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Targeting CXCR4 abrogates resistance to trastuzumab by blocking cell cycle progression and synergizes with docetaxel in breast cancer treatment

BACKGROUND: Although trastuzumab and other HER2-targeted therapies have significantly improved survival in patients with HER2 overexpressed or amplified (HER2+) breast cancer, a significant proportion of patients do not respond or eventually develop clinical resistance. Strategies to reverse trastuz...

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Autores principales: Liu, Shuying, Xie, Shelly M., Liu, Wenbin, Gagea, Mihai, Hanker, Ariella B., Nguyen, Nguyen, Raghavendra, Akshara Singareeka, Yang-Kolodji, Gloria, Chu, Fuliang, Neelapu, Sattva S., Hanash, Samir, Zimmermann, Johann, Arteaga, Carlos L., Tripathy, Debasish
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Journal Experts 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9949251/
https://www.ncbi.nlm.nih.gov/pubmed/36824840
http://dx.doi.org/10.21203/rs.3.rs-2388864/v1
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author Liu, Shuying
Xie, Shelly M.
Liu, Wenbin
Gagea, Mihai
Hanker, Ariella B.
Nguyen, Nguyen
Raghavendra, Akshara Singareeka
Yang-Kolodji, Gloria
Chu, Fuliang
Neelapu, Sattva S.
Hanash, Samir
Zimmermann, Johann
Arteaga, Carlos L.
Tripathy, Debasish
author_facet Liu, Shuying
Xie, Shelly M.
Liu, Wenbin
Gagea, Mihai
Hanker, Ariella B.
Nguyen, Nguyen
Raghavendra, Akshara Singareeka
Yang-Kolodji, Gloria
Chu, Fuliang
Neelapu, Sattva S.
Hanash, Samir
Zimmermann, Johann
Arteaga, Carlos L.
Tripathy, Debasish
author_sort Liu, Shuying
collection PubMed
description BACKGROUND: Although trastuzumab and other HER2-targeted therapies have significantly improved survival in patients with HER2 overexpressed or amplified (HER2+) breast cancer, a significant proportion of patients do not respond or eventually develop clinical resistance. Strategies to reverse trastuzumab resistance remain a high clinical priority. We were the first to report the role of CXCR4 in trastuzumab resistance. The present study aims to explore the therapeutic potential of targeting CXCR4 and better understand the associated mechanisms. METHODS: Immunofluorescent staining, confocal microscopy analysis, and immunoblotting were used to analyze CXCR4 expression. BrdU incorporation assays and flow cytometry were used to analyze dynamic CXCR4expression. Three-dimensional co-culture (tumor cells/ breast cancer-associated fibroblasts / human peripheral blood mononuclear cells) or antibody-dependent cellular cytotoxicity assay was used to mimic human tumor microenvironment, which is necessary for testing therapeutic effect of CXCR4 inhibitor or trastuzumab. The FDA-approved CXCR4 antagonist AMD3100, trastuzumab, and docetaxel chemotherapy were used to evaluate therapeutic efficacy in vitro and in vivo. Reverse phase protein array and immunoblotting were used to discern the associated molecular mechanisms. RESULTS: Using multiple cell lines and patient breast cancer samples we confirmed CXCR4 drives trastuzumab resistance in HER2+ breast cancer and further demonstrated that the increased CXCR4 expression in trastuzumab-resistant cells is associated with cell cycle progression with a peak in the G2/M phases. Blocking CXCR4 with AMD3100 inhibits cell proliferation by downregulating mediators of G2-M transition, leading to G2/M arrest and abnormal mitosis. Using multiple trastuzumab-resistant cell lines and an in vivo established trastuzumab-resistant xenograft mouse model, we demonstrated that targeting CXCR4 with AMD3100 suppresses tumor growth in vitro and in vivo, and synergizes with docetaxel. CONCLUSIONS: Our findings support CXCR4 as a novel therapeutic target and a predictive biomarker for trastuzumab resistance in HER2+ breast cancer.
