bluuebunny/crossref_metadata_2025_split huggingface dataset:
203-OR: VGLL4 and MENIN Function as TEAD1 Corepressors to Block Pancreatic ß-Cell Proliferation
Authors: LI FENG , LIU RUYA , NEGI VINNY , YANG PING , LEE JEONGKYUNG K. , MOULIK MOUSUMI , YECHOOR VIJAY 
Published: May 2022

β-cell mass is significantly reduced especially in the late stage of diabetes. Hence, increasing β-cell proliferation to increase β-cell mass remains a desirable goal for diabetes treatment. The mechanisms that regulate β cell proliferation and concurrent maintenance of mature function is not well understood. Hippo-TEAD pathways regulated proliferation in many tissues. Our data show that TEAD1 (Hippo) pathway activities were significantly reduced in mature β cell. TEAD1 knockout results in the cell-autonomous increase of β cell proliferation, while loss of TAZ has not change in β cell proliferation, suggesting that TEAD1 corepressors, rather than coactivators, play a dominant role in β-cells. Improved Split-GFP system and yeast two-hybrid platform were also adapted to screen TEAD1 cofactors in β cells. Our study showed that MENIN can bind to TEAD1 directly and the pocket area of TEAD1 is critical for this binding. VGLL4 and MENIN work as TEAD1 corepressors to inhibit the expression of target genes in β-cells, including FZD7 and CCN2, which leads to a restriction of β cell proliferation. We also found that TEAD1 can regulate TEAD1, and this auto-inhibition contributes to TEAD1 pathway inhibition with β cell maturity. Our study sheds light on the mechanisms involved in β cell maintenance of mature function and proliferation and offer the mechanistic basis for modulating TEAD pathway to increase proliferation in β cells.

Disclosure
F.Li: None. R.Liu: None. V.Negi: None. P.Yang: None. J.K.Lee: None. M.Moulik: None. V.Yechoor: n/a.
212-LB: Decreasing Drp1 Expression in Mouse Liver Activates the Mitochondrial Integrated Stress Response and Exacerbates NASH
Authors: STEFFEN JANOS , NGO JENNIFER , WANG SHENG-PING , KRAMER HENNING F. , NORQUAY LISA D. , POCAI ALESSANDRO , SHIRIHAI ORIAN , LIESA MARC 
Published: May 2022

Elevated Drp1-mediated mitochondrial fission was proposed to promote NAFLD, as inhibition of hepatic Drp1 in early life prevented high-fat diet induced liver steatosis in mice. However, whether Drp1-knockdown (Drp1si) can reverse established NASH in adulthood is unknown. Hepatocyte-restricted delivery of siRNA conjugated to N-acetyl-galactosamine (GalNac) is a safe and FDA-approved approach to selectively manipulate hepatic gene expression and treat human liver disease. Here, we show that hepatic Drp1si in adult healthy mice decreased body weight by 12% and induced liver damage, as shown by the elevation in circulating transaminases, liver inflammation, fibrosis and necrosis (p&lt;0.001) . Decreased body weight was largely explained by Gdf15 actions decreasing food intake by 11%, as Gdf15 receptor (Gfral) KO prevented the decrease in food intake and mitigated the weight loss caused by Drp1-KD. An activation of the Atf4- integrated stress response (ISR) in liver explained the 4x fold increase in hepatic Gdf15 expression. Drp1si increased both ER stress and the activity of the mitochondrial protease Oma1, two processes that converge in increasing EIf2alpha phosphorylation and Atf4 expression. While NASH induced by GAN diet did not activate Oma1 by itself, Drp1si in mice with NASH was sufficient to activate Oma1 and exacerbate NASH-induced ER-stress. These led to higher ISR activation, exacerbated inflammation, fibrosis and necrosis.
In conclusion, our study supports that increased Drp1 activity is an adaptative mechanism counteracting NASH that prevents ISR overactivation and mitigates ER stress to limit fibrosis, inflammation, and necrosis. Our study argues against blocking Drp1 in hepatocytes to combat NASH.

