Lethal α–thalassaemia created by gene targeting in mice and its genetic rescue

Pászty, Chris; Mohandas, Narla; Stevens, Mary; Loring, Jeanne F.; Liebhaber, Stephen A.; Brion, Catherine M.; Rubin, Edward M.

doi:10.1038/ng0995-33

Cited by 87 publications

(44 citation statements)

References 33 publications

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“…They express 45% of all α-globins as human α and 73% of all β-globins as human β S . Group 4 animals express the NY1 transgene and the γ M transgene (NY1KO-γM or α H β S γM α KK β KK ) and are homozygous for the mouse α-globin knockout [19] and the mouse β-globin knockout [20] as previously described [16]. These mice express 100% human α, 80% β S , and 20% human γ.…”

Section: Transgenic Micementioning

confidence: 99%

Heme degradation and oxidative stress in murine models for hemoglobinopathies: Thalassemia, sickle cell disease and hemoglobin C disease

Nagababu

Fabry

Nagel

et al. 2008

Blood Cells, Molecules, and Diseases

102

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Red blood cells with abnormal hemoglobins (Hb) are frequently associated with increased hemoglobin autoxidation, accumulation of iron in membranes, increased membrane damage and a shorter red cell life span. The mechanisms for many of these changes have not been elucidated. We have shown in our previous studies that hydrogen peroxide formed in association with hemoglobin autoxidation reacts with hemoglobin and initiates a cascade of reactions that results in heme degradation with the formation of two fluorescent emission bands and the release of iron. Heme degradation was assessed by measuring the fluorescent band at ex 321 nm. A 5.6 fold increase in fluorescence was found in red cells from sickle transgenic mice that expressed exclusively human globins when compared to red cells from control mice. When sickle transgenic mice co-express the γM transgene, that expresses HbF and inhibits polymerization, heme degradation is decreased. Mice expressing exclusively hemoglobin C had a 6.9 fold increase in fluorescence compared to control. Heme degradation was also increased 3.5 fold in β-thalassemic mice generated by deletion of murine β major . Membrane bound IgG and red cell metHb were highly correlated with the intensity of the fluorescent heme degradation band. These results suggest that degradation of the heme moiety in intact hemoglobin and/or degradation of free heme by peroxides are higher in pathological RBCs. Concomitant release of iron appears to be responsible for the membrane damage that leads to IgG binding and the removal of red cells from circulation.

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Section: Transgenic Micementioning

confidence: 99%

Heme degradation and oxidative stress in murine models for hemoglobinopathies: Thalassemia, sickle cell disease and hemoglobin C disease

Nagababu

Fabry

Nagel

et al. 2008

Blood Cells, Molecules, and Diseases

102

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“…22,39,43,60,82,85,86 In ovine SCNT placentae at term (147 days of gestation) with fetuses delivered alive and healthy, the number of capillaries per mm 2 (50.7/mm 2 ), evaluated by means of image analysis on semithin sections of resin-embedded samples, was lower than in control placentae (503/mm 2 ), as was the capillary total area per villous area (0.01 versus 0.05). 81 Moreover, reduced numbers of vessels (Figs. 1, 2) were associated with relative immaturity of vessels of the placental villi, as determined by reduced or no immunohistochemical expression of von Willebrand factor and calponin (Figs.…”

Section: Morphologic Pathology Of Scnt Placentaementioning

confidence: 99%

“…Candidates are encoded mainly by pregnancy-related genes and include glycoproteins, growth factors, hormones, and hormone-like factors. 81 Proteins involved in the regulation of placentogenesis, fetogenesis, mammogenesis, steroidogenesis, and immune system activity, 2 such as chorionic somatomammotropin hormone 1, bovine pregnancy-associated glycoproteins 1 and 9, and bovine prolactin-related protein 1, are deregulated throughout the pregnancy in SCNT placentae. 39,82 The reduced expression of these proteins might be responsible for inadequate preparation of the birth canal at parturition and poor mammary gland development in cows carrying SCNT embryos.…”

Section: Pathogenesismentioning

confidence: 99%

Review Paper: A Review of the Pathology of Abnormal Placentae of Somatic Cell Nuclear Transfer Clone Pregnancies in Cattle, Sheep, and Mice

et al. 2008

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Abstract. Cloning of cattle, sheep, and mice by somatic cell nuclear transfer (SCNT) can result in apparently healthy offspring, but the probability of a successful and complete pregnancy is less than 5%. Failures of SCNT pregnancy are associated with placental abnormalities, such as placentomegaly, reduced vascularisation, hypoplasia of trophoblastic epithelium, and altered basement membrane. The pathogenesis of these changes is poorly understood, but current evidence implicates aberrant reprogramming of donor nuclei by the recipient oocyte cytoplast, resulting in epigenetic modifications of key regulatory genes essential for normal placental development. The purpose of this review is to provide an overview of the anatomic pathology of abnormal placentae of SCNT clones and to summarize current knowledge concerning underlying pathogenetic mechanisms.

