bluuebunny/crossref_metadata_2025 huggingface dataset:
1360-P: Calorie Restriction Enhanced Glycogen Metabolism to Compensate for Lipid Insufficiency
Authors: HU LILI , XIA XINYI , GU YUNJIE , YIN JUN
Published: May 2022

Backgrounds: It's well known that calorie restriction (CR) brings striking benefits in metabolic improvement with complex and unclear mechanism. In this study, we aimed to investigate the specific manifestation and possible mechanisms of metabolism in CR mice.

Methods: B6 mice were randomly assigned to three groups: (1) chow diet (control) , (2) chow diet with 40% calorie restriction (CR) , (3) high-fat diet (HFD) . The metabolism status and related examination were performed after 12 weeks of feeding.

Results: In this study, we found that CR increased fasting blood glucose and plasma insulin, as well as reduced insulin sensitivity reflected by insulin tolerance test. Comparing to the control mice, the decrease of phosphorylation levels of IRS-1 including serine307 and tyrosine632 residues, and AKT (threonine308) after insulin rejection further confirmed that CR induced insulin resistance. Different from insulin resistance induced by HFD, CR improved glucose tolerance and reduced gluconeogenesis. Meanwhile, CR led to absolute lipid reduction, manifested in decreased plasma lipid, liver lipid deposition and fat mass. In contrast, liver glycogen increased dramatically. Fasting glycogen was twice as much as that of HFD group and 20 times as that of control group. Furthermore, the CR increased p-PYGL, p-GSK3β and decreased p-GS notably indicating active glycogen synthesis and decomposition. Subsequently, we found that CR increased plasma corticosterone and decreased plasma c-reactive protein (CRP) compared with control and HFD groups, while plasma corticosterone and CRP in HFD were higher than those in control group. Additionally, CR lowered levels of orexigenic neuropeptides and elevated levels of anorexigenic neuropeptides.

Conclusions: Our study suggested that CR induced lipid insufficiency and stress from starvation, resulting in physiological insulin resistance and enhanced glycogen metabolism, including increased glycogen synthesis, glycogenolysis and glycogen content.

Disclosure

L.Hu: None. X.Xia: None. Y.Gu: None. J.Yin: None.

Funding

National Natural Science Foundation of China (No. 82070885) ,Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Grant Support (No. 20172025)
1368-P: Skeletal Muscle Mitochondrial Dysfunction Contributes to Increased GDF15 Levels in Aged Mice with Insulin Resistance and Sarcopenia
Authors: CHEN JEFFREY , BELLO FIONA , KASTROLL JAKE , VANDEVENDER AMBER , PANGBURN MARTHA , SILVA AGUSTIN A. GIL , SIPULA IAN J. , ALDER JONATHAN , JURCZAK MICHAEL J.
Published: May 2022

Growth differentiation factor 15 (GDF15) is a secreted protein whose cellular expression increases in response to a variety of stimuli, including mitochondrial dysfunction. Plasma GDF15 levels increase with advanced age and positively associate with sarcopenia. The prevalence of type 2 diabetes (T2D) increases with age and the incidence of sarcopenia is elevated in patients with T2D. Despite these established associations, the relationship between skeletal muscle mitochondrial function and changes in GDF15 expression, as well as how they may contribute to sarcopenia and T2D in the context of aging, remain poorly defined. We hypothesized that mitochondrial dysfunction in skeletal muscle would be positively associated with increased GDF15 expression and plasma levels in aged mice, and that muscle function and insulin sensitivity would be impaired also. Lean mass as a percentage of body weight was significantly reduced in aged (52 months) male C57BL6 compared with young (4 months) mice. Skeletal muscle function and mobility, assessed as peak and max sustained speeds achieved during voluntary wheel running, were significantly impaired in aged mice. Hyperinsulinemic euglycemic clamps demonstrated impaired insulin sensitivity in skeletal muscle, adipose and liver of aged mice. High-resolution respirometry studies using permeabilized muscle fibers detected significant reductions in the respiratory control ratio (RCR) in aged mice that was due to increased state 4 or leak respiration. Muscle GDF15 gene expression was significantly elevated in old mice, as were plasma levels of circulating GDF15. Our data suggests that skeletal muscle is a significant contributor to increased plasma GDF15 levels seen during aging, and that changes in mitochondrial function may contribute to increased skeletal muscle GDF15 expression. Elevated GDF15 may therefore serve as a strong predictor of mitochondrial pathology and subsequent risk of T2D in aging.

Disclosure

J.Chen: None. F.Bello: None. J.Kastroll: None. A.Vandevender: None. M.Pangburn: None. A.A.Gil silva: None. I.J.Sipula: None. J.Alder: None. M.J.Jurczak: None.
1370-P: Effects of Concomitant High-Intensity Interval Training or Resistance Exercise Training on Muscle Protein Synthesis, Body Composition, and Metabolic Markers following 6-Week VLCD among Healthy Overweight/Obese Men
Authors: AZIZ MUHAMMAD HARIZ ABDUL , RAMZAN IMRAN , GHARAHDAGHI NIMA , BROOK MATTHEW , WILKINSON DANIEL J. , PHILLIPS BETHAN E. , SMITH KEN , ATHERTON PHILIP J. , IDRIS ISKANDAR R.
Published: May 2022

Background: Very Low-Calorie Diet (VLCD) is effective for rapid weight loss but is associated with lean mass (LM) loss. Resistance exercise training (RET) and high-intensity interval training (HIIT) are known to promote muscle hypertrophy or improvements in cardiopulmonary fitness (CPF) . We therefore explored the effects of VLCD with or without RET or HIIT on body composition, metabolic markers, and CPF.

Methods: Participants were randomly allocated into VCLD only (n=10; 46±8y; 104±12kg; BMI 32±4, mean±SD) , VLCD+RET (n= 8; 40±8y;99k±10kg; BMI 32±2) or VLCD+HIIT (n=8; 46±11y; 101±12kg; BMI 33±3) . All groups had six weeks of VLCD. Exercise groups had supervised training three times/week. Pre- and post-interventions assessment included DXA scan, lipid profiles, insulin sensitivities, blood pressure, cardiopulmonary exercise test and biopsy of vastus lateralis muscle for muscle protein synthesis (MPS) using deuterium oxide technique.

