bluuebunny/crossref_metadata_2025 huggingface dataset:
Circulating Exosomal miRNA Signature in Gestational Diabetes Mellitus Influences Glucose Metabolism in Placental Cells
Authors: NAIR SOUMYALEKSHMI , ORMAZABAL VALESKA , JAYABALAN NANTHINI , GUANZON DOMINIC , LAI ANDREW , DAVID MCINTYRE H. , LAPPAS MARTHA , SALOMON CARLOS
Published: June 2018

There is increasing evidence that miRNA, which are enriched in small nanovesicles called exosomes, are important regulators of gene expression. In this study, we tested the hypothesis that circulating exosomes from womem with gestational diabetes mellitus (GDM) carry a specific set of miRNAs which regulate genes associated with placental glucose metabolism. Exosomes were isolated from plasma obtained from normal glucose tolerant women (NGT; n=12) and women with GDM (n=12) pregnancies at delivery by differential and buoyant density centrifugation. Exosomal RNA was extracted and an Illumina TrueSeq Small RNA kit was used to construct a small RNA library. Gene target identification and gene ontology analysis for miRNAs was performed using the Ingenuity pathway analysis (IPA). The effect of exosomes on glucose metabolism in placental cells was assessed using a human glucose metabolism array. In plasma, a total of 44 miRNAs were upregulated in exosomes from women with GDM compared to NGT pregnant women (p<0.005). IPA showed that exosomal miRNAs isolated from women with GDM regulates the expression of placental genes associated with the glycolytic pathway including 6-phosphofructokinase and phosphopyruvate hydratase. Interestingly, exosomes from women with GDM increases the expression of placental genes associated with glycolysis and decrease the expression of genes associated with pentose phosphate pathway. After integration of the miRNA and mRNA data, we identified 74 differentially expressed exosomal miRNAs associated with the modulation of 10 potential target mRNAs in placental cells, which are associated with glucose metabolism. This data suggests that circulating exosomes under diabetic conditions might modulate the placental metabolic state to enhance glycogen metabolism in GDM.

Disclosure

S. Nair: None. V. Ormazabal: None. N. Jayabalan: None. D. Guanzon: None. A. Lai: None. H. McIntyre: Speaker's Bureau; Self; Novo Nordisk Inc.. Research Support; Self; Danish Diabetes Academy. M. Lappas: None. C. Salomon: None.
Association of Urinary Normetanephrine Levels with Increased Insulin Resistance in a General Population
Authors: MURABAYASHI MASAYA , DAIMON MAKOTO , MURAKAMI HIROSHI , KAMBA AYA , MIZUSHIRI SATORU
Published: June 2018

Since activation of the sympathetic nervous system is associated with both impaired insulin secretion and insulin resistance, or namely with diabetes, evaluation of such activation in ordinary clinical settings appears to be important. Urinary levels of catecholamine metabolite, normetanephrine (NM) may be such a marker. Therefore, we here examined association of urinary NM levels with insulin secretion and resistance evaluated by HOMA indices in a general population. From 1,149 participants of the 2016 population-based Iwaki study of Japanese, 1,037 individuals with fasting blood glucose levels, 64-140 to assess HOMA indices precisely were enrolled in the study (gender (M/F): 407/630; age: 53.4±15.5). The Iwaki study is a health promotion study of people over 20 years old to prevent lifestyle-related diseases and prolong their lifespans, and, the participants were those who believe themselves as healthy. Their urinary NM levels were 0.20±0.mg/grCr. Univariate linear regression analyses showed a significant association of urinary NM levels with HOMA-R (β= 0.1508, p <0.0001) and HOMA-β (β= -0.1354, p<0.0001). Adjustment with age, gender, and multiple factors correlated with HOMA indices (Body mass index, Triglyceride, Hypertension, %fat, HbA1c) showed a significant association with HOMA-R (β=0.1167, p= 0.0012) but not with HOMA-β (β=0.011, p =0.782). Further, analysis with subjects stratified into 2 groups (risk and non-risk) using the optimal cut-off value of NM to predict insulin resistance (HOMA-R>1.6) determined by ROC analysis (0.215 mg/grCr) showed those at risk had an OR of 2.36 (CI: 1.49-3.74) after adjustment for the factors listed above. Increased urinary NM levels seem to be a marker for insulin resistance, or namely risk for diabetes in general.

