Increased retention of functional mitochondria in mature sickle red blood cells is associated with increased sickling tendency, hemolysis and oxidative stress

Sofia Esperti (1, 2, 3) , Elie Nader (1, 2) , Antoine Stier (4, 5) , Camille Boisson (1, 2) , Romain Carin (1, 2) , Muriel Marano (6) , Mélanie Robert (1, 2, 3) , Marie Martin (1) , Françoise Horand (3) , Agnes Cibiel (3) , Céline Renoux (1, 2, 7) , Robin van Bruggen (8) , Colin Blans (8) , Yesim Dargaud (6) , Philippe Joly (1, 2, 7) , Alexandra Gauthier (1, 2, 9) , Solène Poutrel (1, 2, 10) , Marc Romana (2, 11) , Damien Roussel (4) , Philippe Connes (1, 2)

1 LIBM - Laboratoire Interuniversitaire de Biologie de la Motricité
2 Labex Gr-Ex - Laboratoire d'Excellence : Biogenèse et pathologies du globule rouge
3 ERYTECH Pharma
4 E2C - Équipe 4 - Écophysiologie, Comportement, Conservation
5 IPHC - Institut Pluridisciplinaire Hubert Curien
6 Hémostase & thrombose
7 Laboratoire de Biochimie et de Biologie Moléculaire
8 Department of Molecular Hematology, Sanquin Research and Landsteiner Laboratory
9 Institut d’Hématologique et d’Oncologique Pédiatrique
10 Service de Médecine Interne [Hôpital Edouard Herriot - HCL]
11 Université de Paris, UMR_S1134, BIGR, Inserm, F-75015 Paris, France; Institut National de la transfusion sanguine, F-75015 Paris, France.

Abnormal retention of mitochondria in mature red blood cells (RBC) has been recently reported in sickle cell anemia (SCA) but their functionality and their role in the pathophysiology of SCA remain unknown. The presence of mitochondria within RBC was determined by flow cytometry in 61 SCA patients and ten healthy donors. Patients were classified according to the percentage of mature RBC with mitochondria contained in the whole RBC population: low (0-4%), moderate (>4% and <8%), or high level (>8%). RBC rheological, hematological, senescence and oxidative stress markers were compared between the three groups. RBC senescence and oxidative stress markers were also compared between mature RBC containing mitochondria and those without. The functionality of residual mitochondria in sickle RBC was measured by high-resolution respirometry assay and showed detectable mitochondrial oxygen consumption in sickle mature RBC but not in healthy RBC. Increased levels of mitochondrial reactive oxygen species were observed in mature sickle RBC when incubated with Antimycin A versus without. In addition, mature RBC retaining mitochondria exhibited greater levels of reactive oxygen species compared to RBC without mitochondria, as well as greater Ca2+, lower CD47 and greater phosphatidylserine exposure. Hematocrit and RBC deformability were lower, and the propensity of RBC to sickle under deoxygenation was higher, in the SCA group with a high percentage of mitochondria retention in mature RBC. This study showed the presence of functional mitochondria in mature sickle RBC, which could favor RBC sickling and accelerate RBC senescence, leading to increased cellular fragility and hemolysis.

Domaines: Sciences de l'environnement

Sofia Esperti, Elie Nader, Antoine Stier, Camille Boisson, Romain Carin, et al.. Increased retention of functional mitochondria in mature sickle red blood cells is associated with increased sickling tendency, hemolysis and oxidative stress. Haematologica, 2023, 108 (11), pp.3086-3094. ⟨10.3324/haematol.2023.282684⟩. ⟨hal-04277331⟩