Poly(adenosine diphosphate ribose) polymerase inhibitors induce autophagy-mediated drug resistance in ovarian cancer cells, xenografts, and patient-derived xenograft models.

Abstract

BACKGROUND:

Poly(adenosine diphosphate ribose) polymerase (PARP) inhibitors exhibit promising activity against ovarian cancers, but their efficacy can be limited by acquired drug resistance. This study explores the role of autophagy in regulating the sensitivity of ovarian cancer cells to PARP inhibitors.

METHODS:

Induction of autophagy was detected by punctate LC3 fluorescence staining, LC3I to LC3II conversion on Western blot analysis, and electron microscopy. Enhanced growth inhibition and apoptosis were observed when PARP inhibitors were used with hydroxychloroquine, chloroquine (CQ), or LYS05 to block the hydrolysis of proteins and lipids in autophagosomes or with small interfering RNA against ATG5 or ATG7 to prevent the formation of autophagosomes. The preclinical efficacy of the combination of CQ and olaparib was evaluated with a patient-derived xenograft (PDX) and the OVCAR8 human ovarian cancer cell line.

RESULTS:

Four PARP inhibitors (olaparib, niraparib, rucaparib, and talazoparib) induced autophagy in a panel of ovarian cancer cells. Inhibition of autophagy with CQ enhanced the sensitivity of ovarian cancer cells to PARP inhibitors. In vivo, olaparib and CQ produced additive growth inhibition in OVCAR8 xenografts and a PDX. Olaparib inhibited PARP activity, and this led to increased reactive oxygen species (ROS) and an accumulation of γ-H2AX. Inhibition of autophagy also increased ROS and γ-H2AX and enhanced the effect of olaparib on both entities. Treatment with olaparib increased phosphorylation of ATM and PTEN while decreasing the phosphorylation of AKT and mTOR and inducing autophagy.

CONCLUSIONS:

PARP inhibitor-induced autophagy provides an adaptive mechanism of resistance to PARP inhibitors in cancer cells with wild-type BRCA, and a combination of PARP inhibitors with CQ or other autophagy inhibitors could improve outcomes for patients with ovarian cancer.

• Authors: Janice M Santiago-O'Farrill, S John Weroha, Xiaonan Hou, Ann L Oberg, Ethan P Heinzen, Matthew J Maurer, Lan Pang, Philip Rask, Ravi K Amaravadi, Sarah E Becker, Ignacio Romero, Ma Jesús Rubio, Xavier Matias-Guiu, Maria Santacana, Antonio Llombart-Cussac, Andrés Poveda, Zhen Lu, Robert C Bast Jr
• Journal: Cancer (Cancer) Vol. 126 Issue 4 Pg. 894-907 (02 15 2020) ISSN: 1097-0142 [Electronic] United States
• PMID: 31714594 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: © 2019 American Cancer Society.

Chemical References

• Antineoplastic Agents
• Indazoles
• Phthalazines
• Piperazines
• Piperidines
• Poly(ADP-ribose) Polymerase Inhibitors
• Chloroquine
• talazoparib
• Poly(ADP-ribose) Polymerases
• niraparib
• olaparib

Topics

• Animals
• Antineoplastic Agents (pharmacology)
• Apoptosis (drug effects)
• Autophagy (drug effects)
• Cell Line, Tumor
• Chloroquine (pharmacology)
• Drug Resistance, Neoplasm (drug effects)
• Drug Synergism
• Female
• Humans
• Indazoles (pharmacology)
• Mice, Nude
• Mice, SCID
• Ovarian Neoplasms (drug therapy, metabolism, pathology)
• Phthalazines (pharmacology)
• Piperazines (pharmacology)
• Piperidines (pharmacology)
• Poly(ADP-ribose) Polymerase Inhibitors (pharmacology)
• Poly(ADP-ribose) Polymerases (metabolism)
• Xenograft Model Antitumor Assays (methods)