Disclosure

M. Murabayashi: None. M. Daimon: None. H. Murakami: None. A. Kamba: None. S. Mizushiri: None.
Advanced Nonalcoholic Steatohepatitis Is Associated with Low Serum Lipoprotein(a) Concentrations
Authors: KONISHI KANAKO , MIYAKE TERUKI , SENBA HIDENORI , YAMAMOTO SHIN , FURUKAWA SHINYA , WATANABE TAKAO , KOIZUMI YOHEI , YOSHIDA OSAMU , HIROOKA MASASHI , KUMAGI TERU , ABE MASANORI , MATSUURA BUNZO , HIASA YOICHI
Published: June 2018

Background: Nonalcoholic fatty liver disease (NAFLD), a complication frequently coexisting with diabetes, is at high risk of progression to nonalcoholic steatohepatitis (NASH), cirrhosis, liver failure, and hepatocellular carcinoma. Advanced NASH and advanced hepatitis C have poor prognoses, and therapeutic intervention is limited. In contrast, NAFLD may be related to serum lipoprotein(a) (Lp[a]) levels. However, the association between serum Lp(a) levels and progression of NAFLD to NASH remains unknown. We therefore investigated this issue among patients with biopsy-proven NAFLD.

Methods: We enrolled 136 patients with biopsy-proven NAFLD (nonalcoholic fatty liver [NAFL], 20 patients; stage 1, 29 patients; stage 2, 32 patients; stage 3, 26 patients; stage 4, 29 patients), and classified them into the advanced NASH group (stages 3 and 4) and the non-advanced group (NAFL, and stages 1 and 2). Serum Lp(a) levels were measured using the turbidimetric latex agglutination method, and data were divided into tertiles.

Results: In total, 76.5% and 47.2% of the patients were diabetic in the advanced NASH group and the non-advanced group, respectively. Serum Lp(a) levels in the advanced NASH group were significantly lower than those in the non-advanced group (P = 0.009). Multivariate analysis, adjusted for age, sex, body mass index, diabetes mellitus, dyslipidemia, and hypertension, showed that lower serum Lp(a) levels were significantly associated with a higher prevalence of advanced NASH; the adjusted odds ratios (95% confidence intervals) for advanced NASH in relation to serum Lp(a) levels of ≤6.1, 6.2-13, and ≥13.1 mg/dL were 1.00 (reference), 0.333 (0.14-0.791), and 0.341 (0.124-0.937), respectively (P for trend = 0.004).

Conclusions: It was concluded that lower levels of serum Lp(a) could be a significant risk factor for advanced NASH and that serum Lp(a) levels may be a useful tool for distinguishing between NASH and NAFL.

Disclosure

K. Konishi: None. T. Miyake: None. H. Senba: None. S. Yamamoto: None. S. Furukawa: None. T. Watanabe: None. Y. Koizumi: None. O. Yoshida: None. M. Hirooka: None. T. Kumagi: None. M. Abe: None. B. Matsuura: None. Y. Hiasa: None.
Parental Type 2 Diabetes and Obesity Development from Childhood to Early Adulthood
Authors: NIELSEN JANNIE , HULMAN ADAM , CUNNINGHAM SOLVEIG A.
Published: June 2018

Due to shared genes and environment, the home setting may be an effective context to provide multiple individuals at risk of type 2 diabetes (T2D) with preventive efforts. The aim of this study is to investigate whether having a parent with T2D affects the BMI development from childhood through adolescence and into early adulthood.

This analysis used the Panel Study of Income Dynamics (PSID), a nationally longitudinal study of families in the U.S. We used information on diabetes status in parents, household socio-economic characteristics and sex, age and anthropometrics in children and adolescent from 2001-2015 (n=5981). Body mass index (BMI) trajectories in children were fitted using mixed-effects models. Random intercept and slope were included to account for the within-person correlation arising from the repeated measures in the dataset. We used age as the time axis and modeled change with linear, quadratic and cubic terms to account for the non-linear change in BMI.