Results: Significant reduction in body weight (-11.1±3 kg, -10.8±4kg, -11.7±4kg) , fat mass (-6.8±2kg, -6.9±2kg, -7.7±3kg, and LM (-4.3±2kg, -3.9±2kg, -4±1kg) were seen for VLCD only, VLCD+RET and VLCD+HIIT respectively (P<0.0001) , with no between-groups difference. MPS rate following interventions were 0.8%±0.2d-1, 0.9%±0.2d-1 and 1.2%±0.2d-1 (statistically higher in VLCD+HIIT compared to VLCD only; P=0.004) . Significant improvements were seen in metabolic markers (insulin sensitivities, lipid profiles and blood pressure) in all groups, with no inter-group difference. No intra- and inter-groups differences were seen for VO2max or anaerobic threshold.

Conclusion: VLCD with or without exercise induced marked weight loss, improved insulin sensitivities, and metabolic markers despite LM loss. HIIT was associated with significant improvement in MPS and may be used as an adjunct to preserve VLCD induced LM loss.

Disclosure

M.Abdul aziz: None. I.Ramzan: None. N.Gharahdaghi: None. M.Brook: None. D.J.Wilkinson: None. B.E.Phillips: None. K.Smith: None. P.J.Atherton: None. I.R.Idris: None.
1372-P: Hyperglycemia Impacts Mouse Skeletal Muscle in a Sex-Specific Manner
Authors: KNAUB LESLIE , POTT GREGORY , HULETT NICHOLAS A. , HULL SARA , SCALZO REBECCA L. , REUSCH JANE
Published: May 2022

Functional Exercise Capacity (FEC) , a universal predictor of cardiovascular and all-cause mortality, is lower in adults and youth with diabetes for reasons that are poorly understood. Skeletal muscle is key determinant of FEC. We hypothesized that skeletal muscle capillary and mitochondrial density are lower with hyperglycemia and respond to exercise training. To test this hypothesis we generated a model of short-term hyperglycemia (HG) with inducible beta cell deletion of glucokinase (GKlox/lox;PdxCreER) . Male (M) and female (F) HG mice and euglycemic littermate controls (EG) completed 3 weeks of treadmill training (Ex) or remained sedentary (Sed) . The gastrocnemius was collected 24 hours after the final exercise bout. Expression of COX1 (mitochondrial marker) , Dystrophin (muscle fiber sarcolemma boundary marker) , and CD31 (capillary marker) was detected by immunofluorescence. Sirtuin 3 (SIRT3) , an NAD+ dependent deacetylase (mitochondrial homeostasis) was measured using the JESS system. (Fig 1) Mitochondrial density (mitochondria per fiber as quantified by fluorescence intensity units per fiber [FIU/fiber]) was significantly lower in HG vs. EG (p<0.0001) and differed by sex (p=0.0048 sex x glycemia interaction) . Ex resulted in greater mitochondrial density (p=0.0265) independent of sex and glycemia. Capillary density and SIRT3 were unchanged. Sex-differences in muscle response to HG warrant further investigation.

Disclosure

L.Knaub: None. G.Pott: None. N.A.Hulett: None. S.Hull: None. R.L.Scalzo: None. J.Reusch: Advisory Panel; Medtronic.

Funding

VA Merit (BX002046)
138-LB: Improving Type 2 Diabetes Medication Prescribing
Authors: RIOLES NICOLE , DESIMONE MARISA , HESS EMILIE J. , IWUMMADU CHINONSO V. , OSPELT EMMA L. , ROMPICHERLA SAKETH , GREENFIELD MARGARET , TSAI SANDRA
Published: May 2022

Background: For people with type 2 diabetes, implementing best practice guidelines for diabetes and cardiovascular comorbidities is important to reduce the risks of complications. Two adult medical clinics used population health EMR data reports to increase medication prescribing for statins, ACE & ARB, SGLT2 & GLP-1.

Methods: The clinics tracked measures and used data analytics to identify a cohort population and medication needs for 6241 patients. Table 1 reflects data from a one-year pilot. Patients with a diagnosis of type 2 diabetes, duration of disease for at least one year, ages 18-75, with at least one in-person or telehealth visit were included.

Results: Three medication prescribing baselines were tracked to improve type 2 diabetes and heart disease management. Prescribing rates of SGLT2 and GLP1 increased by 2%, ACE-I and ARB use increased by 1% and statins rates remained at 73%.

Conclusion: The clinics used data to identify the cohort and prescribe necessary medications. Partnering with clinical pharmacists helped to support patients, educate providers, and standardize medication prescribing workflows. Planning care so that the teams consistently identified appropriate medications for diabetes and cardiovascular disease management were clinical priorities. While the goal rate of medication prescribing has not been achieved yet, new reporting workflows are now incorporated, setting up the clinics for continued improvement.

Disclosure

N. Rioles: None. M. Desimone: None. E. J. Hess: None. C. V. Iwummadu: None. E. L. Ospelt: None. S. Rompicherla: None. M. Greenfield: None. S. Tsai: None.

Funding

Helmsley Charitable Trust
138-OR: Earlier Onset of Type 2 Diabetes Increases Genetic Risk of Cardiovascular Complication
Authors: LEE HYUNSUK , CHOI JAE WON , KIM NA YEON , LEE SEUNGGEUN , PARK KYONG SOO , KWAK SOO HEON
Published: May 2022

Earlier age at diagnosis of type 2 diabetes (T2D) is associated with increased risk of cardiovascular disease (CVD) . However, genetic risk factors associated with accelerated CVD in people with early onset T2D remain elusive. We aimed to investigate if the impact of polygenic risk for developing CVD differs according to age at diagnosis of T2D.

Using prospective cohorts of UK Biobank (T2D, N=15,915) and Seoul National University Hospital (SNUH) T2D cohort (N=1,228) , we compared the incidence of newly developed CVD among four groups based on age at diagnosis of T2D (30-39, 40-49, 50-59, and 60-69 years of age) using time to event analysis. Polygenic risk scores (PRSs) for T2D and coronary artery disease (CAD) were calculated by PRS-CS, Bayesian regression using continuous shrinkage priors, using 1.1 million SNPs each from summary statistics of DIAGRAM and CARDIoGRAMplusC4D Consortium.

