Journal: Nature Communications

Article Title: Base-editing corrects metabolic abnormalities in a humanized mouse model for glycogen storage disease type-Ia

doi: 10.1038/s41467-024-54108-1

Figure Lengend Snippet: Newborn (NB) huR83C mice, non-fasted, were treated with a high dose of BEAM-301 (301H) at 1.5 mg/kg. At age 3 weeks, the phenotype of the resulting NB-301H mice (NB-301H-3W) was evaluated and compared to age-matched unedited huR83C (huR83C-3W), wild-type (mR83-3W), and heterozygote (mR83/huR83C-3W) mice. a Liver and kidney microsomal G6Pase-α activity. Liver (mR83-3W, n = 5; mR83/huR83C-3W, n = 11; huR83C-3W, n = 8; NB-301H-3W, n = 6) and kidney (mR83-3W, n = 6; mR83/huR83C-3W, n = 10; huR83C-3W, n = 8; NB-301H-3W, n = 6). b Restoration of hepatic G6Pase-α activity as a function of base editing efficiency in NB-301H-3W mice ( n = 6) along with on-target and bystander values of liver base editing. c Fasting blood glucose levels in control (mR83-3W and mR83/huR83C-3W; n = 5) and NB-301H-3W ( n = 5) mice. d Histochemical analysis of liver and kidney G6Pase-α activity in control (mR83-3W and mR83/huR83C-3W, n = 6), untreated (huR83C-3W, n = 6), and NB-301H-3W ( n = 6) mice. Each image represents an individual mouse. The arrow indicates the kidney cortex. Scale bar = 100 µm. The numbers represent hepatic G6Pase-α activity expressed in the mice. e Body weight (BW), liver weight (LW)/BW, and kidney weight (KW)/BW values of control (mR83-3W and mR83/huR83C-3W, n = 25), untreated (huR83C-3W, n = 17), and NB-301H-3W ( n = 6) mice. f Size comparison of mR83 and huR83C mice, showing growth retardation of the huR83C mice at age 3 weeks. g Restoration of hepatic G6Pase-α activity as a function of LW/BW values and hepatic levels of glycogen, triglyceride, and G6P in the NB-301H-3W mice ( n = 6). Statistics were performed using a two-tailed unpaired T test. Data are presented as Mean values ± SEM, and individual data points for each animal are displayed. * denotes p < 0.05, ** denotes p value < 0.005.

Article Snippet: A human cDNA encoding the open reading frame for G6PC1 - c.247C > T ( G6PC1 -R83C) were inserted into exon 1 of the mouse G6pc gene at the ATG start codon (Taconic Biosciences) in a way that created a premature STOP codon in the coding sequence of the mouse G6pc exon 1.

Techniques: Activity Assay, Control, Comparison, Two Tailed Test

Journal: Nature Communications

Article Title: Base-editing corrects metabolic abnormalities in a humanized mouse model for glycogen storage disease type-Ia

doi: 10.1038/s41467-024-54108-1

Figure Lengend Snippet: Newborn (NB) huR83C mice, non-fasted, were treated with a high dose of BEAM-301 (301H) at 1.5 mg/kg. At age 3 weeks, biochemical phenotype of the edited mice was analyzed and compared to the age matched unedited huR83C. The 3-week-old mR83 and mR83/huR83C littermates displaying a wild-type phenotype were used as the controls. a Blood glucose levels (control-3W, n = 24; huR83C-3W, n = 16; NB-301H-3W, n = 6), serum cholesterol levels (control-3W, n = 16; huR83C-3W, n = 7; NB-301H-3W, n = 6), serum triglyceride levels (control-3W, n = 24; huR83C-3W, n = 16; NB-301H-3W, n = 6), serum lactate and uric acid levels (control-3W, n = 16; huR83C-3W, n = 8; NB-301H-3W, n = 6). b Liver glucose, triglyceride, lactate, glycogen, and G6P levels in control ( n = 16), huR83C-3W ( n = 8), and NB-301H-3W ( n = 6) mice. c Hematoxylin and eosin (H&E)-staining of liver and kidney sections in mR83/huR83C-3W ( n = 6), huR83C-3W ( n = 6), and NB-301H-3W ( n = 6) mice. The liver and kidney in all experimental animals were examined. A single image from an individual mouse, representative of the results in all mice, is shown to illustrate the results. Scale bar = 20 µm. The numbers represent hepatic G6Pase-α activity expressed in the mice. d Oil Red O staining of liver and kidney sections in mR83/huR83C-3W ( n = 6), huR83C-3W ( n = 6), and NB-301H-3W ( n = 6) mice. The liver and kidney in all experimental animals were examined. A single image from an individual mouse, representative of the results in all mice, is shown to illustrate the results. Scale bar = 20 μm. The numbers represent hepatic G6Pase-α activity expressed in the mice. Statistics were performed using a two-tailed unpaired T test. Data are presented as Mean values ± SEM, and individual data points for each animal are displayed. * denotes p < 0.05, ** denotes p value < 0.005.

