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6127.Full.Pdf Published OnlineFirst August 13, 2019; DOI: 10.1158/1078-0432.CCR-19-0448 Translational Cancer Mechanisms and Therapy Clinical Cancer Research The CHK1 Inhibitor Prexasertib Exhibits Monotherapy Activity in High-Grade Serous Ovarian Cancer Models and Sensitizes to PARP Inhibition Kalindi Parmar1,2, Bose S. Kochupurakkal1,2, Jean-Bernard Lazaro1,2, Zhigang C. Wang3, Sangeetha Palakurthi4, Paul T. Kirschmeier4, Chunyu Yang1,2, Larissa A. Sambel1,2, Anniina Farkkil€ a€1,2, Elizaveta Reznichenko1,2, Hunter D. Reavis1,2, Connor E. Dunn1,2, Lee Zou5, Khanh T. Do6,7, Panagiotis A. Konstantinopoulos6,7, Ursula A. Matulonis6,7, Joyce F. Liu6,7, Alan D. D'Andrea1,2, and Geoffrey I. Shapiro1,6,7 Abstract Purpose: PARP inhibitors are approved for the treatment of Results: Prexasertib monotherapy demonstrated antitu- high-grade serous ovarian cancers (HGSOC). Therapeutic mor activity across the 14 PDX models. Thirteen models were resistance, resulting from restoration of homologous recom- resistant to olaparib monotherapy, including 4 carrying bination (HR) repair or replication fork stabilization, is a BRCA1 mutation. The combination of olaparib with prexa- pressing clinical problem. We assessed the activity of prexa- sertib was synergistic and produced significant tumor growth sertib, a checkpoint kinase 1 (CHK1) inhibitor known to cause inhibition in an olaparib-resistant model and further aug- replication catastrophe, as monotherapy and in combination mented the degree and durability of response in the olaparib- with the PARP inhibitor olaparib in preclinical models sensitive model. HGSOC cell lines, including those with of HGSOC, including those with acquired PARP inhibitor acquired PARP inhibitor resistance, were also sensitive to resistance. prexasertib, associated with induction of DNA damage and Experimental Design: Prexasertib was tested as a single replication stress. Prexasertib also sensitized these cell lines agent or in combination with olaparib in 14 clinically to PARP inhibition and compromised both HR repair and annotated and molecularly characterized luciferized replication fork stability. HGSOC patient-derived xenograft (PDX) models and in a Conclusions: Prexasertib exhibits monotherapy activity in panel of ovarian cancer cell lines. The ability of prexasertib PARP inhibitor–resistant HGSOC PDX and cell line models, to impair HR repair and replication fork stability was reverses restored HR and replication fork stability, and syner- also assessed. gizes with PARP inhibition. 1Center for DNA Damage and Repair, Dana-Farber Cancer Institute, Boston, Introduction 2 Massachusetts. Department of Radiation Oncology, Dana-Farber Cancer Insti- High-grade serous ovarian cancer (HGSOC) is the most 3 tute, Boston, Massachusetts. Department of Cancer Biology, Dana-Farber frequent and aggressive subtype of epithelial ovarian cancer Cancer Institute, Boston, Massachusetts. 4Belfer Center for Applied Cancer Science, Dana-Farber Cancer Institute, Boston, Massachusetts. 5Department of (EOC), with high recurrence rates despite initial responses to Pathology, Massachusetts General Hospital Cancer Center and Harvard Medical platinum-based chemotherapy (1, 2). Approximately 50% of School, Charlestown, Massachusetts. 6Department of Medical Oncology, the HGSOCs exhibit genetic and epigenetic alterations in gene Dana-Farber Cancer Institute, Boston, Massachusetts. 7Department of Medicine, members of the homologous recombination (HR) DNA repair Brigham and Women's Hospital and Harvard Medical School, Boston, pathway including BRCA1/2 and other Fanconi Anemia/BRCA Massachusetts. pathway genes (3, 4). Based on the synthetic lethal interaction Note: Supplementary data for this article are available at Clinical Cancer of defective HR repair and PARP inhibition, HR-deficient Research Online (http://clincancerres.aacrjournals.org/). EOCs are highly sensitive to PARP inhibitors (5). Currently, Corresponding Authors: Geoffrey I. Shapiro, Dana-Farber Cancer Institute, olaparib, rucaparib, and niraparib are all FDA-approved for Mayer 446, 450 Brookline Avenue, Boston, MA 02215. Phone: 617-632-4942; HGSOC (6); in the recurrent disease setting, these agents have Fax: 617-632-1977; E-mail: [email protected]; and Alan D. been primarily used in patients with tumors harboring BRCA D'Andrea, Dana-Farber Cancer Institute, Harvard Institutes of Medicine 243, alterations with substantial response rates. Additionally, these 450 Brookline Avenue, Boston, MA 02215. Phone: 617-632-2080; Fax: 617-632- 6069; E-mail: [email protected] agents are now commonly considered in the maintenance setting, after a response to a platinum-based chemotherapy, Clin Cancer Res 2019;25:6127–40 where PARP inhibition has significantly increased progression- doi: 