bluuebunny/crossref_metadata_2025 huggingface dataset:
2259-PUB: Aging, Systolic Hypertension, Obesity, and Hyperuricemia Increase the Cardiovascular Risk in Patients with Type 2 Diabetes Mellitus in Northwest China
Authors: ZHAO QINGBIN
Published: June 2020

Background: Diabetes has become an epidemic in developing countries. The incidence of adverse cardiovascular events is high and the control rate is low in Chinese patients with type 2 diabetes mellitus (T2DM), particularly in low-income areas of China. The aim of this study was to identify the risk factors for acute coronary syndrome (ACS) in patients with T2DM in Northwest China.

Methods: A total of 456 patients with T2DM admitted to The First Affiliated Hospital of Xi’an Jiaotong University from January 2018 to January 2019 were included in the study. Among these, 186 individuals newly diagnosed with ACS were categorized as the case group and 270 patients without ACS as the control group. Demographic characteristics and serum biomarkers between the two groups were compared using t-test or Chi-square test. Multivariate logistic regression was used to analyze the risk factors for ACS in T2DM.

Results: The proportion of elderly patients was higher in the case group than in the control group (30.65% vs. 15.56%, p <0.001). Similarly, the case group had a higher proportion of patients with hypertension history (66.67% vs. 34.81%, p <0.001), substandard systolic blood pressure (SBP) (44.09% vs. 21.48%, p <0.001), hyperuricemia (34.41% vs. 11.11%, p <0.001), and higher body mass index (BMI) (26.64 ± 3.96 vs. 24.87 ± 3.53, p <0.001) compared to the control group. The multivariate logistic regression analysis showed that age ≥ 65 years, SBP ≥ 140 mm Hg, overweight or obesity, and hyperuricemia were all risk factors for ACS in T2DM, and their OR (95% CI) were 2.283 (1.371-3.802), 2.067 (1.322-3.233), 2.019 (1.501-2.716), and 3.108 (1.831-5.275), respectively.

Conclusion: Aging, systolic hypertension, obesity and hyperuricemia increase the risk of ACS in T2DM in Northwest China. Thus, weight loss and lowering of SBP and of blood uric acid levels may reduce the risk of ACS in T2DM.

Disclosure

Q. Zhao: None.

Funding

Shaanxi Province Science Technology of China (2019KW-079)
2270-PUB: Advanced Diabetes Does Not Necessarily Increase Risk of Liver Disease
Authors: MINHEM MOHAMAD A. , NAKSHABANDI AHMAD , CERASALE MATTHEW
Published: June 2020

Objective: The association between uncontrolled diabetes mellitus (DM) and liver disease (LD) has been established. The impact of different etiologies of LD on specific DM complications (Cx) is not well known. The aim of this study is to determine if the frequency of DM complications are related to specific LD etiologies.

Methods: This is a cross-sectional study of the 2016 National Inpatient Sample database. Adult patients with type 2 DM were studied. ICD-10 CM codes were used to identify DM complications and different LD etiologies. Univariate analyses and multivariate logistic regressions were performed.

Results: 1,538,604 patients met inclusion criteria. The prevalence of DM complications was 26.9%. Based on multivariate analysis results (Table 1), kidney Cx were associated with increased viral hepatitis. Ophthalmic Cx were preferentially associated with an increased prevalence of hepatitis B but not C. Skin and circulatory Cx were associated with less reported in all LD, while neurological Cx were associated more with reported LD.

Conclusion: Among inpatients, the prevalence of DM Cx differ by LD, with more significant differences noted for patients with Hepatitis etiologies. Our analysis is limited with retrospective nature and should be supported with future prospective studies.

Disclosure

M.A. Minhem: None. A. Nakshabandi: None. M. Cerasale: None.
2274-PUB: Prevalence and Risk Factors of Sexual Dysfunction in Romanian Women with Type 2 Diabetes Mellitus and Chronic Autoimmune Thyroiditis
Authors: GHERBON ADRIANA , FRANDES MIRELA , TIMAR ROMULUS
Published: June 2020

Background: Female sexual dysfunction (FSD) represents one of the chronic complications of diabetes, as the male sexual dysfunction, but less studied.

Objective: Assessment of the prevalence and risk factors associated with sexual dysfunction in Romanian women with type 2 diabetes mellitus (T2DM) and chronic autoimmune thyroiditis (CAT).

Material and Methods: The study sample included 132 Romanian women with T2DM and CAT. The presence of sexual disfunction was established using two validated questionnaires: Female Sexual Function Index, and Female Sexual Distress Scale-revised. CAT diagnosis was confirmed by the presence of antithyroid antibodies (antiperoxidase and antithyroglobulin antibodies).

Results: We found that 47% of women presented sexual dysfunctions: 25.8% loss of libido, 21.2% orgasm, 17.4% lubrication, 18.9% arousal, 8.3% pain, and 17.4% satisfaction problems. The analysis revealed that age was a significant risk factor for FSD, along with DM duration and CAT duration. The presence of dyslipidemia as well as a ponderal status of overweight or obese, and also the glycemic control parameters such as average glycemic level and HbA1c were significant risk factors for FSD.

Conclusions: Women with T2DM and CAT are more likely to present FSD and should be aware of additional risk factors.

Disclosure

A. Gherbon: None. M. Frandes: None. R. Timar: None.
2277-PUB: Factors Effecting Diabetes Patients’ Perception of Diet: An Extended Analysis of Facts and Fads
Authors: ERUKULAPATI RAVI S. , RM MANIKANDAN , DASH KALPANA , DAS SAMBIT , REDDY HARINI , S VENKATARAMAN , PUNJANI ARSHAD , YALAMANCHILI SANDHYA , TIPPISETTY SUREKHA , KOLUKULA VAMSI K. , KRISHNA SESHADRI G.
Published: June 2020

Aim: To assess the awareness of dietary constituents, perception of diet in those with diabetes.