Disclosure
J. Steffen: Employee; Janssen Research & Development, LLC. J. Ngo: None. S. Wang: None. H. F. Kramer: Employee; Janssen Research & Development, LLC. L. D. Norquay: Employee; Janssen Pharmaceuticals, Inc. A. Pocai: Employee; Johnson & Johnson. O. Shirihai: None. M. Liesa: None.
221-LB: Normalisation of Glycemic and Extraglycemic Parameters with T2DM Remission—Insights from RCT Enabled by Whole-Body Digital Twin Technology
Authors: SHAMANNA PARAMESH , SHAH LISA , MOHAMED MALUK , MOHAMMED JAHANGIR , POON TERRENCE , DHARMALINGAM MALA , VADAVI ARUN , SABOO BANSHI , DAMODHARAN SURESH , KESHAVAMURTHY ASHOK , THAJUDEEN MOHAMED , BHONSLEY SUCHITRA , JOSHI SHASHANK R. 
Published: May 2022

We analyzed the efficacy of Twin Precision Treatment (TPT) : a novel whole-body digital twin enabled precision nutrition that measures weight, physical activity, sleep, and sensor glucose values, for achieving remission, at six months among 199 people with T2D enrolled in RCT. The mean age was 43 years (±8.7) ; diabetes duration was 3.7 years (±2.7) . 83.9% (167/199) achieved diabetes remission (HbA1c &lt; 6.5, with the absence of glucose-lowering pharmacotherapy, for at least 3 months) . We looked at the change in the number and percentage of patients who were within the normal range for the biochemical, glycemic metrics, metabolic markers, and BMI, waist circumference, and ASCVD risk score (Figure) . There was a significant increase in the number of people having normal ranges at 90 and 180 days compared to baseline. The parameters for which ≥ 90% in the normal range at 180 days were A1C, FPG, GMI, CV, NAFLD FS, MICR, ASCVD risk score, and hsCRP. The proportion of normalisation rates were 60-90% for HOMA2B, HOMA2IR, BMI, waist circumference, TG/HDL, sdLDL, NAFLD LFS, TIR. Our study demonstrates the diverse metabolic benefits which can be obtained by whole-body digital twin enabled precision nutrition in addition to diabetes remission.

Disclosure
P. Shamanna: None. A. Keshavamurthy: None. M. Thajudeen: None. S. Bhonsley: None. S. R. Joshi: Advisory Panel; AstraZeneca, Consultant; Biocon, Franco India, Glenmark Pharmaceuticals, Marico, Sanofi, Twinhealth, Zydus Cadila, Speaker's Bureau; Abbott, Serdia Pharmaceuticals (India) Pvt. Ltd. L. Shah: None. M. Mohamed: None. J. Mohammed: None. T. Poon: None. M. Dharmalingam: None. A. Vadavi: None. B. Saboo: None. S. Damodharan: None.
223-LB: Interactive Effects of Dietary Iron and Chronic Hypoxia on Glucose Homeostasis in a T2D Mouse Model
Authors: HARRISON ALEXANDRIA V. , MCCLAIN DONALD A. , SINK SANDY T. , LORENZO FELIPE 
Published: May 2022

