Clinical and molecular characterization of a re-established line of sheep exhibiting hemophilia A

C. D. Porada, C. Sanada, C. R. Long, J. A. Wood, J. DesaI, N. Frederick, L. Millsap, C. Bormann, S. L. Menges, C. Hanna, G. Flores-Foxworth, T. Shin, M. E. Westhusin, W. Liu, H. Glimp, E. D. Zanjani, J. N. Lozier, V. Pliska, G. Stranzinger, H. JoergD. C. Kraemer, G. Almeida-Porada

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Background: Large animal models that accurately mimic human hemophilia A (HA) are in great demand for developing and testing novel therapies to treat HA. Objectives: To re-establish a line of sheep exhibiting a spontaneous bleeding disorder closely mimicking severe human HA, fully characterize their clinical presentation, and define the molecular basis for disease. Patients/methods: Sequential reproductive manipulations were performed with cryopreserved semen from a deceased affected ram. The resultant animals were examined for hematologic parameters, clinical symptoms, and responsiveness to human FVIII (hFVIII). The full coding region of sheep FVIII mRNA was sequenced to identify the genetic lesion. Results and conclusions: The combined reproductive technologies yielded 36 carriers and 8 affected animals. The latter had almost non-existent levels of FVIII:C and extremely prolonged aPTT, with otherwise normal hematologic parameters. These animals exhibited bleeding from the umbilical cord, prolonged tail and nail cuticle bleeding time, and multiple episodes of severe spontaneous bleeding, including hemarthroses, muscle hematomas and hematuria, all of which responded to hFVIII. Inhibitors of hFVIII were detected in four treated animals, further establishing the preclinical value of this model. Sequencing identified a premature stop codon and frame-shift in exon 14, providing a molecular explanation for HA. Given the decades of experience using sheep to study both normal physiology and a wide array of diseases and the high homology between human and sheep FVIII, this new model will enable a better understanding of HA and facilitate the development and testing of novel treatments that can directly translate to HA patients.

Original languageEnglish (US)
Pages (from-to)276-285
Number of pages10
JournalJournal of Thrombosis and Haemostasis
Volume8
Issue number2
DOIs
StatePublished - Feb 1 2010

Fingerprint

Hemophilia A
Sheep
Hemorrhage
Hemarthrosis
Reproductive Techniques
Bleeding Time
Nonsense Codon
Umbilical Cord
Hematuria
Nails
Semen
Hematoma
Tail
Exons
Animal Models
Muscles
Messenger RNA
Therapeutics

All Science Journal Classification (ASJC) codes

  • Hematology

Cite this

Porada, C. D., Sanada, C., Long, C. R., Wood, J. A., DesaI, J., Frederick, N., ... Almeida-Porada, G. (2010). Clinical and molecular characterization of a re-established line of sheep exhibiting hemophilia A. Journal of Thrombosis and Haemostasis, 8(2), 276-285. https://doi.org/10.1111/j.1538-7836.2009.03697.x
Porada, C. D. ; Sanada, C. ; Long, C. R. ; Wood, J. A. ; DesaI, J. ; Frederick, N. ; Millsap, L. ; Bormann, C. ; Menges, S. L. ; Hanna, C. ; Flores-Foxworth, G. ; Shin, T. ; Westhusin, M. E. ; Liu, W. ; Glimp, H. ; Zanjani, E. D. ; Lozier, J. N. ; Pliska, V. ; Stranzinger, G. ; Joerg, H. ; Kraemer, D. C. ; Almeida-Porada, G. / Clinical and molecular characterization of a re-established line of sheep exhibiting hemophilia A. In: Journal of Thrombosis and Haemostasis. 2010 ; Vol. 8, No. 2. pp. 276-285.
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abstract = "Background: Large animal models that accurately mimic human hemophilia A (HA) are in great demand for developing and testing novel therapies to treat HA. Objectives: To re-establish a line of sheep exhibiting a spontaneous bleeding disorder closely mimicking severe human HA, fully characterize their clinical presentation, and define the molecular basis for disease. Patients/methods: Sequential reproductive manipulations were performed with cryopreserved semen from a deceased affected ram. The resultant animals were examined for hematologic parameters, clinical symptoms, and responsiveness to human FVIII (hFVIII). The full coding region of sheep FVIII mRNA was sequenced to identify the genetic lesion. Results and conclusions: The combined reproductive technologies yielded 36 carriers and 8 affected animals. The latter had almost non-existent levels of FVIII:C and extremely prolonged aPTT, with otherwise normal hematologic parameters. These animals exhibited bleeding from the umbilical cord, prolonged tail and nail cuticle bleeding time, and multiple episodes of severe spontaneous bleeding, including hemarthroses, muscle hematomas and hematuria, all of which responded to hFVIII. Inhibitors of hFVIII were detected in four treated animals, further establishing the preclinical value of this model. Sequencing identified a premature stop codon and frame-shift in exon 14, providing a molecular explanation for HA. Given the decades of experience using sheep to study both normal physiology and a wide array of diseases and the high homology between human and sheep FVIII, this new model will enable a better understanding of HA and facilitate the development and testing of novel treatments that can directly translate to HA patients.",
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Porada, CD, Sanada, C, Long, CR, Wood, JA, DesaI, J, Frederick, N, Millsap, L, Bormann, C, Menges, SL, Hanna, C, Flores-Foxworth, G, Shin, T, Westhusin, ME, Liu, W, Glimp, H, Zanjani, ED, Lozier, JN, Pliska, V, Stranzinger, G, Joerg, H, Kraemer, DC & Almeida-Porada, G 2010, 'Clinical and molecular characterization of a re-established line of sheep exhibiting hemophilia A', Journal of Thrombosis and Haemostasis, vol. 8, no. 2, pp. 276-285. https://doi.org/10.1111/j.1538-7836.2009.03697.x

