Generation of HLA-deficient platelets from hematopoietic progenitor cells
Constança Figueiredo
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover; the Institute for Transfusion Medicine, University Hospital Essen, Essen; and the German Red Cross Blood Services NSTOB, Institute Springe, Springe, Germany.
Search for more papers by this authorLilia Goudeva
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover; the Institute for Transfusion Medicine, University Hospital Essen, Essen; and the German Red Cross Blood Services NSTOB, Institute Springe, Springe, Germany.
Search for more papers by this authorPeter A. Horn
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover; the Institute for Transfusion Medicine, University Hospital Essen, Essen; and the German Red Cross Blood Services NSTOB, Institute Springe, Springe, Germany.
Search for more papers by this authorBritta Eiz-Vesper
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover; the Institute for Transfusion Medicine, University Hospital Essen, Essen; and the German Red Cross Blood Services NSTOB, Institute Springe, Springe, Germany.
Search for more papers by this authorConstança Figueiredo
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover; the Institute for Transfusion Medicine, University Hospital Essen, Essen; and the German Red Cross Blood Services NSTOB, Institute Springe, Springe, Germany.
Search for more papers by this authorLilia Goudeva
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover; the Institute for Transfusion Medicine, University Hospital Essen, Essen; and the German Red Cross Blood Services NSTOB, Institute Springe, Springe, Germany.
Search for more papers by this authorPeter A. Horn
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover; the Institute for Transfusion Medicine, University Hospital Essen, Essen; and the German Red Cross Blood Services NSTOB, Institute Springe, Springe, Germany.
Search for more papers by this authorBritta Eiz-Vesper
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover; the Institute for Transfusion Medicine, University Hospital Essen, Essen; and the German Red Cross Blood Services NSTOB, Institute Springe, Springe, Germany.
Search for more papers by this authorCF designed and performed the experiments and wrote the manuscript, LG and BEV aided in performing experiments and analyzed and interpreted data, PAH analyzed and interpreted data, RB designed parts of the study and contributed to paper writing, and AS designed the study and wrote the manuscript.
Abstract
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.
REFERENCES
- 1 Gulliksson H, AuBuchon JP, Vesterinen M, Sandgren P, Larsson S, Pickard CA, Herschel I, Roger J, Tracy JE, Langweiler M; Biomedical Excellence for Safer Transfusion Working Party of the International Society of Blood Transfusion. Storage of platelets in additive solutions: a pilot in vitro study of the effects of potassium and magnesium. Vox Sang 2002; 82: 131-6.
- 2 Blumberg N, Heal JM, Liesveld JL, Phillips GL, Rowe JM. Platelet transfusion and survival in adults with acute leukemia. Leukemia 2007; 22: 631-5.
- 3 Nevo S, Fuller AK, Zahurak ML, Hartley E, Borinsky ME, Vogelsang GB. Profound thrombocytopenia and survival of hematopoietic stem cell transplant patients without clinically significant bleeding, using prophylactic platelet transfusion triggers of 10 x 10(9) or 20 x 10(9) per L. Transfusion 2007; 47: 1700-9.
- 4 Stephan F, Montblanc J, Cheffi A, Bonnet F. Thrombocytopenia in critically ill surgical patients: a case-control study evaluating attributable mortality and transfusion requirements. Crit Care 1999; 3: 151-8.
- 5 Rebulla P. Refractoriness to platelet transfusion. Curr Opin Hematol 2002; 9: 516-20.
- 6 Kiefel V, Konig C, Kroll H, Santoso S. Platelet alloantibodies in transfused patients. Transfusion 2001; 41: 766-70.
- 7 Laundy GJ, Bradley BA, Rees BM, Younie M, Hows JM. Incidence and specificity of HLA antibodies in multitransfused patients with acquired aplastic anemia. Transfusion 2004; 44: 814-25.
- 8 Matsunaga T, Tanaka I, Kobune M, Kawano Y, Tanaka M, Kuribayashi K, Iyama S, Sato T, Sato Y, Takimoto R, Takayama T, Kato J, Ninomiya T, Hamada H, Niitsu Y. Ex vivo large-scale generation of human platelets from cord blood CD34+ cells. Stem Cells 2006; 24: 2877-87.
- 9 Norol F, Vitrat N, Cramer E, Guichard J, Burstein SA, Vainchenker W, Debili N. Effects of cytokines on platelet production from blood and marrow CD34+ cells. Blood 1998; 91: 830-43.
- 10 Fujimoto TT, Kohata S, Suzuki H, Miyazaki H, Fujimura K. Production of functional platelets by differentiated embryonic stem (ES) cells in vitro. Blood 2003; 102: 4044-51.
- 11 Nishikii H, Eto K, Tamura N, Hattori K, Heissig B, Kanaji T, Sawaguchi A, Goto S, Ware J, Nakauchi H. Metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells. J Exp Med 2008; 205: 1917-27.
- 12 Takayama N, Nishikii H, Usui J, Tsukui H, Sawaguchi A, Hiroyama T, Eto K, Nakauchi H. Generation of functional platelets from human embryonic stem cells in vitro via ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors. Blood 2008; 111: 5298-306.
- 13 Abbas-Terki T, Blanco-Bose W, Deglon N, Pralong W, Aebischer P. Lentiviral-mediated RNA interference. Hum Gene Ther 2002; 13: 2197-201.
- 14 Taulli R, Accornero P, Follenzi A, Mangano T, Morotti A, Scuoppo C, Forni PE, Bersani F, Crepaldi T, Chiarle R, Naldini L, Ponzetto C. RNAi technology and lentiviral delivery as a powerful tool to suppress Tpr-Met-mediated tumorigenesis. Cancer Gene Ther 2005; 12: 456-63.