The micronutrient iron is a risk factor for type II diabetes (T2D) . The effects of iron are, in part, mediated by the iron-sensing hypoxia inducible factor (HIF) pathway. The interactions of iron and hypoxia in the progression of T2D are not fully understood. Mice were fed a fast-food (FF) diet supplemented with either normal iron (35 mg/kg) or high iron (2000 mg/kg) under normoxia and hypoxia. Hypoxic mice had a significant reduction in weight compared to normoxic mice (by 16% for 35mg-Fe diet and 14% for 2000mg-Fe diet, p&lt;0.001) . There was an interactive effect of iron and hypoxia on fasting glucose and glucose tolerance (p=0.016, p=0.41 respectively) in which the combination of hypoxia and normal iron yielded the best improvement. For insulin-mediated glucose uptake mechanisms, insulin sensitivity was observed with high iron and also under hypoxia when using the homeostatic model assessment (HOMA-IR) . Direct measurement using the insulin tolerance test showed no additional sensitivity was observed with hypoxia, suggesting non-insulin mediated glucose uptake (NIMGU) mechanisms. Indirect calorimetry demonstrates that hypoxia and iron interact to increase carbohydrate utilization and energy expenditure (p&lt;0.001 and p=0.005, respectively) . Measured eWAT tissue mass showed significant reduction in tissue mass for hypoxic mice. Transcriptional profile of mouse eWAT shows significant upregulation of classic HIF target genes involved in glycolysis, mitochondrial metabolism, and intracellular iron homeostasis in hypoxic mice and interestingly also in normoxic high iron mice. Lastly, protein expression of HIF regulators FIH1 and PHD2 were significantly reduced (25-40% reduction, p&lt;0.05 for FIH and p&lt;0.01 for PHD2) under hypoxia. We conclude that the observed effects of dietary iron are dependent on oxygen levels, with the best protection against T2D offered by normal iron and hypoxia. Enhanced glucose disposal is due to NIMGU mechanisms, with the two HIF regulatory arms, PHD2 and FIH1, also mediating effects. Results from this study support novel targeted therapies and dietary changes to treat T2D.

Disclosure
A. V. Harrison: None. D. A. Mcclain: None. S. T. Sink: None. F. Lorenzo: None.

Funding
NIDDK Research Service of the Veterans Administration
226-OR: Reduced Fatty Acid Oxidation Prevents Intralipid-Induced Hepatic Insulin Resistance in Humans
Authors: PURNELL JONATHAN Q. , GREGOR ASHLEY N. , CHOI DONGSEOK , BAETSCHER ERIC , ROONEY WILLIAM , HARDING CARY O. , GILLINGHAM MELANIE B. 
Published: May 2022

Mitochondrial handling of excess lipid substrate may be an important determinant of insulin resistance. Animal models support insulin signaling defects resulting from both defective fatty acid oxidation (FAO) with subsequent intra-cellular lipid accumulation on the one hand and enhanced FAO flux on the other. To explore the relationship between FAO and insulin sensitivity, we measured the change in intra-hepatic lipids (IHL) by MRS and hepatic insulin sensitivity during a 5-hour hyperinsulinemic euglycemic clamp with co-infusion of either a control glycerol solution versus a 20% intralipid solution, in random order and separated by 4 months. The study cohort included 18 subjects with an inherited defect in FAO (FAOD; including Carnitine Palmitoyltransferase 2, Long chain 3-Hydroxyacyl-CoA Dehydrogenase, Very Long-chain or Medium-chain Acyl-CoA Dehydrogenase Deficiencies) and 18 matched normal healthy controls. Endogenous glucose production (EGP) before the start of the insulin infusion was similar between FAOD and controls. In controls, insulin suppressed EGP during the glycerol control clamp but this effect was significantly reduced during intralipid infusion (P&lt;0.05, reduced hepatic insulin sensitivity) . In FAOD subjects, insulin suppressed EGP similarly during the glycerol and intralipid clamps (no impairment in insulin action) . Untargeted lipidomics /metabolomics of plasma measured at the end of the intralipid clamp demonstrated significant elevations of plasma acylcarnitines and some triglyceride species but lower production of ketones, and several phospholipids in subjects with FAOD compared to controls. Neither increased IHL nor elevated plasma acylcarnitines in FAOD subjects were associated with hepatic insulin resistance. In fact, impaired FAO protected against intralipid-induced hepatic insulin resistance suggesting that increased FAO flux rather than impaired mitochondrial function is a major determinant of insulin resistance in the liver.

