Proof of principle for transfusion of in vitro–generated red blood cells

Giarratana, Marie‐Catherine; Rouard, Hélène; Dumont, Agnès; Kiger, Laurent; Safeukui, Innocent; Pennec, Pierre‐Yves Le; François, Sabine; Trugnan, Germain; Peyrard, Thierry; Marie, Tiffany; Jolly, Séverine; Hebert, Nicolas; Mazurier, Christelle; Mario, Nathalie; Harmand, Laurence; Lapillonne, Hélène; Devaux, Jean-Yves; Douay, Luc

doi:10.1182/blood-2011-06-362038

Cited by 300 publications

(395 citation statements)

References 44 publications

(42 reference statements)

Supporting

Mentioning

364

Contrasting

Unclassified

Order By: Relevance

“…15 We recently established the proof of principle for transfusion of in vitro generated enucleated RBC in humans. 31 The approach we now report provides an alternative to functional normalization of sickle cell RBC for auto transfusion purposes in the hiPSC model. 5,11,15 In conclusion, using the model of hematopoiesis, we show for the first time that hiPSC can achieve terminal maturation, in vitro in terms of enucleation and in vivo in terms of synthesis of adult Hb.…”

Section: Discussionmentioning

confidence: 99%

Human induced pluripotent stem cells can reach complete terminal maturation: in vivo and in vitro evidence in the erythropoietic differentiation model

et al. 2012

View full text Add to dashboard Cite

The online version of this article has a Supplementary Appendix. BackgroundHuman induced pluripotent stem cells offer perspectives for cell therapy and research models for diseases. We applied this approach to the normal and pathological erythroid differentiation model by establishing induced pluripotent stem cells from normal and homozygous sickle cell disease donors. Design and MethodsWe addressed the question as to whether these cells can reach complete erythroid terminal maturation notably with a complete switch from fetal to adult hemoglobin. Sickle cell disease induced pluripotent stem cells were differentiated in vitro into red blood cells and characterized for their terminal maturation in terms of hemoglobin content, oxygen transport capacity, deformability, sickling and adherence. Nucleated erythroblast populations generated from normal and pathological induced pluripotent stem cells were then injected into non-obese diabetic severe combined immunodeficiency mice to follow the in vivo hemoglobin maturation. ResultsWe observed that in vitro erythroid differentiation results in predominance of fetal hemoglobin which rescues the functionality of red blood cells in the pathological model of sickle cell disease. We observed, in vivo, the switch from fetal to adult hemoglobin after infusion of nucleated erythroid precursors derived from either normal or pathological induced pluripotent stem cells into mice. ConclusionsThese results demonstrate that human induced pluripotent stem cells: i) can achieve complete terminal erythroid maturation, in vitro in terms of nucleus expulsion and in vivo in terms of hemoglobin maturation; and ii) open the way to generation of functionally corrected red blood cells from sickle cell disease induced pluripotent stem cells, without any genetic modification or drug treatment. ABSTRACT© F e r r a t a S t o r t i F o u n d a t i o n

show abstract

Section: Discussionmentioning

confidence: 99%

Human induced pluripotent stem cells can reach complete terminal maturation: in vivo and in vitro evidence in the erythropoietic differentiation model

et al. 2012

View full text Add to dashboard Cite

show abstract

“…Giarratana et al have demonstrated that red cells can be generated efficiently starting from HSPCs derived from human peripheral blood. 7 The cells produced in this study were mature reticulocytes, they expressed adult hemoglobin, they bound oxygen in a reversible manner, and their rheological properties were similar to those of reticulocytes released in vivo into the circulation of healthy individuals. Giarratana and colleagues also showed for the first time that cultured RBCs persisted for several weeks after transfusion into a human volunteer.…”

mentioning

confidence: 98%

Flicking the switch: adult hemoglobin expression in erythroid cells derived from cord blood and human induced pluripotent stem cells

