Journal: bioRxiv

Article Title: Antisense oligonucleotides against monoacylglycerol acyltransferase 1 ( Mogat1 ) improve glucose metabolism independently of Mogat1

doi: 10.1101/2020.08.05.238535

Figure Lengend Snippet: Constitutive liver-specific Mogat1 deletion does not improve insulin sensitivity in mice. Male Mogat1 fl/fl mice and littermate Mogat1 fl/fl albumin Cre+ mice were fed a LFD or a HFD starting at eight weeks of age for 16 weeks. Mice were fasted for 4 hours prior to sacrifice and tissue collection. A: HFD increased bodyweight in both groups. B,C: HFD fed mice have impaired glucose and insulin tolerance compare to LFD groups. D-G: Mogat1 gene expression is reduced in knockout liver and primary hepatocytes without significant compensation of Mogat2. H,I: HFD increased liver weight and TAG content in both genotypes. J: MGAT activity was not affected by diet or genotype. Data are expressed as means ± S.E.M. # p < 0.05 gene effect, † p < 0.05 diet effect; n = 5-10 for mouse studies, n = 3-5 female mice for primary hepatocyte isolations.

Article Snippet: For hepatic Mogat1 overexpression, eight-week-old male C57BL6/J mice were given LFD or HFD for six weeks, then administered AAV8-TBG-eGFP or AAV8-TBG-mouse- Mogat1 by retro-orbital injection (Vector Biolabs, VB1743, custom refseq# BC106135).

Techniques: Gene Expression, Knock-Out, Activity Assay

Journal: bioRxiv

Article Title: Antisense oligonucleotides against monoacylglycerol acyltransferase 1 ( Mogat1 ) improve glucose metabolism independently of Mogat1

doi: 10.1101/2020.08.05.238535

Figure Lengend Snippet: Acute liver-specific deletion of Mogat1 does not improve glucose or insulin tolerance in HFD fed mice. Male Mogat1 fl/fl mice were fed a HFD starting at eight weeks of age. After 16 weeks mice were given a retro-orbital injection of AAV8-TBG-eGFP or Cre recombinase (2 x 10 11 GC per mouse) and remained on diet for an additional 3 weeks. Mice were fasted for 4 hours prior to sacrifice and tissue collection. A: Mogat1 knockdown did not affect body weight. B,C: Acute liver-specific knockout did not improve glucose or insulin tolerance. D: Mogat1 knockout reduced Mogat1 expression without increasing Mogat2 expression in liver. E-F: Liver weights and TAG content were not difference among groups. G: Hepatic MGAT activity was not reduced by acute Mogat1 knockdown in liver. Data are expressed as means ± S.E.M. # p < 0.05 gene effect; n = 9.

Article Snippet: For hepatic Mogat1 overexpression, eight-week-old male C57BL6/J mice were given LFD or HFD for six weeks, then administered AAV8-TBG-eGFP or AAV8-TBG-mouse- Mogat1 by retro-orbital injection (Vector Biolabs, VB1743, custom refseq# BC106135).

Techniques: Injection, Knockdown, Knock-Out, Expressing, Activity Assay

Journal: bioRxiv

Article Title: Antisense oligonucleotides against monoacylglycerol acyltransferase 1 ( Mogat1 ) improve glucose metabolism independently of Mogat1

doi: 10.1101/2020.08.05.238535

Figure Lengend Snippet: Whole-body deletion of Mogat1 causes weight gain and insulin intolerance on a HFD. Male wild-type (WT) and littermate Mogat1 whole body knockout (MOKO) mice were fed a LFD or a HFD starting at eight weeks of age for 16 weeks. Mice were fasted for 4 hours prior to sacrifice and tissue collection. A: Mogat1 knockout mice gain more weight on a HFD than littermate WT controls. B: ECHO MRI indicates MOKO mice have increased whole body mass, while the heterozygous mice had an intermediate phenotype compared to WT controls. C: Glucose tolerance (dosed on lean mass) was not significantly change in MOKO mice. D,E: HFD fed MOKO mice had significantly impaired insulin tolerance and plasma insulin levels compared to WT controls. F: Mogat1 gene expression was not detectable in MOKO mice. G,H: Liver weight but not TAG was increased in MOKO mice fed a HFD compared to LFD controls. I: MGAT activity was unaffected by either diet or genotype. Data are expressed as means ± S.E.M. # p < 0.05 gene effect, † p < 0.05 diet effect; n = 6-7.

Article Snippet: For hepatic Mogat1 overexpression, eight-week-old male C57BL6/J mice were given LFD or HFD for six weeks, then administered AAV8-TBG-eGFP or AAV8-TBG-mouse- Mogat1 by retro-orbital injection (Vector Biolabs, VB1743, custom refseq# BC106135).