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“…Production of HbAS3, Knock-out Transgenic Sickle Mice-Mice that express HbAS3 exclusively were produced by breeding the HbAS3 mice with mouse ␣-and ␤-globin gene knock-out animals (26,27). These animals were subsequently bred with knock-out transgenic sickle mice (28) to obtain mice that were homozygous for mouse ␣-and ␤-globin gene knock-outs and contained the human LCR ␣, LCR ␥-␤ S , and LCR ␤ AS3 transgenes.…”

Section: Analysis and Purification Of Recombinant Humanmentioning

confidence: 99%

A Recombinant Human Hemoglobin with Anti-sickling Properties Greater than Fetal Hemoglobin

Levasseur

Ryan

Reilly

et al. 2004

Journal of Biological Chemistry

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A new recombinant, human anti-sickling ␤-globin polypeptide designated ␤ AS3 (␤Gly 16 3 Asp/␤Glu 22 3 Ala/␤Thr 87 3 Gln) was designed to increase affinity for ␣-globin. The amino acid substitutions at ␤22 and ␤87 are located at axial and lateral contacts of the sickle hemoglobin (HbS) polymers and strongly inhibit deoxyHbS polymerization. The ␤16 substitution confers the recombinant ␤-globin subunit (␤ AS3 ) with a competitive advantage over ␤ S for interaction with the ␣-globin polypeptide. Transgenic mouse lines that synthesize high levels of HbAS3 (␣ 2 ␤ AS3 2 ) were established, and recombinant HbAS3 was purified from hemolysates and then characterized. HbAS3 binds oxygen cooperatively and has an oxygen affinity that is comparable with fetal hemoglobin. Delay time experiments demonstrate that HbAS3 is a potent inhibitor of HbS polymerization. Subunit competition studies confirm that ␤ AS3 has a distinct advantage over ␤ S for dimerization with ␣-globin. When equal amounts of ␤ S -and ␤ AS3 -globin monomers compete for limiting ␣-globin chains up to 82% of the tetramers formed is HbAS3. Knock-out transgenic mice that express exclusively human HbAS3 were produced. When these mice were bred with knock-out transgenic sickle mice the ␤ AS3 polypeptides corrected all hematological parameters and organ pathology associated with the disease. Expression of ␤ AS3 -globin should effectively lower the concentration of HbS in erythrocytes of patients with sickle cell disease, especially in the 30% percent of these individuals who coinherit ␣-thalassemia. Therefore, constructs expressing the ␤ AS3 -globin gene may be suitable for future clinical trials for sickle cell disease.Sickle cell disease (SCD) 1 results from an A to T transversion at the sixth codon of the human ␤-globin gene on chromosome 11 (1, 2). The mutation of a single DNA base leads to the substitution of a valine for a glutamic acid in the ␤-globin polypeptide of sickle hemoglobin (HbS). The positioning of a hydrophobic residue at this position permits an interaction with a hydrophobic pocket on another hemoglobin tetramer (see Fig. 1). This interaction allows deoxy-HbS to polymerize in an entropy-driven process (3-5). The polymerization of deoxyHbS leads to erythrocyte deformation from a biconcave morphology into the sickle shapes for which SCD is named.Polymerization of deoxy-HbS is effectively inhibited by fetal hemoglobin (HbF), and individuals who are homozygous for the sickle mutation but also express high levels of HbF are typically asymptomatic (6 -8). The level of HbF necessary to significantly reduce the symptoms of SCD ranges from about 20 to 25%; however, data showing an enhanced red cell survival with as little as 9% HbF have been reported (9). The efficacy of HbF in inhibiting HbS polymerization suggests that transduction of fetal globin genes into hematopoietic stem cells might be an effective strategy for sickle cell disease gene therapy. However, high levels of ␥-globin gene expression are difficult to achieve in adult erythroid cells even...

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Lethal α–thalassaemia created by gene targeting in mice and its genetic rescue

Cited by 87 publications

References 33 publications

Heme degradation and oxidative stress in murine models for hemoglobinopathies: Thalassemia, sickle cell disease and hemoglobin C disease

Heme degradation and oxidative stress in murine models for hemoglobinopathies: Thalassemia, sickle cell disease and hemoglobin C disease

Review Paper: A Review of the Pathology of Abnormal Placentae of Somatic Cell Nuclear Transfer Clone Pregnancies in Cattle, Sheep, and Mice

A Recombinant Human Hemoglobin with Anti-sickling Properties Greater than Fetal Hemoglobin

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