Remodeling of the Major Pig Xenoantigen by N-Acetylglucosaminyltransferase III in Transgenic Pig

Miyagawa, Shuji; Murakami, Hiroshi; Takahagi, Yoichi; Nakai, Rie; Yamada, Mako; Murase, Ayako; Koyota, Souichi; Koma, Masaru; Matsunami, Katsuyoshi; Fukuta, Daisuke; Fujimura, Tadao; Shigehisa, Tamotsu; Okabe, Masaru; Nagashima, Hiroshi; Shirakura, R; Taniguchi, Naoyuki

doi:10.1074/jbc.m104359200

Cited by 105 publications

(56 citation statements)

References 54 publications

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“…These results suggest that GTKO porcine organs effectively avoid hyperacute rejection following pig-to-human xenotransplantation and that further effective genetic modification is necessary in addition to GTKO. In fact, Miyagawa et al [30] reported that the H-D antigen could be a xenoantigen following removal of the αGal antigen. Moreover, Sharma et al [11] reported that knocking out only the GT gene does not completely remove the αGal antigen.…”

Section: Discussionmentioning

confidence: 99%

Effects of Recloning on the Efficiency of Production of .ALPHA.1,3-Galactosyltransferase Knockout Pigs

Fujimura¹,

Murakami²,

Kurome

et al. 2008

Journal of Reproduction and Development

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Abstract. Obtaining sufficient transgenic cells via selective cultivation of genetically manipulated somatic cells is difficult due to the limited number of cell divisions. Additionally, if irreversible mutations in a cell's chromosomes occur during selective cultivation and the cell is used as the nuclear donor, somatic cell nuclear transfer (SCNT) embryos often exhibit abnormal development. On the other hand, a SCNT method in which fetal cells derived from SCNT embryos are used as the nuclear donor (recloning method) is an effective technique for obtaining large quantities of transgenic cells. In this study, we compared the in vivo development rate of SCNT embryos produced from porcine α1-3 galactosyltransferase gene knockout (GTKO) cells by a recloning method with that of SCNT embryos produced without recloning from porcine GTKO cells (direct method). In the direct method, 557 and 462 cloned embryos were produced using two types of activation methods, the two-step activation (TA) method and the delayed activation (DA) method, and then transferred into 6 and 4 recipients, respectively, but no piglets were born from these recipients. In the recloning method, 956 and 1038 cloned embryos were produced using the TA and DA methods, respectively, and then transferred to 8 and 7 recipients, respectively. Two piglets were born from one recipient in the TA group and 6 piglets were born from 3 recipients in the DA group. This report indicates that the recloning method improved the developmental capacity of SCNT embryos reconstructed with gene-targeted somatic cells.

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Section: Discussionmentioning

confidence: 99%

Effects of Recloning on the Efficiency of Production of .ALPHA.1,3-Galactosyltransferase Knockout Pigs

Fujimura¹,

Murakami²,

Kurome

et al. 2008

Journal of Reproduction and Development

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“…The Mgat3 gene was not detected by Northern analysis of RNA from WW6 embryonic stem cells or by lectin histochemistry, but E-PHA binding was induced in neurite extensions by in vitro differentiation of embryoid bodies. 4 When Mgat3 T37⁄T37 mice in either a mixed or 129 SvJ background were suspended upside down by the tail, they retracted their hind and fore limbs toward the trunk, and in some cases, they rolled upward into a ball (Fig. 5).…”

Section: Mgat3mentioning

confidence: 99%

“…LEC10 cells are ϳ15-fold more resistant to ricin and ϳ10-fold more sensitive to the toxicity of the erythroagglutinin from Phaseolus vulgaris (E-PHA) compared with wild-type CHO cells, reflecting dramatic changes in binding of these lectins to N-linked Gal residues of cell-surface glycoproteins. Similarly, the ectopic expression of an Mgat3 cDNA reduces the expression of terminal ␣3-Gal residues, a key determinant in xenotransplantation (4,5). The regulated expression of the Mgat3 gene could therefore control the binding of animal lectins like galectins to cell-surface N-glycans with a bisecting GlcNAc and thereby modulate cellular interactions.…”

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confidence: 99%

Truncated, InactiveN-Acetylglucosaminyltransferase III (GlcNAc-TIII) Induces Neurological and Other Traits Absent in Mice That Lack GlcNAc-TIII