In UK Biobank, people diagnosed as T2D between age 30-39 had 5.8 (95% CI 4.04-8.42) fold increased risk of CVD compared to those who developed T2D between 60-69. Higher T2D PRS was associated with earlier onset of T2D but not with risk of myocardial infarction (MI) . On the other hand, higher CAD PRS was associated with increased risk of MI but not with earlier age at diagnosis of T2D. The effect of CAD PRS on MI was largest in people diagnosed between age 30-39 and its effect size decreased as the age at diagnosis increased: HR 2.34 (1.47-3.71) for age 30-39, 1.42 (1.20-1.68) for 40-49, 1.39 (1.25-1.54) for 50-59, and 1.28 (1.11-1.48) for 60-69. Interaction analysis between age at diagnosis and CAD PRS showed additional 10% increase in risk of MI for year earlier onset of T2D (P=0.0056) . These findings were replicated in the SNUH T2D cohort, which is a hospital based cohort of East Asians.

In summary, people with earlier onset T2D had higher risks for CVD, and the impact of CAD PRS for incident MI increased as age at diagnosis decreased. Genetic information in people with early onset T2D could be used to identify those with increased risk of CVD.

Disclosure

H.Lee: None. J.Choi: None. N.Kim: None. S.Lee: None. K.Park: None. S.Kwak: None.
1381-P: GDF15 Deficiency in Brown Adipose Tissue Does Not Affect Thermogenesis, but Exacerbates Diet-Induced Obesity
Authors: JENA JAYASHREE , KATO KEVIN , MARTI ALEX A. , PEREIRA RENATA
Published: May 2022

Growth and differentiation factor 15 (GDF15) is induced in brown adipocytes in response to thermogenic stimuli, as well as high-fat feeding in mice. However, whether brown adipose tissue (BAT) -derived GDF15 is required to regulate BAT thermogenesis or energy homeostasis is unclear. In the present study, we sought to test the hypothesis that BAT-derived GDF15 is necessary to mediate thermogenesis and systemic adaptations to diet-induced obesity (DIO) . We generated mice lacking GDF15 specifically in BAT by crossing Gdf15 floxed mice with mice harboring the Cre recombinase under the control of the Ucp1 promoter (GDF15 BAT KO) . GDF15 BAT KO mice were treated with the beta adrenoreceptor agonist CL316,243 (CL) for 5 days to induce thermogenesis, or fed a high-fat diet (60% fat content) for 12 weeks to induce DIO. CL treatment promoted a significant induction in Gdf15 mRNA levels in the BAT of WT mice, which, as expected, was prevented in KO mice. Nonetheless, serum GDF15 levels were similarly induced by CL in both WT and KO mice. CL-induced activation of thermogenic genes in BAT, such as Ucp1 and Dio2, was comparable between genotypes, and core body temperature was unchanged between WT and KO mice. After 12 weeks of high-fat feeding, KO mice had significantly increased body weight and fat mass relative to WT mice. However, food intake, locomotor activity and energy expenditure were unchanged between groups. Respiratory exchange ratio was significantly higher in KO mice, indicating a relative preference for carbohydrate as fuel. Although glucose tolerance was unchanged between genotypes, insulin sensitivity was impaired in KO mice. Taken together, our data indicates that BAT-derived GDF15 does not contribute to GDF15 circulating levels in response to CL treatment and is largely dispensable for CL-induced BAT thermogenesis. However, BAT-derived GDF15 seems to be required to regulate diet-induced weight gain and insulin sensitivity.

Disclosure

J.Jena: None. K.Kato: None. A.A.Marti: None. R.Pereira: None.

Funding

NIHDK125405
1391-P: Reduced DMPC and PMPC in Lung Surfactant Promotes SARS-CoV-2 Infection in Obesity
Authors: DU KANG , SUN LING , LUO ZICHEN , CAO YANG , SHAN JINJUN , YANG QIN
Published: May 2022

Background: Obesity is an established risk factor for higher SARS-CoV-2 viral loads, severe COVID-pneumonia requiring hospitalization, and worse outcomes. However, the underlying mechanisms for the increased risk are not well understood. SARS-CoV-2 is a respiratory virus with the primary route of entry through lungs, where the Spike protein of SARS-CoV-2 binds to ACE2 receptor on pneumocytes. Lung surfactant produced by type II pneumocytes plays a major role in respiratory defense against infections. Surfactant predominantly contains lipids especially phosphatidylcholines (PC) and obesity is characterized by aberrant lipid metabolism. We hypothesized that altered lipid composition in lung surfactant in obesity may promote SARS-CoV-2 infection, leading to severe COVID-disease.

Methods: Lipidomic analysis of lung tissue and bronchoalveolar lavage fluid (BALF) was performed using LC-MS/MS. The effects of PCs on SARS-CoV-2 pseudovirus infection were studied in HEK293T cells with ACE2 overexpression and in Vero-E6 cells with endogenous ACE2 expression.

Results: Lipidomic analysis revealed that myristic acid containing dimyristoyl-PC (DMPC) and palmitoylmirystoyl-PC (PMPC) were commonly reduced in lung tissue and BALF from high fat diet-induced obese mice. DMPC and PMPC markedly inhibited wild type and D614G mutant SARS-CoV-2 infection in HEK293T-ACE2 and Vero-E6 cells. Feeding obese mice with trimyristin, the triglycerides of myristic acid, increased DMPC and PMPC in lung surfactant. Lipid extract from BALF of trimyristin-treated obese mice reduced wild type and D614G mutant SARS-CoV-2 infection. The inhibitory effects of DMPC and PMPC on SARS-CoV-2 infection were reversed by cholesterol.

Conclusions: The reduced DMPC and PMPC in lung surfactant contributes to the increased SARS-CoV-2 infection. Increasing DMPC and PMPC in lung surfactant may be an innovative strategy for preventing and treating severe COVID-disease in obesity.

Disclosure

K.Du: None. L.Sun: None. Z.Luo: None. Y.Cao: None. J.Shan: None. Q.Yang: None.

