Towards manufactured red blood cells for the treatment of inherited anemia

Abstract

Patients with inherited anemia and hemoglobinopathies (such as sickle cell disease and β-thalassemia) are treated with red blood cell (RBC) transfusions to alleviate their symptoms. Some of these patients may have rare blood group types or go on to develop alloimmune reactions, which can make it difficult to source compatible blood in the donor population. Laboratory-grown RBC represent a particularly attractive alternative which could satisfy an unmet clinical need. The challenge, however, is to produce - from a limited number of stem cells - the 2x1012 RBC required for a standard adult therapeutic dose. Encouraging progress has been made in RBC production from adult stem cells under good manufacturing practice. In 2011, the Douay group conducted a successful proof-of-principle mini-transfusion of autologous manufactured RBC in a single volunteer. In the UK, a trial is planned to assess whether manufactured RBC are equivalent to RBC produced naturally in donors, by testing an allogeneic mini-dose of laboratory-grown manufactured RBC in multiple volunteers. This review discusses recent progress in the erythroid culture field as well as opportunities for further scaling up of manufactured RBC production for transfusion practice.

Reticulocyte Maturation

“…Later during differentiation, orthochromatic erythroblasts stain for TfR in the region adjacent to the extruding nucleus where intracellular vesicles accumulate prior to enucleation [ 70 , 77 ]. The inhibition of endocytosis by MitMAB (a dynamin inhibitor) on mouse spleen derived erythroblasts prevented the accumulation of TfR on the plasma membrane at the enucleation site, hampering enucleation and hemoglobin synthesis [ 78 , 79 ]. During hemoglobinization, TfR follows the endosomal recycling pathway; however, once hemoglobin synthesis is complete TfR expression is downregulated to prevent iron-mediated toxicity [ 80 ].…”

Role of Nuclear Receptors in Controlling Erythropoiesis

“…This is due to NRs’ ability to both activate or repress transcription depending on the presence/absence of their ligand, on the cellular context, on the cellular niche and on the plethora of interacting proteins/signaling pathways. Despite this complexity, some agonists acting on NRs are already in use: the synthetic glucocorticoid dexamethasone and thyroid hormone are currently used to expand erythroid cultures, a precondition to reach the final goal to obtain large numbers of cultured red blood cells for transfusion [ 140 ]. Dexamethasone is also used in clinics to expand erythroid progenitors in DBA patients, although with heterogeneous results.…”

The Function of Ion Channels and Membrane Potential in Red Blood Cells: Toward a Systematic Analysis of the Erythroid Channelome

“…We anticipate to bring automated high throughput patch-clamp recordings to the clinic and to the cell biology laboratories combined with the opportunity of bioreactor based in vitro erythropoiesis and the expertise in molecular biology to perform genetic manipulations. Recently, bioreactor based erythropoiesis progressed considerably (Hansen et al, 2019;Heshusius et al, 2019;Connor et al, 2021;Pellegrin et al, 2021), which showed the importance to understand channel activity (Piezo1) in this process (Aglialoro et al, 2021), especially since the cells in bioreactors are due to the nature of these reactors under constant flow or shear stress. Erythropoiesis under physiological conditions in the bone marrow occurs rather in the absence of flow, but crawling over macrophages may cause shear stress.…”