A knock-out model of paroxysmal nocturnal hemoglobinuria: Pig-a- hematopoiesis is reconstituted following intercellular transfer of GPI-anchored proteins

D. E. Dunn, J. Yu, S. Nagarajan, M. Devetten, F. F. Weichold, M. E. Medof, N. S. Young, J. M. Liu

Research output: Contribution to journalArticlepeer-review

75 Scopus citations

Abstract

We created a "knock-out" embryonic stem cell via targeted disruption of the phosphatidylinositol glycan class A (Pig-a) gene, resulting in loss of expression of cell surface glycosyl phosphatidylinositol-anchored proteins and reproducing the mutant phenotype of the human disease paroxysmal nocturnal hemoglobinuria. Morphogenesis of Pig-a- embryoid bodies (EB) in vitro was grossly aberrant and, unlike EB derived from normal embryonic stem cells, Pig-A- EB produced no secondary hematopoietic colonies. Chimeric EB composed of control plus Pig-A- cells, however, appeared normal, and hematopoiesis from knock-out cells was reconstituted. Transfer in situ of glycosyl phosphatidylinositol-anchored proteins from normal to knock-out cells was demonstrated by two-color fluorescent analysis, suggesting a possible mechanism for these functional effects. Hematopoietic cells with mutated PIG-A genes in humans with paroxysmal nocturnal hemoglobinuria may be subject to comparable pathophysiologic processes and amenable to similar therapeutic protein transfer.

Original languageEnglish (US)
Pages (from-to)7938-7943
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number15
DOIs
StatePublished - Jul 23 1996

Keywords

  • Bone marrow failure syndromes
  • Developmental biology
  • Embryoid bodies
  • Embryonic stem cells
  • Paroxysmal nocturnal hemoglobinuria

ASJC Scopus subject areas

  • General

Fingerprint Dive into the research topics of 'A knock-out model of paroxysmal nocturnal hemoglobinuria: Pig-a<sup>-</sup> hematopoiesis is reconstituted following intercellular transfer of GPI-anchored proteins'. Together they form a unique fingerprint.

Cite this