Cantù

Philipsen

2014

Haematologica

View full text Add to dashboard Cite

“…Conjugation of an imaging probe (i.e., a radioisotope), together with such a targeting moiety also could be used for diagnostic purposes. Further advances in the in vitro generation of human RBCs and genetic engineering of their precursors will provide a robust platform for the use of this surface-engineering method, possibly in conjunction with cytosolic modification, in clinical applications (11,(21)(22)(23)(24)(25)(26). Moreover, the established safety of blood transfusions inspires confidence that these engineered RBCs indeed will find use in humans.…”

Section: Sortagging and Cytosolic Modification Of Terminally Differenmentioning

confidence: 99%

Engineered red blood cells as carriers for systemic delivery of a wide array of functional probes

Shi

Kundrat

Pishesha³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

181

159

View full text Add to dashboard Cite

We developed modified RBCs to serve as carriers for systemic delivery of a wide array of payloads. These RBCs contain modified proteins on their plasma membrane, which can be labeled in a sortase-catalyzed reaction under native conditions without inflicting damage to the target membrane or cell. Sortase accommodates a wide range of natural and synthetic payloads that allow modification of RBCs with substituents that cannot be encoded genetically. 2 with a favorable surface-to-volume ratio; and (vi) the absence of a nucleus, mitochondria, and any DNA. Thus, any modification made to the DNA of RBC precursors is eliminated upon their enucleation and cannot lead to abnormal growth or tumorigenicity after their transfusion into a recipient.Engineered RBCs have been generated using encapsulation (2-4), by noncovalent attachment of foreign peptides, or through installation of proteins by fusion to a monoclonal antibody specific for an RBC surface protein (5, 6).However, modified RBCs have limitations if intended for application in vivo. Encapsulation allows the entrapment of sizable quantities of material but does so at the expense of disrupting plasma membrane integrity, with a concomitant reduction in circulatory half life of the modified RBCs. Osmosis-driven entrapment limits the chemical nature of materials that can be encapsulated successfully, the site of release is difficult to control, and encapsulated enzymes are functional only at the final destination, compromising reusability at other sites (5, 6). Targeting of cargo to RBCs by fusion to an RBC-specific antibody, (e.g., anti-glycophorin antibody), has limitations because this mode of attachment to the RBC is noncovalent and dissociates readily, thus reducing circulatory half life and mass of cargo available for delivery (5, 6). Other developments that exploit RBCs for targeted delivery include nanoparticles enveloped by an RBC-mimicking membrane and RBC-shaped polymers (1). The short in vivo survival rate of these RBC-inspired carriers (∼7 d maximum) may limit their therapeutic utility.There is a need to develop new methodology for engineering RBCs so that they can carry a wide variety of useful cargoes to specific locations in the body. We describe an approach that involves minimal modification of the RBCs, with preservation of plasma membrane integrity. The method involves sortase-mediated site-specific covalent attachment of payloads to specific RBC surface proteins.Bacterial sortases are transpeptidases capable of modifying suitably modified proteins in a covalent and site-specific manner (7,8). Sortase A from Staphylococcus aureus recognizes an LPXTG motif positioned close to the substrate's C terminus and cleaves between T and G to form a covalent acyl-enzyme intermediate. This intermediate is resolved by a nucleophilic N-terminal glycine residue on an appropriately designed probe (9) with concomitant formation of a peptide bond between substrate and probe. Conversely, a protein may be labeled at its N terminus by extending it with suitably exposed g...

show abstract

Proof of principle for transfusion of in vitro–generated red blood cells

Cited by 300 publications

References 44 publications

Human induced pluripotent stem cells can reach complete terminal maturation: in vivo and in vitro evidence in the erythropoietic differentiation model

Human induced pluripotent stem cells can reach complete terminal maturation: in vivo and in vitro evidence in the erythropoietic differentiation model

Flicking the switch: adult hemoglobin expression in erythroid cells derived from cord blood and human induced pluripotent stem cells

Engineered red blood cells as carriers for systemic delivery of a wide array of functional probes

Contact Info

Product

Resources

About