Article Snippet: A human cDNA encoding the open reading frame for G6PC1 - c.247C > T ( G6PC1 -R83C) were inserted into exon 1 of the mouse G6pc gene at the ATG start codon (Taconic Biosciences) in a way that created a premature STOP codon in the coding sequence of the mouse G6pc exon 1.

Techniques: Control, Staining, Activity Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Base-editing corrects metabolic abnormalities in a humanized mouse model for glycogen storage disease type-Ia

doi: 10.1038/s41467-024-54108-1

Figure Lengend Snippet: Newborn (NB) and 3-week-old (3W) huR83C mice, non-fasted, were treated with 301H (BEAM-301 at 1.5 mg/kg) and the phenotype of the resulting NB-301H-8W and 3W-301H-8W mice was analyzed at age 8 weeks. The sex-matched mR83-8W and mR83/huR83C-8W littermates displaying a wild-type phenotype were used as the controls. a Liver and kidney microsomal G6Pase-α activity in mR83-8W ( n = 8), mR83/huR83C-8W ( n = 9), NB-301H-8W ( n = 8), and 3W-301H-8W ( n = 9) mice. b Restoration of hepatic G6Pase-α activity as a function of base editing efficiency along with on-target and bystander values of liver base editing. NB-301H-8W ( n = 8); 3W-301H-8W ( n = 9). c Fasting blood glucose levels in control ( n = 23), NB-301H-8W ( n = 8), and 3W-301H-8W ( n = 9) mice. d BW, LW/BW, and KW/BW values of control ( n = 23), NB-301H-8W ( n = 8), and 3W-301H-8W ( n = 9) mice. e Restoration of hepatic G6Pase-α activity as a function of LW/BW values and hepatic levels of glycogen and G6P in the 301H-8W mice ( n = 17), including NB-301H-8W ( n = 8) and 3W-301H-8W ( N = 9) mice. f Blood glucose and serum cholesterol, triglyceride, lactate, and uric acid levels in control ( n = 17), NB-301H-8W ( n = 8), and 3W-301H-8W ( n = 9) mice. g Liver glucose, glycogen, triglyceride, lactate, and G6P levels in control ( n = 17), NB-301H-8W ( n = 8), and 3W-301H-8W ( n = 9) mice. Statistics were performed using a two-tailed unpaired T test. Data are presented as Mean values ± SEM, and individual data points for each animal are displayed. * denotes p < 0.05, ** denotes p value < 0.005.

Article Snippet: A human cDNA encoding the open reading frame for G6PC1 - c.247C > T ( G6PC1 -R83C) were inserted into exon 1 of the mouse G6pc gene at the ATG start codon (Taconic Biosciences) in a way that created a premature STOP codon in the coding sequence of the mouse G6pc exon 1.