Methods: A cross-sectional, objective, qualitative questionnaire-based assessment was performed among outpatient diabetes patients who visited tertiary diabetes care centers in Metropolitan cities of India. The questionnaire was answered by diabetes patients in a simple ’Yes’ or ’No’ format about awareness of various foods. The questions were about diet, quantity that could be consumed, and myths about various food products consumed routinely. Patients’ perception of diet was categorized into ’correct’ and ’incorrect’ depending on their response to each question in a 10-point questionnaire. A score of ’1’ was given for each ’correct’ answer and ’0’ for an ’incorrect’ answer. Maximum score one can get is 10/10 and a minimum of 0/10.

Results: Total 2295 patients with diabetes were assessed. Patients included were in age range 9 to 90 years with median age of 55 years. Males, females were 62% and 38% respectively. A mean score of 3.8 and median score of 4 was achieved. 2.2% patients achieved maximum correct answer score of 9/10 and 5.9% patients had minimum score 0/10. 44.8% patients scored <4, 45.3% scored 4-6 and 9.9% scored >6. Socioeconomic status, education, HbA1c levels, occupation, Insulin use, duration of diabetes were the factors that contributed to better level of awareness about diet (p <0.001). Age <40 years, male gender, type 1 diabetes, previous diet counseling, exercise patterns, family history of diabetes, unmarried, weight <40kgs, history of no alcohol usage and being non-smoker, were associated with better scores, although not statistically significant.

Conclusion: This study gives valuable insights into the lack of awareness about daily food products and the factors associated with the diet perceptions in diabetic patients. This facilitates health care providers to work on areas that could facilitate improving awareness about dietary practices in those with diabetes.

Disclosure

R.S. Erukulapati: None. M. Rm: None. K. Dash: None. S. Das: None. H. Reddy: None. V. S: None. A. Punjani: None. S. Yalamanchili: None. S. Tippisetty: None. V.K. Kolukula: None. S.G. Krishna: None.
229-OR: The Gut Microbiome in Adults with Prediabetes and Effect of Dietary Glycemic Index
Authors: UTZSCHNEIDER KRISTINA , NEUHOUSER MARIAN L. , NEWTON KATHERINE M. , BREYMEYER KARA , HULLAR MEREDITH A.
Published: June 2020

Compositional and functional differences in gut bacteria exist between adults with normal glucose tolerance, IGT and type 2 diabetes and can change with dietary factors. We compared the gut microbiome between adults with impaired fasting glucose (IFG), impaired glucose tolerance (IGT) and IFG+IGT and evaluated the impact of dietary glycemic index (GI) on the gut microbiome. The study enrolled 53 adults (mean±SEM: age 53.0±1.4 y, BMI 32.5±0.9 kg/m2) with IFG (n=20), IGT (n=10) or both IFG+IGT (n=23). Stool samples were collected in RNAlater after 2 weeks on a control, moderate GI diet (GI 55-58) and again after a 4 week dietary intervention of a low GI (GI<35, n=18) or high GI diet (GI>70, n=18 on placebo and n=17 on the antioxidant N-acetyl-cysteine 1200 mg po BID). Diets had the same macronutrient composition (55%carbohydrate/30% fat/15% protein) but more fiber in the LGI (65.1±2.2 g/d) vs. HGI diet (25.8±0.8 g/d). The stool microbiome was analyzed using 16S rRNA gene sequencing (V1-V3).

Results: α diversity was significantly higher in IGT vs. IFG+IGT (7.2±0.08 vs. 7.0±0.06, p<0.05). Linear regression analysis adjusted for age, sex, BMI, race and sequencing plate showed 3 genera differed across groups on the control diet: Fusicatenibacter (p=0.01) and Lachnospiraceae UCG-008 (p=0.02) were lower in IGT vs. IFG+IGT; Oxalobacter was reduced in IFG (p=0.04) and enriched in IGT (p=0.004) vs. IFG+IGT. After 4 weeks on LGI or HGI diets α diversity did not differ and NAC had no effect on the microbiome. Adjusted regression analysis showed only the genus Anaerostipes differed, being higher in the LGI group (p<0.0001).

Conclusions: Differences in microbiota were observed between IGT and IFG+IGT. These associations may contribute to poorer metabolic function in IFG+IGT, but would need further testing to determine cause and effect. The LGI diet was associated with higher Anaerostipes, a butyrate producing bacteria with beneficial anti-inflammatory effects. This may be related to the higher fiber content of the LGI diet.

Disclosure

K. Utzschneider: Other Relationship; Self; Medtronic. M.L. Neuhouser: None. K.M. Newton: None. K. Breymeyer: None. M.A. Hullar: None.

Funding

National Institutes of Health (R01DK092568, P30DK017047, UL1TR002319, P30CA015704, KL2TR002317, TL1TR002318)
2297-PUB: The Shape of Glucose Response Curve during Oral Glucose Tolerance Tests (OGTT) and Long-Term Near-Normoglycemia Remission in African Americans Presenting with Hyperglycemic Crises
Authors: COTTEN LUCIA , STEFANOVSKI DARKO , OLADEJO OMOLADE , UMPIERREZ GUILLERMO E. , VELLANKI PRIYATHAMA
Published: June 2020

Approximately 70% of obese African American (AA) patients with new-onset diabetic ketoacidosis (DKA) and severe hyperglycemia (SH) achieve near-normoglycemia remission (defined as HbA1c <7%, fasting blood glucose [BG]< 130 mg/dl, and off insulin for 1 month) with intensive insulin treatment. The shape of glucose response curve during OGTT has been shown predictive of future glycemic failure and beta(β)-cell function in adolescents with diabetes and adults. Biphasic (Bφ) curves showing better glycemic control and β-cell function compared to monophasic (Mφ) or incessant rise (IR) curves. We hypothesize that in AA patients with new-onset DKA and SH, Bφ shape at remission is associated with lower hyperglycemia relapse compared to Mφ or IR curves. Seventy AA patients underwent a 75g two-hour OGTT after near-normoglycemia remission and at end of study (median follow-up, 392 days). Insulin sensitivity (Si, oral minimal model) and β-cell function (AUCi: incremental area under the curve of insulin), and disposition index (Di= Si x AUCi) were compared among the different OGTT response curves. Hyperglycemia relapse-free survival (fasting BG > 130 mg/dl, 2 random BG > 180 mg/dl or HbA1c > 7%) was calculated with Kaplan-Meier curve. At remission, initial OGTT showed 56% of patients with Mφ, 16% with Bφ, and 29% with IR curve shape. Baseline characteristics did not differ. Even though, more patients in the Bφ group (88%) sustained remission than the Mφ (57%) and IR (53%), hyperglycemia relapse-free survival did not differ by curve shape (log rank, p=0.26). On initial OGTT, Si (p=0.09), AUCi (p=0.12) and Di (p=0.06) did not differ between curve shapes. At last OGTT, Si (p=0.43), AUCi (0.47) and Di (p=0.22) did not differ. In obese AA patients with new-onset DKA and SH, the shape of the glucose response curve during OGTT does not predict long-term glycemic control or β-cell function.

