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WEE1 Inhibition in Combination With Targeted Agents and Standard Chemotherapy in Preclinical Models of Pancreatic Ductal Adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal cancer with high incidences of p53 mutations. AZD1775 (adavosertib, previously MK-1775) is a small molecule WEE1 inhibitor that abrogates the G2M checkpoint and can potentially synergize with DNA damaging therapies commonly used in PDAC trea...

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Autores principales: Hartman, Sarah J., Bagby, Stacey M., Yacob, Betelehem W., Simmons, Dennis M., MacBeth, Morgan, Lieu, Christopher H., Davis, S. Lindsey, Leal, Alexis D., Tentler, John J., Diamond, Jennifer R., Eckhardt, S. Gail, Messersmith, Wells A., Pitts, Todd M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8044903/
https://www.ncbi.nlm.nih.gov/pubmed/33869031
http://dx.doi.org/10.3389/fonc.2021.642328
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author Hartman, Sarah J.
Bagby, Stacey M.
Yacob, Betelehem W.
Simmons, Dennis M.
MacBeth, Morgan
Lieu, Christopher H.
Davis, S. Lindsey
Leal, Alexis D.
Tentler, John J.
Diamond, Jennifer R.
Eckhardt, S. Gail
Messersmith, Wells A.
Pitts, Todd M.
author_facet Hartman, Sarah J.
Bagby, Stacey M.
Yacob, Betelehem W.
Simmons, Dennis M.
MacBeth, Morgan
Lieu, Christopher H.
Davis, S. Lindsey
Leal, Alexis D.
Tentler, John J.
Diamond, Jennifer R.
Eckhardt, S. Gail
Messersmith, Wells A.
Pitts, Todd M.
author_sort Hartman, Sarah J.
collection PubMed
description Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal cancer with high incidences of p53 mutations. AZD1775 (adavosertib, previously MK-1775) is a small molecule WEE1 inhibitor that abrogates the G2M checkpoint and can potentially synergize with DNA damaging therapies commonly used in PDAC treatment. The purpose of this study was to identify combination partners for AZD1775, including standard chemotherapy or targeted agents, in PDAC preclinical models. Low powered preliminary screens demonstrated that two of the four PDX models responded better to the combinations of AZD1775 with irinotecan or capecitabine than to either single agent. Following the screens, two full powered PDAC PDX models of differing p53 status were tested with the combinations of AZD1775 and irinotecan or capecitabine. The combinations of AZD1775 and SN38 or 5-FU were also tested on PDAC cell lines. Cellular proliferation was measured using an IncuCyte Live Cell Imager and apoptosis was measured using a Caspase-Glo 3/7 assay. Flow cytometry was conducted to measure alterations in cell cycle distribution. Western blot analysis was used to determine the effects of the drug combinations on downstream effectors. In PDX models with mutated p53 status, there was significant tumor growth inhibition from the combination of AZD1775 with irinotecan or capecitabine (P ≤ 0.03), while PDX models with wild type p53 did not show anti-tumor synergy from the same combinations (P ≥ 0.08). The combination of AZD1775 with SN38 or 5-FU significantly decreased proliferation in all PDAC cell lines, and enhanced apoptosis in multiple cell lines. Cell cycle distribution was disrupted from the combination of AZD1775 with SN38 or 5-FU which was recorded as G2M arrest and decreased G1 phase. AZD1775 inhibited phospho-CDC2 and increased the expression of γH2AX that was either maintained or enhanced after combination with SN38 or 5-FU. The combination of AZD1775 with irinotecan/SN38 or capecitabine/5-FU showed anti-tumor effects in vivo and in vitro in PDAC models. These results support further investigation for these combination strategies to enhance outcomes for PDAC patients.