Techniques: Activity Assay, Control, Two Tailed Test

Journal: Nature Communications

Article Title: Base-editing corrects metabolic abnormalities in a humanized mouse model for glycogen storage disease type-Ia

doi: 10.1038/s41467-024-54108-1

Figure Lengend Snippet: Newborn (NB) and 3-week-old (3W) huR83C mice, non-fasted, were treated with a low dose of BEAM-301 (301L) at 0.75 mg/kg and the phenotype of the resulting NB-301L-8W and 3W-301L-8W mice was analyzed at age 8 weeks. The sex-matched mR83 and mR83/hR83C littermates displaying a wild-type phenotype were used as the controls. a Liver microsomal G6Pase-α activity in control ( n = 17), NB-301L-8W ( n = 9), and 3W-301L-8W ( n = 8) mice. b Restoration of hepatic G6Pase-α activity as a function of base editing efficiency along with on-target and bystander values of liver base editing in NB-301L-8W ( n = 9) and 3W-301L-8W ( n = 8) mice. c Fasting blood glucose levels in control ( n = 17), NB-301L-8W ( n = 9), and 3W-301L-8W ( n = 8) mice. d BW, LW/BW, and KW/BW values of control ( n = 23), NB-301L-8W ( n = 9), and 3W-301L-8W ( n = 8) mice. e Restoration of hepatic G6Pase-α activity as a function of LW/BW values and hepatic levels of glycogen and G6P in the edited 301L-8W ( n = 17) mice, including NB-301L-8W ( n = 9) and 3W-301L-8W ( n = 8) mice. f Blood glucose and serum cholesterol, triglyceride, lactate, and uric acid levels in control ( n = 17), NB-301L-8W ( n = 9), and 3W-301L-8W ( n = 8) mice. g Liver glucose, glycogen, triglyceride, lactate, and G6P levels in control ( n = 17), NB-301L-8W ( n = 9), and 3W-301L-8W ( n = 8) mice. Statistics were performed using a two-tailed unpaired T test. Data are presented as Mean values ± SEM, and individual data points for each animal are displayed. * denotes p < 0.05, ** denotes p value < 0.005.

Article Snippet: A human cDNA encoding the open reading frame for G6PC1 - c.247C > T ( G6PC1 -R83C) were inserted into exon 1 of the mouse G6pc gene at the ATG start codon (Taconic Biosciences) in a way that created a premature STOP codon in the coding sequence of the mouse G6pc exon 1.

Techniques: Activity Assay, Control, Two Tailed Test

Journal: Nature Communications

Article Title: Base-editing corrects metabolic abnormalities in a humanized mouse model for glycogen storage disease type-Ia

doi: 10.1038/s41467-024-54108-1

Figure Lengend Snippet: Newborn (NB) huR83C mice, non-fasted, were treated with 301H (BEAM-301 at 1.5 mg/kg) and the phenotype of the NB-301H-dosed mice (NB-301H-53W, n = 19) was evaluated at 53 weeks of age using sex-matched wild-type littermates (mR83-53W, n = 16) as the controls. a Liver microsomal G6Pase-α activity. b Restoration of hepatic G6Pase-α activity as a function of base editing efficiency along with on-target and bystander values of liver base editing. c Fasting blood glucose levels. d BW, LW/BW, and KW/BW values. e Restoration of hepatic G6Pase-α activity as a function of LW/BW values and hepatic levels of glycogen and G6P in the edited mice. f Blood glucose and serum cholesterol, triglyceride, lactate, and uric acid levels. g Liver glucose, glycogen, triglyceride, lactate, and G6P levels. Statistics were performed using a two-tailed unpaired T test. Data are presented as Mean values ± SEM, and individual data points for each animal are displayed. * denotes p < 0.05, ** denotes p value < 0.005.

Article Snippet: A human cDNA encoding the open reading frame for G6PC1 - c.247C > T ( G6PC1 -R83C) were inserted into exon 1 of the mouse G6pc gene at the ATG start codon (Taconic Biosciences) in a way that created a premature STOP codon in the coding sequence of the mouse G6pc exon 1.