Disclosure

L. Cotten: None. D. Stefanovski: None. O. Oladejo: None. G.E. Umpierrez: None. P. Vellanki: Consultant; Self; Boehringer Ingelheim Pharmaceuticals, Inc.

Funding

National Institutes of Health (K08DK0830361, K23DK113241-01A1, UL1TR002378, 1P30DK111024-01)
2304-PUB: Increased Glucagon Expression in Diabetes: The Role of miR-320a
Authors: JO SEONGHO , XU GUANLAN , JING GU , SHALEV ANATH
Published: June 2020

Glucagon, secreted from the α-cells of pancreatic islets, plays an important, yet underappreciated role in glucose homeostasis. As the major counter-regulatory hormone to insulin, it helps maintain normal blood glucose levels and prevent life-threatening hypoglycemia. However, while the regulation of glucagon has been shown to be perturbed in subjects with diabetes leading to inadequately high levels and worsening of the hyperglycemia, the molecular mechanisms involved in this abnormal regulation are still unknown. We therefore first investigated the possibility that microRNAs might be involved in the regulation of glucagon expression. Indeed, analysis of the glucagon 3’UTR revealed two potential miR-320a binding sites in human and one potential site in mouse and rat, respectively. In addition, we performed luciferase reporter assays and found that miR-320a directly binds to the 3’ UTR of the human glucagon gene. To further determine the role of miR-320a in the regulation of glucagon gene expression, we performed gain and loss of function studies with miR-320a in αTC1-6 cells. Glucagon gene expression was reduced in response to miR-320a overexpression, whereas inhibition of miR-320a upregulated glucagon gene expression. Interestingly, consistent with its role in the regulation of glucagon, miR-320a is more highly expressed in α-cells than beta cells and its expression was decreased by high glucose. Moreover, this was associated with an increase in glucagon gene expression in mouse αTC 1-6 cells as well as human islets. Finally, these findings are not only consistent with the results in response to high glucose, but also demonstrate the pathophysiological relevance of this phenomenon. Taken together, our data suggest that reduced miR-320a expression may contribute to the increase in glucagon gene expression in type 2 diabetes. These findings indicate for the first time that glucagon gene expression is under the control of a microRNA and provide novel insight into the abnormal regulation of glucagon in diabetes.

Disclosure

S. Jo: None. G. Xu: None. G. Jing: None. A. Shalev: None.
231-OR: Phase I Study Investigating the PD, PK, and Safety of AT247 in Comparison with Novorapid and Fiasp
Authors: PIEBER THOMAS R. , AUGUSTIN THOMAS , MAGNES CHRISTOPH , GERRING DAVID J. , JEZEK JAN , HOWELL SARAH J. , ZAKRZEWSKI LEON , LAWRENCE FIONA J. , SVEHLIKOVA EVA
Published: June 2020

Background: AT247, an ultra-rapid acting formulation of insulin aspart designed for faster absorption following s.c. injection, will lead to better postprandial glycaemic control and is key to the development of closed-loop pump systems.

Method: Plasma glucose and serum insulin concentrations were measured in 19 adult male participants with T1DM following a single s.c. dose (0.3 U/Kg) of insulin in a randomised, double-blind, cross over euglycaemic clamp study.

Results: Data presented as median; range. AT247 had a faster onset of glucose lowering effect than NovoRapid® and Fiasp® (onset of action: 17; 12-30 vs. 37; 20-88 vs. 23;16-50 mins). The early glucose lowering effect was greater for AT247 than NovoRapid® and Fiasp® (AUCGIR 0-60min: 212; 63-465 vs. 49; 0-303 vs. 91; 11-300 mg/kg; GIR=glucose infusion rate) (Fig 1A). Similarly, greater initial insulin exposure was seen for AT247 compared to NovoRapid® and Fiasp® (AUC Insulin 0-60min: 111; 48-205 vs. 41; 12-147 vs. 66; 25-188 mU*h/L). In addition, offset of exposure occurred earlier for AT247 than NovoRapid® and Fiasp® (time to late 50% Cmax Insulin: 173; 89-414 vs. 212; 106-389 vs. 221;106-441 mins). (Fig 1B). Comparisons are statistically significant with p<0.05. No safety signals were detected for AT247.

Conclusion: AT247 has a superior (left-shifted) time-action and time-concentration profile as compared to both NovoRapid® and Fiasp®.

Disclosure

T.R. Pieber: Advisory Panel; Self; ADOCIA, Arecor, AstraZeneca, Eli Lilly and Company, Novo Nordisk A/S, Sanofi. Research Support; Self; AstraZeneca, Novo Nordisk A/S. Speaker’s Bureau; Self; Novo Nordisk A/S, Roche Diagnostics K.K. T. Augustin: None. C. Magnes: None. D.J. Gerring: None. J. Jezek: None. S.J. Howell: None. L. Zakrzewski: None. F.J. Lawrence: None. E. Svehlikova: None.
2314-PUB: The Uric Acid Is Differentiated by Thyroid Hormones Levels in Obese Patients with Acanthosis Nigricans
Authors: ZHANG JUNYI , LI GUIFANG , CHENG XIAOYUN , WANG XINGCHUN , JAYACHANDRAN MUTHUKUMARAN , HUANG YUEYE , QU SHEN
Published: June 2020

Purpose: To investigate the serum levels of uric acid (UA) in obese acanthosis nigricans (AN) patients with different thyroid hormones (THs) levels within the normal range.

