bluuebunny/crossref_metadata_2025 huggingface dataset:
1623-P: Prevalence and Clinical Determinants of Obesity in Type 1 Diabetes Mellitus
Authors: TSUSHIMA YUMIKO , KASHYAP SANGEETA , ZHOU KEREN
Published: June 2020

The prevalence of obesity in adults with type 1 diabetes mellitus (T1DM) is rising and is known to increase cardiovascular (CV) risk and mortality. However, the clinical features of adiposity are poorly described and discovery of novel associations may shed light on areas of possible intervention. We performed a retrospective cross-sectional analysis of 625 adults diagnosed with T1DM who were seen in Endocrinology between 2015-2018. These patients were stratified into 5 weight categories (normal weight, overweight, and obesity classes 1-3) then compared across various comorbidities, medication use, and metabolic parameters. Patients had a mean age of 35.2 ± 15.3 years, 53% were female, and >25% had a BMI ≥ 30. HbA1c did not significantly increase with weight. CV risk factors were more prevalent in patients with obesity (p<0.05) (Table 1). The anti-obesity medication, phentermine, was only prescribed in 3/172 patients and antidiabetic agents associated with weight loss were rarely applied to overweight individuals. Obesity combined with T1DM was associated with higher rates of metabolic disturbances and risk factors for CV disease. The sparse use of phentermine suggests suboptimal treatment of obesity in this population. Therefore, increased attention towards optimization of weight management for patients with obesity and T1DM is needed in an effort to reduce the comorbidities seen in this study.

Disclosure

Y. Tsushima: None. S. Kashyap: Other Relationship; Self; GI Dynamics Inc. K. Zhou: None.
1625-P: Association of Patient- and Disease-Related Factors with Glycemic Target Achievement in Type 1 Diabetes (T1D) in the SAGE Study
Authors: SEUFERT JOCHEN , IKEGAMI HIROSHI , BRETTE SANDRINE , PILORGET VALERIE , LAUAND FELIPE , WILMOT EMMA G. , RENARD ERIC , GAGLIARDINO JUAN J.
Published: June 2020

Most people with T1D do not achieve HbA1c targets, but this may vary with patient- and disease-related factors.

SAGE was a multinational, cross-sectional study using data from medical records and interviews of eligible people with T1D for ≥1 year, aged ≥26 years (N=3858). The relationship between each group of selected factors (sociodemographics, diabetes complications and comorbidities, T1D treatment, structure and process of medical care) and HbA1c target achievement (both general [HbA1c <7 %] and individualized) were evaluated.

Overall, 24.3% of people achieved HbA1c <7 %; 20.9% achieved individualized targets. For both targets, better achievement was associated with: lower body mass index, diastolic blood pressure, and total daily insulin dose; ≥1 symptomatic hypoglycemic event (<54 mg/dL) within the last 3 months; higher levels of education; and having health insurance (Table). Poorer target achievement was associated with ≥1 severe hyperglycemic event leading to diabetic ketoacidosis within the last 6 months, microvascular complications, and dyslipidemia. Older people were more likely to achieve individualized targets while younger people were more likely to achieve HbA1c <7 %.

HbA1c target achievement remains a challenge but SAGE identified factors that appear to be associated with HbA1c target achievement, offering opportunities for intervention.

Disclosure

J. Seufert: Research Support; Self; Sanofi. Speaker’s Bureau; Self; Sanofi. H. Ikegami: None. S. Brette: Other Relationship; Self; Sanofi. V. Pilorget: Employee; Self; Sanofi. F. Lauand: Employee; Self; Sanofi. E.G. Wilmot: Advisory Panel; Self; Abbott, Dexcom, Inc., Eli Lilly and Company, Insulet Corporation, Medtronic. Speaker’s Bureau; Self; Abbott, Eli Lilly and Company, Insulet Corporation, Novo Nordisk Inc., Sanofi. E. Renard: Board Member; Self; Abbott, Insulet Corporation, Novo Nordisk A/S, Roche Diabetes Care. Consultant; Self; Air Liquide, Eli Lilly and Company, Sanofi. Research Support; Self; Dexcom, Inc., Tandem Diabetes Care. J.J. Gagliardino: None.
1637-P: Role of mTORC1 Regulation in the T1D Organ Donor Pancreas
Authors: BRUGGEMAN BRITTANY S. , BEACHY DAWN , JACOBSEN LAURA M. , NICK HARRY S. , ATKINSON MARK A. , SCHATZ DESMOND , WASSERFALL CLIVE
Published: June 2020

Current evidence suggests that alteration in cellular metabolism leads to pancreatic β-cell dysfunction and loss in type 1 diabetes (T1D). The mammalian target of rapamycin, mTOR is a key regulator of cell survival and growth in response to intracellular energy levels, nutrients, and growth factor signaling. Dysregulated mTOR complex 1 (mTORC1) has been implicated in β-cell loss and dysfunction in T2D but has not been well studied in T1D. We hypothesized that genes associated with three pathways involved in the regulation of mTORC1 would display altered expression in T1D organ donor pancreata compared to nondiabetic controls. We isolated total RNA from 30 T1D and 30 unaffected control human organ donor pancreata obtained from the Network for Pancreatic Organ donors with Diabetes (nPOD) program and performed real-time qPCR on 26 genes related to these pathways, all of which demonstrated significant (p<0.05) induction. In relation to cellular energy balance, we observed induction of the AMPK-related genes. Concerning nutrient availability and the GCN2/ATF4 pathway, genes associated with all Sestrins and the GATOR 1 and GATOR 2 complexes were induced. In relation to growth factor signaling, components of the PI3K/AKT pathway were induced (Table 1). Both AMPK and PI3K/AKT pathways converge on TSC2, which had the largest degree of induction observed (22.4x). To conclude, our results clearly indicate dysregulation of mTORC1 in the T1D donor pancreas.