Funding

National Institutes of Health (R01DK121146) ;Pacific Life Foundation Fellowship to UC Irvine Diabetes Center;Chinese National Natural Science Foundation (No.81774156)
1392-P: Gut Microbiota Contributes to the Beneficial Metabolic Effects of Gelesis Hydrogel Technology
Authors: SILVESTRI ALESSANDRA , GIL-GOMEZ ANTONIO , CHIQUETTE ELAINE , DEMITRI CHRISTIAN , JONES BRYAN W. , SANNINO ALESSANDRO , RESCIGNO MARIA
Published: May 2022

Gelesis' Oral superabsorbent hydrogels (OSH) are crosslinked cellulose-based materials that expand upon hydration in the gastrointestinal tract and are engineered to mimic the mechanical properties of masticated vegetables. One OSH prototype, denoted Gel-B, was tested in both preventative and treatment settings in murine models of diet-induced obesity. Gel-B administration blunted weight gain, reversed gut atrophy, improved metabolic parameters (glucose and insulin tolerance tests; GTT and ITT) and restored barrier function. Here, we investigated the functional role of the gut microbiome to partially explain these protective metabolic effects.

Metabolic disease was induced in both Donor (n=24) and Recipient (n=24) C57BL6/J mice via consumption of a high fat and high cholesterol/carbohydrate (HFHCC) diet for weeks (wks) . Donor mice continued on HFHCC or were treated with HFHCC+Gel-B (2 or 4%) for an additional 6 wks. After 2 wks treatment, Donor fecal samples were processed every other day for intestinal microbiota transplantation (IMT) . Recipient mice continued HFHCC and received IMT derived from either HFHCC or Gel-B (2 or 4%) treated Donors via gavage every other day for 4 wks.

At 4 wks, the HFHCC-corrected % body weight difference in Recipients was -10.8% and -9.6% for the 2% Gel-B-derived IMT (p=0.02) , and 4% Gel-B-derived IMT (p= 0.02) respectively. Glucose excursions, AUC 0-2h from GTT, were significantly reduced in both Recipient groups (p=0.for both 2% and 4% Gel-B-derived IMT) . The AUC 0-2h from ITT were significantly increased in the 4% Gel-B-derived IMT Recipients (p=0.04) and trended for the 2% Gel-B-derived IMT group (p=0.13) .

The present study demonstrates for the first time that IMT from Gel-B treated Donors confers beneficial metabolic effects in HFHCC treated mice. Future experiments will define Gel-B associated microbiota and examine engraftment and durability of IMT.

Disclosure

A.Silvestri: None. A.Gil-gomez: None. E.Chiquette: Employee; Gelesis. C.Demitri: Stock/Shareholder; Gelesis. B.W.Jones: Employee; Gelesis. A.Sannino: None. M.Rescigno: Research Support; Gelesis.
1393-P: Cardiac and Skeletal Muscles Exhibit Discordant Flux Adaptations as a Result of Western Diet–Induced Obesity
Authors: HASENOUR CLINTON M. , RAHIM MOHSIN , BEDNARSKI TOMASZ , BANERJEE DEVEENA R. , YOUNG JAMEY
Published: May 2022

The metabolic complications of obesity are distributed throughout the body, yet studies assessing its impact on metabolic fluxes have primarily focused on single tissues. Here we use a 13C tracer approach to simultaneously quantify metabolic flux dysregulation in the liver, skeletal muscle, and heart induced by Western diet (WD) feeding in a mouse model of obesity. Male wildtype (WT) and Mc4r-/- mice (KO) were fed chow until 8wks of age, when ∼50% of the KO cohort transitioned to WD (KO+WD) for 8wks. KO and KO+WD had increased adiposity, hepatic steatosis, HOMA-IR, and reduced glucose tolerance relative to WT by 16wks of age; however, only KO+WD had extensive liver fibrosis. Stable- and radio-labeled isotopes were infused and blood was sampled from fasted, conscious, and unrestrained mice (n ≥ 6 per group) through jugular vein and carotid artery catheters, respectively. Metabolic fluxes were estimated by fitting an integrated mathematical model of liver, skeletal muscle, and cardiac metabolism to 13C enrichment measurements of plasma and tissue metabolites taken at the end of the infusion. Endogenous glucose production was elevated in both KO cohorts, with WD further increasing hepatic gluconeogenic and citric acid cycle (CAC) fluxes. Glycolytic, anaplerotic, and CAC fluxes were lower in the skeletal muscle of KO and KO+WD compared to WT, which coincided with a decrease and increase in gene expression of glycolytic and β-oxidation enzymes, respectively. In the heart, however, WD feeding increased glycolytic and CAC fluxes as well as the gene expression of glycolytic and β-oxidation enzymes. Thus, cardiac and skeletal muscles exhibit discordant flux adaptations in obese, WD-fed mice, as WD accelerates aerobic glycolysis and terminal oxidation in the heart but not skeletal muscle. We expect the multi-organ fluxomics platform described here to broadly benefit research to understand systemic metabolism in health and disease.

Disclosure

C.M.Hasenour: Employee; Eli Lilly and Company. M.Rahim: None. T.Bednarski: None. D.R.Banerjee: None. J.Young: Consultant; Pfizer Inc., Research Support; Merck & Co., Inc., Stock/Shareholder; Metalytics, Inc.

Funding

National Institutes of Health (R01DK106348, U01CA235508, T32DK101003)
1394-P: Peri-Islet Glial-Cell Expansion Occurs in Response to High-Fat Diet-Induced Obesity and Hyperglycemia
Authors: NIU JINGJING , MOSS JENNIFER B. , ROSARIO WILFREDO , ARUMUGAM RAMAMANI , HACKNEY DARYL J. , HULL-MEICHLE REBECCA L. , TONG JENNY
Published: May 2022