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spelling pubmed-80449032021-04-15 WEE1 Inhibition in Combination With Targeted Agents and Standard Chemotherapy in Preclinical Models of Pancreatic Ductal Adenocarcinoma Hartman, Sarah J. Bagby, Stacey M. Yacob, Betelehem W. Simmons, Dennis M. MacBeth, Morgan Lieu, Christopher H. Davis, S. Lindsey Leal, Alexis D. Tentler, John J. Diamond, Jennifer R. Eckhardt, S. Gail Messersmith, Wells A. Pitts, Todd M. Front Oncol Oncology Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal cancer with high incidences of p53 mutations. AZD1775 (adavosertib, previously MK-1775) is a small molecule WEE1 inhibitor that abrogates the G2M checkpoint and can potentially synergize with DNA damaging therapies commonly used in PDAC treatment. The purpose of this study was to identify combination partners for AZD1775, including standard chemotherapy or targeted agents, in PDAC preclinical models. Low powered preliminary screens demonstrated that two of the four PDX models responded better to the combinations of AZD1775 with irinotecan or capecitabine than to either single agent. Following the screens, two full powered PDAC PDX models of differing p53 status were tested with the combinations of AZD1775 and irinotecan or capecitabine. The combinations of AZD1775 and SN38 or 5-FU were also tested on PDAC cell lines. Cellular proliferation was measured using an IncuCyte Live Cell Imager and apoptosis was measured using a Caspase-Glo 3/7 assay. Flow cytometry was conducted to measure alterations in cell cycle distribution. Western blot analysis was used to determine the effects of the drug combinations on downstream effectors. In PDX models with mutated p53 status, there was significant tumor growth inhibition from the combination of AZD1775 with irinotecan or capecitabine (P ≤ 0.03), while PDX models with wild type p53 did not show anti-tumor synergy from the same combinations (P ≥ 0.08). The combination of AZD1775 with SN38 or 5-FU significantly decreased proliferation in all PDAC cell lines, and enhanced apoptosis in multiple cell lines. Cell cycle distribution was disrupted from the combination of AZD1775 with SN38 or 5-FU which was recorded as G2M arrest and decreased G1 phase. AZD1775 inhibited phospho-CDC2 and increased the expression of γH2AX that was either maintained or enhanced after combination with SN38 or 5-FU. The combination of AZD1775 with irinotecan/SN38 or capecitabine/5-FU showed anti-tumor effects in vivo and in vitro in PDAC models. These results support further investigation for these combination strategies to enhance outcomes for PDAC patients. Frontiers Media S.A. 2021-03-25 /pmc/articles/PMC8044903/ /pubmed/33869031 http://dx.doi.org/10.3389/fonc.2021.642328 Text en Copyright © 2021 Hartman, Bagby, Yacob, Simmons, MacBeth, Lieu, Davis, Leal, Tentler, Diamond, Eckhardt, Messersmith and Pitts https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Oncology
Hartman, Sarah J.
Bagby, Stacey M.
Yacob, Betelehem W.
Simmons, Dennis M.
MacBeth, Morgan
Lieu, Christopher H.
Davis, S. Lindsey
Leal, Alexis D.
Tentler, John J.
Diamond, Jennifer R.
Eckhardt, S. Gail
Messersmith, Wells A.
Pitts, Todd M.
WEE1 Inhibition in Combination With Targeted Agents and Standard Chemotherapy in Preclinical Models of Pancreatic Ductal Adenocarcinoma
title WEE1 Inhibition in Combination With Targeted Agents and Standard Chemotherapy in Preclinical Models of Pancreatic Ductal Adenocarcinoma
title_full WEE1 Inhibition in Combination With Targeted Agents and Standard Chemotherapy in Preclinical Models of Pancreatic Ductal Adenocarcinoma
title_fullStr WEE1 Inhibition in Combination With Targeted Agents and Standard Chemotherapy in Preclinical Models of Pancreatic Ductal Adenocarcinoma
title_full_unstemmed WEE1 Inhibition in Combination With Targeted Agents and Standard Chemotherapy in Preclinical Models of Pancreatic Ductal Adenocarcinoma
title_short WEE1 Inhibition in Combination With Targeted Agents and Standard Chemotherapy in Preclinical Models of Pancreatic Ductal Adenocarcinoma
title_sort wee1 inhibition in combination with targeted agents and standard chemotherapy in preclinical models of pancreatic ductal adenocarcinoma
topic Oncology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8044903/
https://www.ncbi.nlm.nih.gov/pubmed/33869031
http://dx.doi.org/10.3389/fonc.2021.642328
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