A genetic strategy to control expression of human blood group antigens in red blood cells generated in vitro

Abstract

Background: The ability to generate red blood cells of a chosen blood group phenotype would be a major advance in transfusion when considering low- and high-frequency blood group antigens.

Study design and methods: Cord blood CD34+ cells undergoing erythroid differentiation in vitro were genetically manipulated with human immunodeficiency virus Type 1-derived lentiviral vectors expressing hUT-B1 cDNA (overexpression strategy) or bicistronic vectors expressing both enhanced green fluorescent protein and a short-hairpin RNA (shRNA) designed to silence SLC14A1(JK) gene that encodes hUT-B1 protein (silencing strategy). Resulting cell populations were analyzed by fluorescent-activated cell sorting and gel affinity column assay.

Results: When transduced with hUT-B1 cDNA lentiviral vectors encoding JK*B and JK*A alleles, respectively, CD34+ cell-derived erythroid populations from Jk(a+b-) and Jk(a-b+) donors exhibited a Jk(a+b+) phenotype different from the original phenotype. In concomitant tests, Jk(a+b+) donor cells transduced with lentiviral vectors carrying a shRNA designed to interfere with hUT-B1 transcription showed a marked decrease in hUT-B1 expression and were assessed as null for Jk antigen by a routine assay.

Conclusion: In this work focusing on the Kidd blood group system that relies on expression of hUT-B1 glycoprotein under the Jk(a) or Jk(b) antigenic configurations, we demonstrated that hematopoietic progenitors could be genetically modified to exhibit a chosen Kidd phenotype. Beyond production of atypical Kidd phenotypes, this genetic strategy could allow generation of rare blood phenotypes from hematopoietic stem cells regardless of initial donor phenotype. Potential applications for genetically modified blood include production of control samples for immunohematologic testing and for resolution of antibody detection in multiply transfused patients.