Sensitization of colorectal cancer to irinotecan therapy by PARP inhibitor rucaparib
Intended to explore synthetic lethality and develop better combinatorial regimens, we screened colorectal cancer (CRC) cells using poly ADP-ribose (PAR) polymerase (PARP) inhibitors and cytotoxic agents. We studied four PARP inhibitors and three DNA-damaging agents, and their combinations using sulforhodamine B assay. Rucaparib demonstrated the greatest synergy with irinotecan, followed by olaparib and PJ34. Rucaparib and irinotecan was further subjected to detailed examination to determine combination index (CI) and underlying mechanism of action. Effectiveness and sequence dependence of this combination were assessed in microsatellite stable (MSS) and unstable (MSI) CRC and HCT116 isogenic cell lines. The degree of cell cycle arrest and apoptosis was determined by FACS. In vivo studies were performed to confirm efficacy of this combination. PAR levels in MSI and PARP expression in MSI and MSS cell lines were diminished upon combinatorial treatment. HCT116 isogenic cells revealed the importance of p21, p53 and PTEN in exerting synergy. In MSI cells, administration of rucaparib prior to irinotecan enhanced cytotoxicity compared to other strategies explored. FACS revealed S-phase arrest and increased late-stage apoptosis in MSS, and G2-M arrest and total and early-stage apoptosis in MSI cells. In in vivo murine xenograft models, a significant reduction in tumor volume and expression of Ki67, pancytokeratin and RPS6KB1, and increase in expression of caspase 3 were observed with the combination. In conclusion, among the various combinations studied, rucaparib plus irinotecan was the most synergistic one. Alterations in cell cycle arrest and apoptosis were dependent on MSI status in CRC cells.
KeywordsPARP Rucaparib Irinotecan Colorectal cancer Combinatorial Synergy
S. Goel is supported by a K-12 award from the National Cancer Institute of the National Institutes of Health 1K12CA132783-01A1, and an Advanced Clinical Research Award (ACRA) in colon cancer, by the ASCO (now Conquer) Cancer Foundation. The authors would like to thank Dr. Tanya Dragic from Department of Microbiology & Immunology, Dr. Balazs Halmos from Department of Medicine and Dr. Thomas J. Ow from Department of Pathology, Albert Einstein College of Medicine/Montefiore Medical Center for their helpful advice on various technical issues examined in this paper, and Dr. Dhanonjoy C. Saha from Office of Grant Support, Albert Einstein College of Medicine, for his advice and comments. The authors also greatly appreciate the expert advice/assistance of Hillary Guzik, Analytical Imaging Facility, Albert Einstein College of Medicine in microscopy/IHC studies. The imaging was conducted in the Analytical Imaging Facility, which is funded by the NCI Cancer Grant P30CA013330.
The work was supported by the K-12 award from the National Cancer Institute of the National Institutes of Health 1K12CA132783-01A1, and an Advanced Clinical Research Award (ACRA) in colon cancer, by the ASCO (now Conquer) Cancer Foundation to Dr. Sanjay Goel.
Compliance with ethical standards
Conflict of interest
Titto Augustine declares that he has no conflict of interest. Radhashree Maitra declares that she has no conflict of interest. Jinghang Zhang declares that she has no conflict of interest. Jay Nayak declares that he has no conflict of interest. Sanjay Goel declares that he has no conflict of interest.
This article does not contain any studies with human participants. All applicable institutional guidelines (by Institutional Animal Care and Use Committee) for the care and use of animals were followed.
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