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spelling pubmed-99492512023-02-24 Targeting CXCR4 abrogates resistance to trastuzumab by blocking cell cycle progression and synergizes with docetaxel in breast cancer treatment Liu, Shuying Xie, Shelly M. Liu, Wenbin Gagea, Mihai Hanker, Ariella B. Nguyen, Nguyen Raghavendra, Akshara Singareeka Yang-Kolodji, Gloria Chu, Fuliang Neelapu, Sattva S. Hanash, Samir Zimmermann, Johann Arteaga, Carlos L. Tripathy, Debasish Res Sq Article BACKGROUND: Although trastuzumab and other HER2-targeted therapies have significantly improved survival in patients with HER2 overexpressed or amplified (HER2+) breast cancer, a significant proportion of patients do not respond or eventually develop clinical resistance. Strategies to reverse trastuzumab resistance remain a high clinical priority. We were the first to report the role of CXCR4 in trastuzumab resistance. The present study aims to explore the therapeutic potential of targeting CXCR4 and better understand the associated mechanisms. METHODS: Immunofluorescent staining, confocal microscopy analysis, and immunoblotting were used to analyze CXCR4 expression. BrdU incorporation assays and flow cytometry were used to analyze dynamic CXCR4expression. Three-dimensional co-culture (tumor cells/ breast cancer-associated fibroblasts / human peripheral blood mononuclear cells) or antibody-dependent cellular cytotoxicity assay was used to mimic human tumor microenvironment, which is necessary for testing therapeutic effect of CXCR4 inhibitor or trastuzumab. The FDA-approved CXCR4 antagonist AMD3100, trastuzumab, and docetaxel chemotherapy were used to evaluate therapeutic efficacy in vitro and in vivo. Reverse phase protein array and immunoblotting were used to discern the associated molecular mechanisms. RESULTS: Using multiple cell lines and patient breast cancer samples we confirmed CXCR4 drives trastuzumab resistance in HER2+ breast cancer and further demonstrated that the increased CXCR4 expression in trastuzumab-resistant cells is associated with cell cycle progression with a peak in the G2/M phases. Blocking CXCR4 with AMD3100 inhibits cell proliferation by downregulating mediators of G2-M transition, leading to G2/M arrest and abnormal mitosis. Using multiple trastuzumab-resistant cell lines and an in vivo established trastuzumab-resistant xenograft mouse model, we demonstrated that targeting CXCR4 with AMD3100 suppresses tumor growth in vitro and in vivo, and synergizes with docetaxel. CONCLUSIONS: Our findings support CXCR4 as a novel therapeutic target and a predictive biomarker for trastuzumab resistance in HER2+ breast cancer. American Journal Experts 2023-02-14 /pmc/articles/PMC9949251/ /pubmed/36824840 http://dx.doi.org/10.21203/rs.3.rs-2388864/v1 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use.
spellingShingle Article
Liu, Shuying
Xie, Shelly M.
Liu, Wenbin
Gagea, Mihai
Hanker, Ariella B.
Nguyen, Nguyen
Raghavendra, Akshara Singareeka
Yang-Kolodji, Gloria
Chu, Fuliang
Neelapu, Sattva S.
Hanash, Samir
Zimmermann, Johann
Arteaga, Carlos L.
Tripathy, Debasish
Targeting CXCR4 abrogates resistance to trastuzumab by blocking cell cycle progression and synergizes with docetaxel in breast cancer treatment
title Targeting CXCR4 abrogates resistance to trastuzumab by blocking cell cycle progression and synergizes with docetaxel in breast cancer treatment
title_full Targeting CXCR4 abrogates resistance to trastuzumab by blocking cell cycle progression and synergizes with docetaxel in breast cancer treatment
title_fullStr Targeting CXCR4 abrogates resistance to trastuzumab by blocking cell cycle progression and synergizes with docetaxel in breast cancer treatment
title_full_unstemmed Targeting CXCR4 abrogates resistance to trastuzumab by blocking cell cycle progression and synergizes with docetaxel in breast cancer treatment
title_short Targeting CXCR4 abrogates resistance to trastuzumab by blocking cell cycle progression and synergizes with docetaxel in breast cancer treatment
title_sort targeting cxcr4 abrogates resistance to trastuzumab by blocking cell cycle progression and synergizes with docetaxel in breast cancer treatment
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9949251/
https://www.ncbi.nlm.nih.gov/pubmed/36824840
http://dx.doi.org/10.21203/rs.3.rs-2388864/v1
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