Towards a transgenic mouse model of sickle cell disease: hemoglobin SAD.

Abstract

In order to obtain a transgenic mouse model of sickle cell disease, we have synthesized a novel human beta-globin gene, beta SAD, designed to increase the polymerization of the transgenic human hemoglobin S (Hb S) in vivo. beta SAD (beta S-Antilles-D Punjab) includes the beta 6Val substitution of the beta S chain, as well as two other mutations, Antilles (beta 23Ile) and D Punjab (beta 121Gln) each of which promotes the polymerization of Hb S in human. The beta SAD gene and the human alpha 2-globin gene, each linked to the beta-globin locus control region (LCR) were co-introduced into the mouse germ line. In one of the five transgenic lines obtained, SAD-1, red blood cells contained 19% human Hb SAD (alpha 2 human 1 beta 2SAD) and mouse-human hybrids in addition to mouse hemoglobin. Adult SAD-1 transgenic mice were not anemic but had some abnormal features of erythrocytes and slightly enlarged spleens. Their erythrocytes displayed sickling upon deoxygenation in vitro. SAD-1 neonates were anemic and many did not survive. In order to generate adult mice with a more severe sickle cell syndrome, crosses between the SAD progeny and homozygous for beta-thalassemic mice were performed. Hemoglobin SAD was increased to 26% in beta-thal/SAD-1 mice which exhibited: (i) abnormal erythrocytes with regard to shape and density; (ii) an enlarged spleen and a high reticulocyte count indicating an increased erythropoiesis; (iii) mortality upon hypoxia; (iv) polymerization of hemolysate similar to that obtained in human homozygous sickle cell disease; and (v) anemia and mortality during development.

• Authors: M Trudel, N Saadane, M C Garel, J Bardakdjian-Michau, Y Blouquit, J L Guerquin-Kern, P Rouyer-Fessard, D Vidaud, A Pachnis, P H Roméo
• Journal: The EMBO journal (EMBO J) Vol. 10 Issue 11 Pg. 3157-65 (Nov 1991) ISSN: 0261-4189 [Print] England
• PMID: 1915288 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Hemoglobin, Sickle
• hemoglobin SAD
• Globins
• DNA
• Trypsin
• Oxygen

Topics

• Anemia, Sickle Cell (mortality, physiopathology)
• Animals
• Chromatography, High Pressure Liquid
• DNA (genetics)
• Disease Models, Animal
• Electrophoresis, Polyacrylamide Gel
• Erythrocyte Indices
• Globins (genetics)
• Hemoglobin, Sickle (genetics, metabolism)
• Isoelectric Focusing
• Mice
• Mice, Transgenic
• Oxygen (metabolism)
• Peptide Mapping
• Phenotype
• Polymerase Chain Reaction
• Trypsin