Methods: We investigated 834 obese patients (385 females and 449 males) with euthyroid function in the department of endocrinology of shanghai 10th people’s hospital from 2011 to 2019 with the median age of 29 years (interquartile range 22-36 years) for this study. 343 patients were diagnosed of AN. The cut-offs for thyroid-stimulating hormone (TSH), free triiodothyronine (FT3) and free thyroxine (FT4) were 2mIU/L, 16mIU/L and 5mIU/L, respectively.

Results: In male obese patients, AN patients with high TSH and FT4 had higher levels of UA compared to those with high TSH and low FT4 (523.20±120.98 vs. 477.56±86.92umol/L, P=0.036) and showed a correlation with fasting C-peptide (FCP) (r=0.330, P=0.029). The TSH, FT3, and FT4 alone did not showed any predictive value in male patients. In female patients, high FT3 was associated with higher levels of UA (388.47±100.95 vs. 366.33±82.40umol/L, P=0.019). However, the predictive value of FT3 in female was obtained only in low TSH subgroup (low TSH+ high FT3 vs. low TSH+ low FT3, P=0.033) not in high TSH patients. The levels of fasting insulin, 2h-insulin, FCP, 2h C-peptide, total cholesterol, triglyceride and testosterone were in correlation with higher UA levels in female obese patients with low TSH and high FT3(all P<0.05).

Conclusion: The metabolic characteristics of UA are found to differ in obese patients with euthyroid function, in which AN plays an important role for male obese patients with high TSH and high FT4. But in the case of female, a combination of THs instead of AN showed it’s unique in UA metabolism.

Disclosure

J. Zhang: None. G. Li: None. X. Cheng: None. X. Wang: None. M. Jayachandran: None. Y. Huang: None. S. Qu: None.

Funding

National Key Research and Development Program of China (2018YFC1314100); National Natural Science Foundation of China (81970677); Shanghai Pujiang Program (2019PJD040); Effect of ApoC3 High Expression on Islet Cells and Related Molecular Mechanism (03.05.18.002)
236-OR: Molecular and Biological Properties of Insulin Icodec, a New Insulin Analog Designed to Give a Long Half-Life Suitable for Once-Weekly Dosing
Authors: NISHIMURA ERICA , KJELDSEN THOMAS , HUBALEK FRANTISEK , GLENDORF TINE , STIDSEN CARSTEN E. , HANSEN BO F. , PEDERSEN THOMAS A. , LUETZEN ANNE , PRIDAL LONE , MADSEN PETER
Published: June 2020

Insulin icodec is a novel insulin analog with a terminal elimination half-life of ∼196 hours currently in clinical development, designed to be able to cover a full week’s basal insulin requirements with a single subcutaneous injection. To attain such a long half-life, the insulin molecule was modified to achieve an albumin-bound circulating depot of icodec which acts just as human insulin (HI) but is more slowly cleared.Addition of a C20 fatty diacid containing side chain at B29K via a hydrophilic linker imparts strong but reversible albumin binding (10x that of insulin detemir). Three amino acid substitutions (A14E, B16H and B25H) ensure reduced enzymatic degradation of icodec and contribute to attenuating insulin receptor (IR) binding and clearance, further prolonging the half-life. In vitro studies demonstrated that icodec is a specific and full agonist of the human IR, and that it displays the same dose-dependent mode of action as HI, exemplified by its ability to phosphorylate the IR and stimulate intracellular signaling pathways (phospho-AKT and -ERK). Functional assays have demonstrated that icodec elicits the same pattern of metabolic effects as HI, including glucose uptake and lipogenesis in fat cells, and stimulation of glycogen synthesis in liver cells. The affinity of icodec for the IGF-1 receptor was found to be proportionately lower than its binding to the IR. The in vitro mitogenic effect of icodec in primary human mammary cells (HMEC), as well as in mammary and colon carcinoma cells (MCF-7 and COLO 205) was found to be low relative to that of HI.

In conclusion, icodec is a new insulin analog designed to achieve a slowly cleared, albumin-bound circulating depot which results in a long half-life suited for once weekly injections, covering the basal insulin requirements for a full week. *Insulin icodec (proposed INN).

Disclosure

E. Nishimura: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. Stock/Shareholder; Spouse/Partner; Novo Nordisk A/S. T. Kjeldsen: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. Stock/Shareholder; Spouse/Partner; Novo Nordisk A/S. F. Hubalek: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. T. Glendorf: Employee; Spouse/Partner; Novo Nordisk A/S. Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Spouse/Partner; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. C.E. Stidsen: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. B.F. Hansen: Stock/Shareholder; Self; Novo Nordisk A/S. T.A. Pedersen: Employee; Self; Novo Nordisk A/S. A. Luetzen: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. L. Pridal: None. P. Madsen: Employee; Self; Novo Nordisk A/S.
237-LB: Cellular Heterogeneity in Brown, Dermal, and Bone Marrow Adipose Depots
Authors: POWELL NOAH D. , SHARMA RITA , HUANG JUN , LEE KEVIN Y.
Published: June 2020

Recent studies have shown that heterogeneity among adipocytes exists even within a single white adipose tissue depot. Previous studies from our laboratory have identified three functionally distinct subpopulations of adipocytes with different gene expression and metabolic properties. These adipocyte subpopulations differentially contribute to the individual white adipose tissue depots and can be distinguished by their specific expression of three marker genes: Wilms Tumor 1 (Wt1), Transgelin (Tagln), and Myxovirus 1 (Mx1). In this study, we extend these observations by determining the contributions of these adipocyte subpopulations to other adipose tissue depots including the interscapular brown, dermal, and bone marrow adipose tissue depots. Lineage tracing analysis of Wt1-cre and Tagln-cre crossed to a dual-fluorescent reporter mice which mark type 1 and type 2 preadipocytes and adipocytes, respectively, was performed in two to four month old mice and preadipocytes and adipocytes were quantified by FACS analysis and confocal microscopy. In all depots analyzed in this study, little to no type 1 preadipocytes or adipocytes were present in any of these depots. On the other hand, a significant number of type 2 preadipocytes and adipocytes were found in all depots. In dermal tissue, type 2 adipocytes comprised 9±2% of adipocytes and 45±7% of preadipocytes. Type 2 adipocytes significantly contributed to the bone marrow adipose tissue as 51±4% of adipocytes present and 83±3% of preadipocytes were type 2 cells. Lastly, in brown adipose tissue, type 2 cells comprised 15±1% of adipocytes and 10±3% of preadipocytes. Further studies investigating the metabolism and thermogenic properties of these type 2 brown adipocytes is currently being performed, as a more thorough understanding these adipocyte subpopulations may lead to novel, targeted treatments for obesity and diabetes.