Techniques: Knock-Out, Clinical Proteomics, Gene Expression, Activity Assay

Journal: bioRxiv

Article Title: Antisense oligonucleotides against monoacylglycerol acyltransferase 1 ( Mogat1 ) improve glucose metabolism independently of Mogat1

doi: 10.1101/2020.08.05.238535

Figure Lengend Snippet: Hepatic Mogat1 overexpression increases liver TAG and MGAT activity in mice fed a LFD. Male C57BL6/J mice were fed LFD or a HFD starting at eight weeks of age. After 6 weeks of diet mice were injected (retro-orbitally) with AAV8-TGB-eGFP- or AAV8-TGB-Mogat1 (2 x 10 11 GC per mouse) and remained on diet for an additional 10 weeks. Mice were fasted for 4 hours prior to sacrifice and tissue collection. A: HFD fed mice gained weight compare to LFD fed mice in both treatment groups. B,C: Mogat1 overexpression did not impair glucose or insulin tolerance. D-F: Mogat1 and GFP gene expression was significantly increased in AAV8-Mogat1 and AAV8-GFP treated mice, respectively without a change in Mogat2 gene expression. G: Liver weight was unaffected by any treatment. H,I: AAV8-Mogat1 overexpression increased both liver TAG and MGAT activity in LFD mice. J: Western blot analysis indicated Mogat1 and eGFP protein increased in membrane fractions of AAV8 treated livers. Data are expressed as means ± S.E.M. # p < 0.05 gene effect, † p < 0.05 diet effect; n = 8-10.

Article Snippet: For hepatic Mogat1 overexpression, eight-week-old male C57BL6/J mice were given LFD or HFD for six weeks, then administered AAV8-TBG-eGFP or AAV8-TBG-mouse- Mogat1 by retro-orbital injection (Vector Biolabs, VB1743, custom refseq# BC106135).

Techniques: Over Expression, Activity Assay, Injection, Gene Expression, Western Blot, Membrane

Journal: bioRxiv

Article Title: Antisense oligonucleotides against monoacylglycerol acyltransferase 1 ( Mogat1 ) improve glucose metabolism independently of Mogat1

doi: 10.1101/2020.08.05.238535

Figure Lengend Snippet: shRNA mediated Mogat1 knockdown did not improve glucose or insulin tolerance on a HFD. Male C57Bl/6J mice were fed a 10% LFD or a 60% HFD starting at eight weeks of age. After 16 weeks of diet mice were injected (retro-orbitally) with AAV8-U6-shRNA Scramble or shRNAs targeted against Mogat1 (2 x 10 11 GC per mouse) and remained on diet for an additional three weeks. Mice were fasted for 4 hours prior to sacrifice and tissue collection. A: HFD fed mice gained weight compare to LFD Scramble treated controls. B,C: HFD fed mice had impaired glucose and insulin tolerance compared to LFD Scramble control treated mice. D,E: Liver weight and TAG were increased by the HFD but unaffected by Mogat1 shRNA treatment. F,G: Mogat1 ShRNA treatment reduced Mogat1 expression but did not affect Mogat2 expression. Data are expressed as means ± S.E.M. # p < 0.05 from HFD shRNA Scramble control mice, † P < 0.05 from LFD shRNA Scramble controls; n = 7-10.

Article Snippet: For hepatic Mogat1 overexpression, eight-week-old male C57BL6/J mice were given LFD or HFD for six weeks, then administered AAV8-TBG-eGFP or AAV8-TBG-mouse- Mogat1 by retro-orbital injection (Vector Biolabs, VB1743, custom refseq# BC106135).

Techniques: shRNA, Knockdown, Injection, Control, Expressing, Mouse Assay

Journal: bioRxiv

Article Title: Antisense oligonucleotides against monoacylglycerol acyltransferase 1 ( Mogat1 ) improve glucose metabolism independently of Mogat1

doi: 10.1101/2020.08.05.238535

Figure Lengend Snippet: Mogat1 antisense oligonucleotide (ASO, sequence 1) treatment improves insulin sensitivity in HFD fed mice. Male C57Bl/6J mice were fed a LFD or a HFD starting at eight weeks of age. After 16 weeks of diet, mice were injected (intraperitoneally) twice weekly with ASOs targeted against Mogat1 or scramble control (25 mg/Kg) for three weeks. Mice were fasted for 4 hours prior to sacrifice and tissue collection. A: HFD fed mice gained more weight compare to LFD fed controls. B,C: Control ASO treated mice had significantly higher blood glucose and plasma insulin than LFD Control ASO treated mice and HFD Mogat1 ASO treated mice. D: Mogat1 ASO treatment significantly reduced hepatic expression of Mogat1, but not Mogat2 as measured by RT-qPCR. E: Mogat1 ASO treatment increased liver weight (%BW) on HFD. F: Liver triglycerides (TAG) were measured enzymatically and were increased by the HFD. G: Hepatic gene expression of Gpase, Pepck, and Pygl were reduced in the HFD Mogat1 ASO treated mice. H: Glycogen was extracted and measure enzymatically and increased in HFD Mogat1 ASO treated mice. Data are expressed as means ± S.E.M. # p < 0.05 ASO effect, † p < 0.05 diet effect; n = 7-10.

