Knockout-transgenic mouse model of sickle cell disease

Science

Volumn 278, Issue 5339, 1997, Pages 873-876

  Ryan, T M ^a   Ciavatta, D J ^a   Townes, T M ^a  

^a UNIVERSITY OF ALABAMA AT BIRMINGHAM   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ALPHA GLOBIN; BETA GLOBIN; DNA FRAGMENT; HEMOGLOBIN S;

ANIMAL CELL; ANIMAL EXPERIMENT; ANIMAL MODEL; ARTICLE; BLOOD SMEAR; BLOOD VESSEL OCCLUSION; ERYTHROCYTE; GENE EXPRESSION; HEMOLYTIC ANEMIA; MOUSE; NONHUMAN; PRIORITY JOURNAL; SICKLE CELL ANEMIA; THROMBOSIS; TRANSGENIC MOUSE;

ANEMIA, SICKLE CELL; ANIMALS; CHROMATOGRAPHY, HIGH PRESSURE LIQUID; CROSSES, GENETIC; DISEASE MODELS, ANIMAL; ERYTHROCYTES; GLOBINS; HEMOGLOBIN, SICKLE; HEMOGLOBINS; HUMANS; MICE; MICE, INBRED C57BL; MICE, KNOCKOUT; MICE, TRANSGENIC;

ANIMALIA; MUS MUSCULUS;

EID: 0030725279     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.278.5339.873     Document Type: Article

References (45)