At baseline, (mean age 12.2 years) children with a parent with T2D were 1.2 years older, had a 3.0 kg/m2 higher BMI than children with no parental T2D (p<0.01 for all tests). The differences occurred with age: There was no significant difference in BMI between the two groups up to 8 years of age. However, at age 8, children with parental diabetes had a 0.41 kg/m2 (0.20-0.61) faster annual increase in BMI than the non-exposed group’s, whose annual increase was 1.03 kg/m2 (0.94-1.11). The rate of annual BMI increase became smaller with age. A maximal separation of 2.04 kg/m2 (1.53-2.55) was reached at 15 years of age, which remained relatively stable in early adulthood.

Children with a parent with T2D as compared to children without parental T2D exhibit a faster increasing BMI trajectory that leads to significant differences in adolescent BMI. This difference persists in early adulthood, when most children no longer live with their parents. Once a parent is diagnosed with T2D, obesity and diabetes prevention may be a priority for the children as well.

Disclosure

J. NIelsen: None. A. Hulman: None. S.A. Cunningham: None.
Residual Beta-Cell Function in Long Duration Type 1 Diabetes (T1D)
Authors: GUBITOSI-KLUG ROSE , BRAFFETT BARBARA , HITT SUSAN M. , ARENDS VALERIE , STEFFES MICHAEL , SAENGER AMY K. , LACHIN JOHN , PALMER JERRY P.
Published: June 2018

We investigated residual beta cell function in participants from the DCCT/EDIC study with an average of 35 (range 27, 48) years duration of T1D. Between 2015-2017 after 22-24 years of follow-up in EDIC, a 4-hour mixed meal tolerance test was administered to 944 participants and 7 timed plasma specimens were collected and assayed for C-peptide using a chemiluminescent immunoassay (Roche). We defined a significant post-stimulus response as a peak C-peptide concentration >0.03 nmol/L, based on an earlier DCCT study which demonstrated that the risk of microvascular disease progression was markedly higher among participants who entered the trial with values below this concentration. Overall, 71 (7.5%) participants were classified as “responders” with a median peak C-peptide of 0.10 nmol/L (IQR 0.06, 0.15). Among the 873 participants classified as “non-responders”, 46 had detectable peak C-peptide between 0.003 (limit of detection) and 0.029 nmol/L. Responders were slightly older than non-responders (58.5 ± 6.2 vs. 56.5 ± 6.8 y; p=0.02), yet had similar age of onset of diabetes (22.5 ± 7.0 vs. 21.4 ± 7.8 y) and duration of diabetes (35.9 ± 5.1 vs. 35.0 ± 4.9 y). Compared to non-responders, responders had lower HbA1c values (8.6 ± 1.7 vs. 8.9 ± 1.5%; p=0.01) and higher stimulated C-peptide (0.22 ± 0.14 vs. 0.11 ± 0.11 nmol/L; p<0.0001) at DCCT baseline; and had lower insulin requirements both at DCCT baseline (0.54 ± 0.18 vs. 0.67 ± 0.24 units/kg/day; p<0.0001) and throughout DCCT follow-up (0.61 ± 0.20 vs. 0.69 ± 0.19 units/kg/day; p=0.0008). Significantly fewer responders experienced severe hypoglycemia [requiring assistance] throughout the DCCT/EDIC study (45% of responders vs. 70% of non-responders; p<0.0001). Both the initial episode and recurrent episodes of severe hypoglycemia were less frequent with increasing C-peptide concentrations.

In conclusion, beta cell function persists in some very long duration T1D patients and is associated with clinically meaningful reductions in the frequency of severe hypoglycemia.

Disclosure

R. Gubitosi-Klug: None. B. Braffett: None. S.M. Hitt: None. V. Arends: None. M. Steffes: None. A.K. Saenger: Advisory Panel; Self; Alere Inc. J. Lachin: Board Member; Self; AbbVie Inc., Celgene Corporation. J.P. Palmer: None.
Time-to-Peak C-Peptide and Peak Glucose during an Oral Glucose Tolerance Test (OGTT) Are Independent Risk Factors for Type 1 Diabetes (T1D) in Antibody-Positive (Ab+) Individuals
Authors: ISMAIL HEBA M. , XU PING , EVANS-MOLINA CARMELLA , PALMER JERRY P. , SOSENKO JAY
Published: June 2018