Disclosure

N.D. Powell: None. R. Sharma: None. J. Huang: None. K.Y. Lee: None.
241-OR: Causal Gene Candidates for Type 2 Diabetes Based on Protein-Coding Variants in 127,676 Individuals
Authors: DORNBOS PETER , RAFFIELD LAURA , YIN XIANYONG , FLANNICK JASON
Published: June 2020

Genome-wide association studies (GWAS) have identified >250 genetic loci associated with type 2 diabetes (T2D), but, currently, few associations have been translated to causal genes. To directly implicate genes in T2D pathogenesis, we analyzed coding variants from the largest whole exome sequencing (WES) study to date, consisting of 36,652 cases and 91,024 controls from the Accelerating Medicines Partnership-T2D-GENES, Cohorts for Heart and Aging Research in Genomic Epidemiology, Myocardial Infarction Genetics, and NHLBI Trans-Omics for Precision Medicine consortia. Through single-variant and gene-level meta-analysis, we identify three evidence tiers of T2D-susceptibility genes. First, 11 genes harbor exome-wide significant missense associations (p≤4.3x10-7) and four genes have significant gene-level associations (p≤2.5x10-6). Two of these associations are novel: a missense mutation in MET (p=2.33x10-7), a gene with no previous genetic link to T2D, and, while presence of misdiagnosed MODY cases cannot be excluded, a series of rare alleles in GCK providing the first exome-wide significant association with common T2D. Second, we apply a gene-level Bayesian statistical approach to identify 29 genes with posterior probabilities (PPA) of ≥50% of explaining signals at GWAS loci. Twenty-one of these genes lie within loci where no causal gene is known, including SGP7 (PPA=0.85), a zinc metalloprotease that contains a previously reported independent common missense variant associated with T2D. Finally, we extend this Bayesian approach to another 1,069 genes previously studied in mice. This analysis identifies 17 genes with gene-level association PPA ≥50%, including 11 genes with no prior human genetic link to T2D such as ONECUT1 (PPA=0.54) which is exclusively expressed in liver and pancreas. Our results further illustrate the value of WES in identifying genes that may be future T2D drug targets. All results presented will be available through the AMP-T2D Knowledge Portal.

Disclosure

P. Dornbos: None. L. Raffield: None. X. Yin: None. J. Flannick: None.
245-OR: CIS-Regulatory Architecture of Human ESC-Derived Hypothalamic Neuron Differentiation Aids in Variant-to-Gene Mapping of Type 2 Diabetes Related Traits
Authors: PAHL MATTHEW C. , LITTLETON SHERIDAN H. , DOEGE CLAUDIA , HODGE KENYAITA M. , LEONARD MICHELLE , LU SUMEI , HAMMOND REZA , BOEHM KEITH , LASCONI CHIARA , SU CHUN , CHESI ALESSANDRA , PIPPIN JAMES A. , JOHNSON MATTHEW , WELLS ANDREW D. , VOIGHT BENJAMIN F. , LEIBEL RUDOLPH , GRANT STRUAN F. , COUSMINER DIANA L.
Published: June 2020

Dysregulation of hypothalamus-mediated metabolic homeostasis influences risk factors for T2D, such as BMI, pubertal timing and sleep. However, the gene regulatory landscape of the hypothalamus remains unclear. We characterized the physical relationships among gene promoters and their corresponding cis-regulatory elements in human embryonic stem cell (ESC)-derived hypothalamic cells by integrating RNA-seq, ATAC-seq and promoter-focused Capture C profiles. We conducted this effort in an in vitro model of hypothalamic neurons using human ESCs at 3 stages of differentiation: ESCs, hypothalamic progenitors (HPs) and hypothalamic-like neurons (HNs). We identified 87,170 open chromatin regions (OCRs) contacting promoters in at least 1 stage of HN differentiation. Interestingly, we observed that regardless of gene expression pattern during differentiation, overall accessibility of interacting OCRs first increased during the transition to HP, then decreased following differentiation to HN. In order to relate these findings to GWAS data, we tested for enrichment of SNPs within promoter-interacting OCRs. We found significant associations with BMI, sleep traits, age at menarche (AAM) and major depressive disorder (MDD). We mapped these trait-associated variants to their physically interacting genes. We identified 2 putative effector genes for AAM (RPS26 and SUOX at chr12q13.2) and 1 for BMI (DHRS11), which were also supported by colocalized hypothalamus eQTLs in GTEx. Additionally, HyPrColoc analyses revealed several putative effector genes associated with >1 trait, including ZRANB2 for AAM, BMI and MDD, and FEZF1 for AAM and BMI, the latter being known to harbor Mendelian mutations that impair puberty. Taken together, our analyses provide insight into cis-regulatory architecture during hypothalamic development and how it may contribute to phenotypes associated with T2D risk.

Disclosure

M.C. Pahl: None. S.H. Littleton: None. C. Doege: None. K.M. Hodge: None. M. Leonard: None. S. Lu: None. R. Hammond: None. K. Boehm: None. C. Lasconi: None. C. Su: None. A. Chesi: None. J.A. Pippin: None. M. Johnson: None. A.D. Wells: None. B.F. Voight: None. R. Leibel: None. S.F. Grant: None. D.L. Cousminer: None.