- 15 Kock N, Kasmieh R, Weissleder R, Shah K. Tumor therapy mediated by lentiviral expression of shBcl-2 and S-TRAIL. Neoplasia 2007; 9: 435-42.
- 16 Figueiredo C, Seltsam A, Blasczyk R. Class-, gene-, and group-specific HLA silencing by lentiviral shRNA delivery. J Mol Med 2006; 84: 425-37.
- 17 Italiano JE, Jr, Lecine P, Shivdasani RA, Hartwig JH. Blood platelets are assembled principally at the ends of proplatelet processes produced by differentiated megakaryocytes. J Cell Biol 1999; 147: 1299-312.
- 18 Ungerer M, Peluso M, Gillitzer A, Massberg S, Heinzmann U, Schulz C, Münch G, Gawaz M. Generation of functional culture-derived platelets from CD34+ progenitor cells to study transgenes in the platelet environment. Circ Res 2004; 95: e36-44.
- 19 Giarratana MC, Kobari L, Lapillonne H, Chalmers D, Kiger L, Cynober T, Marden MC, Wajcman H, Douay L. Ex vivo generation of fully mature human red blood cells from hematopoietic stem cells. Nat Biotechnol 2005; 23: 69-74.
- 20 Baek EJ, Kim HS, Kim S, Jin H, Choi TY, Kim HO. In vitro clinical-grade generation of red blood cells from human umbilical cord blood CD34+ cells. Transfusion 2008; 48: 2235-45.
- 21 Dorn I, Lazar-Karsten P, Boie S, Ribbat J, Hartwig D, Driller B, Kirchner H, Schlenke P. In vitro proliferation and differentiation of human CD34+ cells from peripheral blood into mature red blood cells with two different cell culture systems. Transfusion 2008; 48: 1122-32.
- 22 Neildez-Nguyen TM, Wajcman H, Marden MC, Bensidhoum M, Moncollin V, Giarratana MC, Kobari L, Thierry D, Douay L. Human erythroid cells produced ex vivo at large scale differentiate into red blood cells in vivo. Nat Biotechnol 2002; 20: 467-72.
- 23 Choi ES, Nichol JL, Hokom MM, Hornkohl AC, Hunt P. Platelets generated in vitro from proplatelet-displaying human megakaryocytes are functional. Blood 1995; 85: 402-13.
- 24 Lu SJ, Feng Q, Park JS, Vida L, Lee BS, Strausbauch M, Wettstein PJ, Honig GR, Lanza R. Biologic properties and enucleation of red blood cells from human embryonic stem cells. Blood 2008; 112: 4475-84.
- 25 Olivier EN, Qiu C, Velho M, Hirsch RE, Bouhassira EE. Large-scale production of embryonic red blood cells from human embryonic stem cells. Exp Hematol 2006; 34: 1635-42.
- 26 Bagnis C, Chapel S, Chiaroni J, Bailly P. A genetic strategy to control expression of human blood group antigens in red blood cells generated in vitro. Transfusion 2009.
- 27 Shanwell A, Sallander S, Olsson I, Gulliksson H, Pedajas I, Lerner R. An alloimmunized, thrombocytopenic patient successfully transfused with acid-treated, random-donor platelets. Br J Haematol 1991; 79: 462-5.
- 28 Bertolini F, Porretti L, Corsini C, Rebulla P, Sirchia G. Platelet quality and reduction of HLA expression in acid-treated platelet concentrates. Br J Haematol 1993; 83: 525-7.
- 29 Novotny VM, Doxiadis IN, Van Doorn R, Brand A. The kinetics of HLA Class I elution and the relevance for the use of HLA-eluted platelet transfusions. Br J Haematol 1996; 95: 416-22.
- 30 Castro E, Muncunill J, Barea L, Gonzalez R, Fernandez-Villalta MJ. Acid elution of platelets HLA-Class I antigens in the treatment of a refractory patient. Br J Haematol 1998; 100: 245-6.
- 31 Novotny VM, Huizinga TW, Van Doorn R, Briet E, Brand A. HLA Class I-eluted platelets as an alternative to HLA-matched platelets. Transfusion 1996; 36: 438-44.
- 32 Figueiredo C, Horn PA, Blasczyk R, Seltsam A. Regulating MHC expression for cellular therapeutics. Transfusion 2007; 47: 18-27.
- 33 Cosgrove LJ, Vaughan HA, Tjandra JJ, Thurlow PJ, McKenzie IF. HLA (Class I) antigens on platelets are involved in platelet function. Immunol Cell Biol 1988; 66: 69-77.
- 34 Rubinstein E, Urso I, Boucheix C, Carroll RC. Platelet activation by cross-linking HLA Class I molecules and Fc receptor. Blood 1992; 79: 2901-8.
- 35 Baum C, Dullmann J, Li Z, Fehse B, Meyer J, Williams DA, Von Kalle C. Side effects of retroviral gene transfer into hematopoietic stem cells. Blood 2003; 101: 2099-114.
- 36 Cavazzana-Calvo M, Fischer A. Gene therapy for severe combined immunodeficiency: are we there yet? J Clin Invest 2007; 117: 1456-65.
- 37 Ott MG, Schmidt M, Schwarzwaelder K, Stein S, Siler U, Koehl U, Glimm H, Kühlcke K, Schilz A, Kunkel H, Naundorf S, Brinkmann A, Deichmann A, Fischer M, Ball C, Pilz I, Dunbar C, Du Y, Jenkins NA, Copeland NG, Lüthi U, Hassan M, Thrasher AJ, Hoelzer D, Von Kalle C, Seger R, Grez M. Correction of X-linked chronic granulomatous disease by gene therapy, augmented by insertional activation of MDS1-EVI1, PRDM16 or SETBP1. Nat Med 2006; 12: 401-9.
Citing Literature