1. V. M. Ingram, Nature 178, 792 (1956); ibid. 180, 326 (1957).
2. V. M. Ingram, Nature 178, 792 (1956); ibid. 180, 326 (1957).
3. E. A. Padlan and W. E. Love, J. Biol. Chem. 260, 8280 (1985); B. C. Wishner, K. B. Ward, E. E. Lattman, W. E. Love, J. Mol. Biol. 98, 179 (1975); W. G. Wood, C. Bunch, S. Kelly, Y. Gunn, G. Breckon, Nature 313, 320 (1985); R. H. Crepeau, G. Dykes, R. Garrell, S. J. Edelstein, ibid 274, 616 (1978); G. W. Dykes, R. H. Crepeau, S. J. Edelstein, J. Mol. Biol. 130, 451 (1979); J. Hofrichter, P. D. Ross, W. A. Eaton, Proc. Natl. Acad Sci. U.S.A. 71, 4864 (1974); W. A. Eaton and J. Hofrichter, Blood 70, 1245 (1987).
4. E. A. Padlan and W. E. Love, J. Biol. Chem. 260, 8280 (1985); B. C. Wishner, K. B. Ward, E. E. Lattman, W. E. Love, J. Mol. Biol. 98, 179 (1975); W. G. Wood, C. Bunch, S. Kelly, Y. Gunn, G. Breckon, Nature 313, 320 (1985); R. H. Crepeau, G. Dykes, R. Garrell, S. J. Edelstein, ibid 274, 616 (1978); G. W. Dykes, R. H. Crepeau, S. J. Edelstein, J. Mol. Biol. 130, 451 (1979); J. Hofrichter, P. D. Ross, W. A. Eaton, Proc. Natl. Acad Sci. U.S.A. 71, 4864 (1974); W. A. Eaton and J. Hofrichter, Blood 70, 1245 (1987).
5. E. A. Padlan and W. E. Love, J. Biol. Chem. 260, 8280 (1985); B. C. Wishner, K. B. Ward, E. E. Lattman, W. E. Love, J. Mol. Biol. 98, 179 (1975); W. G. Wood, C. Bunch, S. Kelly, Y. Gunn, G. Breckon, Nature 313, 320 (1985); R. H. Crepeau, G. Dykes, R. Garrell, S. J. Edelstein, ibid 274, 616 (1978); G. W. Dykes, R. H. Crepeau, S. J. Edelstein, J. Mol. Biol. 130, 451 (1979); J. Hofrichter, P. D. Ross, W. A. Eaton, Proc. Natl. Acad Sci. U.S.A. 71, 4864 (1974); W. A. Eaton and J. Hofrichter, Blood 70, 1245 (1987).
6. E. A. Padlan and W. E. Love, J. Biol. Chem. 260, 8280 (1985); B. C. Wishner, K. B. Ward, E. E. Lattman, W. E. Love, J. Mol. Biol. 98, 179 (1975); W. G. Wood, C. Bunch, S. Kelly, Y. Gunn, G. Breckon, Nature 313, 320 (1985); R. H. Crepeau, G. Dykes, R. Garrell, S. J. Edelstein, ibid 274, 616 (1978); G. W. Dykes, R. H. Crepeau, S. J. Edelstein, J. Mol. Biol. 130, 451 (1979); J. Hofrichter, P. D. Ross, W. A. Eaton, Proc. Natl. Acad Sci. U.S.A. 71, 4864 (1974); W. A. Eaton and J. Hofrichter, Blood 70, 1245 (1987).
7. E. A. Padlan and W. E. Love, J. Biol. Chem. 260, 8280 (1985); B. C. Wishner, K. B. Ward, E. E. Lattman, W. E. Love, J. Mol. Biol. 98, 179 (1975); W. G. Wood, C. Bunch, S. Kelly, Y. Gunn, G. Breckon, Nature 313, 320 (1985); R. H. Crepeau, G. Dykes, R. Garrell, S. J. Edelstein, ibid 274, 616 (1978); G. W. Dykes, R. H. Crepeau, S. J. Edelstein, J. Mol. Biol. 130, 451 (1979); J. Hofrichter, P. D. Ross, W. A. Eaton, Proc. Natl. Acad Sci. U.S.A. 71, 4864 (1974); W. A. Eaton and J. Hofrichter, Blood 70, 1245 (1987).
8. E. A. Padlan and W. E. Love, J. Biol. Chem. 260, 8280 (1985); B. C. Wishner, K. B. Ward, E. E. Lattman, W. E. Love, J. Mol. Biol. 98, 179 (1975); W. G. Wood, C. Bunch, S. Kelly, Y. Gunn, G. Breckon, Nature 313, 320 (1985); R. H. Crepeau, G. Dykes, R. Garrell, S. J. Edelstein, ibid 274, 616 (1978); G. W. Dykes, R. H. Crepeau, S. J. Edelstein, J. Mol. Biol. 130, 451 (1979); J. Hofrichter, P. D. Ross, W. A. Eaton, Proc. Natl. Acad Sci. U.S.A. 71, 4864 (1974); W. A. Eaton and J. Hofrichter, Blood 70, 1245 (1987).
9. E. A. Padlan and W. E. Love, J. Biol. Chem. 260, 8280 (1985); B. C. Wishner, K. B. Ward, E. E. Lattman, W. E. Love, J. Mol. Biol. 98, 179 (1975); W. G. Wood, C. Bunch, S. Kelly, Y. Gunn, G. Breckon, Nature 313, 320 (1985); R. H. Crepeau, G. Dykes, R. Garrell, S. J. Edelstein, ibid 274, 616 (1978); G. W. Dykes, R. H. Crepeau, S. J. Edelstein, J. Mol. Biol. 130, 451 (1979); J. Hofrichter, P. D. Ross, W. A. Eaton, Proc. Natl. Acad Sci. U.S.A. 71, 4864 (1974); W. A. Eaton and J. Hofrichter, Blood 70, 1245 (1987).
10. C. T. Noguchi and A. N. Schechter, Blood 58, 1057 (1981); G. M. Brittenham, A. N. Schechter, C. T. Noguchi, ibid. 65, 183 (1985).