Funding

American Diabetes Association (1-17-PDF-077 to D.L.C.); National Institutes of Health (K99HD099330-01); National Center for Research Resources (UL1RR024134); National Center for Advancing Translational Sciences (UL1TR000003); Institute for Translational Medicine and Therapeutics; Children’s Hospital of Philadelphia
249-LB: Effects of Oxidative Phosphorylation Complex Defects on Beta-Cell Biology
Authors: LANG ANNA L. , CAICEDO ALEJANDRO
Published: June 2020

Background: The insulin-secreting pancreatic beta cell is a highly metabolic cell type relying on mitochondrial metabolism for stimulus-secretion coupling. Specifically, the ATP-producing oxidative phosphorylation (OXPHOS) pathway is central to insulin secretion. Although disruptions in ATP production occur in beta cell dysfunction, the exact mechanisms remain unknown. We have generated beta cell specific OXPHOS complex inducible knock out mouse models of Complex I, III, and IV to determine the direct effects OXPHOS complexes on beta cell function. We hypothesize that individual complexes contribute distinctly to beta cell function.

Methods: Eight-week-old, male OXPHOSf/f - MIPcreERT mice were administered tamoxifen (150mg/kg) or vehicle by I.P. injection for 5 consecutive days. Non-fasting glycemia and body weight were measured one week prior and once per week up to 5 weeks after induction. Glucose tolerance was assessed by glucose tolerance tests (IP-GTT) 1 week before and 3 weeks after induction for all groups. Mice were anesthetized with ketamine/xylazine and perfused with sterile PBS followed by 4% PFA. Pancreata were dissected, embedded in OCT, and cut at 40µM sections for immunofluorescent staining of insulin, glucagon, and somatostatin.

Results: No changes in body weight were observed throughout the experimental period for any group. Complex III knockout animals developed severe hyperglycemia and significant glucose intolerance at 3 weeks after induction. Although the other models display a minor trend towards hyperglycemia starting at 5 weeks after induction, they retain their ability to regulate glucose at 3 weeks as seen by IP-GTT. Interestingly, no changes in islet cytoarchitecture are observed in any of the groups at the 3-week timepoint.

Conclusions: Mice with Complex III deficient beta cells show a robust diabetic phenotype compared to Complex I and IV models. These results support the hypothesis that specific OXPHOS complexes contribute differentially to beta cell function.

Disclosure

A.L. Lang: None. A. Caicedo: None.

Funding

National Institute of Environmental Health Sciences (R21ES025673-01)
251-OR: Cardiac Autoantibodies (AB) Predict Cardiovascular Disease (CVD) in T1D and Are Associated with Cytokine Signatures Linked to CVD Risk
Authors: SOUSA GIOVANE R. , NIEWCZAS MONIKA , LIPES MYRA A.
Published: June 2020

CVD is the leading cause of mortality in T1D; however, only ∼50% of this risk is explained by traditional risk factors. Recent studies show that subjects with T1D in the DCCT developed cardiac autoimmunity, defined by the presence of ≥2 Ab. We tested the hypothesis that some of the unexplained CVD risk in T1D is mediated by Ab-linked cytokine pathways. We measured Ab in serum from 892 post-DCCT EDIC subjects (mean age 49 and T1D duration 28 years), with no CVD events at baseline (Figure). Cumulative incidence of a first CVD event in subjects with 0, 1, and >2 Ab was 2.0%, 8.9%, and 43%, respectively, over 4-year median follow-up. In Cox regression models, ≥2 Ab predicted CVD events with minimal attenuation by known risk factors. Using a 42-plex array to measure baseline serum cytokines in subjects with ≥2 Ab and matched controls without Ab, the ≥2 Ab group showed 3.4-fold odds (95% CI, 1.6-6.1; P < 0.0001)of having a 1 tertile increase in a proinflammatory component comprising IFN-γ, TNF-α, G-CSF, MDC, IL1-RA, IL-8, and EGF. Risk of CVD events was also independently associated with IFN-γ, TNF-α, and G-CSF levels (HR, 5.4; 95% CI, 1.5-18.9; P = 0.009).

In conclusion, in this cohort with longstanding T1D, cardiac Ab identified subjects at high risk for CVD events, potentially through Ab-linked cytokine pathways. These findings raise the possibility to immunologically predict and prevent CVD in T1D.

Disclosure

G.R. Sousa: None. M. Niewczas: None. M.A. Lipes: None.

Funding

National Institutes of Health (DK1036909)
290-OR: Diabetes Progression in IFG-Receptor Deficient NOD Mice Is Associated with Increased CD8+ T Lymphocyte Sensitivity to Common Gamma Chain Cytokines
Authors: KAY THOMAS , JHALA GAURANG , KRISHNAMURTHY BALASUBRAMANIAN , THOMAS HELEN E.
Published: June 2020

IFN-γ is a potential drug target in type 1 diabetes and this is part of the rationale for trialing JAK inhibitors (JAKi) for treatment of new onset T1D. It regulates migration and adhesion of leucocytes, as well as expression of MHC class I on islet beta cells. However, neither genetic deficiency of IFN-γ nor its receptor in NOD mice modifies disease progression. To understand this paradox we characterised islet antigen-specific T cells in NOD mice lacking the IFN-γ receptor (Ifngr) generated directly in NOD mouse embryos using CRISPR/Cas9 gene editing. Islet antigen-specific CD8+ T cells (proinsulin B15-23 and IGRP206-214) and CD4+ T cells (proinsulin B10-23and BDC2.5 mimotope) were quantified using tetramer enrichment and flow cytometry. At 125 days of age, insulin-specific (177±41 cells in wild type and 1799±1123 cells in Ifngr mutant, p=0.036) and IGRP-specific CD8+ T cells (2524±735 cells in wild type and 30598±10426 cells in Ifngr mutant, p<0.0001) were present in significantly greater numbers in Ifngr mutant NOD mice. There were no differences in antigen-specific CD4+ T cells. Islet infiltrating T cells from Ifngr mutant mice had increased responses to common gamma chain (γc) cytokines, prevented by a JAKi, and reduced expression of suppressor of cytokine signalling-1 (Socs1) mRNA. Our data suggest that as well as its pro-inflammatory effects, IFN-γ homeostatically controls the expansion of antigen-specific CD8+ T cells by regulating their response to γc cytokines, by mechanisms that include regulation of SOCS-1 expression. Expansion of antigen-specific CD8+ T cells is likely to explain the lack of protection from diabetes in Ifngr mutant NOD mice. JAKi therapy experimentally controlled this increased responsiveness to γc cytokines. Our data show mechanisms underlying protection from diabetes in NOD mice by JAKi and the lack of protection by blocking IFN-γ alone.