10.1158/1078-0432.CCR-19-0448 free survival and the chemotherapy-free interval for many Ó2019 American Association for Cancer Research. patients (7, 8). www.aacrjournals.org 6127 Downloaded from clincancerres.aacrjournals.org on September 23, 2021. © 2019 American Association for Cancer Research. Published OnlineFirst August 13, 2019; DOI: 10.1158/1078-0432.CCR-19-0448 Parmar et al. derived xenograft (PDX) models, and in a panel of ovarian cancer Translational Relevance cell lines. Prexasertib showed broad preclinical monotherapy The CHK1 inhibitor prexasertib is currently in clinical activity and was synergistic with PARP inhibition in several development as a single agent and in combination with both models, including those representing BRCA1-mutant, PARP cytotoxic and targeted agents. In high-grade serous ovarian inhibitor–resistant disease, where there was reversal of both cancer (HGSOC), prexasertib monotherapy has demonstrated restored HR repair and of replication fork stability. activity in patients with BRCA wild-type, platinum-resistant disease. Here, we tested the activity of prexasertib in both Materials and Methods BRCA wild-type and BRCA1-mutated, PARP inhibitor–resis- Establishment and treatment of PDXs tant HGSOC patient-derived xenograft (PDX) models and cell Tumor ascites was collected from patients with suspected or lines. These preclinical models were routinely sensitive to established ovarian cancer at the Brigham and Women's Hospital prexasertib, likely related to induction of DNA damage and or the Dana-Farber Cancer Institute (DFCI) under IRB-approved replication stress. Additionally, prexasertib impaired both protocols conducted in accordance with the Declaration of homologous recombination repair and replication fork sta- Helsinki and the Belmont Report. Written informed consent was bility, addressing 2 major mechanisms of resistance to PARP obtained from patients when required. Fourteen ascites-derived inhibition. Consequently, prexasertib was synergistic with ovarian PDX models were established, luciferized, and propagat- olaparib in multiple models. These data suggest that prexa- ed at the DFCI and have been described previously (30). All sertib, either as monotherapy or in combination with olaparib, animal studies were performed in accordance with DFCI Institu- has broad activity against HGSOCs, including those harboring tional Animal Care and Use Committee guidelines per DFCI- BRCA1 mutation with acquired PARP inhibitor resistance, approved animal protocols. Treatment studies in mice bearing informing additional ovarian cancer populations for clinical PDX tumors were performed as described previously (30, 31). trials. Approximately 2 to 10 Â 106 luciferized PDX cells derived from mouse ascites were injected intraperitoneally into 8-week-old female NSG mice. Tumors were typically established 1 to 2 weeks after implantation. Tumor burden was measured by biolumines- With more common PARP inhibitor use, acquired resistance cence imaging (BLI) using a Xenogen IVIS-200 system (Xenogen). has emerged as an unmet medical need. The major mechanisms of Mice were randomized into treatment groups and disseminated PARP inhibitor resistance include restoration of HR repair or disease progression in the peritoneal cavity was measured serially stabilization and protection of replication forks (9–12). There is once per week by BLI. a critical need for combinations utilizing agents that can inhibit Prexasertib (mesylate monohydrate salt of LY2606368; HR and/or reverse replication fork stability and thereby sensitize Eli Lilly) was formulated in 20% Captisol (CyDex Inc.) pH 4.0 HGSOCs to PARP inhibition. Such approaches may also extend and administered subcutaneously at 8 mg/kg twice daily for the use of PARP inhibitors to include HR-proficient, BRCA wild- 3 days, followed by 4 days of rest and repeated for 2 additional type tumors. cycles. Olaparib (ChemExpress) was formulated in PBS contain- Because of the endogenous DNA damage caused by HR defi- ing 10% DMSO and 10% (w/v) 2-hydroxy-propyl-b-cyclodextrin ciency, HGSOCs tumors require an intact ataxia telangiectasia (Sigma-Aldrich) and administered orally at 100 mg/kg daily for and Rad3-related (ATR)–checkpoint kinase 1 (CHK1) pathway to 3 weeks. Tumor-bearing mice were monitored for survival and allow time for alternative DNA repair pathways to function and tumor burden for approximately 100 to 150 days. For pharma- maintain genome integrity. Additionally, HGSOCs often dem- codynamic studies, mice were euthanized at specific time points onstrate genomic alterations suggestive of replicative stress. For and tumor cells were harvested from ascites. example, 20% of HGSOCs may express high levels of cyclin fi E (4, 13), often as a result of gene
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