Bhattacharyya

Bhaumik

Raju³

et al. 2002

Journal of Biological Chemistry

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N-Acetylglucosaminyltransferase III (GlcNAc-TIII), the product of the Mgat3 gene, transfers the bisecting GlcNAc to the core mannose of complex N-glycans. The addition of this residue is regulated during development and has functional consequences for receptor signaling, cell adhesion, and tumor progression. Mice homozygous for a null mutation at the Mgat3 locus (Mgat3 ⌬ ) or for a targeted mutation in the Mgat3 gene (previously called Mgat3 neo , but herein renamed Mgat3 T37 because the allele generates inactive GlcNAc-TIII of ϳ37 kDa) were found to exhibit retarded progression of liver tumors. Matrix-assisted laser desorption/ionization time-offlight mass spectrometry of neutral N-glycans from kidneys revealed no significant differences, and both mutants showed the expected lack of N-glycan species with an additional GlcNAc. However, the two mutants differed in several biological traits. Mgat3 T37/T37 homozygotes in a mixed or 129SvJ background were retarded in growth rate and exhibited an altered leg clasp reflex, an altered gait, and defective nursing behavior. Pups abandoned by Mgat3 T37/T37 mothers were rescued by wildtype foster mothers. None of these Mgat3 T37/T37 traits were exhibited by Mgat3 ⌬/⌬ mice or by heterozygous mice carrying the Mgat3 T37 mutation. Similarly, no dominant-negative effect was observed in Chinese hamster ovary cells expressing truncated GlcNAc-TIII in the presence of wild-type GlcNAc-TIII. However, compound heterozygotes carrying both the Mgat3 T37 and Mgat3 ⌬ mutations exhibited a marked leg clasp reflex, indicating that in the absence of wild-type GlcNAc-TIII, truncated GlcNAc-TIII causes this phenotype. The Mgat3 gene was expressed in brain at embryonic day 10.5 and thereafter and in neurons of adult cerebellum. The mutant Mgat3 gene was also highly expressed in Mgat3 T37/T37 brain. This may be the basis of the unexpected neurological phenotype induced by truncated, inactive GlcNAc-TIII in the mouse.The N-glycans of mammalian glycoproteins vary widely in structure, but the biological significance of this variation is largely unknown. A well studied example is the bisecting GlcNAc. This residue is transferred to the ␤-linked Man of the core of N-glycans by the glycosyltransferase termed N-acetylglucosaminyltransferase III (GlcNAc-TIII 1 ; EC 2.4.1.144) (1), the product of the Mgat3 gene (2). The presence of the bisecting GlcNAc alters the lectin binding properties of a cell, a fact initially revealed by the gain-of-function Chinese hamster ovary (CHO) glycosylation mutant LEC10, which expresses GlcNAc-TIII (3). LEC10 cells are ϳ15-fold more resistant to ricin and ϳ10-fold more sensitive to the toxicity of the erythroagglutinin from Phaseolus vulgaris (E-PHA) compared with wild-type CHO cells, reflecting dramatic changes in binding of these lectins to N-linked Gal residues of cell-surface glycoproteins. Similarly, the ectopic expression of an Mgat3 cDNA reduces the expression of terminal ␣3-Gal residues, a key determinant in xenotransplantation (4, 5). The regulated expression o...

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“…Xenograft rejection comprises several steps, including hyperacute rejection, delayed xenograft rejection and cellular rejection [86]. To overcome these rejection reactions, multilayered genetic modification is required [87][88][89][90][91][92][93][94][95][96][97][98][99]. In other words, the complex mechanisms of xenograft rejection [91,92,100] must be countered by multiple gene modifications.…”

Section: Development Of Genetically Modified Pigs For Xenotransplantamentioning

confidence: 99%

Application of Genetically Modified and Cloned Pigs in Translational Research

Matsunari

Nagashima

2009

Journal of Reproduction and Development

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Abstract. Pigs are increasingly being recognized as good large-animal models for translational research, linking basic science to clinical applications in order to establish novel therapeutics. This article reviews the current status and future prospects of genetically modified and cloned pigs in translational studies. It also highlights pigs specially designed as disease models, for xenotransplantation or to carry cell marker genes. Finally, use of porcine somatic stem and progenitor cells in preclinical studies of cell transplantation therapy is also discussed. Key words: Cloned pig, Gene knockout, Transgenic pig, Translational research (J. Reprod. Dev. 55: [225][226][227][228][229][230] 2009) t present, pigs are not only considered important livestock but are also essential large-animal models in various types of biomedical research [1][2][3][4][5][6]. Furthermore, due to recent technological advances in genetic modification and somatic cell cloning, the range of applications for porcine models has increased markedly [7][8][9][10][11][12][13][14]. The present review focuses on development of genetically modified and cloned pigs and their use in translational studies.Translational research plays an important role as a bridge between basic science outcomes and the clinical realm, leading to development of novel therapeutics. However, disparate findings for rodent models and humans have hindered the translation of rodent data into actionable technologies for patients [15]. In contrast, pigs have many anatomical and physiological similarities to humans, including their cardiovascular systems, omnivorous gastrointestinal tracts and central nervous systems. Their physical sizes are also more comparable to that of humans, rendering pigs more appropriate for development of new surgical or endoscopic techniques.In the future, more advanced translational research may be conducted by improving the clinical relevance of pig models through genetic engineering. This article reviews the current status and future prospects of genetically modified pigs in preclinical studies of stem and progenitor cell therapy, the development of disease models and xenotransplantation.

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Remodeling of the Major Pig Xenoantigen by N-Acetylglucosaminyltransferase III in Transgenic Pig

Cited by 105 publications

References 54 publications

Effects of Recloning on the Efficiency of Production of .ALPHA.1,3-Galactosyltransferase Knockout Pigs

Effects of Recloning on the Efficiency of Production of .ALPHA.1,3-Galactosyltransferase Knockout Pigs

Truncated, InactiveN-Acetylglucosaminyltransferase III (GlcNAc-TIII) Induces Neurological and Other Traits Absent in Mice That Lack GlcNAc-TIII

Application of Genetically Modified and Cloned Pigs in Translational Research

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