Erythrocyte ion content and dehydration modulate maximal Gardos channel activity in KCNN4 V282M/+ hereditary xerocytosis red cells

Am J Physiol Cell Physiol. 2019 Aug 1;317(2):C287-C302. doi: 10.1152/ajpcell.00074.2019. Epub 2019 May 15.


Hereditary xerocytosis (HX) is caused by missense mutations in either the mechanosensitive cation channel PIEZO1 or the Ca2+-activated K+ channel KCNN4. All HX-associated KCNN4 mutants studied to date have revealed increased current magnitude and red cell dehydration. Baseline KCNN4 activity was increased in HX red cells heterozygous for KCNN4 mutant V282M. However, HX red cells maximally stimulated by Ca2+ ionophore A23187 or by PMCA Ca2+-ATPase inhibitor orthovanadate displayed paradoxically reduced KCNN4 activity. This reduced Ca2+-stimulated mutant KCNN4 activity in HX red cells was associated with unchanged sensitivity to KCNN4 inhibitor senicapoc and KCNN4 activator Ca2+, with slightly elevated Ca2+ uptake and reduced PMCA activity, and with decreased KCNN4 activation by calpain inhibitor PD150606. The altered intracellular monovalent cation content of HX red cells prompted experimental nystatin manipulation of red cell Na and K contents. Nystatin-mediated reduction of intracellular K+ with corresponding increase in intracellular Na+ in wild-type cells to mimic conditions of HX greatly suppressed vanadate-stimulated and A23187-stimulated KCNN4 activity in those wild-type cells. However, conferral of wild-type cation contents on HX red cells failed to restore wild-type-stimulated KCNN4 activity to those HX cells. The phenotype of reduced, maximally stimulated KCNN4 activity was shared by HX erythrocytes expressing heterozygous PIEZO1 mutants R2488Q and V598M, but not by HX erythrocytes expressing heterozygous KCNN4 mutant R352H or PIEZO1 mutant R2456H. Our data suggest that chronic KCNN4-driven red cell dehydration and intracellular cation imbalance can lead to reduced KCNN4 activity in HX and wild-type red cells.

Keywords: dehydrated stomatocytosis; ionophore; potassium channel; red blood cell; senicapoc.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anemia, Hemolytic, Congenital / blood*
  • Anemia, Hemolytic, Congenital / diagnosis
  • Anemia, Hemolytic, Congenital / genetics
  • Calcium Signaling
  • Case-Control Studies
  • Erythrocyte Indices
  • Erythrocytes / metabolism*
  • Genetic Predisposition to Disease
  • Humans
  • Hydrops Fetalis / blood*
  • Hydrops Fetalis / diagnosis
  • Hydrops Fetalis / genetics
  • Intermediate-Conductance Calcium-Activated Potassium Channels / blood*
  • Intermediate-Conductance Calcium-Activated Potassium Channels / genetics
  • Ion Channels / blood
  • Ion Channels / genetics
  • Membrane Potentials
  • Mutation, Missense
  • Osmotic Fragility
  • Phenotype
  • Potassium / blood*


  • Intermediate-Conductance Calcium-Activated Potassium Channels
  • Ion Channels
  • KCNN4 protein, human
  • PIEZO1 protein, human
  • Potassium

Supplementary concepts

  • Xerocytosis, hereditary