Disclosure

B.S. Bruggeman: None. D. Beachy: None. L.M. Jacobsen: None. H.S. Nick: None. M.A. Atkinson: None. D. Schatz: None. C. Wasserfall: None.
1646-P: Role Of Microbiota In Diabetes-Associated Colorectal Cancer
Authors: NOURELDEIN MOHAMED H. , EID ASSAAD A.
Published: June 2020

Diabetes changes the microbiota of the host, a condition known as dysbiosis. Dysbiosis have emerged to be an important contributing factor for the pathogenesis of diabetes and diabetes-associated colorectal cancer. In this study we tangled some of the molecular mechanisms that links diabetes to CRC that are in tune relation with dysbiosis. To test our hypothesis, MKRmice that can spontaneously develop type 2 diabetes were used in our work. For CRC induction, another subset of mice was treated with azoxymethane (AOM) and dextran sulfate sodium. (DSS), to identify the link between both diabetes and CRC. In parallel experiments, a subset of control mice was depleted of their microbiota after weaning, then mice were inoculated with fecal microbial transplant (FMT) from diabetic mice and mice with CRC. After 5 weeks of transplantation, all mice groups were treated with AOM and DSS. Further, a subset of the diabetic mice and diabetic mice with CRC were treated with probiotics or butyrate. At the end of the treatment, 16S rRNA sequencing was performed to identify different microbial communities in the fecal samples. Besides, at sacrifice, blood was collected, and colons were harvested for molecular, and biochemical analysis. Our preliminary results show that diabetes is associated with a defined microbial signature characterized by reduction of butyrate-forming bacteria which is associated with gastrointestinal complications.These changes are reversed upon treatment with probiotics or butyrate. Inoculation of control mice with diabetic microbiota and cancer microbiota resulted in the development of more aggressive CRC. Our data also show that IL-1βand NOX4 are over-expressed in diabetic mice as a result of activation of HDAC3 in colon tissue. Collectively our data suggest that diabetes is associated with dysbiosis characterized by lower abundance of butyrate-forming bacteria leading to less butyrate production and activation of HDAC3 resulting in over-expression of IL-1βand NOX4 leading to CRC.

Disclosure

M.H. Noureldein: None. A.A. Eid: None.
1648-P: Identification and Functional Analysis of E59Q Mutation in NEUROD1 Gene in a Chinese MODY Family
Authors: LIU LIMEI , CHEN YATING , ZHANG JUAN , JIANG YANYAN , LU MING , GE XIAOXU , JIA WEIPING
Published: June 2020

Objectives: In contrast to the common MODY2, MODY3 and MODY5, MODY6 is a relatively rare subtype of MODY. To investigate whether NEUROD1 is responsible for Chinese MODY, we screened its mutations in MODY pedigrees and explored the potential pathogenic mechanisms.

Methods: PCR direct sequencing was performed to screen NEUROD1 mutations in 32 Chinese MODY probands who were negative for MODY2, MODY3 and MODY5 genes. The functional significances of newly identified mutations were analyzed by studies on clinical phenotypes, pathophysiology and three-dimensional (3D) structure.

Results: 1) E59Q (c.175 G>C, p.Glu59Gln), a heterozygous mutation in NEUROD1 gene, was identified in one of the MODY families. 2) Glu59 residue in NeuroD1 is highly conserved across mammalian species. 3) Four diabetic patients (proband and her son, brother and sister) carring the mutation were thinner with BMI of 21.0 (20.3-21.2). In comparison with unaffected relatives (n = 6), E59Q carriers showed obviously decreased insulin secretion at fasting and postprandial status (both p<0.05). The proband’s father with E59Q mutation was normal glucose tolerance, suggesting non-penetrants. 4) The E59Q mutation was not detected in other probands or in the 201 nondiabetic control subjects. 5) The negatively charged Glu59 forms hydrogen bonds and strong salt bridges with the surrounding positively charged Arg54 and Lys88, and the main chains of Glu59 forms hydrogen bonds with Gly56 and Asp61, respectively. In the E59Q mutation, the salt bridges of Gln59 side chain disrupts, forming the new hydrogen bonds between Gln59 and Arg54.

Conclusions: NEUROD1-E59Q mutation clearly affected the interaction between E59 residue and its surrounding amino acids and thus changed the molecular conformation of N-terminal in NeuroD1. The abnormal conformation of NeuroD1 may decrease the E59Q mutant binding to the insulin promoter and activity of insulin transcription, therefore causing MODY6 subtype with defected insulin secretion.

Disclosure

L. Liu: None. Y. Chen: None. J. Zhang: None. Y. Jiang: None. M. Lu: None. X. Ge: None. W. Jia: None.

Funding

National Natural Science Foundation of China (81970686, 81770791, 81471012, 81270876); Shanghai Jiao Tong University (YG2019ZDA08); Shanghai Leading Talent (SLJ15055)
1656-P: FGF-21 Receptor Variants Demonstrate Severe Insulin Resistance in a Mouse Model of Insulin Mediated Pseudoacromegaly
Authors: STONE STEPHEN I. , LUPU TRAIAN , URANO FUMIHIKO , ORNITZ DAVID M.
Published: June 2020

Insulin mediated pseduoacromegaly (IMPA) is a rare insulin resistance syndrome characterized by tall stature, acanthosis nigricans, obesity, and acromegaloid features. Subjects with IMPA have normal growth hormone and IGF-1. However, their insulin levels are markedly elevated. To date, no molecular or genetic etiology has been demonstrated as a definitive cause of IMPA. We previously described an adolescent female diagnosed with IMPA. Exome sequencing identified 2 rare, predicted deleterious, variants in Fibroblast Growth Factor Receptor 1 (FGFR1) and beta-Klotho (KLB) which form the receptor for fibroblast growth factor 21 (FGF-21). Due to FGF-21’s role in insulin signaling, we hypothesized that these mutations may be contributing to our proband’s condition. Our study aimed to knock-in these FGFR1/KLB variants into a mouse model using CRISPR/Cas9 gene editing technology. After embryo microinjection, we were able to generate 4 pups mosaic for the missense mutations in FGFR1 or KLB. The resulting founder mice were backcrossed to wild type mice, and then bred to recapitulate the same digenic geneotype found in our adolescent proband with IMPA. When exposed to a high fat diet, double mutant (FGFR1/KLB) mice demonstrate hyperglycemia (158.0 compared to 125.3 mg/dL, p=<0.0001). At 9 weeks-old, the double mutant mice weighed 40.9 g compared to 25.4 g (p=0.0177). These mice underwent a 2 g/kg intraperitoneal glucose tolerance test. While fasting, both the double mutant and wild type mice had normal glucose (87 and 76 mg/dL). However fasting insulin was elevated in the double mutant mice (1246.1 to 231.5 pg/mL). 30 minutes post glucose, the double mutant mice demonstrated hyperglycemia (414 to 274 mg/dL) and marked hyperinsulinemia (2,358.0 to 884.3 pg/mL). Although further characterization of these mice is required, thus far, these studies suggest that our proband’s digenic variants in FGFR1/KLB are likely to be pathogenic and are a potential cause of IMPA.