Clinical and molecular characterization of a re-established line of sheep exhibiting hemophilia A. / Porada, C. D.; Sanada, C.; Long, C. R.; Wood, J. A.; DesaI, J.; Frederick, N.; Millsap, L.; Bormann, C.; Menges, S. L.; Hanna, C.; Flores-Foxworth, G.; Shin, T.; Westhusin, M. E.; Liu, W.; Glimp, H.; Zanjani, E. D.; Lozier, J. N.; Pliska, V.; Stranzinger, G.; Joerg, H.; Kraemer, D. C.; Almeida-Porada, G.

In: Journal of Thrombosis and Haemostasis, Vol. 8, No. 2, 01.02.2010, p. 276-285.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Clinical and molecular characterization of a re-established line of sheep exhibiting hemophilia A

AU - Porada, C. D.

AU - Sanada, C.

AU - Long, C. R.

AU - Wood, J. A.

AU - DesaI, J.

AU - Frederick, N.

AU - Millsap, L.

AU - Bormann, C.

AU - Menges, S. L.

AU - Hanna, C.

AU - Flores-Foxworth, G.

AU - Shin, T.

AU - Westhusin, M. E.

AU - Liu, W.

AU - Glimp, H.

AU - Zanjani, E. D.

AU - Lozier, J. N.

AU - Pliska, V.

AU - Stranzinger, G.

AU - Joerg, H.

AU - Kraemer, D. C.

AU - Almeida-Porada, G.

PY - 2010/2/1

Y1 - 2010/2/1

N2 - Background: Large animal models that accurately mimic human hemophilia A (HA) are in great demand for developing and testing novel therapies to treat HA. Objectives: To re-establish a line of sheep exhibiting a spontaneous bleeding disorder closely mimicking severe human HA, fully characterize their clinical presentation, and define the molecular basis for disease. Patients/methods: Sequential reproductive manipulations were performed with cryopreserved semen from a deceased affected ram. The resultant animals were examined for hematologic parameters, clinical symptoms, and responsiveness to human FVIII (hFVIII). The full coding region of sheep FVIII mRNA was sequenced to identify the genetic lesion. Results and conclusions: The combined reproductive technologies yielded 36 carriers and 8 affected animals. The latter had almost non-existent levels of FVIII:C and extremely prolonged aPTT, with otherwise normal hematologic parameters. These animals exhibited bleeding from the umbilical cord, prolonged tail and nail cuticle bleeding time, and multiple episodes of severe spontaneous bleeding, including hemarthroses, muscle hematomas and hematuria, all of which responded to hFVIII. Inhibitors of hFVIII were detected in four treated animals, further establishing the preclinical value of this model. Sequencing identified a premature stop codon and frame-shift in exon 14, providing a molecular explanation for HA. Given the decades of experience using sheep to study both normal physiology and a wide array of diseases and the high homology between human and sheep FVIII, this new model will enable a better understanding of HA and facilitate the development and testing of novel treatments that can directly translate to HA patients.

AB - Background: Large animal models that accurately mimic human hemophilia A (HA) are in great demand for developing and testing novel therapies to treat HA. Objectives: To re-establish a line of sheep exhibiting a spontaneous bleeding disorder closely mimicking severe human HA, fully characterize their clinical presentation, and define the molecular basis for disease. Patients/methods: Sequential reproductive manipulations were performed with cryopreserved semen from a deceased affected ram. The resultant animals were examined for hematologic parameters, clinical symptoms, and responsiveness to human FVIII (hFVIII). The full coding region of sheep FVIII mRNA was sequenced to identify the genetic lesion. Results and conclusions: The combined reproductive technologies yielded 36 carriers and 8 affected animals. The latter had almost non-existent levels of FVIII:C and extremely prolonged aPTT, with otherwise normal hematologic parameters. These animals exhibited bleeding from the umbilical cord, prolonged tail and nail cuticle bleeding time, and multiple episodes of severe spontaneous bleeding, including hemarthroses, muscle hematomas and hematuria, all of which responded to hFVIII. Inhibitors of hFVIII were detected in four treated animals, further establishing the preclinical value of this model. Sequencing identified a premature stop codon and frame-shift in exon 14, providing a molecular explanation for HA. Given the decades of experience using sheep to study both normal physiology and a wide array of diseases and the high homology between human and sheep FVIII, this new model will enable a better understanding of HA and facilitate the development and testing of novel treatments that can directly translate to HA patients.

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