Dimeric erythropoietin fusion protein with enhanced erythropoietic activity in vitro and in vivo

被引:45
作者
Dalle, B
Henri, A
Rouyer-Fessard, P
Bettan, M
Scherman, D
Beuzard, Y
Payen, E
机构
[1] Univ Paris, Hop St Louis, Lab Therapie Gen Hematopoiet, Inst Hematol, F-75475 Paris 10, France
[2] ENSCP Aventis Gencell, CNRS, UMR 7001, Lab Chim Bioorgan & Biotechnol Mol & Cellulaire, Vitry Sur Seine, France
关键词
D O I
10.1182/blood.V97.12.3776
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
High doses of recombinant human erythropoietin (rhEpo) are required for the treatment of chronic anemia. Thus, it is clear that therapy for chronic anemia would greatly benefit from an erythropoietin derivative with increased erythropoietic activity rather than the native endogenous hormone. In this report, the activity of a human Epo-Epo dimer protein, obtained by recombinant technology, is described and compared with its Epo monomer counterpart produced under identical conditions. Although monomer Epo and dimer Epo-Epo had similar pharmacokinetics in normal mice, the increase in hematocrit value was greater with the dimer than with the monomer, Moreover, in clonogenic assays using CD34(+) human hematopoietic cells, the human dimer induced a 3- to 4-fold-greater proliferation of erythroid cells than the monomer, Controlled secretion of dimeric erythropoietin was achieved in beta -thalassemic mice by in vivo intramuscular electrotransfer of a mouse Epo-Epo plasmid containing the tetO element and of a plasmid encoding the tetracycline controlled transactivator tTA, Administration of tetracycline completely inhibited the expression of the mEpo dimer, On tetracycline withdrawal, expression of the Epo-Epo dimer resumed, thereby resulting in a large and sustained hematocrit increase in beta -thalassemic mice. No immunologic response against the dimer was apparent in mice because the duration of the hematocrit increase was similar to that observed with the monomeric form of mouse erythropoietin. (C) 2001 by The American Society of Hematology.
引用
收藏
页码:3776 / 3782
页数:7
相关论文
共 42 条
[31]   Erythropoietin receptor activation by a ligand-induced conformation change [J].
Remy, I ;
Wilson, IA ;
Michnick, SW .
SCIENCE, 1999, 283 (5404) :990-993
[32]   A gene therapy approach to regulated delivery of erythropoietin as a function of oxygen tension [J].
Rinsch, C ;
Regulier, E ;
Deglon, N ;
Dalle, B ;
Beuzard, Y ;
Aebischer, P .
HUMAN GENE THERAPY, 1997, 8 (16) :1881-1889
[33]   THE EXPRESSION OF FUNCTIONAL ERYTHROPOIETIN RECEPTORS ON AN INTERLEUKIN-3 DEPENDENT CELL-LINE [J].
SAKAGUCHI, M ;
KOISHIHARA, Y ;
TSUDA, H ;
FUJIMOTO, K ;
SHIBUYA, K ;
KAWAKITA, M ;
TAKATSUKI, K .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1987, 146 (01) :7-12
[34]   A MODIFIED TETRACYCLINE-REGULATED SYSTEM PROVIDES AUTOREGULATORY, INDUCIBLE GENE-EXPRESSION IN CULTURED-CELLS AND TRANSGENIC MICE [J].
SHOCKETT, P ;
DIFILIPPANTONIO, M ;
HELLMAN, N ;
SCHATZ, DG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (14) :6522-6526
[35]   Erythropoietin can promote erythroid progenitor survival by repressing apoptosis through Bcl-X(L) and Bcl-2 [J].
Silva, M ;
Grillot, D ;
Benito, A ;
Richard, C ;
Nunez, G ;
FernandezLuna, JL .
BLOOD, 1996, 88 (05) :1576-1582
[36]   Human erythropoietin dimers with markedly enhanced in vivo activity [J].
Sytkowski, AJ ;
Lunn, ED ;
Davis, KL ;
Feldman, L ;
Siekman, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (03) :1184-1188
[37]   An erythropoietin fusion protein comprised of identical repeating domains exhibits enhanced biological properties [J].
Sytkowski, AJ ;
Lunn, ED ;
Risinger, MA ;
Davis, KL .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (35) :24773-24778
[38]  
VEDOVATO M, 1993, HAEMATOLOGIA, V25, P19
[39]  
WEINBERG RS, 1993, BLOOD, V81, P2591
[40]  
WEN DY, 1994, J BIOL CHEM, V269, P22839