Differential effects of adenylyl cyclase-protein kinase A cascade on shear-induced changes of sickle cell deformability

Clin Hemorheol Microcirc. 2019;73(4):531-543. doi: 10.3233/CH-190563.

Abstract

Background: Erythrocyte deformability is impaired in sickle cell disease (SCD). The regulation of cytoskeletal protein organization plays a key role in erythrocyte deformability. The activation of adenylyl cyclase (AC)/cAMP/Protein kinase A (PKA) signaling pathway was associated with increased deformability in healthy erythrocytes, however the role of this pathway in SCD is unknown.

Objective: We evaluated mechanical responses of sickle red blood cells under physiological levels of shear stress and the possible link between their deformability and AC/cAMP/PKA signaling pathway.

Methods: The shearing of sickle red blood cells at physiological level (5 Pa) and the measurement of deformability were performed by a laser assisted optical rotational cell analyzer (LORRCA).

Results: Red blood cell deformability increased of 2.5-6.5% by blocking the activity of phosphodiesterase with Pentoxifylline (10μM) (p < 0.05). The inhibition of AC with SQ22536 (100μM) produced more significant rise in deformability (+4.8-12%, p < 0.01). No significant change was observed by the inhibition of PKA with H89 (10μM).

Conclusion: Pentoxifylline and SQ22536 increased the deformability of sickle red blood cells under fluid shear stress. Modulation of the AC/cAMP/PKA pathway could have the potential to be an effective therapeutic approach for SCD through shear-induced improvements of RBC deformability.

Keywords: Protein kinase a; adenylyl cyclase; phosphodiesterase; red blood cell deformability; sickle cell disease.

MeSH terms

  • Adenylyl Cyclases / metabolism*
  • Adult
  • Anemia, Sickle Cell / blood*
  • Cyclic AMP-Dependent Protein Kinases / metabolism*
  • Erythrocytes / metabolism
  • Erythrocytes, Abnormal / metabolism*
  • Female
  • Humans
  • Male
  • Young Adult

Substances

  • Cyclic AMP-Dependent Protein Kinases
  • Adenylyl Cyclases