Article Snippet: For hepatic Mogat1 overexpression, eight-week-old male C57BL6/J mice were given LFD or HFD for six weeks, then administered AAV8-TBG-eGFP or AAV8-TBG-mouse- Mogat1 by retro-orbital injection (Vector Biolabs, VB1743, custom refseq# BC106135).

Techniques: Sequencing, Injection, Control, Clinical Proteomics, Expressing, Quantitative RT-PCR, Gene Expression

Journal: bioRxiv

Article Title: Antisense oligonucleotides against monoacylglycerol acyltransferase 1 ( Mogat1 ) improve glucose metabolism independently of Mogat1

doi: 10.1101/2020.08.05.238535

Figure Lengend Snippet: Mogat1 ASO treatment improves glucose tolerance in whole-body Mogat1 null mice on a HFD. Male wild-type (WT) and littermate Mogat1 whole body knockout (MOKO) mice were fed a HFD starting at eight weeks of age. After 16 weeks of diet, mice were injected (intraperitoneally) twice weekly with ASOs targeted against Mogat1 or scramble control (25 mg/Kg) for three weeks. Mice were fasted for 4 hours prior to sacrifice and tissue collection. A,B: Mogat1 ASO treatment improves glucose tolerance despite Mogat1 expression. C,D: Mogat1 ASO treatment causes hepatomegaly and increases liver TAG in wild-type mice on HFD. E: Mogat1 gene expression was reduced by ASO treatment and nearly absent in MOKO livers. F-I: Gene expression markers of interferon signaling are increased in Mogat1 ASO treated MOKO mice on HFD. Data are expressed as means ± S.E.M. # p < 0.05 from Scramble ASO † p < 0.05 from WT controls; n = 5-7.

Article Snippet: For hepatic Mogat1 overexpression, eight-week-old male C57BL6/J mice were given LFD or HFD for six weeks, then administered AAV8-TBG-eGFP or AAV8-TBG-mouse- Mogat1 by retro-orbital injection (Vector Biolabs, VB1743, custom refseq# BC106135).

Techniques: Knock-Out, Injection, Control, Expressing, Gene Expression, Allele-specific Oligonucleotide

Journal: bioRxiv

Article Title: Antisense oligonucleotides against monoacylglycerol acyltransferase 1 ( Mogat1 ) improve glucose metabolism independently of Mogat1

doi: 10.1101/2020.08.05.238535

Figure Lengend Snippet: Type I interferon receptor (INFR-1) neutralizing antibody prevents inflammation without effecting glucose tolerance in mice fed a HFD. Male C57BL/6J mice were fed a HFD starting at eight weeks of age. After 16 weeks of diet, mice were injected (intraperitoneally) twice weekly with ASOs targeted against Mogat1 (sequence 2) or scramble control (25 mg/Kg) with an INFR-1 neutralizing antibody (INFR-Ab) or IgG control (250 ug per mouse twice a week for weeks 1&2, and 500 ug per mouse 3 times for week 3). Mice were fasted for 4 hours prior to sacrifice and tissue collection. A,B: Mogat1 ASO treatment improves glucose tolerance despite INFR-1 antibody treatment. C,D: Liver weight and TAG were unaffected by any treatment E: Liver Mogat1 gene expression was reduced by Mogat1 ASO treatment compared to controls. F-I: Liver gene expression markers of interferon signaling are increased in Mogat1 ASO treated mice and inhibited by INFR-Ab. J: Epididymal Mogat1 gene expression was decreased by Mogat1 ASO treatment. K-N: Epididymal gene expression markers of interferon signaling are inhibited by INFR-Ab. O-Q: Expression of adipose beiging genes were increased by Mogat1 ASO and inhibited by INFR-Ab treatment. Data are expressed as means ± S.E.M. # p < 0.05 from Scramble ASO † p < 0.05 from IgG controls; n = 5.

Article Snippet: For hepatic Mogat1 overexpression, eight-week-old male C57BL6/J mice were given LFD or HFD for six weeks, then administered AAV8-TBG-eGFP or AAV8-TBG-mouse- Mogat1 by retro-orbital injection (Vector Biolabs, VB1743, custom refseq# BC106135).

Techniques: Injection, Sequencing, Control, Gene Expression, Expressing, Allele-specific Oligonucleotide