Further characterization of factor VIII-deficient mice created by gene targeting: RNA and protein studies

Bi, Lei; Sarkar, Rita; Naas, Thierry; Lawler, A. M.; Shumaker, SL; Bedian,; Kazazian, H. H.

doi:10.1182/blood.v88.9.3446.bloodjournal8893446

Cited by 166 publications

(124 citation statements)

References 15 publications

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“…There was bleeding and synovial hyperplasia recorded in the left knee joints of some mice in the absence of induced trauma. As previously described [12,13], these mice, although lacking coagulation FVIII, do not manifest spontaneous bleeding and as reported by us, do not develop synovitis in the absence of bleeding [14,15]. However, there are at least two extraneous factors that could confound the data in control joints.…”

Section: Discussionsupporting

confidence: 80%

Prevention of haemarthrosis in a murine model of acute joint bleeding

Valentino¹,

Hakobyan²,

Kazarian³

et al. 2009

Haemophilia

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Preservation of normal joint function in patients with haemophilia is a goal of modern therapy. Regular injections of anti-haemophilic factor concentrate reduce the risk of joint bleeding, the optimal regimen for which remains under investigation. The goals of the experiment described here are: (i) to assess the capacity of a murine model of severe haemophilic arthropathy to predict the likelihood of success of a test product to prevent joint bleeding and the complications that follow and (ii) to compare the effectiveness of recombinant human activated factor VII (rFVIIa) to recombinant human factor VIII (rFVIII) to prevent acute joint bleeding in the mouse model of haemarthrosis. Mice lacking expression of FVIII received a single intravenous injection of human rFVIII (280 U kg(-1)), rFVIIa (10 mg kg(-1)) or vehicle prior to blunt trauma injury to the knee joint. Mice receiving rFVIII and rFVIIa developed less injury-induced joint bleeding, swelling and loss of range of motion compared to mice pretreated with vehicle. Despite the reduction in clinical symptoms, synovial hyperplasia was evident in all groups after 7 days although less pronounced in mice receiving rFVIII and rFVIIa. The data under these experimental conditions demonstrate: (i) that this model can be used to evaluate novel therapies designed to prevent joint bleeding (prophylaxis) and (ii) both rFVIII and rFVIIa reduced acute haemarthrosis but did not completely prevent synovitis, the sequelae of blood induced joint injury.

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Section: Discussionsupporting

confidence: 80%

Prevention of haemarthrosis in a murine model of acute joint bleeding

Valentino¹,

Hakobyan²,

Kazarian³

et al. 2009

Haemophilia

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“…The Animal Care and Use Committee at Albert Einstein College of Medicine approved the studies. Hemophilia A mice were originally produced by inserting a neomycin gene into the 3¢-end of the 16th exon of FVIII gene and were in the 129-C57BL/6J background [2]. Donor C57BL/6J-TgN(MT-nLacZ)204Bri mice and 129-Gt(ROSA)26 Sor mice were from Jackson Labs (Bar Harbor, ME, USA) and wild-type C57BL/6 mice were from the National Cancer Institute (Bethesda, MD, USA).…”

Section: Animalsmentioning

confidence: 99%

“…Recipient animals were not treated with FVIII either prior to or subsequent to cell transplantation. The tail-clip challenge was designed to analyze hemophilia A mice 1 week after cell transplantation by cutting 1 cm of the tail and monitoring animal survival over 24 h [2].…”

Section: Animalsmentioning

confidence: 99%

“…Maintenance of FVIII levels in people with hemophilia A requires repeated administration of products derived from pooled plasma with inactivation of pathogens, although some risk of disease transmission remains, and use of recombinant FVIII is expensive. Although the gene therapy approach has shown promise in a FVIII knockout mouse model [2], progress in this area has been relatively slow [3][4][5][6][7][8][9].…”

Section: Introductionmentioning

confidence: 99%

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Transplantation of endothelial cells corrects the phenotype in hemophilia A mice

Kumaran

Benten

Follenzi

et al. 2005

Journal of Thrombosis and Haemostasis

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Summary. Background: The deficiency of factor VIII, a co‐factor in the intrinsic coagulation pathway results in hemophilia A. Although FVIII is synthesized largely in the liver, the specific liver cell type(s) responsible for FVIII production is controversial. Objective: This study aimed to determine the cellular origin of FVIII synthesis and release in mouse models. Methods: We transplanted cells into the peritoneal cavity of hemophilia A knockout mice. Plasma FVIII activity was measured using a Chromogenix assay 2–7 days after cell transplantation, and phenotypic correction was determined with tail‐clip challenge 7 days following cell transplantation. Transplanted cells were identified by histologic and molecular assays. Results: Untreated hemophilia A mice, as well as mice treated with the hepatocyte‐enriched fraction, showed extensive mortality following tail‐clip challenge. In contrast, recipients of unfractionated liver cells (mixture of hepatocytes, liver sinusoidal endothelial cells (LSEC), Kupffer cells, and hepatic stellate cells) or of the cell fraction enriched in LSECs survived tail‐clip challenge (P < 0.001). FVIII was secreted in the blood stream in recipients of unfractionated liver cells, LSECs and pancreatic islet‐derived MILE SVEN 1 (MS1) endothelial cells. Although transplanted hepatocytes maintained functional integrity in the peritoneal cavity, these cells did not produce detectable plasma FVIII activity. Conclusions: The assay of cell transplantation in the peritoneal cavity showed that endothelial cells but not hepatocytes produced phenotypic correction in hemophilia A mice. Therefore, endothelial cells should be suitable additional targets for cell and gene therapy in hemophilia A.

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“…Animals FIX knockout C57Bl/6 mice were purchased from Jackson Laboratories [8]. The FVIII knockout mouse strain was produced at AstraZeneca using a C57Bl/6 mice background essentially as described by Bi et al [9]. Both the FVIII and the FIX knockouts were bred on a C57Bl/6 background (substrain 6JolaHsd) from Harlan BV, the Netherlands.…”

Section: Methodsmentioning

confidence: 99%