Peak C-peptide (C-P) and glucose levels are known predictors of T1D, but T1D prediction by times to those peak levels has not been studied. We thus assessed T1D prediction by times to peak C-P or glucose during OGTTs in 709 Ab+ Diabetes Prevention Trial-type 1 participants with 2-hour OGTTs [mean age: 13.9±9.6 years; BMI percentile (%ile): 59.3±29.0; 56% male]. Log rank testing and Cox regression with adjustments for age, sex, BMI %ile, proband relation, HLA, and Ab number were used. Risks were higher for time to peak C-P >60 mins [HR: 1.96 (CI: 1.49-2.63), p<0.001)] and for time to peak glucose >30 mins [HR: 2.22 (1.67-2.94), p<0.001)]. With time to peak C-P and peak C-P as covariates, T1D associations were greater for time to peak C-P (X2=22.8 vs. 9.3, both p<0.001). Time to peak glucose and peak glucose associations also remained significant (both p<0.001). Findings were validated in Ab+ relatives in the TrialNet Pathway to Prevention study (Figure); those with a peak C-P >60 mins or a peak glucose >30 mins had higher risk (both p<0.001).

In conclusion, Ab+ relatives with delayed peak C-P or glucose are at higher T1D risk, independent of peak levels. Moreover, the time to peak C-P appears to be more predictive than the peak level. Times to peak C-P and glucose have potential use as predictors of T1D and as markers for progression.

Disclosure

H.M. Ismail: None. P. Xu: None. C. Evans-Molina: None. J.P. Palmer: None. J. Sosenko: None.
Reduced Rate of Acute Complications with Tubeless Insulin Pump Use—Data from 2,640 Patients in the German/Austrian DPV Registry
Authors: BIESTER TORBEN , DANNE THOMAS , BOLLOW ESTHER , SCHWANDT ANKE , HEIDTMANN BETTINA , RAMI-MERHAR BIRGIT , HAAK THOMAS , HOLL REINHARD W.
Published: June 2018

Little is known about the rates of acute complications when switching from MDI or traditional insulin pumps to tubeless insulin pump therapy. The German/Austrian Diabetes patients follow-up-registry (DPV) includes 2691 patients with type 1 (n=2640), type 2 (n=17) and LADA/other (n=34) diabetes types treated for 1.3±1.3 years with a tubeless insulin pump (Omnipod® Insulin Management System, Insulet Corp, Billerica, MA) since 2013. Patients with type 1 diabetes had a mean age of 12.3 year [IQR 9.6-15.0], diabetes duration 2.5 year [0.7-5.9], HbA1c 7.7% [6.8-8.3]. The prior treatment modality was 59.3% MDI, 37.5% other pump and 2.4% tubeless pump as initial therapy (0.3% unknown). Discontinuation of tubeless pump treatment in patients with type 1 diabetes was 7.3% and occurred after 0.7±0.9 mo. Despite the typical age-dependent increase in HbA1c in this primarily adolescent population, the rate of severe hypoglycemia (Level 3), hypoglycemic coma and DKA decreased between 29% to 76% with Omnipod System use compared to the year prior to treatment switch in the total cohort and type 1 patients with 3 year follow-up (n=835) (Table).

In conclusion, analysis of the DPV registry shows that use of the Omnipod System has high treatment retention rates in type 1 diabetes and is associated with low rates of severe hypoglycemia and DKA in this age-group prone to acute complications.Three Year Outcomes with Omnipod System UseVariable (mean)Total CohortPatients with 3 yr Follow-upPost-OmnipodPost-OmnipodPre-Omnipod (prior year)1 yr (n=2167)2 yr (n=1351)3 yr (n=848)Pre-Omnipod (prior year)1 yr (n=835)2 yr (n=835)3 yr (n=835)Age, yr13.914.714.414.611.512.413.514.4BMI-SDS0.440.540.560.600.430.500.550.61Insulin dose, U/kg/d0.820.760.790.820.790.770.790.83HbA1c, %7.697.597.857.947.557.497.817.97Severe hypoglycemia (Level 3) patients affected per year, %5.25.54.33.75.77.24.73.6Severe hypoglycemia (coma) patients affected per year, %1.51.41.00.51.71.81.40.4DKA patients affected per year, %5.83.33.02.16.63.63.02.2