Disclosure

S.I. Stone: Research Support; Self; National Institutes of Health. Speaker’s Bureau; Self; Rhythm Pharmaceuticals. T. Lupu: None. F. Urano: Stock/Shareholder; Self; CytRx Corporation. Other Relationship; Self; AETAS Pharma, Amylyx Pharmaceuticals, Mitochon, National Center for Advancing Translational Sciences, Novus Biologicals?. D.M. Ornitz: None.

Funding

National Institutes of Health (R21HD098872); Washington University (P30DK020579); National Center for Advancing Translational Sciences (UL1TR002345)
1658-P: A Novel Mutation in Hepatocyte Nuclear Factor-1 Beta (HNF1B) in Maturity-Onset Diabetes of the Young 5 (MODY5)
Authors: FUJITA YUKI , HYO TAKANORI , MATSUBARA MIHO , HAMAMOTO YOSHIYUKI , TANAKA DAISUKE , SEINO YUTAKA
Published: June 2020

Maturity-onset diabetes of the young (MODY) is a group of dominantly inherited non-autoimmune diabetes characterized by early onset. Delayed diagnosis or misdiagnosis as type 1 or 2 diabetes is common, but early and correct diagnosis of MODY is important because it significantly alters the management of both the patient and his or her relatives. MODY5 is caused by mutations in the HNF1B, associated with renal and pancreatic abnormalities. Recently, we had identified a novel pathogenic mutation in IVS3-1G>C of HNF1B, in a Japanese MODY family. A 30-year-old male presented to our office with a diagnosis of NIDDM at age 15; autoimmune antibodies were negative, glycemia was initially managed with basal-bolus insulin. Abdominal ultrasound showed bilateral hyperechogenic kidneys but unlike with the reported typical abnormalities in MODY5, only a few renal cysts were observed, and pancreas was intact. An initial diagnosis of early-onset type 2 diabetes was made. However, the case subsequently showed high probability of MODY, when he reported a strong family history of early-onset diabetes in his sister, his father and paternal grandmother. Genetic testing performed in him, his sister and his father revealed shared unreported gene mutation g.37731831C>G (GRCH38 Chr17) in the HNF1B gene suggesting a diagnosis of MODY5. The mutation locates in the splicing site which may result in the production of abnormal proteins or failure of protein synthesis. Our MODY5 case was unique in the point that despite the mild renal morphological abnormalities, the proband had renal dysfunction and microalbuminuria from the early stage of disease, evoking the importance of considering the possibility of monogenic diabetes even in scarce structural abnormalities but having atypical diabetes with strong family history. It also suggests the necessity of further investigation of the exact mechanism of the development of diabetes and the cause of abnormalities in the kidney by this certain mutation.

Disclosure

Y. Fujita: None. T. Hyo: Speaker’s Bureau; Self; ARKRAY, Kissei Pharmaceutical Co., Ltd., LifeScan, Inc., Medtronic, Novo Nordisk Inc. M. Matsubara: None. Y. Hamamoto: Research Support; Self; Sumitomo Dainippon Pharma Co., Ltd. Speaker’s Bureau; Self; Novo Nordisk Inc. D. Tanaka: None. Y. Seino: Research Support; Self; ARKRAY, Bayer Yakuhin, Ltd., Boehringer Ingelheim International GmbH, Eli Lilly Japan K.K., Merck Sharp & Dohme Corp., Nippon Boehringer Ingelheim Co. Ltd., Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited, Terumo Medical Corporation. Speaker’s Bureau; Self; Becton, Dickinson and Company, Eli Lilly Japan K.K., Kao Corporation, Merck Sharp & Dohme Corp., Nippon Boehringer Ingelheim Co. Ltd., Novo Nordisk Inc., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited.
1671-P: Pathogenesis of Obesity-Driven Metabolic Sequelae Is Associated with Th17 Metabolic Switch and Inflammatory Hepatic Th17 Cells
Authors: MORENO-FERNANDEZ MARIA E. , OATES JARREN , DAMEN MICHELLE S. , DOLL JESSICA R. , STANKIEWICZ TRACI E. , DIVANOVIC SENAD
Published: June 2020

Nonalcoholic fatty liver disease (NAFLD) is a major global health issue. Lack of effective therapies to NAFLD is attributed to limited knowledge of disease etiopathogenesis. Emerging evidence reveal key contribution of the Th17 axis in NAFLD. However, the characteristics and mechanisms underlying the pathogenic potential of Th17 cells in NAFLD remain unknown. Here, we uncover a unique population of inflammatory hepatic CXCR3+Th17 cells (ihTh17) that contribute to NAFLD progression. Longitudinal hepatic accrual of ihTh17 cells correlated with NAFLD severity in mice and was dependent on obesity and CXCR3 axis activation. ihTh17 cells, compared to conventional CXCR3-Th17 cells, exhibited increased open chromatin marks in enhancer regions of inflammatory and metabolic genes, increased glycolytic skewing and metabolic capacity, and were able to simultaneously produce an array of pro inflammatory cytokines that directly promoted hepatocellular damage. Adoptive transfer of ihTh17 cells accelerated and exacerbated NAFLD pathogenesis in vivo. Modulation of the metabolic state of ihTh17 cells was sufficient to reverse ihTh17-centric inflammatory vigor and NAFLD severity. Importantly, ihTh17 cell hepatic accrual and inflammatory capacity, CXCR3 axis activation, and hepatic expression of glycolysis-associated genes were conserved in humans with NAFLD and positively correlated with disease severity. In sum, we show that the obesity-associated environment regulates Th17 cell metabolism and competence towards a unique inflammatory hepatic Th17 fate in turn contributing to NAFLD progression. Notably, modulation of these pathways holds potential for development of novel therapeutic strategies to NAFLD.

