BACKGROUND: Exposure to allogeneic blood products often leads to the development of human leukocyte antigen (HLA) antibodies. Refractoriness to platelet (PLT) transfusion caused by alloimmunization against HLA Class I antigens constitutes a significant clinical problem. STUDY DESIGN AND METHODS: We developed an RNA interference (RNAi)-based approach to silence the expression of HLA Class I molecules on PLTs derived from CD34+ progenitor cells. A lentiviral-based system was used to express short-hairpin RNA (shRNA) targeting β2-microglobulin (β2m) transcripts in CD34+ progenitor cells. Differentiation to PLTs was performed by incubating progenitor cells in the presence of thrombopoietin and interleukin-3. RESULTS: The transduction of RNAi cassettes containing the sequences for shRNAs targeting β2m caused up to 85% reduction of progenitor cells HLA Class I antigen expression, which was maintained in the culture-derived PLTs. The HLA-deficient PLTs derived from HLA-silenced CD34+ cells proved to be fully functional in in vitro tests when compared to peripheral blood-derived PLTs. CONCLUSIONS: Our data show that in vitro generating HLA Class I-deficient PLTs from hematopoietic progenitor cells prove to be feasible. As malignancy risks associated with insertional mutagenesis are not to be expected in anucleated PLTs, provision of HLA-deficient PLTs from large-scale production units may become reality in the management of patients suffering from PLT transfusion refractoriness.
BACKGROUND: Exposure to allogeneic blood products often leads to the development of human leukocyte antigen (HLA) antibodies. Refractoriness to platelet (PLT) transfusion caused by alloimmunization against HLA Class I antigens constitutes a significant clinical problem. STUDY DESIGN AND METHODS: We developed an RNA interference (RNAi)-based approach to silence the expression of HLA Class I molecules on PLTs derived from CD34+ progenitor cells. A lentiviral-based system was used to express short-hairpin RNA (shRNA) targeting β2-microglobulin (β2m) transcripts in CD34+ progenitor cells. Differentiation to PLTs was performed by incubating progenitor cells in the presence of thrombopoietin and interleukin-3. RESULTS: The transduction of RNAi cassettes containing the sequences for shRNAs targeting β2m caused up to 85% reduction of progenitor cells HLA Class I antigen expression, which was maintained in the culture-derived PLTs. The HLA-deficient PLTs derived from HLA-silenced CD34+ cells proved to be fully functional in in vitro tests when compared to peripheral blood-derived PLTs. CONCLUSIONS: Our data show that in vitro generating HLA Class I-deficient PLTs from hematopoietic progenitor cells prove to be feasible. As malignancy risks associated with insertional mutagenesis are not to be expected in anucleated PLTs, provision of HLA-deficient PLTs from large-scale production units may become reality in the management of patients suffering from PLT transfusion refractoriness.
Authors: Zaruhi Karabekian; Hao Ding; Gulnaz Stybayeva; Irina Ivanova; Narine Muselimyan; Amranul Haque; Ian Toma; Nikki G Posnack; Alexander Revzin; David Leitenberg; Michael A Laflamme; Narine Sarvazyan Journal: Tissue Eng Part A Date: 2015-09-10 Impact factor: 3.845
Authors: Ann-Kathrin Börger; Dorothee Eicke; Christina Wolf; Christiane Gras; Susanne Aufderbeck; Kai Schulze; Lena Engels; Britta Eiz-Vesper; Axel Schambach; Carlos A Guzman; Nico Lachmann; Thomas Moritz; Ulrich Martin; Rainer Blasczyk; Constança Figueiredo Journal: Mol Med Date: 2016-05-16 Impact factor: 6.354
Authors: Constança Figueiredo; Dirk Wedekind; Thomas Müller; Stefanie Vahlsing; Peter A Horn; Axel Seltsam; Rainer Blasczyk Journal: Biomed Res Int Date: 2013-10-09 Impact factor: 3.411