Disclosure

T. Kay: None. G. Jhala: None. B. Krishnamurthy: None. H.E. Thomas: None.
291-OR: Deletion within the Helicase 1 Domain of MDA5 Dampens Immune Cell Activation and Delays Type 1 Diabetes
Authors: BLUM SAMUEL I. , CHEN YI-GUANG , TSE HUBERT M.
Published: June 2020

Antiviral responses are associated with triggering type 1 diabetes (T1D) including the synthesis of reactive oxygen species, proinflammatory cytokines, and autoreactive T cell activation. We previously demonstrated that Coxsackievirus B3 (CVB3) infection of Non-Obese Diabetic (NOD) mice accelerates T1D, partly due to the expression of melanoma differentiation-associated protein 5 (MDA5) antiviral sensor. Genetic mutations within human IFIH1, the gene encoding MDA5, are correlated with T1D susceptibility potentially due to increased IFN-α/β synthesis. However, the role of MDA5 in immune cell activation during T1D is unknown. By utilizing mice that lack (NOD.Ifih1KO) or contain a deletion within the helicase 1 domain of MDA5 (NOD.Ifih1ΔHel1), we can test the hypothesis that reduced MDA5 signaling delays T1D by decreasing IFN-α/β synthesis and debilitating islet-infiltrating macrophage and T cell effector responses. We observed that NOD and NOD.Ifih1KO mice develop T1D similarly, but NOD.Ifih1ΔHel1 were significantly protected from spontaneous and CVB3-accelerated T1D. During spontaneous T1D at 12 weeks of age, pancreatic macrophages from NOD.Ifih1ΔHel1 mice have a significant reduction in MHC-II (∼41%) and CD86 (∼43%) expression compared to NOD and NOD.Ifih1KO mice. CVB3-infected NOD.Ifih1ΔHel1 mice exhibited a significant decrease in pancreatic IFN-α (∼54%) and IFN-β (∼67%) synthesis at day 3 post-infection compared to NOD mice. Pancreatic NOD.Ifih1ΔHel1 CD4 and CD8 T cells had reduced T cell activation via CD25 (∼76 and ∼57%) and PD-1 (∼65 and ∼100%) synthesis, respectively, compared to NOD and NOD.Ifih1KO T cells following CVB3 infection. The Ifih1ΔHel1 mutation, but not the Ifih1KO mutation dampened IFN-α/β synthesis, reduced macrophage and T cell activation, and delayed both spontaneous and CVB3-accelerated T1D. Future studies will examine immune differences between NOD.Ifih1ΔHel1 and NOD.Ifih1KO mice that delay or promote autoimmunity, respectively.

Disclosure

S.I. Blum: None. Y. Chen: None. H.M. Tse: None.

Funding

American Diabetes Association (7-12-CD-11 to H.M.T.); National Institute of Diabetes and Digestive and Kidney Diseases (DK099550)
299-OR: Loss of Insulin and IGF1 Receptors in Muscle Impairs Complex-I Dependent Mitochondrial Bioenergetics and Supercomplex Formation via Foxo Transcription Factors
Authors: BHARDWAJ GOURAV , PENNIMAN CHRISTIE M. , BELTRAN PABLO A. SUAREZ , JENA JAYASHREE , FOSTER COLLIN M. , PORO KENNEDY , JUNCK TAYLOR , HINTON ANTENTOR , SOUVENIR RHONDA , FUQUA JORDAN , MORALES PABLO E. , BRAVO-SAGUA ROBERTO , LIRA VITOR A. , ABEL EVAN D. , ONEILL BRIAN T.
Published: June 2020

Diabetes is associated with decreased muscle strength and mitochondrial metabolism which can lead to disability. Recently, we reported that insulin receptor (IR) and IGF-1 receptor (IGF1R) prevent muscle atrophy via suppression of FoxOs, but how FoxOs interact with mitochondrial function remains unclear. To test whether IR/IGF1R suppression of FoxOs coordinates muscle strength with mitochondrial function, we measured mitochondrial bioenergetics and RNA-Seq in mice lacking IR, IGF1R, and/or FoxO1/3/4 in muscle. Perinatal muscle-specific deletion of IR (MIRKO) showed a mild 20% decrease in pyruvate-driven respiratory capacity in soleus permeabilized fibers, whereas succinate/Rotenone respiration was not different. Deletion of both IR/IGF1R (MIGIRKO) showed a marked 60% decrease in pyruvate respiration and ATP production in soleus fibers. These mitochondrial abnormalities were reversed in muscle-specific IR/IGF1R and FoxO1/3/4 quintuple knockout (QKO), suggesting FoxO activation mediates the mitochondrial decline. FoxOs are known inducers of autophagy, but mitophagy quantification using MitoTIMER or mitoKeima biosensors was not altered in Day 21 Tamoxifen-inducible IR/IGF1R KO in adult muscle (IND-IGIRKO) or by acute in-vitro deletion of IR/IGF1R primary myotubes. Complex-I activity was decreased 50-70% in MIGIRKO and Day 21 IND-IGIRKO muscle, but other complex activities were unchanged. Furthermore, 14 complex-I subunits were decreased in MIGIRKO by RNA-Seq, and 13 of these were reversed in QKO. IND-IGIRKO also showed a decrease in pyruvate respiration, ATP production, complex-I activity, and complex-I/supercomplex content by blue-native gels 21 days after tamoxifen. Thus, loss of IR or both IR/IGF1R in muscle decreases complex-I driven mitochondrial respiration and supercomplex assemblies, at least in part by FoxO-mediated downregulation of mitochondrial genes.