Disclosure

M.E. Moreno-Fernandez: None. J. Oates: None. M.S. Damen: None. J.R. Doll: None. T.E. Stankiewicz: None. S. Divanovic: Consultant; Self; Janssen Pharmaceuticals, Inc., Pfizer Inc.

Funding

American Diabetes Association (1-19-PMF-019 to M.E.M-F.); National Institutes of Health (R01DK099222, R01DK099222-02S1P30, DK078392)
1677-P: Higher Frequency of HLA DRB1*08:03 and DRB1*04:01 Observed in A-beta+ Ketosis-Prone Type 2 Diabetes in Japanese Subjects
Authors: SHIMADA AKIRA , SATOMURA ATSUSHI , SUZUKI SEIYA , HAISA AKIFUMI , OIKAWA YOICHI
Published: June 2020

Ketosis-prone type 2 diabetes, especially islet-associated autoantibody negative and beta-cell function reserved (A-beta+; KPD) group, is recognized as atypical diabetes. Because we detected insulin peptide reactive T cells in these KPD subjects, we realized that KPD possesses characteristics of type 1 diabetes (ADA meeting, 2017). Therefore, in this study, we examined HLA types. After institutional review board permission was obtained, written informed consent and blood samples were obtained from the enrolled Japanese subjects (KPD: 22 males and 0 female, mean onset age 36.4 years of age; acute onset type 1 diabetes (AT1D): 21 males and 22 females, mean onset age 36.3 years of age; slowly progressive type 1 diabetes (SP): 10 males and 10 females, mean onset age 49.0 years of age). Then the HLA allele frequency was compared to normal controls (http://hla.or.jp/med/frequency_search/ja/allele/search/R/). The frequencies of HLA DRB1*04:05 and DRB1*09:01, which are known susceptible alleles in Japanese type 1 diabetes, were not higher in KPD (13.6%, 27.3%, respectively) subjects as compared to controls (13.5%, 14.3%, respectively), whereas were significantly higher in AT1D (41.9%, 53.5%, respectively; both p<0.001) and SP subjects (45.0%, 40.0%, respectively; both p<0.002). However, interestingly, the frequency of HLA DRB1*08:03 was significantly higher in KPD (34.8%, p<0.001) subjects as compared to controls (8.3%), whereas it was not the case both in AT1D (4.7%) and SP (10.0%) subjects. Moreover, the frequency of HLA DRB1*04:01 which is known susceptible alleles in Caucasian type 1 diabetes was significantly higher in KPD (18.2%, p<0.001) subjects as compared to controls (0.9%), whereas it was not the case both in AT1D (2.3%) and SP (5.0%) subjects.

In conclusion, although KPD subjects possess similar insulin peptide T cell responses to type 1 diabetes, different genetic background seems to exist probably leading to different clinical phenotype.

Disclosure

A. Shimada: Advisory Panel; Self; Astellas Pharma Inc. Speaker’s Bureau; Self; Eli Lilly Japan K.K., Novo Nordisk Inc., Sanofi-Aventis. A. Satomura: None. S. Suzuki: None. A. Haisa: None. Y. Oikawa: None.

Funding

Saitama Medical University
1682-P: Cotransplantation of Islets with Adipose-Derived Stem Cells Synergistically Reduced Blood Glucose in STZ-Induced Diabetic Rats
Authors: CHEN XIAO-DONG , LI SHENGXIAN , WANG HANZHOU
Published: June 2020

Mesenchymal stem cells (MSCs) not only serve as a cell reservoir, replacing damaged cells, but also as a “drug store”, providing trophic factors that exert multiple potent cellular effects, such as anti-apoptotic, anti-fibrotic, anti-inflammatory, and immunomodulatory. There is increasing evidence that MSC trophic factors are able to improve the ability of β-cells to secrete insulin in vitro and promote islet graft survival in vivo. Here, we report that co-transplantation of freshly isolated islets with adipose-derived mesenchymal stem cells (Ad-MSCs) results in a rapid, significant reduction in hyperglycemia in streptozotocin (STZ)-induced diabetic rats. Lewis rats (n=30) were injected intraperitoneally with STZ (80 mg/kg); by day 3 post-injection, 24 animals (∼80%) achieved elevated blood glucose levels that stabilized at ∼300 mg/dL. On day 14, STZ-treated rats received the following transplants via hepatic portal vein infusion: PBS (negative controls), Lewis (isografts) or Fischer (allografts) islets alone or combined with Lewis rat Ad-MSCs. Initially, co-transplantation of islets (isografts or allografts) with Ad-MSCs dramatically reduced blood glucose levels compared to transplantation of islets alone (∼60% vs. ∼30%). However, only isografts combined with Ad-MSCs promoted the retention of transplanted islet function, resulting in the maintenance of blood glucose levels at ∼47% of the pre-transplantation level (=100%) through 70 days post-transplantation. Morphologically, these co-transplants in liver were larger in size, had more capillaries, and produced more insulin than islets alone. Although islets (iso- or allo-grafts) co-transplanted with Ad-MSCs appeared to provide an accelerated normalization of blood glucose levels, no evidence was found that allograft rejection was reduced. We conclude that co-transplantation improved islet viability more, with a prolonged anti-hyperglycemic effect, than islets alone.

Disclosure

X. Chen: None. S. Li: None. H. Wang: None.

Funding

San Antonio Medical Foundation
1691-P: Alcohol-Related Liver Disease as Independent Risk Factor for Posttransplant Diabetes Mellitus in Liver Recipients
Authors: CAMPOS MARIANA B. , BOIN ILKA D. , NETO ARNALDO M.
Published: June 2020

Background and Aims: There are few data related to identification of risk factors for post transplant diabetes mellitus (PTDM) in liver transplantation (Tx). The aim of this study was to investigate the risk factors for PTDM.