11. C. T. Noguchi and A. N. Schechter, Blood 58, 1057 (1981); G. M. Brittenham, A. N. Schechter, C. T. Noguchi, ibid. 65, 183 (1985).
12. T. M. Ryan et al., Science 247, 566 (1990).
13. D. R. Greaves et al., Nature 343, 183 (1990).
14. M. E. Fabry et al., Blood 86, 2419 (1995); M. Trudel et al., EMBO J. 10, 3157 (1991); M. Trudel et al., Blood 84, 3189 (1994); M. E. Fabry et al., J. Clin. Invest. 98, 2450 (1996); M. E. Fabry et al., Proc. Natl. Acad. Sci. U.S.A. 89, 12155 (1992); R. A. Popp et al., Blood 89, 4204 (1997).
15. M. E. Fabry et al., Blood 86, 2419 (1995); M. Trudel et al., EMBO J. 10, 3157 (1991); M. Trudel et al., Blood 84, 3189 (1994); M. E. Fabry et al., J. Clin. Invest. 98, 2450 (1996); M. E. Fabry et al., Proc. Natl. Acad. Sci. U.S.A. 89, 12155 (1992); R. A. Popp et al., Blood 89, 4204 (1997).
16. M. E. Fabry et al., Blood 86, 2419 (1995); M. Trudel et al., EMBO J. 10, 3157 (1991); M. Trudel et al., Blood 84, 3189 (1994); M. E. Fabry et al., J. Clin. Invest. 98, 2450 (1996); M. E. Fabry et al., Proc. Natl. Acad. Sci. U.S.A. 89, 12155 (1992); R. A. Popp et al., Blood 89, 4204 (1997).
17. M. E. Fabry et al., Blood 86, 2419 (1995); M. Trudel et al., EMBO J. 10, 3157 (1991); M. Trudel et al., Blood 84, 3189 (1994); M. E. Fabry et al., J. Clin. Invest. 98, 2450 (1996); M. E. Fabry et al., Proc. Natl. Acad. Sci. U.S.A. 89, 12155 (1992); R. A. Popp et al., Blood 89, 4204 (1997).
18. M. E. Fabry et al., Blood 86, 2419 (1995); M. Trudel et al., EMBO J. 10, 3157 (1991); M. Trudel et al., Blood 84, 3189 (1994); M. E. Fabry et al., J. Clin. Invest. 98, 2450 (1996); M. E. Fabry et al., Proc. Natl. Acad. Sci. U.S.A. 89, 12155 (1992); R. A. Popp et al., Blood 89, 4204 (1997).
19. M. E. Fabry et al., Blood 86, 2419 (1995); M. Trudel et al., EMBO J. 10, 3157 (1991); M. Trudel et al., Blood 84, 3189 (1994); M. E. Fabry et al., J. Clin. Invest. 98, 2450 (1996); M. E. Fabry et al., Proc. Natl. Acad. Sci. U.S.A. 89, 12155 (1992); R. A. Popp et al., Blood 89, 4204 (1997).
20. M. D. Rhoda et al., Biochim. Biophys. Acta 953, 208 (1988).
21. D. J. Ciavatta, T. M. Ryan, S. C. Farmer, T. M. Townes, Proc. Natl. Acad. Sci. U.S.A. 92, 9259 (1995).
22. B. Yang et al., ibid., p. 11608.
23. C. Paszty et al., Nature Genet. 11, 33 (1995).
24. G. Stamatoyannopoulos, A. W. Nienhuis, P. W. Majerus, H. Varmus, The Molecular Basis of Blood Diseases (Saunders, Philadelphia, 1994).
25. H. F. Bunn and B. G. Forget, Hemoglobin: Molecular, Genetic, and Clinical Aspects (Saunders, Philadelphia, 1986).
26. R. R. Behringer, T. M. Ryan, R. D. Palmiter, R. L. Brinster, T. M. Townes, Genes Dev. 4, 380 (1990); T. Enver et al., Nature 344, 309 (1990); K. M. Gaensler, M. Kitamura, Y. W. Kan, Proc. Natl. Acad. Sci. U.S.A. 90, 11381 (1993); K. R. Peterson et al., ibid., p. 7593; J. Strouboulis, N. Dillon, F. Grosveld, Genes Dev. 6, 1857 (1992).
27. R. R. Behringer, T. M. Ryan, R. D. Palmiter, R. L. Brinster, T. M. Townes, Genes Dev. 4, 380 (1990); T. Enver et al., Nature 344, 309 (1990); K. M. Gaensler, M. Kitamura, Y. W. Kan, Proc. Natl. Acad. Sci. U.S.A. 90, 11381 (1993); K. R. Peterson et al., ibid., p. 7593; J. Strouboulis, N. Dillon, F. Grosveld, Genes Dev. 6, 1857 (1992).
28. R. R. Behringer, T. M. Ryan, R. D. Palmiter, R. L. Brinster, T. M. Townes, Genes Dev. 4, 380 (1990); T. Enver et al., Nature 344, 309 (1990); K. M. Gaensler, M. Kitamura, Y. W. Kan, Proc. Natl. Acad. Sci. U.S.A. 90, 11381 (1993); K. R. Peterson et al., ibid., p. 7593; J. Strouboulis, N. Dillon, F. Grosveld, Genes Dev. 6, 1857 (1992).
29. R. R. Behringer, T. M. Ryan, R. D. Palmiter, R. L. Brinster, T. M. Townes, Genes Dev. 4, 380 (1990); T. Enver et al., Nature 344, 309 (1990); K. M. Gaensler, M. Kitamura, Y. W. Kan, Proc. Natl. Acad. Sci. U.S.A. 90, 11381 (1993); K. R. Peterson et al., ibid., p. 7593; J. Strouboulis, N. Dillon, F. Grosveld, Genes Dev. 6, 1857 (1992).
