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A three-marker signature identifies senescence in human breast cancer exposed to neoadjuvant chemotherapy

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Abstract

Purpose

Despite the beneficial effects of chemotherapy, therapy-induced senescence (TIS) manifests itself as an undesirable byproduct. Preclinical evidence suggests that tumor cells undergoing TIS can re-emerge as more aggressive divergents and contribute to recurrence, and thus, senolytics were proposed as adjuvant treatment to eliminate senescent tumor cells. However, the identification of TIS in clinical samples is essential for the optimal use of senolytics in cancer therapy. In this study, we aimed to detect and quantify TIS using matched breast cancer samples collected pre- and post-exposure to neoadjuvant chemotherapy (NAC).

Methods

Detection of TIS was based on the change in gene and protein expression levels of three senescence-associated markers (downregulation of Lamin B1 and Ki-67 and upregulation of p16INK4a).

Results

Our analysis revealed that 23 of 72 (31%) of tumors had a shift in the protein expression of the three markers after exposure to NAC suggestive of TIS. Gene expression sets of two independent NAC-treated breast cancer samples showed consistent changes in the expression levels of LMNB1, MKI67 and CDKN2A.

Conclusions

Collectively, our study shows a more individualized approach to measure TIS hallmarks in matched breast cancer samples and provides an estimation of the extent of TIS in breast cancer clinically. Results from this work should be complemented with more comprehensive identification approaches of TIS in clinical samples in order to adopt a more careful implementation of senolytics in cancer treatment.

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Data availability

The patients’ datasets generated during the current work are not publicly available due to patients’ privacy concerns of the institutional review board policies on human tissue data but are available upon request from the corresponding author. The data of the gene expression profiles that were utilized in support the findings of this work are available from the Gene Expression Omnibus (GEO) database (reference numbers: GSE28844 https://doi.org/10.1371/journal.pone.0053983 and GSE21974 https://doi.org/10.3892/or.2011.1392).

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Acknowledgements

The authors would like to thank the Cell Therapy Center (University of Jordan) for granting access to the use of the microscopic imaging facility. The authors would also like to thank Dr. AbdelKader Battah and Dr. Heyam Awad (The School of Medicine, The University of Jordan) for their kind assistance and directions throughout this project. This work was funded by the Master Student Funding Program by The Deanship of Scientific Research, The University of Jordan (Grant no. 26/2020-2021). Work in Dr. Tareq Saleh’s laboratory is supported by Deanship of Scientific Research, The Hashemite University (Grants no. 465/83/2019 and 418/84/2019). All authors are highly thankful to the Researchers Supporting Project number (RSPD-2023R786), King Saud University, Riyadh, Saudi Arabia.

Funding

This work was funded by the Master Student Funding Program by The Deanship of Scientific Research, The University of Jordan (Grant no. 26/2020–2021). Work in Dr. Tareq Saleh’s laboratory is supported by Deanship of Scientific Research, The Hashemite University (Grants no. 465/83/2019 and 418/84/2019). All authors are highly thankful to the Researchers Supporting Project number (RSPD-2023R786), King Saud University, Riyadh, Saudi Arabia.

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  1. Mohammed El-Sadoni and Sofian Al Shboul have contributed equally to this work.

Authors and Affiliations

  1. Department of Pathology, Microbiology and Forensic Medicine, School of Medicine, The University of Jordan, Amman, 11942, Jordan

    Mohammed El-Sadoni, Ahmad Alhesa, Nisreen Abu Shahin & Bilal Azab

  2. Department of Pharmacology and Public Health, Faculty of Medicine, The Hashemite University, Zarqa, 13133, Jordan

    Sofian Al Shboul & Tareq Saleh

  3. Department of Pathology, Royal Medical Services, King Hussein Medical Center, Amman, 11942, Jordan

    Elham Alsharaiah

  4. Cell Therapy Center, The University of Jordan, Amman, 11942, Jordan

    Mohammad A. Ismail & Nidaa A. Ababneh

  5. Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia

    Moureq R. Alotaibi

  6. Department of Pathology and Cell Biology, Columbia University Medical Center, New York, NY, 10032, USA

    Bilal Azab

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  1. Mohammed El-Sadoni
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Contributions

ME-S and AA collected patients’ samples and performed all laboratory experiments including tissue processing and immunostaining. SAS performed data analysis, designed tables and figures, and contributed to the writing. NAS and EA performed pathological assessment including diagnosis and evaluation of protein marker expression. MAI and NAA performed microscopic procedures. Manuscript was revised and edited by BA and MRA. TS was responsible for the experimental design, supervised the work, and wrote the manuscript.

Corresponding author

Correspondence to Tareq Saleh.

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Conflict of interest

The authors declare that all research activities were conducted in the absence of any commercial or financial conflicts of interest.

Ethics approval and consent to participate

All experimental procedures were approved by the Institutional Review Board (IRB) committees at both JRMS (6/2021), the Hashemite University (no. 3/5/2018/2019), and The University of Jordan (no. 237/2021) in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments. Obtaining informed consents for this work was waived by both IRB protocols since all the samples used for this study were surplus (archived) tumor tissue samples, and that patients undergoing surgery or biopsy collection provide informed consent to donate any excess tissue (i.e., beyond that needed for clinical purposes).

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El-Sadoni, M., Shboul, S.A., Alhesa, A. et al. A three-marker signature identifies senescence in human breast cancer exposed to neoadjuvant chemotherapy. Cancer Chemother Pharmacol 91, 345–360 (2023). https://doi.org/10.1007/s00280-023-04523-w

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