Patients and methods: A retrospective study of adult patients submitted to liver Tx. Data for sex, age, BMI, pre and PTDM, Tx cause, presence of hepatocellular carcinoma (HCC), use of immunosuppressants, hepatitis B, hepatitis C (HepC) and liver biopsy were analyzed. Mann-Whitney and Chi Square tests as well as a multivariate logistic regression were performed.

Results: Among 207 consecutive patients, 60.4% had no diabetes (group 1), 18.8% had pre Tx DM (group 2) and 20.8% were classified as PTDM (group 3). The major cause of transplantation was HepC (48.8%). Triglycerides (164 vs. 106mg/dL, p= 0.002) and creatinine (1.24 vs. 1.1mg/dL, p = 0.029) were higher in group 2 than in group 1, probably due to long-standing DM and metabolic syndrome in group 2. Comparisons using the Chi-Square test showed higher frequencies in group 3 than group 1 and 2, respectively, for HepC (65.1% vs. 35.1% and 37.9%, p=0.004), tacrolimus use (95.3% vs. 81.6% and 71.1%, p<0.001) and prednisone use (48.8% vs. 24% and 36.8%, p=0.009). HCC before Tx was more common in group 2 than in groups 1 and 3 (64.1% vs. 40.8% and 41.9%, respectively; p=0.034). Regarding biopsy data, interface activity was more severe in groups 1 and 3 compared to group 2 (62.2% and 64%% vs. 11%; p=0.040, respectively). The independent risk factors for PTDM according to the multivariate logistic model were HepC (OR 4.4; CI 95% 1.73-11.22; p=0.002), alcoholism (OR = 3.7, 95% CI, 1.2-11.5, p = 0.025), the use of tacrolimus (OR = 6.3, 95% CI 1.2-32.5, p = 0.029) and prednisone (OR = 7.2, 95% CI 2.4-21.8, p<0.001).

Conclusion: Alcohol-related liver disease is an independent risk factor for PTDM in liver recipients. Patients with HepC and those exposed to tacrolimus and prednisone are other groups of high risk for PTDM.

Disclosure

M.B. Campos: None. I.D. Boin: None. A.M. Neto: None.
1695-P: Ablation of Adipose CREB3L3 Prevents Browning and Promotes Inflammation
Authors: MCCANN MAXIMILIAN A. , MUNOZ MARCOS D. , GIL VICTORIA , ZHANG KEZHONG , LIEW CHONG WEE
Published: June 2020

The body contains two kinds of fat: white fat, which acts as an energy storage organ, and brown fat, which utilizes energy to generate heat. White fat consists of the subcutaneous and the visceral fat. Accumulation of visceral fat contributes more to metabolic dysfunction due to its increased rate of lipolysis, reduced ability to form brown-like adipocytes known as “beige” cells, and its more inflammatory nature. In this study, we investigated the role that cyclic-AMP Responsive Element Binding Protein 3-like-3 (CREB3L3) plays in adipose tissue. CREB3L3 is an ER-bound transcription factor that has previously been described as liver specific, and an important regulator of lipid metabolism. Upon discovering that CREB3L3 is not only expressed in adipose tissue, but selectively downregulated in the more “metabolically healthy” subcutaneous fat in both obese mice and humans, we investigated how ablating this transcription factor would affect adipose biology. To study this, we created a CREB3L3 fat-specific knockout (fKO) mouse. When challenged with high-fat diet, the fKO mice became 18% heavier than floxed controls. They had significant expansion of their epididymal and inguinal adipose depots, due to a reduction in whole-body energy expenditure and oxygen consumption as measured by indirect calorimetry. The fKO mice are also more resistant to browning during cold exposure and treatment with the β3-adrenergic receptor agonist CL316,243, suggesting impairment of adaptive thermogenesis in the subcutaneous fat. Expansion of the visceral epididymal fat caused the tissue to become more inflammatory as evidenced by an increase in the number of crown-like structures and expression of the macrophage marker F/480 in the fKO tissue. This led to the fKO mice becoming more insulin resistant following high-fat feeding. Together, ablation of CREB3L3 enhances adiposity and insulin resistance during obesity due to a reduction in browning potential and amplified visceral inflammation.

Disclosure

M.A. McCann: None. M.D. Munoz: None. V. Gil: None. K. Zhang: None. C. LIew: None.

Funding

National Institutes of Health (R01DK109015-01); American Heart Association (16PRE30190011)
170-OR: Association of Timing of Physical Activity Bouts with Cardiorespiratory Fitness in Adults with Type 2 Diabetes in the Look AHEAD Study
Authors: QIAN JINGYI , WALKUP MICHAEL P. , CHEN SHYH-HUEI , BRUBAKER PETER H. , BOND DALE , RICHEY PHYLLIS A. , JAKICIC JOHN M. , HU KUN , SCHEER FRANK A. , MIDDELBEEK ROELAND
Published: June 2020

Background: Moderate-to-vigorous physical activity (MVPA) is an important treatment in type 2 diabetes (T2D). Few studies examine the timing of MVPA, as there are no established methods to characterize the MVPA bout timing throughout the day. We determined the timing of MVPA bouts in adults with T2D, and examined whether cardiorespiratory fitness and odds of cardiovascular disease (CVD) history relate to timing of MVPA bouts in men and women.

Methods: Baseline data from Look AHEAD participants (n=2153, 57% women) were analyzed. Cardiorespiratory fitness assessed by VO2max test was expressed as METs. MVPA bouts (≥3METs in ≥10mins bouts) were determined using the hip-worn RT3 accelerometer for 7 days. Participants were categorized into six groups based on the time of day with the major amount of MVPA bouts (MET-min): ≥50% of MVPA bouts during the same time window (Early morning, Late morning, Afternoon, or Evening, N=214-280/group), <50% of MVPA bouts in any time category (Mixed; reference group, N=436), and ≤1 day with MVPA bouts per week (Inactive, N=624).

