β-Cell Knockout of SENP1 Reduces Responses to Incretins and Worsens Oral Glucose Tolerance in High-Fat Diet–Fed Mice

Abstract

Lin et al. Supplementary FiguresSupplementary Figure 1. Additional in vivo glucose homeostasis assessment of male and female pSENP1-KO mice on CD. (A) Fasting insulin; (B) fasting glucose, (C) body weight, and (D) IP insulin tolerance of pSENP1-WT, -HET and -KO male mice on CD (n=6-22 mice). (E) Fasting insulin, (F) fasting glucose, (G) body weight, and (H) IP insulin tolerance of pSENP1-WT, -HET and -KO female mice on CD (n=7-17 mice). AUC -area under the curve. Data are mean ± SEM and were compared with student t test, one-way or two-way ANOVA followed by Bonferroni post-test. Unless stated otherwise, *-p<0.05, indicated comparison between pSENP1-WT and -KO. Supplementary Figure 2. In vivo glucose homeostasis assessment of female pSENP1-KO mice following HFD. (A) OGTT and (B) associated plasma insulin responses, (C) IP insulin tolerance, (D) body weight, (E) fasting glucose, and (F) fasting insulin of pSENP1-WT, -HET and -KO female mice following HFD (n=9-14 mice). Supplementary Figure 3. Islet morphometry analysis of female pSENP1-KO mice after HFD. (A) Representative immunostaining, -cell mass, islet number, and islet size distribution of female pSENP1-WT (n = 5 mice, 15 sections and 189 islets) and pSENP1-KO (n = 6 mice, 18 sections and 197 islets) following HFD. Insulin (green), glucagon (red), and nuclei (blue). Scale bar=100 µm. Data are presented as mean ± SEM. Lin et al. -cell SENP1 limits oral glucose intolerance 2 Supplementary Figure 4. Additional in vivo glucose homeostasis assessment of βSENP1-KO male mice. (A) IP insulin tolerance of male SENP1-WT, -HET, and -KO mice following HFD (n=15, 8, 8 mice). (B) Delta area under the curve (AUC) of IP insulin tolerance tests of male SENP1-WT, -HET, and -KO mice on CD and following HFD (n=6, 6, 6, 15, 8, 8 mice). (C) Fasting insulin of male SENP1-WT, -HET, and -KO mice on CD and following HFD (n=6, 4, 5, 13, 17, 14). (D) fasting glucose (n=15-22 mice) levels and (E) Body weight (n=18-23 mice) in male SENP1-WT and -KO mice on CD and following HFD. Data are presented as mean ± SEM. AUC -area under the curve. Data are mean ± SEM and were compared with student t test, one-way or two-way ANOVA followed by Bonferroni post-test. Unless stated otherwise, *-p<0.05, indicated comparison between βSENP1-WT and -KO. Supplementary Figure 5. In vivo glucose homeostasis assessment of βSENP1-KO female mice. (A) Fasting insulin (n=6, 8, 4, 13, 16, 14 mice), (B) fasting glucose (n=21, 22, 20, 21, 21, 22), and (C) body weights (n=30, 23, 25, 20, 22, 22) of female SENP1-WT, -HET, and -KO mice on CD and following HFD. (D) OGTT (n=11, 12, 12 mice), (E) IPGTT (n=10, 11, 9 mice), and (F) IP insulin tolerance (n=6, 9, 8) of female βSENP1-WT, -HET and -KO mice on CD. (G) OGTT (n=11, 13, 11 mice) and (H) associated plasma insulin response (n=7, 7, 8 mice) of female SENP1-WT, -HET, and -KO mice following HFD. (I) IPGTT (n=8, 9, 10 mice), and (J) associated plasma insulin response (n=6, 9, 6 mice) of female SENP1-WT, -HET, and -KO mice following HFD. (K) IP insulin tolerance (n...

Zmiz1 is required for mature β-cell function and mass expansion upon high fat feeding

“…The worsened glucose intolerance in response to oral compared to IP glucose suggests an implication of incretin response, which is known to be enhanced following HFD (Ahrén et al, 2008; Gupta et al, 2017; Yamane et al, 2016). This is consistent with our previous observations in HFD-mouse models (Lin et al, 2021).…”

“…While Zmiz1 interacts with the androgen receptor (AR), male mice with specifical deletion of the Ar in β-cells showed no obvious defect in islet mass or mass expansion upon metabolic stress (Navarro et al, 2016). The HFD-fed female mice were more resistant to the development of insulin resistance and glucose intolerance, consistent with previous reports that female mice are protected against HFD-induced metabolic changes (Lin et al, 2021; Pettersson et al, 2012), and as such islet mass expanded very little in the female controls.…”

“…Isolated islets were hand-picked or subjected to purification using Histopaque Gradient Centrifugation (Smith, Lin, et al, 2018) and were cultured overnight. Glucose-stimulated insulin secretion was determined as previously described (Lin et al, 2021).…”

Loss of SUMO-specific protease 2 causes isolated glucocorticoid deficiency by blocking adrenal cortex zonal transdifferentiation

“…As such, alterations of this posttranslational modification have been associated with chronic pathologies such as cancers, degenerative and metabolic diseases [13]. From a physiological perspective, conditional gene invalidation of SUMO enzymes has provided functional evidence that altering the cellular SUMOylation rheostat by installing either a deficit (targeting E2 conjugating enzyme) or an excess (targeting SENP deconjugases), has an impact on critical functions such as cardiac and neuronal physiology [22,46], intestinal [18] and uterine stem cells homeostasis [47], adipose differentiation and metabolism [17,48], pancreatic β-cell secretion [49], macrophages polarisation [50] and immune tolerance [19]. However, how this rheostat can be challenged by or adapted to physiological demand remains unclear.…”

Zmiz1 is required for mature β-cell function and mass expansion upon high fat feeding

“…Isolated islets were hand-picked or subjected to purification using Histopaque Gradient Centrifugation [ 19 ] and were cultured overnight. Glucose-stimulated insulin secretion was determined as previously described [ 20 ].…”