Peri-islet glial cells (PIGCs) are positioned to control neuronal-endocrine-vascular cell communication within the islet, but their role in islet cell function is not known. Hypothalamic glial cells become reactive in response to high fat diet (HFD) feeding and are implicated in the pathogenesis of obesity and type 2 diabetes. We hypothesize that metabolic stress such as diet-induced obesity and hyperglycemia promote the expansion of PIGCs, similar to what has been observed in the central nervous system. To test this hypothesis, we used the glial cell marker, glial fibrillary acidic protein (GFAP) , to identify PIGCs and compared GFAP immunoreactivity in pancreas from the following groups. First, pancreas was harvested from HFD- or chow fed control C57BL/6J male mice after 2 days, 2 weeks or 8 weeks on diet (n=6-8/group) . Second, pancreas was taken from 12-wk old db/db male mice and their control littermates with or without empagliflozin (EMPA) treatment (20mg/kg/d for 6 weeks; n=4-7/group) . Islet PIGC area was quantified as GFAP-positive area/total islet area x100%. Mann-Whitney test and one-way ANOVA were used for data analysis. We found that HFD feeding increased mouse pancreatic peri-islet GFAP expression relative to chow controls at 2 days (HFD 1.4±0.2%, chow 0.8±0.1%; p=0.02) , 2 weeks (HFD 3.5±0.4%, chow 1.2±0.1%; p<0.001) and 8 weeks (HFD 5.4±0.3%, chow 1.8±0.2%; p=0.002) in association with body weight gain and impaired glucose tolerance. Similar results were observed in obese and hyperglycemic db/db mouse vs. control littermates (db/db 5.0±1.4%, control 1.2±0.4%; p=0.004) . There was a trend for EMPA treatment to reduce islet PIGC area in the db/db mice (EMPA 2.0±0.5, VEH 5.0±1.4%, p=0.08) .

In conclusion, these findings indicate that islet gliosis is observed in two obese and hyperglycemic mouse models and raise the possibility that pancreatic ‘gliosis’ may have a relevant role in islet-cell function regulation under metabolic challenge.

Disclosure

J.Niu: None. J.B.Moss: None. W.Rosario: Employee; Merck & Co., Inc. R.Arumugam: None. D.J.Hackney: None. R.L.Hull-meichle: Research Support; Casma Therapeutics. J.Tong: None.

Funding

National Institutes of Health/National Institute of Diabetes and Digestive and Kidney Diseases (R01DK097550) ;Department of Veterans Affairs (IBX004063)
1398-P: Lactobacillus rhamnosus Attenuates Depressive-Like Behavior in High-Fat Diet Fed Mice and Regulates Tyrosine Hydroxylase
Authors: SCHELL MAREIKE , WARDELMANN KRISTINA , HAUFFE ROBERT , RATH MICHAELA , CHOPRA SIMRAN , KLEINRIDDERS ANDRE
Published: May 2022

Prevalence of depression is higher in diabetic patients compared to healthy humans due to a shared pathogenesis including insulin resistance and inflammation. These factors are also linked to intestinal dysbiosis. Interestingly, preventive intake of probiotic lactobacilli has been shown to improve dysbiosis along with mood and metabolism. Yet, a potential therapeutic role and underlying mechanism of Lactobacillus rhamnosus GG (LGG, ATCC 53103) to improve emotional behavior in diet-induced obesity is unknown.

Male C57BL/6N mice were fed a low-fat diet (LFD, 10% kcal from fat) or high-fat diet (HFD, 45% kcal from fat) for 6 weeks and followed by daily oral gavage of vehicle or 1x108 CFU of LGG. Dark-Light Box Test was used to evaluate anxiety, Mousetail Suspension Test (MST) and Splash Test were used to assess depressive-like behavior and motivation for self-care. Cecal microbiome composition was analyzed using 16S rRNA Sequencing. Plasma and CSF were collected for metabolomic analysis, and gene expression was analyzed using qPCR.

We observed that 12 weeks of HFD feeding increased anxiety and depressive-like behavior. Strikingly, LGG decreased specifically depressive-like behavior in the MST which was confirmed by the Splash Test. Metagenomics analysis revealed that HFD but not LGG alters cecal microbiome composition, indicating a transient effect. Moreover, HFD feeding significantly altered plasma lipid metabolism whereas LGG decreased branched chain amino acids which associated with improved emotional behavior. In dopaminergic brain regions, LGG restored the HFD-induced decrease of tyrosine hydroxylase, the rate-limiting enzyme in dopamine synthesis, while not altering enzymes responsible for dopamine degradation.

Our data indicate that LGG attenuates specifically depressive-like behavior in HFD-fed mice with a molecular signature of restored dopamine synthesis.

Disclosure

M.Schell: Other Relationship; Novozymes A/S, Denmark. K.Wardelmann: None. R.Hauffe: None. M.Rath: None. S.Chopra: None. A.Kleinridders: Other Relationship; Novozymes.

Funding

German Ministry of Education and Research (BMBF) and the State of Brandenburg (DZD grant 82DZD00302 and BMBF grant 031B0569) .
1399-P: Quantitation of the Energy Cost of Physical Activity in Mice
Authors: SKOP VOJTECH , GUO JUEN , LIU NAILI , HALL KEVIN D. , GAVRILOVA OKSANA , REITMAN MARC L.
Published: May 2022

Mice are widely used for diabetes and obesity research, but the quantitative contribution of physical activity to mouse energy metabolism is poorly understood. The energy cost of activity (PAEE) is typically determined by regression of total energy expenditure (TEE) versus physical activity. Confounding factors are 1) that muscle-produced heat reduces cold induced thermogenesis by brown fat and 2) that core body temperature (Tb) increases during physical activity. Minimizing cold induced thermogenesis addresses the first problem. Prior studies used housing at 30°C, but 30°C is now known to be below thermoneutrality in the dark phase. Furthermore, the Tb increase accompanying physical activity increases energy expenditure by non-activity processes; this was not considered previously. To account for these factors, we studied mice by indirect calorimetry at an ambient temperature of 35 °C while measuring Tb by telemetry and physical activity by beam break. We developed a model, partitioning TEE into PAEE and resting energy expenditure (REE) . Significant features of the model are i) physical activity measurements are aligned to the measured TEE using the time constant of the indirect calorimetry system, ii) the REE term is adjusted for the measured Tb using a Q10 (van’t Hoff coefficient) of 3.0, and iii) the REE term is scaled for body mass. In this model, the within mouse PAEE was robustly fit by linear regression, yielding PAEE of 22.5 ± 0.8 (light) and 21.7 ± 1.0 (dark) cal/1000 beam breaks (n=17) . The correlations between measured and model-calculated TEE were r = 0.888 ± 0.013 (light) and 0.888 ± 0.0 (dark) . Under usual housing conditions (ambient temperature of 23 °C) , the average physical activity was 1390 ± 130 (light) and 4940 ± 390 (dark) beam breaks/h. Based on these data, PAEE accounts for 8.6 ± 0.9 % (light) and 21.7 ± 1.8 % (dark) of TEE. This rigorous determination of PAEE advances understanding the role of physical activity in mouse energy metabolism, improving the predictive value of the mouse as a model for human disorders.