Results: Cardiorespiratory fitness differed by timing of MVPA bout category (P<0.0001). In a multivariate adjusted model, the association of timing of MVPA bouts with fitness varied by sex (P=0.002). In women, the evening group had higher fitness (β 0.35 [0.04, 0.66] METs). After further adjustment for individual total bout-MVPA (MET-min) per week, in men, the early morning group was associated with higher fitness (β 0.46 [0.06, 0.85] METs). Interestingly, in men, but not in women, the early morning group also had higher odds of CVD history (OR 2.4 [1.38, 4.21]).

Conclusion: Higher cardiorespiratory fitness associated with evening MVPA bouts in women, and with early morning MVPA bouts in men independent of total amount of bout-MVPA. Prospective studies are needed to investigate the relationship between timing of MVPA and cardiovascular outcomes to optimize MVPA benefits in adults with T2D.

Disclosure

J. Qian: None. M.P. Walkup: None. S. Chen: None. P.H. Brubaker: None. D. Bond: None. P.A. Richey: None. J.M. Jakicic: Advisory Panel; Self; Weight Watchers International, Inc. K. Hu: None. F.A. Scheer: None. R. Middelbeek: None.
1718-P: Heme Oxygenase-1 Promotes Hepatic Glucose Production via Increasing the Intracellular Ferrous Iron
Authors: LIAO WANG , YANG WANBAO , PAN QUAN , SHEN ZHENG , AI WEIQI , GUO SHAODONG
Published: June 2020

The heme oxygenase-1 (HO1) catalyzes the degradation of heme into biliverdin, ferrous iron and carbon monoxide (CO). Although HO1 has long been considered as an anti-inflammatory molecule, recent studies showed that HO1 disrupts the mitochondrial function in hepatocytes, as well as drives inflammation and insulin resistance in mice and humans. Such different findings from the historical view impelled us to further investigate the roles of HO1 in glucose metabolism. In the present study, we found that overexpression of HO1 promoted hepatic glucose production (HGP) in hepatocytes from the wild type (WT) mice, which was comparable to the treatment of glucagon. Since the regulatory roles of the transcription factor forkhead box O1 (Foxo1) in HGP have been well recognized, we then generated the liver-specific Foxo1 knockout (L-FKO) mice to study the contribution of Foxo1 in mediating the effects of HO1. We found that HO1 overexpression did not have a significant effect in promoting HGP in hepatocytes from the L-FKO mice, suggesting HO1 improved HGP dependent on Foxo1. Besides, HO1 overexpression improved the mRNA levels of Foxo1-targeted gluconeogenic genes and increased the phosphorylation of Foxo1 at Ser273, which is a phosphorylation site integrating into the glucagon signaling. We also found that HO1 overexpression activated p38α and p65/NF-κB in hepatocytes. Further study using siRNA or specific inhibitor demonstrated that p65/NF-κB, but not p38α, mediated the HO1-induced pFoxo1-S273 and HGP in hepatocytes. We then explored the role of HO1-induced ferrous iron in activating p65/NF-κB and promoting HGP. We found that HO1 overexpression in hepatocytes substantially increased the intracellular ferrous iron. The pretreatment of iron chelator deferoxamine mesylate diminished the effects of HO1 overexpression on p65/NF-κB, pFoxo1-S273, and HGP. Taken together, results from this study revealed the mechanisms of HO1 in driving HGP, which is through the generation of ferrous iron.

Disclosure

W. Liao: None. W. Yang: None. Q. Pan: None. Z. Shen: None. W. Ai: None. S. Guo: None.

Funding

National Institutes of Health (R01DK120968)
1729-P: Insulin Resistance Cutoff for High Probability of Blood Glucose Normalization in T2DM Patients with Obesity after Bariatric Surgery
Authors: SKLYANIK IGOR , SHESTAKOVA EKATERINA , STAFEEV IURII , MICHURINA SVETLANA , KARMADONOV ANDREY , VOROTNIKOV ALEXANDER V. , YAHYAEV KAMIL , YURASOV ANATOLIY , MENSHIKOV MIKHAIL , MAYOROV ALEXANDR , PARFYONOVA YELENA V. , SHESTAKOVA MARINA V.
Published: June 2020

There are several models of T2DM remission after bariatric surgery (ABCD, DiaRem, DRS), but none of them includes insulin resistance. Moreover these models are not suitable for early prediction of T2DM remission (within several months after surgery).

Methods: We included 42 patients with T2DM and morbid obesity (BMI 42.3 [38.9; 48.1] kg/m2) after bariatric surgery (16 pts after restrictive procedures and 26 pts after bypass surgery). Insulin resistance was measured by hyperinsulienic euglycaemic test with M-index estimation (mg/kg/min) and HOMA-IR before the surgery. Glycaemia normalization was assessed by self-monitoring of blood glucose (FPG < 110 mg/dl (6.1 mmol/l) and postprandial PG < 140 mg/dl (7.8 mmol/l) in 1 month after surgery) as well as HbA1c < 6.0% (42 mmol/mol) in 3 months after surgery. ROC-curve identified insulin resistance predictive value of blood glucose normalization after bariatric surgery. Statistical analysis was performed using SPSS v.23.0.

Results: All patients had severe insulin resistance at baseline (median M-index before surgery 1.535 mg/kg/min, HOMA-IR 10.0). Within 1 month after surgery 7 patients (16,7%) reached blood glucose normalization, within 3 months - 22 patients (52,4%), within 6 months - 31 patient (73,8%), in 12 months - 35 patients (83,3%). ROC-analysis showed that M-index within 1st month after surgery was predictive of blood glucose normalization; insulin resistance cut-off value was 1.876 mg/kg/min (Youden’s index). HOMA-IR and M-index in 3, 6 and 12 months after surgery didn’t predict glycaemia normalization.

Conclusion: Baseline insulin resistance value defined as M-index > 1.876 mg/kg/min can be used to predict early blood glucose normalization within 1 month after bariatric surgery.

