Generation of red blood cells from stem cells: Achievements, opportunities and perspectives for malaria research

Abstract

Parasites of the genus Plasmodium that cause malaria survive within humans by invasion of, and proliferation within, the most abundant cell type in the body, the red blood cell. As obligate, intracellular parasites, interactions between parasite and host red blood cell components are crucial to multiple aspects of the blood stage malaria parasite lifecycle. The requirement for, and involvement of, an array of red blood cell proteins in parasite invasion and intracellular development is well established. Nevertheless, detailed mechanistic understanding of host cell protein contributions to these processes are hampered by the genetic intractability of the anucleate red blood cell. The advent of stem cell technology and more specifically development of methods that recapitulate in vitro the process of red blood cell development known as erythropoiesis has enabled the generation of erythroid cell stages previously inaccessible in large numbers for malaria studies. What is more, the capacity for genetic manipulation of nucleated erythroid precursors that can be differentiated to generate modified red blood cells has opened new horizons for malaria research. This review summarises current methodologies that harness in vitro erythroid differentiation of stem cells for generation of cells that are susceptible to malaria parasite invasion; discusses existing and emerging approaches to generate novel red blood cell phenotypes and explores the exciting potential of in vitro derived red blood cells for improved understanding the broad role of host red blood cell proteins in malaria pathogenesis.

Basigin mediation of Plasmodium falciparum red blood cell invasion does not require its transmembrane domain or interaction with monocarboxylate transporter 1

“…Generation of modified in vitro derived reticulocytes can be achieved through differentiation of both immortalised erythroblast cell lines such as BEL-A or of primary CD34 + haematopoietic stem cells. Each system has its own respective advantages and disadvantages [ 17 ]. Efficiency of differentiation and high rates of enucleation of primary cells currently facilitate a much greater cost effective yield of leukofiltered reticulocytes, providing more material for experimentation, although this is countered by variable transduction efficiency and reduced control over rescue overexpression levels.…”

Targeting PfProhibitin 2-Hu-Hsp70A1A complex as a unique approach towards malaria vaccine development

“…EPCs are erythroid progenitor cells that are known to support merozoite invasion. 19 Downregulation of Hsp70A1A expression in EPCs was performed using siRNA at different concentrations (100 nM and 250 nM), and Hsp70 expression in these transfected cells was detected by immunofluorescence assay at 72 h post-transfection. Our results indicated a significant reduction in Hsp70A1A protein expression in Hsp70A1A-specific siRNA-transfected cells as compared to untransfected cells (control) ( Figure 4 Ai, ii, iii).…”

Development of pathophysiologically relevant models of sickle cell disease and β-thalassemia for therapeutic studies

“…Our study demonstrates that reticulocytes derived from BEL-A SCM exhibit a similar level of resistance to P. falciparum invasion as SCD patient-derived RBCs. Numerous mechanisms including sickling of infected RBCs, translocation of HbS-specific parasite-growth inhibiting miRNAs, stimulation of heme-oxygenase-1 and lower oxygen states are plausible explanations for HbSS malaria resistance 29 . Despite decades of research, the precise biological mechanism through which the HbSS mutation protects against severe malaria remains elusive 30 , 31 .…”