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Translational Therapeutics

PFKFB3 works on the FAK-STAT3-SOX2 axis to regulate the stemness in MPM

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Abstract

Background

Malignant pleural mesothelioma (MPM) is an aggressive neoplasm and often acquires chemoresistance by increasing stemness in tumour tissue, thereby generating cancer stem cells (CSCs). CSCs escape treatment by deploying metabolic pathways to trigger dormancy or proliferation, also gaining the ability to exit and re-enter the cell cycle to hide their cellular identity.

Methods

We employed various cellular and biochemical assays to identify the role of the glycolytic enzyme PFKFB3, by knocking it down and pharmacologically inhibiting it with PFK158, to determine its anticancer effects in vitro and in vivo by targeting the CSC population in MPM.

Results

Here, we have identified PFKFB3 as a strategic player to target the CSC population in MPM and demonstrated that both pharmacologic (PFK158) and genetic inhibition of PFKFB3 destroy the FAK-Stat3-SOX2 nexus resulting in a decline in conspicuous stem cell markers viz. ALDH, CD133, CD44, SOX2. Inhibition of PFKFB3 accumulates p21 and p27 in the nucleus by decreasing SKP2. Lastly, PFK158 diminishes tumour-initiating cells (TICs) mediated MPM xenograft in vivo.

Conclusions

This study confers a comprehensive and mechanistic function of PFKFB3 in CSC maintenance that may foster exceptional opportunities for targeted small molecule blockade of the TICs in MPM.

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Fig. 1: PFKFB3 inhibition downregulates stem cell markers in MPM.
Fig. 2: PFKFB3 inhibition diminishes the ALDH+ CD133+ cell population.
Fig. 3: PFK158 works on the FAK-Stat3-SOX2 axis.
Fig. 4: MPM cells show the nuclear localisation and function of PFKFB3.
Fig. 5: Enrichment and characterisation of MPM-CSC population.
Fig. 6: Repression of p21/p27 and overexpression of PFKFB3 in spheroid MPM cells regulate the stemness.
Fig. 7: Tumour-initiating stem-like cells (TIC) in human pleural mesothelioma exhibit upregulation of PFKFB3.
Fig. 8: A summarised illustration (BioRender, https://biorender.com) showing the contribution of PFKFB3 in the enrichment of MPM stemness and cell cycle regulation.

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Data availability

The data that support the findings of this study are available from the corresponding author on a reasonable request.

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Acknowledgements

We sincerely acknowledge Dr. Tobias Peikert, Mayo Clinic, Rochester, MN for providing the EMMeso cell line. Our acknowledgement towards the personnel of Microscopy and Cell Analysis Core and Pathology Research Cores Mayo Clinic, Rochester, MN.

Funding

This work is supported (in part) by the Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN (JM and VS) and a generous gift from Samuel and Ilda Conde to JM—Mayo Clinic, Rochester.

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Authors and Affiliations

  1. Department of Experimental Pathology and Laboratory Medicine, Mayo Clinic, Rochester, MN, USA

    Sayantani Sarkar Bhattacharya, Prabhu Thirusangu, Ling Jin, Julie Staub & Viji Shridhar

  2. Department of Medical Oncology, Mayo Clinic, Rochester, MN, USA

    Sayantani Sarkar Bhattacharya & Julian R. Molina

  3. Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA

    Sayantani Sarkar Bhattacharya

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  1. Sayantani Sarkar Bhattacharya
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Contributions

Conceptualisation: SSB and VS; formal analysis: SSB; funding acquisition: VS and JRM; investigation: SSB and VS; methodology: SSB and PT; project administration: VS; supervision: JRM and VS; writing—original draft: SSB; writing—review and editing: SSB, PT, JS and VS.

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Correspondence to Viji Shridhar or Julian R. Molina.

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Sarkar Bhattacharya, S., Thirusangu, P., Jin, L. et al. PFKFB3 works on the FAK-STAT3-SOX2 axis to regulate the stemness in MPM. Br J Cancer 127, 1352–1364 (2022). https://doi.org/10.1038/s41416-022-01867-7

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