Disclosure

I. Sklyanik: None. E. Shestakova: Employee; Spouse/Partner; AstraZeneca. Speaker’s Bureau; Self; Abbott, AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Novartis Pharma K.K., Novo Nordisk A/S, Sanofi-Aventis, Takeda Pharmaceutical Company Limited. I. Stafeev: None. S. Michurina: None. A. Karmadonov: None. A.V. Vorotnikov: None. K. Yahyaev: None. A. Yurasov: None. M. Menshikov: None. A. Mayorov: Speaker’s Bureau; Self; Abbott, LifeScan, Inc., Novo Nordisk A/S, Roche Diabetes Care, Sanofi-Aventis. Y.V. Parfyonova: None. M.V. Shestakova: None.

Funding

Russian Science Foundation (17-15-01435)
173-OR: High Out-of-Pocket Health Care Burden for Medicare Beneficiaries with Diabetes, 1999-2015
Authors: PARK JOOHYUN , WANG YU , LOOK KEVIN A. , LUMAN ELIZABETH , ZHANG PING
Published: June 2020

People with diabetes face higher out-of-pocket (OOP) health care costs than those without diabetes. This burden can lead to low adherence to treatments and adverse health outcomes, particularly among older adults. The OOP burden in older adults with diabetes has not been studied in the U.S. We estimated the OOP burden from 1999-2015 in Medicare beneficiaries with diabetes aged ≥65 using the Medicare Current Beneficiary Survey. OOP costs included premiums, deductibles, copays, coinsurance, and payments not covered by insurance. High OOP burden was defined as a person’s OOP costs ≥ 10% of their income. We estimated the prevalence of high OOP burden in all beneficiaries, by income quartile, and by race/ethnicity. Average OOP costs ranged from $2,864 to $4,212 (2015 $) and the percentage of those with high OOP burden varied from 38% to 46%, with slight increases over time. By income quartile, the average OOP costs were $2,292, $3,661, $4,032, and $4,430 for <25%, 25-50%, 50-75% and >75% groups, with decreases only in <25% group over time. There were large income-related disparities in prevalence of high OOP burden, with the highest in 25-50% group. Non-Hispanic whites had the highest prevalence of high OOP burden. In elderly Medicare beneficiaries with diabetes, OOP burden was high and increased over time, with large income-related disparities. Efforts to reduce both OOP burden and disparities in this population may be needed.

Disclosure

J. Park: None. Y. Wang: None. K.A. Look: None. E. Luman: None. P. Zhang: None.
1731-P: The Mechanism of the Protective Effect of GLP-1 on the Amelioration of Alzheimer’s Disease by Enhancing Brain Insulin Production
Authors: HUANG JIAOJIAO , YANG YAN
Published: June 2020

Background: Growing evidence supported that type 2 diabetes mellitus (T2DM) patients had a greater risk of developing Alzheimer’s disease (AD). Insulin secreted by “β cells” in brain played a critical role in T2DM patients and in AD patients. GLP-1 could improve cognition in AD patients via enhancing brain insulin signaling. We hypothesize the therapeutic effect of GLP-1 in T2DM might be due to promoting the insulin secretion of “β cells” in the nervous system. However, the molecular mechanism is unclear.

Methods: The hippocampus cell- HT-22 cell line and T2DM rats were used in this study. The genes of insulin syntheses, animals’ cognitive function and the phosphorylation of tau protein in hippocampus of T2DM brain were examined.

Results: Firstly, HT22 cell line was intervened by GLP-1 or high glucose or GLP-1+ high glucose. When stimulated by GLP-1 combined with high glucose, IRS1/PI3K/AKT located in insulin signal pathway could be activated and the expression of neurod-1 and ngn3 (insulin synthesis genes) were notably increased. These results indicated that neuronal cell could secrete insulin under a certain condition.

Secondly, RAAV-sh- neurod-1 and RAAV-sh-ngn3 were icv to the T2DM rats to prevent the production of brain insulin. The Barnes Maze test indicated the T2DM rats with peripheral GLP-1 and icv RAAV-sh-nc injection had a more intelligence than those with peripheral GLP-1 and icv RAAV-sh- neurod-1 and RAAV-sh-ngn3.

Thirdly, the T2DM rats with peripheral GLP-1 and icv RAAV-sh- neurod-1 and RAAV-sh-ngn3 had a significantly hyperphsphorylation of tau protein than those rats with peripheral GLP-1 and icv RAAV-sh-nc injection. This result showed that brain insulin could improve cognition by GLP-1 in T2DM rats.

Conclusions: These results suggested that neuronal cells might synthesis and secrete insulin under the condition of high glucose and GLP-1. And GLP-1 could not improve cognitive ability in T2DM when brain insulin synthesized was prevented.

Disclosure

J. Huang: None. Y. Yang: None.

Funding

National Natural Science Foundation of China (81670754)
1733-P: Phosphoproteomic Analysis of Insulin Action Using Human IPS-Derived Myoblasts Reveals Important Interactions between Differences Based on Insulin Sensitivity and Sex
Authors: HAIDER NIDA , LEBASTCHI JASMIN , BATISTA THIAGO M. , KNOWLES JOSH , PAN HUI , DREYFUSS JONATHAN , KAHN C. RONALD
Published: June 2020

To identify disease-specific indicators associated with insulin resistance by using human induced pluripotent stem cells (iPSCs) as a “disease-in-a-dish” model, iPSCs were derived from 20 nondiabetic individuals, with 10 each in the most insulin resistant (IR) or insulin sensitive (IS) quintiles as determined using SSPG measurements. Half of each group was male and half female. Given that muscle is the first site of insulin resistance in type 2 diabetes, iPSCs were differentiated into myoblasts, and global phosphoproteomics was performed without/with insulin stimulation to determine the full spectrum of signaling. Analysis of phosphoproteome revealed altered serine/threonine phosphorylation of the insulin-regulated sites on IRS2, Akt1, mTOR, RPS6 in the IR iPSC-myoblasts indicating defects related to insulin resistance. Unexpectedly, hierarchical clustering of the phosphosites (FDR< 0.05) also revealed alterations in IR males as compared to IS males but not in females. Rather female cells demonstrated a drastically different overall phosphoproteome as compared to the males, indicating the existence of an important sexual dimorphism that dominated over differences in insulin resistance. The top up-regulated phosphosites in the males, such as SRRM1 S697, THRAP3 S535, PRRC2C T1244 (all p<1x10-9 and fold change >5 compared to females) belonged to mRNA splicing pathway and included many targets of mTOR. In contrast, some of the top up-regulated phosphosites in the females included LMNB1 S23, SKP2 S64, MARCKS S167 (p<1x10-7 and fold change >5 compared to males) belonged to cell cycle pathways. These findings indicate that iPSC-derived myoblasts provide a potent approach to understanding differences in phosphorylation networks. These studies have also uncovered previously unrecognized differences in signaling between males and females, which may play a major role in the pathogenesis of disease.