Techniques: Activity Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Base-editing corrects metabolic abnormalities in a humanized mouse model for glycogen storage disease type-Ia

doi: 10.1038/s41467-024-54108-1

Figure Lengend Snippet: Three-week-old (3W) huR83C mice, non-fasted, were treated with 301L (BEAM-301 at 0.75 mg/kg) and the phenotype of the 3W-301L-dosed mice (3W-301L-53W) was evaluated at 53 weeks of age using the sex-matched mR83-53W and mR83/huR83C-53W littermates (Control-53W) as the controls. a Liver microsomal G6Pase-α activity in Control ( n = 14) and 3W-301L-53W ( n = 9) mice. b Restoration of hepatic G6Pase-α activity as a function of base editing efficiency along with on-target and bystander values of liver base editing ( n = 9). c Fasting blood glucose levels in Control-53W ( n = 13) and 3W-301L-53W ( n = 8) mice. d BW, LW/BW, and KW/BW values in Control ( n = 14) and 3W-301L-53W ( n = 9) mice. e Restoration of hepatic G6Pase-α activity as a function of LW/BW values and hepatic levels of glycogen and G6P in the edited mice ( n = 9). f Blood glucose and serum cholesterol, triglyceride, lactate, and uric acid levels in Control ( n = 14) and 3W-301L-53W ( n = 9) mice. g Liver glucose, glycogen, triglyceride, lactate, and G6P levels (nmol/mg) in Control ( n = 14) and 3W-301L-53W ( n = 9) mice. Statistics were performed using a two-tailed unpaired T test. Data are presented as Mean values ± SEM, and individual data points for each animal are displayed. * denotes p < 0.05, ** denotes p value < 0.005.

Article Snippet: A human cDNA encoding the open reading frame for G6PC1 - c.247C > T ( G6PC1 -R83C) were inserted into exon 1 of the mouse G6pc gene at the ATG start codon (Taconic Biosciences) in a way that created a premature STOP codon in the coding sequence of the mouse G6pc exon 1.

Techniques: Control, Activity Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Base-editing corrects metabolic abnormalities in a humanized mouse model for glycogen storage disease type-Ia

doi: 10.1038/s41467-024-54108-1

Figure Lengend Snippet: Three-week-old (3W) huR83C mice, non-fasted, were treated with 301L (BEAM-301 at 0.75 mg/kg) and the biochemical phenotype of the 3W-301L-dosed mice (3W-301L-53W), was evaluated at 53 weeks of age using the sex-matched mR83-53W and mR83/huR83C-53W littermates (Control-53W) as the controls. a BMI values in Control-53W ( n = 14) and 3W-301L-53W ( n = 9) mice. b Insulin tolerance test in Control-53W ( n = 11) and 3W-301L-53W ( n = 8) mice. A reduced insulin dose of 0.25 IU/kg was used because GSD-Ia mice have an increased insulin sensitivity . c ALT and AST values in Control-53W ( n = 12), 3W-301L-53W ( n = 9), mR83-53W ( n = 15) and NB-301H-53W ( n = 17) mice. d Histochemical analysis of liver G6Pase-α activity in Control-53W ( n = 6) and 3W-301L-53W ( n = 6) mice. Each image represents an individual mouse. Scale bar = 100 µm. The numbers represent hepatic G6Pase-α activity expressed in the mice. e Hematoxylin and eosin (H&E) and Oil Red O staining of the liver sections in Control-53W ( n = 6) and 3W-301L-53W ( n = 6). The liver in the six pairs of experimental animals were examined. A single image from an individual mouse, representative of the results in all mice, is shown to illustrate the results. Scale bar = 20 µm. The numbers represent hepatic G6Pase-α activity expressed in the mice. Statistics were performed using a two-tailed unpaired T test. Data are presented as Mean values ± SEM, and individual data points for each animal are displayed. * denotes p < 0.05, ** denotes p value < 0.005.

Article Snippet: A human cDNA encoding the open reading frame for G6PC1 - c.247C > T ( G6PC1 -R83C) were inserted into exon 1 of the mouse G6pc gene at the ATG start codon (Taconic Biosciences) in a way that created a premature STOP codon in the coding sequence of the mouse G6pc exon 1.

Techniques: Control, Activity Assay, Staining, Two Tailed Test