High-level transgene expression in induced pluripotent stem cell-derived megakaryocytes: correction of Glanzmann thrombasthenia. Blood 2014 Jan 30;123(5):753-7
Date
12/18/2013Pubmed ID
24335497Pubmed Central ID
PMC3907760DOI
10.1182/blood-2013-10-530725Scopus ID
2-s2.0-84897024164 (requires institutional sign-in at Scopus site) 47 CitationsAbstract
Megakaryocyte-specific transgene expression in patient-derived induced pluripotent stem cells (iPSCs) offers a new approach to study and potentially treat disorders affecting megakaryocytes and platelets. By using a Gp1ba promoter, we developed a strategy for achieving a high level of protein expression in human megakaryocytes. The feasibility of this approach was demonstrated in iPSCs derived from two patients with Glanzmann thrombasthenia (GT), an inherited platelet disorder caused by mutations in integrin αIIbβ3. Hemizygous insertion of Gp1ba promoter-driven human αIIb complementary DNA into the AAVS1 locus of iPSCs led to high αIIb messenger RNA and protein expression and correction of surface αIIbβ3 in megakaryocytes. Agonist stimulation of these cells displayed recovery of integrin αIIbβ3 activation. Our findings demonstrate a novel approach to studying human megakaryocyte biology as well as functional correction of the GT defect, offering a potential therapeutic strategy for patients with diseases that affect platelet function.
Author List
Sullivan SK, Mills JA, Koukouritaki SB, Vo KK, Lyde RB, Paluru P, Zhao G, Zhai L, Sullivan LM, Wang Y, Kishore S, Gharaibeh EZ, Lambert MP, Wilcox DA, French DL, Poncz M, Gadue PAuthors
Sevasti Koukouritaki PhD Research Scientist II in the Pediatrics department at Medical College of WisconsinDavid A. Wilcox PhD Professor in the Pediatrics department at Medical College of Wisconsin
MESH terms used to index this publication - Major topics in bold
Gene ExpressionHumans
Induced Pluripotent Stem Cells
Megakaryocytes
Membrane Glycoproteins
Platelet Glycoprotein GPIIb-IIIa Complex
Platelet Glycoprotein GPIb-IX Complex
Promoter Regions, Genetic
Thrombasthenia
Transgenes