Disclosure

V.Skop: None. J.Guo: n/a. N.Liu: None. K.D.Hall: None. O.Gavrilova: None. M.L.Reitman: None.

Funding

Intramural NIDDK
1402-P: Endogenous Erythropoietin Is Protective in Mice during High-Fat Diet to Obesity and Cardiac Function while Exogenous Erythropoietin Exacerbates Cardiac Function
Authors: LEE JEEYOUNG , ROGERS HEATHER , NOGUCHI CONSTANCE T.
Published: May 2022

Endogenous Erythropoietin (EPO) provides protection against obesity and heart function. Mice with EPO receptor restricted to erythroid tissue (ΔEPORE mice) become obese, glucose intolerant and insulin resistant. In male mice at 4 months of age, after three weeks of high fat diet feeding, ΔEPORE mice showed an increase in fat mass from to 13 grams compared to fat mass in wild type mice that more than doubled from 4 to grams. EPO treatment (i.p. 3000 U/kg, 3 times/week for three weeks) during high fat diet feeding in wild type mice reduced the increase in fat mass to 3 grams and had a minimal effect on fat mass in ΔEPORE mice. The EPO stimulated increase in hematocrit was comparable in wild type and ΔEPORE mice. Using echocardiography to measure cardiac systolic function and cardiac output after high fat diet feeding, without EPO treatment ejection fraction (%) and fractional shortening (%) of left ventricular contractility were decreased in ΔEPORE mice compared with wild type mice, suggesting that endogenous EPO provided a cardioprotective effect mediated by EPO receptor expression in non-erythroid cells. EPO treatment in wild type mice decreased peak velocity and left ventricular outflow tract velocity but there were no significant changes in ΔEPORE mice, indicating that the decreases in these parameters with EPO administration are independent of EPO stimulated erythropoiesis and mediated by EPO receptor in non-erythroid cells. EPO treatment also decreased ejection fraction and fractional shortening in wild type mice and had no further significant effect in ΔEPORE mice. These data provide evidence that while endogenous EPO may be protective for heart function, high dose EPO administration during high fat diet feeding in wild type mice compromises cardiac function. EPO modulation of heart function requires EPO receptor expression beyond erythroid cells and is distinct from EPO stimulated erythropoiesis.

Disclosure

J.Lee: None. H.Rogers: None. C.T.Noguchi: None.
1406-P: ZP8396, a Novel Amylin Analogue, Induces Weight Loss in DIO Rats with a Formulation Space at Physiological pH
Authors: SKARBALIENE JOLANTA , LUNDQVIST JOAKIM , VILLADSEN JESPER
Published: May 2022

ZP8396, currently in phase 1, is a long-acting, amylin analogue designed to improve solubility and stability and allow for co-formulation with other peptides at physiological pH. Here we present in-vivo efficacy and formulation design space of ZP8396. The acute effects on body weight (BW) loss after a single s.c. injection of ZP8396 (0,5, 3, 30, 300 nmol/kg) were measured in lean rats and monitored for 96 hours after dosing. The ability of ZP8396 (1, 3, and 15 nmol/kg, s.c., every 2nd day for 21 days) to reduce BW was tested in diet-induced obese (DIO) rats. ZP8396, at concentration supporting pharmacological relevant human doses, is stable in an aqueous formulation at neutral pH and shows no aggregation potential. ZP8396 is compatible with different physiological buffer systems and isotonic agents, commonly used for commercial medicinal products (Table 1) . Formulation of ZP8396 at physiological pH induces a significant BW loss in both lean and DIO rats vs. vehicle group. A significant BW loss was observed from start of the dosing and throughout the study period.

In conclusion, ZP8396 potently induces BW loss in lean and DIO rats and allows for aqueous stable formulation at neutral pH which provides opportunities for co-formulation with other peptides for optimal BW loss. Further preclinical characterization of co-formulation of ZP8396 with different peptides for weight management is ongoing.

Disclosure

J.Skarbaliene: Employee; Zealand Pharma A/S, Stock/Shareholder; Zealand Pharma A/S. J.Lundqvist: Employee; Zealand Pharma A/S. J.Villadsen: Employee; Zealand Pharma A/S.
1409-P: Revealing Heterogeneity in Human White Adipose Tissue at Single-Cell Level
Authors: EFTHYMIOU VISSARION , KODANI SEAN , GUPTA ANUSHKA , SHAMSI FARNAZ , STREETS AARON , TSENG YU-HUA , PATTI MARY-ELIZABETH
Published: May 2022

White adipose tissue is characterized by substantial functional heterogeneity based on anatomic location and developmental lineages. Such differences may contribute to depot-dependent differences in adipose regulation of systemic metabolism and the relative impact of adipose distribution (e.g. subcutaneous, SQ, vs. intraabdominal, IA) on risk for type 2 diabetes. While mouse adipose depot heterogeneity at the single-cell level has recently been explored, comparative analysis of human white fat depots is still in its infancy.Hereby, we establish a single-nucleus RNA sequencing (snRNA Seq) method for isolating and sequencing intact single nuclei from frozen human WAT biopsies. Using this approach, we analyzed the transcriptome of individual nuclei from 22 WAT SQ or IA biopsies (SQ, 12 IA, 3 paired) acquired from subjects (16 females, 3 males) with or without type 2 diabetes (T2D) across a wide BMI range (Mean+/-SD = 45.4+/-10.3 kg/m2. Range: 23.6 - 60.9 kg/m2) . We show that both SQ and IA human fat depots demonstrate substantial cell-to-cell heterogeneity, and identify multiple adipose and non-adipose-related cell types resident in both tissues. Our analysis identifies several novel subclusters, including two distinct mature adipocyte subclusters, characterized by low vs. high adiponectin expression and thus potentially contributing to adipose-mediated effects on systemic metabolism. Moreover, we identify a novel cluster present in both SQ and IA depots, with a unique transcriptomic signature marked by DAPK1 and enrichment for hedgehog signaling and endocytosis KEGG terms. Comprehensive cross-sample integrative bioinformatics analysis using the software tools scVI and VISION reveals distinct molecular markers and transcriptomic signatures between depots, between clusters, and across a range of BMI, that may serve as promising targets for understanding the mechanism by which specific adipose-resident cell populations mediate metabolic risk associated with adiposity.

