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PIM3 kinase promotes tumor metastasis in hepatoblastoma by upregulating cell surface expression of chemokine receptor cxcr4

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Abstract

Patients presenting with metastatic hepatoblastoma have limited treatment options and survival rates as low as 25%. We previously demonstrated that Proviral Integration site in Maloney murine leukemia virus 3 (PIM3) kinase promotes tumorigenesis and cancer cell stemness in hepatoblastoma. In this study, we assessed the role of PIM3 kinase in promoting hepatoblastoma metastasis. We utilized a tail vein injection model of metastasis to evaluate the effect of CRISPR/Cas9-mediated PIM3 knockout, stable overexpression of PIM3, and pharmacologic PIM inhibition on the formation of lung metastasis. In vivo studies revealed PIM3 knockout impaired the formation of lung metastasis: 5 out of 6 mice injected with wild type hepatoblastoma cells developed lung metastasis while none of the 7 mice injected with PIM3 knockout hepatoblastoma cells developed lung metastasis. PIM3 overexpression in hepatoblastoma increased the pulmonary metastatic burden in mice and mechanistically, upregulated the phosphorylation and cell surface expression of CXCR4, a key receptor in the progression of cancer cell metastasis. CXCR4 blockade with AMD3100 decreased the metastatic phenotype of PIM3 overexpressing cells, indicating that CXCR4 contributed to PIM3’s promotion of hepatoblastoma metastasis. Clinically, PIM3 expression correlated positively with CXCR4 expression in primary hepatoblastoma tissues. In conclusion, we have shown PIM3 kinase promotes the metastatic phenotype of hepatoblastoma cells through upregulation of CXCR4 cell surface expression and these findings suggest that targeting PIM3 kinase may provide a novel therapeutic strategy for metastatic hepatoblastoma.

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Acknowledgements

The authors wish to thank Vidya Sagar Hanumanthu from the UAB Comprehensive Flow Cytometry Core and Dr. Michael Crowley from the UAB Genomics Core.

Funding

This work was supported in part by funding from National Institutes of Health (NIH) T32 CA091078 Surgical Oncology Research Training Program (LLS); NIH T32 CA229102 Surgical Oncology Research Training Program (JRJ, LVB); NIH 5T32GM008361 MSTP Training Program (CHQ); Vince Lombardi Cancer Research Fund Bart Starr Award, Hyundai Hope on Wheels, Kaul Pediatric Research Foundation, Sid Strong Foundation, Elaine Roberts Foundation, and Open Hands Overflowing Hearts (EAB). This work utilized the University of Alabama at Birmingham (UAB) Comprehensive Flow Cytometry Core (supported by NIH P30 AR048311 and NIH P30 AI27667), and the Comprehensive Genomics and Preclinical Imaging Cores, shared facilities which are supported by the O’Neal Comprehensive Cancer Center (P30 CA013148).

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    Authors and Affiliations

    1. Division of Pediatric Surgery, Department of Surgery, University of Alabama at Birmingham, 35233, Birmingham, AL, USA

      Raoud Marayati, Janet Julson, Laura V. Bownes, Colin H. Quinn, Laura L. Stafman, Andee M. Beierle, Hooper R. Markert, Jerry E. Stewart & Elizabeth A. Beierle

    2. Division of Pediatric Hematology Oncology, Department of Pediatrics, University of Alabama at Birmingham, 35233, Birmingham, AL, USA

      Sara C. Hutchins

    3. Department of Genetics, University of Alabama at Birmingham, 35233, Birmingham, AL, USA

      David K. Crossman

    4. Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham, 35233, Birmingham, AL, USA

      Anita B. Hjelmeland

    5. Department of Pathology, The Children’s Hospital of Alabama, 35233, Birmingham, AL, USA

      Elizabeth Mroczek-Musulman

    6. 1600 7th Ave South Lowder Room 300, 35233, Birmingham, AL, USA

      Elizabeth A. Beierle

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    1. Raoud Marayati
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    Contributions

    R Marayati was involved in study concept and design, development of methodology, data collection, data analysis, and manuscript preparation. J Julson contributed to data analysis and manuscript preparation. LV Bownes, CH Quinn, AM Beierle, HR Markert, SC Hutchins, and JE Stewart contributed with data collection and analysis. LL Stafman developed of the CRISPR/Cas9 PIM3 knockout. DK Crossman performed bioinformatics analyses of the RNA sequencing data. E Mroczek-Musulman was involved in the histology evaluation of lung sections. AB Hjelmeland provided the luciferase labeling of the hepatoblastoma cells. EA Beierle provided senior guidance with study concept and design, data analysis, and manuscript preparation.

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    Correspondence to Elizabeth A. Beierle.

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    Raoud Marayati and Janet Julson contributed equally to the work.

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    Marayati, R., Julson, J., Bownes, L.V. et al. PIM3 kinase promotes tumor metastasis in hepatoblastoma by upregulating cell surface expression of chemokine receptor cxcr4. Clin Exp Metastasis 39, 899–912 (2022). https://doi.org/10.1007/s10585-022-10186-3

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