Morpholino-mediated splice switching of NELFE suppresses hepatocellular carcinoma cell phenotypes in vitro and in vivo.
11 affiliations
- 1Department of Surgery, Thomas Jefferson University, Philadelphia, PA 19107, USA.
- 2Department of Radiology, Thomas Jefferson University, Philadelphia, PA 19107, USA.
- 3Institute of Pathology, University Medical Center Mainz, JGU-Mainz, 55131 Mainz, Germany.
- 4TRON, Translational Oncology at the University Medical Center, JGU-Mainz, 55131 Mainz, Germany.
- 5Department of Pathology and Genome Medicine, Thomas Jefferson University, Philadelphia, PA 19107, USA.
- 6Laboratory of Human Carcinogenesis, National Cancer Institute, Bethesda, MD 20892, USA.
- 7Department of Medical Oncology, Thomas Jefferson University, Philadelphia, PA 19107, USA.
- 8Sidney Kimmel Comprehensive Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA.
- 9Liver Cancer Program, Center for Cancer Research, National Cancer Institute, Bethesda, MD 20892, USA.
- 10Department of Surgery, Pennsylvania State University College of Medicine, Hershey, PA 17033, USA.
- 11Department of Molecular Biology, Cell Biology, and Biochemistry, Brown University, Providence, RI 02912, USA.
Abstract
Hepatocellular carcinoma (HCC) is among the deadliest cancers worldwide, with rising incidence and few targeted therapies, driving high rates of recurrence and poor survival. Negative elongation factor E (NELFE), an RNA-binding subunit of the NELF complex, acts oncogenically when overexpressed and is associated with worse prognosis in HCC, yet no therapies target it. Here, we identify a rare germline intronic variant in the NELFE locus, NM_002904.6:c.943-110C>T (rs79208225) that promotes skipping of NELFE exon 10 and identifies patients with a trend toward longer recurrence-free survival and lower NELFE expression in their tumors. Mechanistically, the variant reduces U2AF2 occupancy at intron 9, likely through an indirect effect on splice-site recognition rather than direct disruption of U2AF2 binding. Recognizing exon 10 as a targetable splice event, we designed morpholino (MO) antisense oligonucleotides against its splice junctions that phenocopied the variant, lowering NELFE protein expression and impairing malignant phenotypes across HCC cell lines spanning diverse etiologies and NELFE expression levels. Delivered by ultrasound-targeted microbubble destruction (UTMD) in orthotopic liver tumors, NELFE MOs significantly reduced tumor burden and extended survival. These findings establish a naturally occurring splicing event as the basis for a localized, splice-switching therapeutic strategy for HCC.
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BibTeX
@article{reynolds2026morpholinomediated,
title = {{Morpholino-mediated splice switching of NELFE suppresses hepatocellular carcinoma cell phenotypes in vitro and in vivo.}},
author = {Laura M Reynolds and Ryan Lamm and Kai Zhang and Anna E Barry-Wolbers and Alvaro Lucci and Brittany N Ruiz and Corinne E Wessner and Trish Nguyen and Kayla M Hammond and Christoph Eckert and Ji-Bin Liu and Pongsakorn Choochuen and Kai Ying Hong and Wei Jiang and Marshonna Forgues and Gaia M Palazzo and Daniel Lin and Adam S Bodzin and Ashesh P Shah and Xin Wei Wang and Sean P Martin and Nicolas L Fawzi and Matthias M Gaida and John R Eisenbrey and Hien Dang},
journal = {Molecular therapy. Nucleic acids},
year = {2026},
month = oct,
doi = {10.1016/j.omtn.2026.103124},
url = {https://europepmc.org/article/MED/42927183},
note = {CC BY},
}
RIS
TY - JOUR
TI - Morpholino-mediated splice switching of NELFE suppresses hepatocellular carcinoma cell phenotypes in vitro and in vivo.
AU - Laura M Reynolds
AU - Ryan Lamm
AU - Kai Zhang
AU - Anna E Barry-Wolbers
AU - Alvaro Lucci
AU - Brittany N Ruiz
AU - Corinne E Wessner
AU - Trish Nguyen
AU - Kayla M Hammond
AU - Christoph Eckert
AU - Ji-Bin Liu
AU - Pongsakorn Choochuen
AU - Kai Ying Hong
AU - Wei Jiang
AU - Marshonna Forgues
AU - Gaia M Palazzo
AU - Daniel Lin
AU - Adam S Bodzin
AU - Ashesh P Shah
AU - Xin Wei Wang
AU - Sean P Martin
AU - Nicolas L Fawzi
AU - Matthias M Gaida
AU - John R Eisenbrey
AU - Hien Dang
PY - 2026
DA - 2026/10/01
JO - Molecular therapy. Nucleic acids
DO - 10.1016/j.omtn.2026.103124
UR - https://europepmc.org/article/MED/42927183
AB - Hepatocellular carcinoma (HCC) is among the deadliest cancers worldwide, with rising incidence and few targeted therapies, driving high rates of recurrence and poor survival. Negative elongation factor E (NELFE), an RNA-binding subunit of the NELF complex, acts oncogenically when overexpressed and is associated with worse prognosis in HCC, yet no therapies target it. Here, we identify a rare germline intronic variant in the NELFE locus, NM_002904.6:c.943-110C>T (rs79208225) that promotes skipping of NELFE exon 10 and identifies patients with a trend toward longer recurrence-free survival and lower NELFE expression in their tumors. Mechanistically, the variant reduces U2AF2 occupancy at intron 9, likely through an indirect effect on splice-site recognition rather than direct disruption of U2AF2 binding. Recognizing exon 10 as a targetable splice event, we designed morpholino (MO) antisense oligonucleotides against its splice junctions that phenocopied the variant, lowering NELFE protein expression and impairing malignant phenotypes across HCC cell lines spanning diverse etiologies and NELFE expression levels. Delivered by ultrasound-targeted microbubble destruction (UTMD) in orthotopic liver tumors, NELFE MOs significantly reduced tumor burden and extended survival. These findings establish a naturally occurring splicing event as the basis for a localized, splice-switching therapeutic strategy for HCC.
SN - 2162-2531
ER -