Disclosure

V.Efthymiou: None. S.Kodani: None. A.Gupta: None. F.Shamsi: None. A.Streets: None. Y.Tseng: Consultant; Cellarity. M.Patti: Consultant; AstraZeneca, Eiger BioPharmaceuticals, Hanmi Pharm. Co., Ltd., MBX Biosciences, Poxel SA, Other Relationship; Fractyl Health, Inc., Xeris Pharmaceuticals, Inc., Research Support; Dexcom, Inc.

Funding

Chan Zuckerberg Initiative (CZI)
1410-P: Changes in Circulating MicroRNA-193b and Long-Term Successful Weight Loss in Response to Dietary and Lifestyle Interventions: The POUNDS Lost and CENTRAL Trials
Authors: HEIANZA YORIKO , XUE QIAOCHU , ROOD JENNIFER , KROHN KNUT K. , MEIR ANAT YASKOLKA , BLÜHER MATTHIAS , KOVACS PETER , SHAI IRIS , BRAY GEORGE , SACKS FRANK , QI LU
Published: May 2022

Circulating microRNAs (miRNAs) have been implicated in controlling whole-body metabolism through intercellular communications. MicroRNA-193b (miR-193b) plays essential roles in white and brown adipocyte adipogenesis and regulating adipokine secretion. We tested whether changes in circulating miR-193b were associated with reductions of adiposity and various fat depots in response to weight-loss dietary and lifestyle interventions in two independent trials. This study included participants of a 2-year weight-loss diet intervention trial (POUNDS Lost) with data on changes in circulating miR-193b-5p from baseline to 6 months after interventions (n=509) . We also assessed changes in circulating miR-193b-5p over 18 months in the CENTRAL weight-loss lifestyle intervention trial (n=111) . At baseline, higher miR-193b was associated with greater adiposity, energy expenditure, ectopic fat depots, and lower adiponectin levels. In response to the interventions, decreases in miR-193b were associated with larger reductions of weight (p <.0001) , waist circumference (p <.0001) , and whole-body total % fat mass (p=0.03) at 6 months in the POUNDS Lost trial. In the CENTRAL trial, decreases in miR-193b were associated with larger reductions of weight (p=0.03) , total (p=0.002) and visceral adipose tissue (p=0.004) , and different fat depots (deep and superficial subcutaneous, and hepatic fat; p <0.for all) . In both trials, every 1 SD decrease of miR-193b was associated with an increased probability for the achievement of successful weight loss (-5% or more weight loss) at the end of interventions (odds ratio [OR]: 1.4; p=0.0in POUNDS Lost; OR 1.9; p=0.02 in CENTRAL) .

In conclusion, changes in circulating miR-193b in response to weight-loss diet/lifestyle interventions were significantly associated with the long-term successful weight loss and reductions of fat depots closely related to the risk of diabetes.

Disclosure

Y.Heianza: None. F.Sacks: None. L.Qi: None. Q.Xue: None. J.Rood: None. K.K.Krohn: None. A.Yaskolka meir: None. M.Blüher: Consultant; AstraZeneca, Lilly, Novo Nordisk A/S, Pfizer Inc., Sanofi, Speaker's Bureau; Bayer AG, Boehringer Ingelheim International GmbH, Novartis AG. P.Kovacs: None. I.Shai: None. G.Bray: None.

Funding

NIH (DK091718, DK100383, DK115679)
1412-P: Preexisting Nonalcoholic Steatohepatitis Attenuates Improvement of Adipose Tissue OXPHOS Capacity after Weight-Loss Surgery
Authors: KAHL SABINE , STRASSBURGER KLAUS , GANCHEVA SOFIYA , SAATMANN NINA , GRANATA CESARE , HERDER CHRISTIAN , PAFILI KALLIOPI , PUETZER-FURMANCZAK JENNIFER , SARABHAI THERESIA , HEILMANN GERONIMO , ESPOSITO IRENE , SCHLENSAK MATTHIAS , RODEN MICHAEL
Published: May 2022

Impaired adipose tissue (AT) function closely associates with obesity, type 2 diabetes and nonalcoholic fatty liver disease (NAFLD) . Mitochondrial oxidative phosphorylation (OXPHOS) capacity of AT improves after weight loss surgery. However, it is not clear if the improvement is affected by the presence of NAFLD before surgery. We examined 52 severely obese people exhibiting no histological liver disease (OBE_CON) , simple steatosis (OBE_NAFL) or steatohepatitis (OBE_NASH) before (0 weeks) , 12 and 52 weeks after bariatric surgery. Whole-body insulin sensitivity was measured by hyperinsulinemic euglycemic clamps. OXPHOS capacity of subcutaneous AT was assessed by high-resolution respirometry. Transcript levels of TNFα in AT were quantified by RT-PCR. Before surgery, OBE_NASH presented with lower insulin sensitivity and higher HbA1c compared to OBE_CON and OBE_NAFL (all p<0.05) whereas OXPHOS capacity in AT was comparable between the groups. Maximum coupled respiration declined at week 12 by 17% from week 0 in OBE_CON only (p<0.05) . From 12 to 52 weeks, AT maximum coupled respiration markedly increased by 82 and 49% in OBE_CON and OBE_NAFL (both p<0.001) but only by 28% in OBE_NASH (p=0.05; maximum coupled OXPHOS at week 52: OBE_CON vs. OBE_NASH p<0.05) . The changes in AT maximum coupled respiration from 12 to 52 weeks correlated negatively with changes in AT TNFα levels in OBE_CON (β=-0.83, p<0.001) and negatively with changes in systemic C-reactive protein levels in OBE_NASH (β=-0.34, p<0.05) . Maximal noncoupled OXPHOS yielded similar changes and correlations.

In conclusion, in severe obesity, weight loss-related improvements in AT OXPHOS capacity may be mediated to a great extent by reduced AT inflammation in people without liver disease; however, with pre-surgery manifestation of NASH and worsening of metabolic control, residual systemic subclinical inflammation may attenuate the improvement of AT OXPHOS capacity after bariatric surgery.

