Combining Multiplexed CRISPR/Cas9-Nickase and PARP Inhibitors Efficiently and Precisely Targets Cancer Cells PMID: 40327605

ADC Targets in Oncology

2026-04-10

Combining Multiplexed CRISPR/Cas9-Nickase and PARP Inhibitors Efficiently and Precisely Targets Cancer Cells

Cancer Res. 2025 Aug 1;85(15):2890-2904. doi: 10.1158/0008-5472.CAN-24-2938.

Soyoung Lee ^# ¹ ² , Kyunghwan Kim ^# ¹ ³ , Hye-Jin Jeong ^# ² , Subin Choi ² , Himchan Cheng ² , Dayoung Kim ² , Soomin Heo ¹ ² , Jinhee Mun ² , Minjong Kim ⁴ , Eunjin Lee ² , Yoon Ji Choi ⁵ , Seon-Gyeong Lee ⁶ , Eun A Lee ¹ , Yewon Jang ¹ , Kayeong Lim ⁷ , Heon Seok Kim ⁸ , Euihwan Jeong ⁶ ⁹ , Seung-Jae Myung ¹⁰ ¹¹ ¹² , Deok-Beom Jung ¹¹ , Chang Sik Yu ¹³ , In Ho Song ¹³ , M Ryan Corces ¹⁴ ¹⁵ ¹⁶ , Joo H Kang ² , Kyungjae Myung ¹ ² , Taejoon Kwon ¹ ² ⁶ ⁹ ¹⁷ , Tae-Eun Park ² , Jinmyoung Joo ¹ ² ¹⁷ , Seung Woo Cho ¹ ² ¹⁷

Affiliations

1. Center for Genomic Integrity, Institute for Basic Science, Ulsan, Republic of Korea.
2. Department of Biomedical Engineering, College of Information and Biotechnology, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea.
3. Department of Chemistry, College of Natural Science, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea.
4. Department of Biological Science, College of Information and Biotechnology, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea.
5. In Vivo Research Center, UNIST Central Research Facilities, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea.
6. Division of Research and Development, CasCure Therapeutics, Seoul, Republic of Korea.
7. Brain Science Institute, Korea Institute of Science and Technology, Seoul, Republic of Korea.
8. Department of Life Science, College of Natural Science, Hanyang University, Seoul, Republic of Korea.
9. CasCure Therapeutics, Ulsan, Republic of Korea.
10. Department of Gastroenterology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea.
11. Digestive Diseases Research Center, University of Ulsan College of Medicine, Seoul, Republic of Korea.
12. EDIS Biotech, Seoul, Republic of Korea.
13. Division of Colon and Rectal Surgery, Department of Surgery, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea.
14. Gladstone Institute of Neurological Disease, San Francisco, California.
15. Gladstone Institute of Data Science and Biotechnology, San Francisco, California.
16. Department of Neurology, University of California San Francisco, San Francisco, California.
17. Graduate School of Health Science and Technology, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea.
# Contributed equally.

Abstract

Triggering Cancer cell death by inducing DNA damage is the primary aim of radiotherapy; however, normal cells are also damaged. In this study, we showed that delivery of only four synthetic guide RNAs with Cas9 Endonuclease efficiently induced simultaneous DNA double-strand breaks, resulting in efficient cell death in a cell type-specific manner. Off-target effects of Cas9 Endonuclease were prevented by using Cas9-nickase to induce DNA single-strand breaks and blocking their repair with PARP inhibitors (PARPi). When recombinant Cas9-nickase protein and multiple synthetic guide RNAs were delivered with PARPis into cultured cells, in vivo xenografts, and patient-derived Cancer organoids via lipid nanoparticles, Cancer cells were unable to tolerate the induced DNA damage even in the presence of a functional BRCA2 gene. This approach has the potential to expand the use of PARPis with verified safety and thus is a potentially powerful tool for personalized genome-based Anticancer therapy.

Significance: Targeting cancer-specific variants with CRISPR/Cas9-nickase induces cancer-specific cell death in combination with DNA repair pathway inhibitors, demonstrating the potential of CRISPR Cancer therapy for treating a broad range of cancers.