Disclosure

T. Biester: Speaker's Bureau; Self; Medtronic, Ypsomed AG. T. Danne: Speaker's Bureau; Self; A. Menarini Diagnostics. Advisory Panel; Self; Abbott, AstraZeneca, Bayer AG, Boehringer Ingelheim GmbH. Speaker's Bureau; Self; Dexcom, Inc.. Research Support; Self; Eli Lilly and Company. Stock/Shareholder; Self; DreaMed Diabetes, Ltd.. Research Support; Self; Insulet Corporation. Speaker's Bureau; Self; Menarini Group. E. Bollow: None. A. Schwandt: None. B. Heidtmann: None. B. Rami-Merhar: Speaker's Bureau; Self; A. Menarini Diagnostics, Eli Lilly and Company, Medtronic MiniMed, Inc., Roche Diabetes Care Health and Digital Solutions. T. Haak: Speaker's Bureau; Self; Abbott. Advisory Panel; Self; Roche Diabetes Care Health and Digital Solutions, Sanofi, Eli Lilly and Company. Speaker's Bureau; Self; Eli Lilly and Company, AstraZeneca. R.W. Holl: None.
A Comprehensive Type 1 Diabetes Genetic Risk Score Is Associated with Type 2 Diabetes in the Framingham Heart Study
Authors: SRINIVASAN SHYLAJA , LEONG AARON , UDLER MIRIAM , PORNEALA BIANCA C. , MEIGS JAMES B. , RICH STEPHEN S. , DUPUIS JOSEE
Published: June 2018

It is often difficult to distinguish between type 1 (T1D) and type 2 diabetes (T2D), particularly in young adulthood. Both T1D and T2D have significant genetic contributions to risk and understanding their genetic overlap may offer clinical insight. We examined whether a T1D genetic risk score (GRS) is associated with clinical features and diagnosis of T2D in the Framingham Heart Study (FHS).

We constructed a T1D GRS based on 79 SNPs from fine mapping results, with SNPs weighted by published effect sizes. The GRS included both HLA (27 SNPs) and non-HLA (52 SNPs) regions, and was applied to 1503 participants with T2D. Multivariable regression models assessed the association of the GRS with age of T2D onset, BMI, HbA1C, HOMA-β and HOMA-IR at the exam of diagnosis. Case-control analysis of 1503 cases and 5649 controls was done using multivariate generalized estimating equations (GEE) logistic models to determine the association between the T1D GRS and T2D risk with age and sex as covariates.

In FHS participants, the mean age was 57.2 y, BMI was 30 kg/m2, and HbA1c 6.05%. The T1D GRS was not associated with T2D age of onset, or BMI, HbA1c, HOMA-β and HOMA-IR after excluding participants on medications for T2D. There was a trend towards lower HOMA-β and HOMA-IR values in the top quintile of the GRS (n=244, mean=2.47, SD= 0.7 for ln(HOMA-β) and mean=2.88, SD=0.8 for ln(HOMA-IR)) when compared to the lowest quintile of the GRS (n=260, mean= 2.53, SD=0.8 for ln(HOMA-β) and mean=2.92, SD=0.8 for ln(HOMA-IR)). The T1D GRS was associated with T2D (β = 0.per SD increase in GRS, SE= 0.03, OR =1.1, P = 0.04). When the non-HLA and HLA components of the GRS were tested separately, the results remained significant only for the non-HLA T1D GRS (β = 0.per SD increase in GRS, SE= 0.03, OR=1.1, P= 0.02).

A T1D GRS of non-HLA SNPs is associated with T2D in the FHS participants. Our further analysis with increased sample size in multi-ethnic populations may refine the impact of the T1D GRS to characterize individuals with T2D.

