Inverted chimeric RNAi molecules synergistically co-target MYC and KRAS in KRAS-driven cancers

Yogitha S Chareddy; Hayden P. Huggins; Snehasudha S Sahoo; Lyla Stanland; Christina Gutierrez-Ford; Kristina M. Whately; Lincy Edatt; Salma H Azam; Matthew C. Fleming; Jonah Im; Alessandro Porrello; Imani Simmons; Jillian L. Perry; Albert A. Bowers; Martin Egli; Chad V. Pecot

doi:10.1172/JCI187204

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Chareddy, Y. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Huggins, H. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Sahoo, S. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Stanland, L. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Gutierrez-Ford, C. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Whately, K. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Edatt, L. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Azam, S. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Fleming, M. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Im, J. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Porrello, A. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Simmons, I. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Perry, J. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Bowers, A. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Egli, M. in: PubMed | Google Scholar

¹Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

²EnFuego Therapeutics, Inc, Morrisville, United States of America

³Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁴Division of Chemical Biology and Medicinal Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁵Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, United States of America

⁶Department of Biochemistry, Vanderbilt University, Nashville, United States of America

Find articles by Pecot, C. in: PubMed | Google Scholar |

J Clin Invest. https://doi.org/10.1172/JCI187204.
Copyright © 2025, Chareddy et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

Published July 31, 2025 - Version history

Mutant KRAS has been implicated in driving a quarter of all cancer types. Although inhibition of the KRAS^G12C mutant protein has shown clinical promise, there is still a need for therapies that overcome resistance and target non-KRAS^G12C mutations. KRAS activates downstream MYC, which is also a challenging-to-drug oncoprotein. We have developed an “inverted” RNAi molecule with the passenger strand of a MYC-targeting siRNA fused to the guide strand of a KRAS-targeting siRNA. The chimeric molecule simultaneously inhibits KRAS and MYC, showing marked improvements in efficacy beyond the individual siRNA components. This effect is mediated by 5’-dT overhangs following endosomal metabolism. The synergistic RNAi activity led to a >10-40-fold improvement in inhibiting cancer viability in vitro. When conjugated to an epidermal growth factor receptor (EGFR)-targeting ligand, the chimeric siRNA was delivered to and internalized by tumor cells. As compared with individual targeting siRNAs, the chimeric design resulted in considerably improved metabolic stability in tumors, enhanced silencing of both oncogenes, and reduced tumor progression in multiple cancer models. This inverted chimeric design establishes proof-of-concept for ligand-directed, dual-silencing of KRAS and MYC in cancer and constitutes an innovative molecular strategy for co-targeting any two genes of interest, which has broad implications.

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Inverted chimeric RNAi molecules synergistically co-target MYC and KRAS in KRAS-driven cancers

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Inverted chimeric RNAi molecules synergistically co-target MYC and KRAS in KRAS-driven cancers

Yogitha S Chareddy,1 Hayden P. Huggins,2 Snehasudha S Sahoo,3 Lyla Stanland,2 Christina Gutierrez-Ford,3 Kristina M. Whately,3 Lincy Edatt,3 Salma H Azam,1 Matthew C. Fleming,4 Jonah Im,3 Alessandro Porrello,3 Imani Simmons,3 Jillian L. Perry,5 Albert A. Bowers,4 Martin Egli,6 and Chad V. Pecot3

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