Disclosure
J.Q.Purnell: Advisory Panel; Novo Nordisk. A.N.Gregor: None. D.Choi: None. E.Baetscher: None. W.Rooney: Consultant; Ultragenix, Research Support; Italofarmco, Revalesio. C.O.Harding: None. M.B.Gillingham: None.

Funding
NIH/NIDDK: R01DK102813NIH/NCATS: UL1TR002369
228-LB: Mitochondrial Architecture Controls Fatty Acid Utilization by Regulating CPT1 Sensitivity to Malonyl-CoA
Authors: NGO JENNIFER , WOOK CHOI DONG , STILES LINSEY , DIVAKARUNI AJIT S. , LIESA MARC , DANIAL NIKA N. , SHIRIHAI ORIAN 
Published: May 2022

Mitochondrial network fragmentation has been associated with metabolic dysregulation, although the underlying mechanisms remain unknown. Herein, using multiple cellular models where the physiological relevance of fatty acid is well-established, we show that mitochondrial fragmentation is highly associated with increased fatty acid oxidation (FAO) rates. Through confocal imaging and stable isotope tracing analysis, forced mitochondrial elongation using Mfn2 over-expression or Drp1 depletion significantly decreased FAO rates, while Mfn2 knockout leads to enhanced FAO rates (P-value =0.0003) , suggesting that mitochondrial morphology regulates fatty acid utilization. To determine the mechanism by which mitochondrial architecture controls fatty acid utilization we studied the effects of mitochondrial fragmentation and elongation on CPT1 activity. Remarkably, we find that CPT1 sensitivity to its main regulator, malonyl-CoA is strongly affected by molecular interventions that inhibit mitochondrial fusion and fission. Notably, the reduction in hepatic FAO rates upon mitochondrial elongation is attributed to a 14 fold increase in CPT1 sensitivity to malonyl-CoA. Consistent with this mechanism we show that the inhibition of FAO imposed by mitochondrial elongation can be overcome using CPT1 pharmacological activator that is malonyl-CoA-independent, but not by the malonyl-CoA-sensitive activator. Furthermore, we tested the functional implications of this mechanism in hepatocytes and beta cells. In hepatocytes we show that forced elongation by the inhibition of fission reduced gluconeogenesis by 30-70%. Whereas in islets, forced mitochondrial fragmentation through molecular inhibition of fusion, resulted in a 2.5 fold increase in insulin secretion at basal glucose concentrations, a phenomenon associated with prediabetes and obesity. Taken together, our study provides a biochemical and mechanistic explanation by which changes to mitochondrial architecture affects fuel utilization and lipid metabolism.

Disclosure
J. Ngo: None. D. Choi: None. L. Stiles: None. A. S. Divakaruni: None. M. Liesa: None. N. N. Danial: None. O. Shirihai: None.

Funding
American Diabetes Association (1-19-IBS-049)
236-LB: Sex Differences in Lipid Storage and Metabolism Underlay Dimorphic Inflammatory Responses in Obesity
Authors: MOHAN RAMKUMAR , VARGHESE MITA , ABRISHAMI SIMIN , SINGER KANAKADURGA 
Published: May 2022