Disclosure

G. Bhardwaj: None. C.M. Penniman: None. P.A. Suarez Beltran: None. J. Jena: None. C.M. Foster: None. K. Poro: None. T. Junck: None. A. Hinton: None. R. Souvenir: None. J. Fuqua: None. P.E. Morales: None. R. Bravo-Sagua: None. V.A. Lira: None. E.D. Abel: None. B.T. ONeill: None.
309-OR: IGF-1 Receptors, Not Insulin Receptors, on Mesangial Cells Are Accelerating Mesangial Expansion and Albuminuria in Streptozotocin-Induced Diabetic Mice
Authors: ISHIKADO ATSUSHI , SHINJO TAKANORI , YOKOMIZO HISASHI , MAEDA YASUTAKA , PARK KYOUNGMIN , QI WEIER , ST-LOUIS RONALD , LI QIAN , KING GEORGE L.
Published: June 2020

Clinical studies have suggested that loss of insulin’s action and excessive IGF-1 levels in the glomeruli elevate the risk for diabetic kidney disease (DKD). To define the potential pathogenic role of IGF-1 and insulin signaling in DKD, we generated mice with selective loss of insulin receptor (IR) or IGF-1 receptor (IGF-1R) in the mesangial cells (MCs) by cross breeding IRflox/flox mice with Sm22α-Cre mice (SMIRKO) or IGF-1Rflox/flox mice with Myh11-Cre mice (SMIGF1RKO), and examined urinary albumin excretion (ACR) and mesangial expansion with 6 month duration of streptozotocin (STZ)-induced diabetes. IGF-1 levels were significantly increased by 1.5 fold in the renal cortex, but not in the plasma of all three types of diabetic mice. Diabetes induced elevation of ACR (2.0 fold), glomerular size and mesangial expansion (2.1 fold), in parallel with increased glomerular expressions of fibrosis-related (Tgfβ1, fibronectin and Col3a) and matrix (Has2) genes by 2-3 fold, significantly, in Wt and SMIRKO mice, but were prevented in diabetic SMIGF1RKO mice. In cultured MCs, IGF-1, at physiological levels, and insulin, only at supra-physiological levels, induced the expressions of Has2 (3.8 fold) and Tgfβ1 (1.7 fold). Deletion of IGF-1R, but not IR, in MCs inhibited both IGF-1 and insulin-induced expressions of Has2. Inhibitor of MAP kinase pathway, but not inhibitor of PI3 kinase reduced insulin or IGF-1-induced expressions of Has2 in MCs. IGF-1-induced phosphorylations of IGF-1R (39 fold), Akt (34 fold) and Erk/MAPK (8 fold) were significantly inhibited in both IR or IGF-1R-deleted MCs. Insulin-induced phosphorylations of IR-β (11 fold) and Akt (27 fold) were also decreased in IR-deleted MCs, but surprisingly increased in IGF-1R-deleted MCs. Thus, MAPK activation by elevated glomerular IGF-1 levels via IGF-1R, but not IR, on MCs, may contribute to the pathogenesis of DKD through induction of Has2 and fibrotic genes.

Disclosure

A. Ishikado: Employee; Self; Sunstar Group. Employee; Spouse/Partner; Sunstar Group. T. Shinjo: None. H. Yokomizo: None. Y. Maeda: None. K. Park: None. W. Qi: None. R. St-Louis: None. Q. Li: None. G.L. King: Research Support; Self; Janssen Pharmaceuticals, Inc.
320-OR: Bariatric Surgery Improves Ca2+ Dynamics across Pancreatic Islets In Vivo
Authors: AKALESTOU ELINA , SUBA KINGA , NORIEGA LIVIA LOPEZ , CHABOSSEAU PAULINE L. , LECLERC ISABELLE , SALEM VICTORIA , RUTTER GUY A.
Published: June 2020

Background: Bariatric surgery improves both insulin sensitivity and secretion in T2D. However, the impact on insulin secretion is difficult to monitor directly, requiring correction for insulin sensitivity. The degree to which surgery impacts β-cell function versus mass, and the time course over which these changes occur, also remain unclear.

Aim: To investigate the effect of bariatric surgery on β-cell function in vivo, we imaged Ca2+ dynamics prospectively at the cellular level in islets engrafted into the anterior eye chamber.

Methods: Ins1Cre mice were crossed to animals that express the genetic calcium indicator GCaMP6f behind a Lox PSTOPLoxP cassette. Isolated islets were engrafted into male C57BL6 mice maintained for 12 weeks on high fat/high sucrose diet (HFD). At pre- (-1) and post-operative weeks 4, 8 and 12, Vertical Sleeve Gastrectomy (VSG) (n=7) and sham (n=6) groups were anaesthetized, and islets imaged in the eye using a Nikon microscope equipped with a spinning disc, 488 nm laser and 20x/1.0 water immersion objective. Islets were scored as positive for wave activity when these were recurrent, had a defined origin and covered >75 % of the image plane. Glucose (3g/kg) tolerance and insulin secretion were assessed in parallel.

Results: The VSG group demonstrated substantial weight loss during the first 4 weeks post-operatively but regained pre-operative weight by week 12. Nonetheless, VSG mice displayed significantly improved glucose tolerance and insulin secretion (p<0.001) when compared to sham mice. VSG improved coordinated Ca2+ activity, with 100% of islets exhibiting enhanced wave readouts 8 weeks post surgery. Over the same time period islet wave activity dropped by 34% in the sham group, falling to zero discernible coordinated islet Ca2+ activity by week 12.

Conclusions: By imaging islet function in the eye in vivo we demonstrate that bariatric surgery leads to a time-dependent increase in glucose-induced Ca2+ dynamics of individual islets, likely to underlie increased insulin secretion.

Disclosure

E. Akalestou: None. K. Suba: None. L. Lopez Noriega: None. P.L. Chabosseau: None. I. Leclerc: Consultant; Spouse/Partner; Sun Pharmaceutical Industries Ltd. Research Support; Spouse/Partner; Servier. V. Salem: None. G.A. Rutter: Consultant; Self; Sun Pharmaceutical Industries Ltd. Research Support; Self; Servier, Sun Pharmaceutical Industries Ltd.

Funding

UK Wellcome Trust (212625/Z/18/Z); Rosetrees Trust (M825); UK Medical Research Council (MR/R022259/1); Harry Keen Diabetes UK Fellowship (BDA 15/0005317); European Union