Disclosure

N. Haider: None. J. Lebastchi: None. T.M. Batista: None. J. Knowles: None. H. Pan: None. J. Dreyfuss: None. C. Kahn: Advisory Panel; Self; ERX Pharmaceuticals, Kaleido Biosciences. Consultant; Self; AntriaBio, Flagship Pioneering, Sana-Cobalt.
1734-P: High Dietary Fat Intake Exacerbates Insulin Resistance in a Type 2 Diabetes Genetic Mouse Model
Authors: REILLY AUSTIN , YAN SHIJUN , LONCHARICH ALEXA J. , REN HONGXIA
Published: June 2020

GLUT4, an insulin-sensitive glucose transporter, plays a major role of postprandial glucose disposal. We previously reported that human GLUT4 promoter-driven insulin receptor knockout (GIRKO) mice have reduced insulin signaling in central and peripheral tissues and develop hyperglycemia, albeit at a lower rate in an age-dependent manner on normal chow diet. Long-term high fat diet (HFD) feeding is well established for causing adiposity gain, increased inflammation, and insulin resistance. We hypothesized that HFD feeding will exacerbate the insulin resistance and result in rapid progression to overt diabetes in GIRKO mice. To test this, we examined the physiological response of euglycemic GIRKO mice to HFD in a time course study. We used indirect calorimetry analysis and found that GIRKO mice showed differences in energy partitioning shortly after HFD feeding started. The majority of GIRKO mice cohorts had 5-fold higher circulating insulin and 1.5-fold elevated blood glucose after 1 month of HFD feeding. In addition to the hyperglycemia and hyperinsulinemia, GIRKO mice also exhibited impaired glucose tolerance and islet hyperplasia, suggesting extreme insulin resistance exceeded the compensatory increase of insulin production. After 3 months on HFD, GIRKO mice had increased relative liver size, elevated hepatic triglycerides, and reduced hepatic glycogen, demonstrating impaired hepatic insulin signaling to stimulate glycogen synthesis and inhibit gluconeogenesis. We also confirmed the increase of key hepatic gluconeogenic gene expression, which likely contributed to hyperglycemia. The rapid progression of metabolic defects of GIRKO mice on HFD is not due to excessive weight or adiposity gain.

In conclusion, our studies of HFD-fed GIRKO mouse will offer insights in the role of the dietary contribution to diabetes disease progression without the confounding factor of increased adiposity.

Disclosure

A. Reilly: None. S. Yan: None. A.J. Loncharich: None. H. Ren: None.

Funding

Indiana University Diabetes and Obesity Research Training Program (T32DK064466); National Institute of Diabetes and Digestive and Kidney Diseases (R01DK120772, R00DK098294); Indiana Diabetes Center (P30DK097512); Indiana Clinical and Translational Sciences Institute (UL1TR002529)
1739-P: Insulin Rapidly Increases Skeletal Muscle Mitochondrial ADP Sensitivity, Mitigating HFD-Induced Mitochondrial Dysfunction
Authors: BRUNETTA HENVER , PETRICK HEATHER L. , NUNES EVERSON A. , HOLLOWAY GRAHAM
Published: June 2020

Introduction: Causality of the association between impairments in mitochondrial function and insulin resistance remains debatable. We have previously demonstrated that mitochondrial ADP sensitivity is impaired in high-fat diet (HFD)-induced insulin resistance, which contributes to bioenergetic imbalance and higher reactive oxygen species (ROS) production. However, insulin regulates several metabolic processes within skeletal muscle, and we tested the hypothesis that insulin could rapidly improve mitochondrial ADP sensitivity, a phenomenon possibly lost in HFD-induced insulin resistance.

Methods: Male and female C57Bl/6 mice were fed a control diet (10% fat) or HFD (60% fat) for 8 weeks. At the end of treatment, insulin was injected intraperitoneally (3 U/kg) twice (0 and 30 minutes). At 60 minutes red gastrocnemius muscle was excised to evaluate mitochondrial bioenergetics, ADP sensitivity, and ROS production.

Results: In control mice, insulin increased respiration by ∼2-fold in the presence of submaximal (100-1000 μM ADP) but not maximal ADP-supported respiration. As a result, insulin decreased the apparent ADP Km from 700 to 380 μM ADP. Despite these respiratory changes, insulin had no effect on mitochondrial maximal succinate-supported ROS emission or the ability of ADP to suppress ROS emission. HFD consumption impaired whole-body glucose intolerance, skeletal muscle insulin resistance (decreased insulin-stimulated Akt Thr-308 and Ser-473 phosphorylation) and impaired mitochondrial submaximal ADP-supported respiration by ∼50% (ADP Km ∼1100 μM). However, despite the reduction in insulin signaling following HFD, insulin increased submaximal ADP-supported respiration by ∼2-fold, decreased the apparent ADP Km to ∼400 μM and improved the ability of ADP to suppress mitochondrial ROS emission.

Conclusion: Insulin rapidly improves mitochondrial ADP sensitivity, mitigating the apparent HFD-induced mitochondrial dysfunction.

Disclosure

H. Brunetta: None. H.L. Petrick: None. E.A. Nunes: None. G. Holloway: None.

Funding

Natural Sciences and Engineering Research Council of Canada (400360); Coordination for the Improvement of Higher Education Personnel