Obesity-induced inflammation is a risk factor for metabolic impairment leading to diabetes and cardiovascular disease. We, and others, have shown that sex differences exist in adipose tissue (AT) and myeloid inflammatory responses to high-fat diet (HFD) and lipolysis. However, the causal cellular mechanisms behind this sexual dimorphism are not well defined. Our objective was to investigate sex differences in lipid storage and metabolism that drive sexually dimorphic inflammatory responses. C57Bl/6J mice were fed a normal diet or 60%kcal%fat (HFD) for 16 weeks. AT macrophages (ATMs) were sorted for RNA sequencing. Animals were gonadectomized (GX) prior to starting HFD and gonadal white AT (GWAT) was collected for lipidomics. To directly model androgen receptor (AR) activation, GX mice were given dihydrotestosterone (DHT) to study AR-specific effects. RNA sequencing identified upregulated genes (Lpl, Ctsb, and Cd36) linked to a metabolically-activated macrophage only in male HFD ATMs. Furthermore, pathways associated with mitochondrial depolarization and fatty acid (FA) metabolism differed significantly by sex. GWAT lipidomics identified shifts in lipid species both by sex and hormone status, and differences were also observed in acylcarnitines and saturation of ceramide species that are implicated in inflammation. In addition, males showed elevated levels of key precursor free FAs (palmitic, linoleic, and arachidonic acid) when compared to female and GX mice. DHT treatment did not completely mimic male pro-inflammatory responses but induced GWAT crown-like structures in both sexes. DHT failed to upregulate inflammatory markers (Il6 and Mcp1) but showed impaired expression of adipogenic genes (Pparg and Pgc1α) in GX mice. Altogether, our data suggest there are sex-specific alterations in GWAT lipid storage and metabolism that may drive sexually dimorphic ATM polarization, which is only partially reversed by AR activation.

Disclosure
R. Mohan: None. M. Varghese: None. S. Abrishami: None. K. Singer: None.

Funding
National Institutes of Health - NIDDK (R01DK115583)
236-OR: Sleep Deprivation Prevents Counterregulatory Adaptation to Recurrent Hypoglycaemia
Authors: MEYHOEFER SVENJA , DEMBINSKI KATHARINA , SCHULTES BERND , JAN BORN , WILMS BRITTA , LEHNERT HENDRIK , HALLSCHMID MANFRED , MEYHÖFER SEBASTIAN M. 
Published: May 2022

Aims/Hypothesis: Attenuated counterregulation after recurrent hypoglycaemia is a major complication of diabetes treatment. Considering previous evidence for the relevance of sleep in metabolic control, we assessed the acute contribution of sleep to the counterregulatory adaptation to recurrent hypoglycaemia.
Methods: Within a balanced cross-over design, 15 healthy, normal-weight male participants aged 18-35 years underwent three hyperinsulinaemic-hypoglycaemic clamps with a glucose nadir of 2.5 mmol/L, under two experimental conditions, sleep and sleep deprivation (SD) . Participants were exposed to two hypoglycaemic episodes, followed by a third hypoglycaemic clamp after one night of regular 8-hour sleep vs. SD. The counterregulatory response of relevant hormones (glucagon, growth hormone (GH) , adrenocorticotropic hormone (ACTH) , cortisol, epinephrine, and norepinephrine) . Moreover, autonomic and neuroglycopenic symptoms were assessed.
Results: SD compared to sleep dampened the adaptation to recurrent hypoglycaemia of epinephrine (p=0.004) , and this pattern also emerged in an overall analysis including epinephrine, GH, and glucagon (p=0.064) . Whereas after regular sleep, counterregulatory responses of epinephrine (p=0.005) , GH (p=0.029) , and glucagon (p=0.009) were attenuated during the third compared to the first clamp, they were preserved after SD (all p&gt;0.225) . Neuroglycopenic and autonomic symptoms during the third compared to the first clamp were likewise reduced after sleep (p=0.0 and p=0.019, respectively) . In SD, neuroglycopenic symptoms increased (p=0.014) and autonomic symptoms remained unchanged (p=0.859) .
Conclusions/Interpretation: The counterregulatory adaptation to recurrent hypoglycaemia is compromised by SD between hypoglycaemic episodes, indicating that sleep is essential for the formation of a neuro-metabolic memory and a potential target of interventions to improve hypoglycaemia unawareness.

Disclosure
S.Meyhoefer: n/a. K.Dembinski: None. B.Schultes: None. B.Jan: None. B.Wilms: None. H.Lehnert: None. M.Hallschmid: None. S.M.Meyhöfer: None.

