Impairment of CFTR activity in cultured epithelial cells upregulates the expression and activity of LDH resulting in lactic acid hypersecretion

Abstract

Mutations in the gene encoding the CFTR chloride channel produce cystic fibrosis (CF). CF patients are more susceptible to bacterial infections in lungs. The most accepted hypothesis sustains that a reduction in the airway surface liquid (ASL) volume favor infections. Alternatively, it was postulated that a reduced HCO3 transport through CFTR leads to a decreased ASL pH, favoring bacterial colonization. The issue is controversial, since recent data from cultured primary cells and CF children showed normal pH values in the ASL. We have reported previously a decreased mitochondrial Complex I (mCx-I) activity in cultured cells with impaired CFTR activity. Thus, we hypothesized that the reduced mCx-I activity could lead to increased lactic acid production (Warburg-like effect) and reduced extracellular pH (pHe). In agreement with this idea, we report here that cells with impaired CFTR function (intestinal Caco-2/pRS26, transfected with an shRNA-CFTR, and lung IB3-1 CF cells) have a decreased pHe. These cells showed increased lactate dehydrogenase (LDH) activity, LDH-A expression, and lactate secretion. Similar effects were reproduced in control cells stimulated with recombinant IL-1β. The c-Src and JNK inhibitors PP2 and SP600125 were able to increase the pHe, although the differences between control and CFTR-impaired cells were not fully compensated. Noteworthy, the LDH inhibitor oxamate completely restored the pHe of the intestinal Caco-2/pRS26 cells and have a significant effect in lung IB3-1 cells; therefore, an increased lactic acid secretion seems to be the key factor that determine a reduced pHe in these epithelial cells.

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Abbreviations

ASL:

Airway surface liquid

CF:

Cystic fibrosis

CFTR:

Cystic fibrosis transmembrane conductance regulator

CFTR-KD:

CFTR knockdown

DMSO:

Dimethyl sulfoxide

IL-1β:

Interleukin-1β

JNK:

JUN N-terminal kinase

LDH:

Lactate dehydrogenase

mCx-I:

Mitochondrial Complex I

pHe:

Extracellular pH

shRNA:

Short hairpin RNA

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Acknowledgements

We thank Professor Diego Battiato and Romina D’Agostino for administrative assistance, and María de los Angeles Aguilar for technical assistance. We also thank Dr. Lutz Birnbaumer for his continuous support to our work and very valuable criticisms. This work was supported by National Agency for the Promotion of Science and Technology (ANPCYT) [grant numbers PICT 2012-1278 to TASC and PICT-2015-1031 to AGV]; National Scientific and Technical Research Council of Argentina (CONICET) [grants PIP-2016 112201-501002-27 and PUE-2016 22920160100129CO to TASC]; and Pontifical Catholic University of Argentina (UCA) to TASC. Fellowships from CONICET to MMC, CM, and MC.

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Correspondence to Ángel G. Valdivieso or Tomás A. Santa-Coloma.

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Valdivieso, Á.G., Clauzure, M., Massip-Copiz, M.M. et al. Impairment of CFTR activity in cultured epithelial cells upregulates the expression and activity of LDH resulting in lactic acid hypersecretion. Cell. Mol. Life Sci. 76, 1579–1593 (2019). https://doi.org/10.1007/s00018-018-3001-y

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Keywords

  • CFTR
  • Cystic fibrosis
  • Lactate
  • Oxamate
  • Extracellular pH
  • IL-1β
  • Inflammation