Disclosure

S.Kahl: None. G.Heilmann: None. I.Esposito: None. M.Schlensak: None. M.Roden: Advisory Panel; Eli Lilly and Company, Research Support; Boehringer Ingelheim International GmbH, Nutricia, Speaker's Bureau; Novo Nordisk. K.Strassburger: None. S.Gancheva: None. N.Saatmann: None. C.Granata: None. C.Herder: Research Support; Sanofi. K.Pafili: None. J.Puetzer-furmanczak: None. T.Sarabhai: None.

Funding

German Federal Ministry of Health; Ministry of Culture and Science of the State North Rhine-Westphalia;German Federal Ministry of Education and Research;European Funds for Regional Development (EFRE-0400191) ;EUREKA Eurostars-2 (E!113230DIA-PEP) ;German Science Foundation (CRC/SFB1116/2 B12; RTG/GRK 2576 vivid, Project 3) ;Schmutzler Stiftung
1419-P: Insulin Resistance Predicts Severe Acute COVID-and Long COVID
Authors: AYHAN EKREM M. , CAO SHU , RAEBER OLIVIA , CHINTHRAJAH SHARON , NADEAU KARI , MCLAUGHLIN TRACEY
Published: May 2022

Background: Obesity and diabetes are known risk factors for severe acute COVID-19. About half of individuals with obesity have insulin resistance (IR) , which may potentiate severe acute disease and/or predispose to long COVID.

Methods: We identified 596 adults from Stanford hospital or clinics confirmed COVID-19+ by rtPCR and categorized according to severity of illness based on the NIH categories1. Pre-COVID predictors including BMI, fasting plasma glucose (FPG) , and triglyceride/HDL-cholesterol ratio (TG/HDL) as a surrogate2 for IR were abstracted from the EMR using the Stanford Research Repository Tools software. Follow-up surveys were administered via REDCap 2-4x in the first month and 1x per month for 1 year thereafter to assess symptom type and duration. Long COVID was defined as symptom duration > 30 days. Metabolic predictors of acute COVID-severity (BMI, FPG, TG/HDL) were evaluated via multiple linear regression adjusted for sex, age, ethnicity, and other metabolic predictors. A logistic regression to predict long COVID included all metabolic predictors along with age, sex, and ethnicity. Models were repeated with a stepwise approach to increase statistical power given the smaller number of participants with complete data.

Results: Participants were 51±18 years of age, 49% female, and 62% racial and ethnic minorities. Mean BMI was 29.5±7.9 kg/m2. NIH illness severity was 7.9% asymptomatic, 37.2% mild, 25.9% moderate, 15.1% severe, and 13.9% critical. Diabetes, lung disease, and hypertension prevalence were 27%, 18%, and 34%, respectively. Mean follow-up was 94±99 and incidence of long COVID was 47.1%. BMI, FPG, and TG/HDL were independently associated with acute COVID-severity in the cohort as a whole (r=0.187, P<0.001; r=0.180, P=0.002; r=0.180, P=0.009) , while only TG/HDL was associated with long COVID (r=0.173, p=0.013) .

Conclusions: Findings suggest that IR, independent of sex, age, ethnicity, obesity and hyperglycemia, confers increased risk for both severe acute COVID-and long COVID.

Disclosure

E.M.Ayhan: None. S.Cao: None. O.Raeber: None. S.Chinthrajah: Advisory Panel; Alladapt Therapeutics, Novartis, Other Relationship; Aimmune, Astellas, Consortium of Food Allergy Research (CoFAR) , DBV Technologies, FARE, MCHRI , NIAID, Regeneron. K.Nadeau: None. T.Mclaughlin: Board Member; January, Inc., Research Support; Merck & Co., Inc., Novo Nordisk, Stock/Shareholder; Eiger BioPharmaceuticals.

Funding

National Institutes of Health (28291) National Institutes of Health (198537) National Institutes of Health (196111) Stanford Diabetes Research Center (128573)
1421-P: Long-Term Type 2 Diabetes Remission after Bariatric Surgery in Patients with and without Liver Steatosis
Authors: LAUTENBACH ANNE , WERNECKE MARIE , MEYHÖFER SEBASTIAN M. , MEYHÖFER SVENJA , STOLL FABIAN D. , ABEREL JENS
Published: May 2022

Background: Nonalcoholic steatohepatitis (NASH) is considered the hepatic manifestation of insulin resistance. Therefore, we aimed to assess the association between biopsy-proven liver steatosis and long-term remission of type 2 diabetes (T2D) 8 years following different bariatric procedures.

Methods: In a retrospective cohort study including 249 patients with and without T2D, the association between biopsy-proven NASH and long-term remission of T2D 8 years following sleeve gastrectomy (SG) and Roux-enY-gastric bypass (RYGB) has been assessed.

Results: Out of 249 patients, 15.3% showed NASH and T2D at the time of surgery. 8 years after surgery, T2D remission was achieved in 44.7% of patients with NASH compared to 76.0% without NASH. Patients with T2D remission were younger, had higher BMI, displayed lower HbA1c and lower preoperative insulin use (p<0.001) . Patients without remission of T2D showed higher steatosis scores (p<0.05) . In a multivariate logistic regression, higher preoperative HbA1c (OR 0.41) , insulin use (OR 0.16) and preexisting liver fibrosis (OR 0.19) decreased the probability of long-term T2D remission (p<0.05) . Liver steatosis, hepatocyte ballooning or lobular inflammation were not significantly associated with T2D remission. Patients without T2D were predominantly female, of younger age (p<0.001) and displayed lower scores of steatosis, hepatocyte ballooning and lobular inflammation (p<0.05) . With regard to type of surgery, there was no significant difference in T2D remission.

Discussion and Conclusion: Our data suggest that long-term remission of T2D after bariatric surgery (BS) is associated with lower preoperative insulin use and lower biopsy-proven steatosis scores in patients with NASH. Furthermore, T2D remission might be less likely in patients with liver fibrosis. Type of surgery did not affect T2D remission. Our results might help identify patients with NAFLD who benefit most from BS with regard to glycemic outcomes long-term.

Disclosure

A.Lautenbach: Advisory Panel; Novo Nordisk, Speaker's Bureau; Boehringer Ingelheim International GmbH, Novo Nordisk. M.Wernecke: None. S.M.Meyhöfer: None. S.Meyhöfer: None. F.D.Stoll: None. J.Aberel: Advisory Panel; Novo Nordisk, Speaker's Bureau; Novo Nordisk.