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Double inhibition of cAMP and mTOR signalling may potentiate the reduction of cell growth in ADPKD cells

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Abstract

Background

ADPKD is a renal pathology caused by mutations of PKD1 and PKD2 genes, which encode for polycystin-1 (PC1) and polycystin-2 (PC2), respectively. PC1 plays an important role regulating several signal transducers, including cAMP and mTOR, which are involved in abnormal cell proliferation of ADPKD cells leading to the development and expansion of kidney cysts that are a typical hallmark of this disease. Therefore, the inhibition of both pathways could potentiate the reduction of cell proliferation enhancing benefits for ADPKD patients.

Methods

The inhibition of cAMP- and mTOR-related signalling was performed by Cl-IB-MECA, an agonist of A3 receptors, and rapamycin, respectively. Protein kinase activity was evaluated by immunoblot and cell growth was analyzed by direct cell counting.

Results

The activation of A3AR by the specific agonist Cl-IB-MECA causes a marked reduction of CREB, mTOR, and ERK phosphorylation in kidney tissues of Pkd1 flox/−: Ksp-Cre polycystic mice and reduces cell growth in ADPKD cell lines, but not affects the kidney weight. The combined sequential treatment with rapamycin and Cl-IB-MECA in ADPKD cells potentiates the reduction of cell proliferation compared with the individual compound by the inhibition of CREB, mTOR, and ERK kinase activity. Conversely, the simultaneous application of these drugs counteracts their effect on cell growth, because the inhibition of ERK kinase activity is lost.

Conclusion

The double treatment with rapamycin and Cl-IB-MECA may have synergistic effects on the inhibition of cell proliferation in ADPKD cells suggesting that combined therapies could improve renal function in ADPKD patients.

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Acknowledgments

We greatly thanks Prof. PC Harris, (Mayo Clinic; Rochester, USA) for providing human normal and cystic kidney epithelial cells.

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Authors and Affiliations

  1. Section of Biochemistry, Molecular Biology and Medical Genetics, Department of Biomedical and Surgical Specialty Sciences, University of Ferrara, via Fossato di Mortara, 74, 44121, Ferrara, Italy

    Lucia de Stephanis, Anna Bonon & Gianluca Aguiari

  2. Section of Pharmacology, Department of Medical Sciences, University of Ferrara, via Fossato di Mortara, 17-19, 44121, Ferrara, Italy

    Katia Varani

  3. Section of Pathological Anatomy and Molecular Diagnostic, Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, via Aldo Moro 8, 44124, Ferrara, Italy

    Giovanni Lanza & Roberta Gafà

  4. Section of Pathology, Oncology and Experimental Biology, Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, via Fossato di Mortara 64/b, 44121, Ferrara, Italy

    Paolo Pinton

  5. Dibit 1 San Raffaele, via Olgettina 60, 20132, Milan, Italy

    Monika Pema & Alessandra Boletta

  6. Department of Internal Medicine, Yale University School of Medicine, New Haven, CT, 06520, USA

    Stefan Somlo

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  1. Lucia de Stephanis
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  2. Anna Bonon
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  10. Gianluca Aguiari
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Correspondence to Gianluca Aguiari.

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The authors have no conflicts of interest to disclose.

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de Stephanis, L., Bonon, A., Varani, K. et al. Double inhibition of cAMP and mTOR signalling may potentiate the reduction of cell growth in ADPKD cells. Clin Exp Nephrol 21, 203–211 (2017). https://doi.org/10.1007/s10157-016-1289-1

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  • DOI: https://doi.org/10.1007/s10157-016-1289-1

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