Ex Vivo Activation of Red Blood Cell Senescence by Plasma from Sickle-Cell Disease Patients: Correlation between Markers and Adhesion Consequences during Acute Disease Events

Abstract

BACKGROUND: Blood transfusion remains a key treatment for managing occlusive episodes and painful crises in sickle-cell disease (SCD). In that clinical context, red blood cells (RBCs) from donors and transfused to patients, may be affected by plasma components in the recipients’ blood. Senescence lesion markers appear on the red cells after transfusion, shortening the RBC lifespan in circulation. In the specific context of SCD, senescence signals can also trigger the occlusive painful events, typical of the disease. This work follows through our previous data that described a RBC senescence process, rapidly detected after challenge with SCD pathological plasmas. In this clinical context, we wanted here to further explore the characteristics and physiologic consequences of AA RBC lesions associated with senescence, as lesions caused by RBCs after transfusion may have adverse consequences for SCD patients. METHODS: Plasma samples from SCD patients, with acute symptoms (n = 20) or steady-state disease (n = 34) were co-incubated with donor AA RBCs from blood units for 24 to 48 h. Specific markers signing RBC senescence were quantified after the incubation with SCD plasma samples. The physiologic in-flow adhesion was investigated on senescent RBCs, an in vitro technic into biochips that mimic adherence of RBCs during the occlusive events of SCD. RESULTS: Senescence markers on AA RBCs, together with their in-flow adhesion to the plasma-bridging protein thrombospondin, were associated with the clinical status of the SCD patients from whom plasma was obtained. In these experiments, the highest values were obtained for SCD acute plasma samples. Adhesion of senescent RBCs into biochips, which is not reversed by a pre-treatment with recombinant Annexin V, can be reproduced with the use of chemical agents acting on RBC membrane channels that regulate either Ca2+ entry or modulating RBC hydration. CONCLUSION: We found that markers on red cells are correlated, and that the senescence induced by SCD plasma provokes the adhesion of RBCs to the vessel wall protein thrombospondin. In-flow adhesion of senescent red cells after plasma co-incubations can be reproduced with the use of modulators of RBC membrane channels; activating the Piezo1 Ca2+ mechanosensitive channel provokes RBC adhesion of normal (non-senescent) RBCs, while blocking the Ca2+-dependent K+ Gardos channel, can reverse it. Clinically modulating the RBC adhesion to vascular wall proteins might be a promising avenue for the treatment of painful occlusive events in SCD.

Hypoxic storage of donor red cells preserves deformability after exposure to plasma from adults with sickle cell disease

“…Donor RBCs may not be able to protect themselves adequately from such an environment, especially toward the end of its storage life, leading to changes in deformability and increased cell death. In support of this hypothesis, donor RBCs incubated in the plasma of patients with SCD have demonstrated increases in surface PS, especially when exposed to the plasma of patients who were in VOC 50,51 . In our pilot study, we further elucidate that the plasma of patients who are hospitalized for VOC demonstrate greater donor red cell lysis post‐incubation, regardless of the storage type.…”

“…In support of this hypothesis, donor RBCs incubated in the plasma of patients with SCD have demonstrated increases in surface PS, especially when exposed to the plasma of patients who were in VOC. 50,51 In our pilot study, we further elucidate that the plasma of patients who are hospitalized for VOC demonstrate greater donor red cell lysis post-incubation, regardless of the storage type.…”

PIEZO1, sensing the touch during erythropoiesis

“…In RBCs from patients with SCD, Yoda1-mediated chemical activation of PIEZO1 triggered phosphatidylserine exposure through calcium entry, but the effect on RBCs lifespan or clearance has not been evaluated [46]. In parallel, the co-incubation of healthy RBCs with plasma from patients with SCD triggered phosphatidylserine exposure, increasing their adhesion to thrombospondin [83]. The adhesion of senescent RBCs has been reproduced by the chemical activation of PIEZO1, but its effect on phosphatidylserine exposure has not been reported.…”