Funding
DFG (German Research Funding Organisation) : TR-SFB654-B1
239-LB: Single-Cell Analysis Identified Inhibitory Neurons Associated with High-Fat Diet in the Bed Nucleus of the Stria Terminalis
Authors: YANG XIUXIU , FLAK JONATHAN , JOHNSON TRAVIS S. 
Published: May 2022

Obesity occurs due to inadequate homeostatic function controlled by the central nervous system (CNS) . Outside of a few brain areas, little information is available on structures, such as the bed nucleus of the stria terminalis (BST) and preoptic area (POA) , that are essential for body weight regulation. To better understand BST and POA mechanisms involved in obesity, we overlayed deep sequencing results collected from obese (by Echo MRI) and diabetic (by glucose tolerance test) mice on high fat diet (HFD) with a previously published single cell RNA-seq data set. We identified neural subtypes using Seurat (Fig. 1A) and deconvoluted the RNA-seq with CIBERSORT. The Inhibitory 1 subtype had a lower proportion in HFD BST than control BST (Fig. 1B, p=0.035) but no difference was found between BST and POA (Fig. 1C, p=0.505) . We performed differential gene expression analysis to identify Inhibitory 1 markers and associated pathways. Aminobutyric acid (GABA) -ergic synapse pathway was enriched (Fig. 1D,E, p&lt; 0.0001) , which is essential to the CNS control of body weight. HFD induced strong transcriptional changes in BST Inhibitory 1 markers between HFD and control diet (Fig. 1F) . One gene of particular interest was Glutathione Peroxidase 3 (GPX3, p&lt;0.0001) . GPX3, localized to the BST and POA (Fig. 1G) , is down-regulated in both prostate and adipose tissues during HFD and implicated in neurodegenerative disease, but not yet been studied in the brain and obesity. Future studies will investigate BST GPX3 in obesity.

Disclosure
X. Yang: None. J. Flak: Research Support; Eli Lilly and Company. T. S. Johnson: Research Support; Eli Lilly and Company.
248-LB: Nrf2 Is Essential for Expanding Neonatal Pancreatic Beta-Cell Mass
Authors: BAUMEL-ALTERZON SHARON , GARCIA-OCANA ADOLFO , SCOTT DONALD 
Published: May 2022

Patients with both major forms of diabetes suffer from insufficient functional β-cell mass. Moreover, the sharp increase in the annual incidence of both type 1 (1.8%) and type 2 (4.8%) diabetes in the pediatric population in the last decades highlights the urgent need for discovering the mechanisms that regulate functional β-cell mass. β-cell proliferation, which reaches its maximal peak during postnatal stages is the major mechanism for increasing β-cell mass in early postnatal ages. We previously found that induction of the antioxidant regulator, Nrf2, is required for adaptive β-cell expansion under hypercaloric conditions via stimulation of β-cell proliferation, increased β-cell survival, and maintenance of β-cell identity in adult mice. Based on these observations, we hypothesize that Nrf2 regulates β-cell proliferation and survival at early stages of life and contribute to maintain normal β-cell mass later in life. Nrf2 expression levels were found to correlate with β-cell proliferation in wild type neonatal mice. Importantly, 28 day-old β-cell specific Nrf2 knockout mice (Ins1Cre-Nrf2lox/lox) exhibited 50% reduction in β-cell mass, 58% reduction in total islet number, 43% decrease in extra-large islets, 46% decrease in large islets and 53% decrease in extra small islets, compared to control mice. No changes in pancreas weight were observed. Additionally, depletion of Nrf2 in β-cells increased β-cell death in 7 (5.8-fold) and 14 (12.5-fold) day-old mice and attenuated β-cell proliferation in 1- and 7-day-old mice. Surprisingly, we also found a marked decrease in islet size in 1- and 7-day-old mice, suggesting that Nrf2 may affect β-cell proliferation, survival and/or development during embryonic stages. We conclude that Nrf2 is required for β-cell mass expansion in postnatal ages by controlling β-cell proliferation and survival.

Disclosure
S. Baumel-alterzon: None. A. Garcia-ocana: Consultant; Sun Pharmaceutical Industries Ltd. D. Scott: None.