathaniel Diehl; Samuel D George; Antje Schaefer; Bjoern Papke; Kathleen Klotz-Noack; Andrew M Waters; Craig M Goodwin; Prson Gautam; Mariaelena Pierobon; Sen Peng; Thomas S K Gilbert; Kevin H Lin; Onur Dagliyan; Krister Wennerberg; Emanuel F Petricoin 3rd; Nhan L Tran; Shripad V Bhagwat; Ramon V Tiu; Sheng-Bin Peng; Laura E Herring; Lee M Graves; Christine Sers; Kris C Wood; Adrienne D Cox; Channing J Der

doi:10.1016/j.celrep.2020.107764

Low-Dose Vertical Inhibition of the RAF-MEK-ERK Cascade Causes Apoptotic Death of KRAS Mutant Cancers

Cell Rep. 2020 Jun 16;31(11):107764. doi: 10.1016/j.celrep.2020.107764.

Authors

Irem Ozkan-Dagliyan¹, J Nathaniel Diehl², Samuel D George³, Antje Schaefer¹, Bjoern Papke⁴, Kathleen Klotz-Noack⁵, Andrew M Waters⁴, Craig M Goodwin⁴, Prson Gautam⁶, Mariaelena Pierobon⁷, Sen Peng⁸, Thomas S K Gilbert⁹, Kevin H Lin¹⁰, Onur Dagliyan¹¹, Krister Wennerberg⁶, Emanuel F Petricoin 3rd⁷, Nhan L Tran¹², Shripad V Bhagwat¹³, Ramon V Tiu¹³, Sheng-Bin Peng¹³, Laura E Herring⁹, Lee M Graves¹, Christine Sers¹⁴, Kris C Wood¹⁰, Adrienne D Cox¹⁵, Channing J Der¹⁶

Affiliations

¹ Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
² Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
³ Department of Computer Science, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
⁴ Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
⁵ Charité Universitätsmedizin Berlin, Institute of Pathology, Laboratory of Molecular Tumor Pathology and Systems Biology, 10117 Berlin, Germany.
⁶ Institute for Molecular Medicine Finland, University of Helsinki, 00290 Helsinki, Finland.
⁷ Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA 20110, USA.
⁸ Departments of Cancer and Cell Biology, Translational Genomics Research Institute, Phoenix, AZ 85004, USA.
⁹ UNC Michael Hooker Proteomics Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
¹⁰ Department of Pharmacology and Cancer Biology, Duke University, Durham, NC 27710, USA.
¹¹ Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA.
¹² Departments of Cancer Biology and Neurosurgery, Mayo Clinic Arizona, Scottsdale, AZ 85259, USA.
¹³ Eli Lilly and Company, Indianapolis, IN 46285, USA.
¹⁴ Charité Universitätsmedizin Berlin, Institute of Pathology, Laboratory of Molecular Tumor Pathology and Systems Biology, 10117 Berlin, Germany; German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany; Berlin Institute of Health (BIH), Anna-Louise-Karsch-Str. 2, 10178 Berlin, Germany.
¹⁵ Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Department of Radiation Oncology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
¹⁶ Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Charité Universitätsmedizin Berlin, Institute of Pathology, Laboratory of Molecular Tumor Pathology and Systems Biology, 10117 Berlin, Germany. Electronic address: [email protected].

Abstract

We address whether combinations with a pan-RAF inhibitor (RAFi) would be effective in KRAS mutant pancreatic ductal adenocarcinoma (PDAC). Chemical library and CRISPR genetic screens identify combinations causing apoptotic anti-tumor activity. The most potent combination, concurrent inhibition of RAF (RAFi) and ERK (ERKi), is highly synergistic at low doses in cell line, organoid, and rat models of PDAC, whereas each inhibitor alone is only cytostatic. Comprehensive mechanistic signaling studies using reverse phase protein array (RPPA) pathway mapping and RNA sequencing (RNA-seq) show that RAFi/ERKi induced insensitivity to loss of negative feedback and system failures including loss of ERK signaling, FOSL1, and MYC; shutdown of the MYC transcriptome; and induction of mesenchymal-to-epithelial transition. We conclude that low-dose vertical inhibition of the RAF-MEK-ERK cascade is an effective therapeutic strategy for KRAS mutant PDAC.

Keywords: ERK; FOSL1; KRAS; MYC; RAF; mesenchymal-to-epithelial transition; pancreatic cancer.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Apoptosis / drug effects
Apoptosis / genetics*
Carcinoma, Pancreatic Ductal / genetics*
Carcinoma, Pancreatic Ductal / metabolism
Carcinoma, Pancreatic Ductal / pathology
Cell Line, Tumor
Humans
MAP Kinase Signaling System / genetics*
Mitogen-Activated Protein Kinase Kinases / genetics
Mitogen-Activated Protein Kinase Kinases / metabolism
Mutation / genetics
Pancreatic Neoplasms / genetics*
Pancreatic Neoplasms / metabolism
Pancreatic Neoplasms / pathology
Protein Kinase Inhibitors / pharmacology
Proto-Oncogene Proteins c-akt / genetics
Proto-Oncogene Proteins c-akt / metabolism
Proto-Oncogene Proteins p21(ras) / genetics*
Proto-Oncogene Proteins p21(ras) / metabolism
Signal Transduction / drug effects
Signal Transduction / genetics

Substances

KRAS protein, human
Protein Kinase Inhibitors
Proto-Oncogene Proteins c-akt
Mitogen-Activated Protein Kinase Kinases
Proto-Oncogene Proteins p21(ras)

Abstract

Publication types

MeSH terms

Substances

Grants and funding