30. R. R. Behringer, T. M. Ryan, R. D. Palmiter, R. L. Brinster, T. M. Townes, Genes Dev. 4, 380 (1990); T. Enver et al., Nature 344, 309 (1990); K. M. Gaensler, M. Kitamura, Y. W. Kan, Proc. Natl. Acad. Sci. U.S.A. 90, 11381 (1993); K. R. Peterson et al., ibid., p. 7593; J. Strouboulis, N. Dillon, F. Grosveld, Genes Dev. 6, 1857 (1992).
31. T. M. Ryan and T. M. Townes, unpublished data; N. A. Roberts, J. Sloane-Stanley, J. A. Sharpe, S. J. Stanworth, W. G. Wood, Blood 89, 713 (1997).
32. T. M. Ryan and T. M. Townes, unpublished data; N. A. Roberts, J. Sloane-Stanley, J. A. Sharpe, S. J. Stanworth, W. G. Wood, Blood 89, 713 (1997).
33. S Fragments were excised from the cosmid vector and injected into C57BI/6 × SJL fertilized mouse eggs. Transgenic founder animals were bred with C57BI/6 × SJL mice to establish lines.
34. T. M. Ryan, D. J. Ciavatta, T. M. Townes, data not shown.
35. _, in preparation.
36. G. R. Serjeant, Br. J. Hemat. 19, 635 (1970).
37. 3H-leucine (0.5 mCi/ml, Amersham), and the pH was adjusted to 7.1. Cells were washed three times with 0.85% saline after incubation and subsequently lysed in 5 mM phosphate, 0.5 mM EDTA, pH 7.4. Hemolysates were subjected to HPLC as described (22). To determine α/non-α ratios, we collected fractions every 20 s, and the radioactivity in each fraction was detected with a Beckman LS 6500 liquid scintillation counter.
38. D. J. Weatherall and J. B. Clegg, The Thalassemia Syndromes (Blackwell, Oxford, UK, 1981).
39. Food and water were withheld for 4 hours and urine was collected from 4-to 4.5-month-old mice. Urine osmolality was measured on a Wescor model 5130C vapor pressure osmometer.
40. Reverse-phase HPLC was performed on a Rainin Dynamax system with a Vydac C4 column (25 cm by 0.46 cm). Buffer A was 10% acetonitrile (ACN)/0.1% trifluoroacetic acid (TFA), and buffer B was 90% ACN/0.1% TFA. A nonlinear gradient for 1 hour was used starting at 35% B to 36% B in the first minute, followed by an increase to 42% over the next 48 min, maintained at 42% B for 6 min, and returned to 35% B in the final 5 min. The flow rate was kept at 1 ml/min, and the effluent was monitored at 214 nm. Individual globin chains were quantitated with Dynamax HPLC Method Manager software.
41. Reticulocytes were counted manually after supravital staining with 1% new methylene blue (Sigma) for 10 min at 37°C.
42. RBC counts were measured on a Coulter Counter (Model MHR). Hemoglobin concentrations were determined at 540 nm by the cyanmethemoglobin method with a kit from Sigma (catalog no. 525-A). PVCs were determined by centrifugation of EDTA-treated blood in a Jorvet J503 centrifuge.
43. Tissues were fixed in 70% alcoholic formalin, embedded in paraffin, sectioned, and stained with hematoxylin-eosin by standard methods.
44. Spleen from a HbS mouse was fixed in 1% buffered glutaraldehyde and then fixed in 1% osmium tetroxide. After dehydration, the sample was embedded in polybed, sectioned, and stained with uranyl acetate and lead citrate. Ultra-thin sections were examined and photographed with the Hitachi-7000 transmission electron microscope.
45. We thank C. Paszty and E. Rubin for the α-globin knockout mice, R. Lindsey for valuable discussions of histopathology, P. Sanders and D. Thornley-Brown for help with urine osmolality measurements, E. Arms of the University of Alabama at Birmingham (UAB) Comprehensive Cancer Center Electron Microscopy Core Facility for help with electron microscopy, and especially J. Prchal for many helpful discussions and for his critical review of the manuscript. We also thank the UAB Transgenic Mouse Facility for production of some of the transgenic mice; the facility is supported by National Cancer Institute grant CA13148. Supported by grants from the NIH National Heart, Lung, ana Blood Institute.