Disclosure

S. Srinivasan: None. A. Leong: None. M. Udler: None. B.C. Porneala: None. J.B. Meigs: None. S.S. Rich: None. J. Dupuis: None.
TOPMed Whole Genome Sequence Combined with Pancreas-Specific Annotation for Rare Variants Tests of Type 2 Diabetes Risk
Authors: MANNING ALISA , MAJARIAN TIM , DE VRIES PAUL S. , WESSEL JENNIFER , MEIGS JAMES B. ,   TOPMED DIABETES WORKING GROUP
Published: June 2018

Genetic studies have shown that common variants associated with type 2 diabetes are enriched in pancreatic-specific regulatory regions. Most whole genome sequence (WGS) studies have used general annotation of regulatory regions not specific to relevant tissues. In contrast, we used functional annotation from pancreatic islets to define rare variant groupings for aggregate tests in coding and non-coding regions in a pilot analysis on chr 10 with 13,200 individuals across six NHLBI TOPMed studies: 2,601 individuals with type 2 diabetes and 10,599 normoglycemic controls with <30X WGS coverage. We identified genes expressed in pancreas and nearby regions showing islet-specific regulatory activity, including open chromatin and chromatin states associated with promoters and enhancers. Variants with minor allele frequency < 1% were aggregated by gene, including the region 5 kbp upstream and downstream of transcribed regions. We only included variants annotated as: "splice acceptor", "splice donor", "splice region", "missense", or "stop gained" within exons expressed in islets, while outside of those exons we included only those variants within “active transcription start sites” and “active enhancer” predicted chromatin states. We performed SKAT association tests, adjusting the association with type 2 diabetes for age, sex, study, and a genetic relatedness matrix to accounted for genetic similarities within families and between populations. Overall, 219 genes (median of 161 variants per gene-region) expressed in islets were included in the analysis: ATP5C1, AVPI1, and YME1L1 reached nominal significance (P<0.01). Chr 10 pilot analyses provide a framework for tailoring rare variant analyses in WGS data through the use of disease-specific functional annotation. WGS analysis in larger samples and additional functional data generated from collaborators in the NIDDK’s AMP-T2D Consortium will extend the genomic spectrum associated with type 2 diabetes risk.

Disclosure

A. Manning: None. T. Majarian: None. P.S. de Vries: None. J. Wessel: Employee; Spouse/Partner; Eli Lilly and Company. J.B. Meigs: None.
ILC1s Alteration in Type 2 Diabetes Patients
Authors: LIU FANGCEN , BI YAN , WANG HONGDONG
Published: June 2018

Type 2 diabetes (T2DM) is promoted by systemic and low-grade inflammation in adipose tissue(AT). Group 1 innate lymphoid cells (ILC1s) regulate inflammatory processes and exacerbate metabolic dysfunction, but their significance in T2DM pathogenesis remains unknown. Here, a total of 41 newly-diagnosed T2DM subjects (age 43.4±1.9 year, BMI 29.3±1.1 kg/m2) and 43 matched non-T2DM subjects (age 42.5±1.0 year, BMI 28.3±1.2 kg/m2) were recruited and peripheral blood mononuclear cells (PBMCs) and visceral AT were obtained for measurement of ILC1s by flow cytometry. The results showed that the rates of circulating ILC1s were dramatically increased in T2DM subjects relative to non-T2DM patients (2.44%±0.47% vs. 0.56%±0.14%, p<0.001). Compared with those in the lowest circulating ILC1s tertile, subjects in the highest tertile had significantly higher HbA1c, fasting blood glucose (FBG) and Homeostasis model assessment of insulin resistance (HOMA-IR). Further correlation analysis revealed a positive correlation between the rates of circulating and adipose ILC1s (r=0.536, p=0.007). Remarkably, adipose ILC1s were significantly increased in T2DM subjects (1.70%±0.58% vs. 0.23%±0.17%, p=0.024). adipose ILC1s were positively associated with HbA1c (r=0.583, p=0.003) and FBG (r=0.494, p=0.014) after adjustment of age and BMI. In linear regression analysis, adipose ILC1s were the major determinants (β=0.629, p=0.004) of the variations of HbA1c. This study for the first time provided direct human evidence that ILC1s abnormalities are critically involved in the development of diabetes, suggesting their potential role as a therapeutic target in the treatment of T2DM.

Disclosure

F. Liu: None. Y. Bi: None. H. Wang: None.
Spontaneous Diabetic FATZO Mice with Intact Leptin Signaling Develop Peripheral Immune Cell Responses Similar to T2D and Obese Patients
Authors: SUN GAO , YE WEIWEI , AN XIAOYU , XIAO YONG-FU , WANG YIXIN
Published: June 2018

Introduction: Type 2 diabetes (T2D) and obesity are strongly associated with peripheral immune cell responses. Subsets of T cells in the circulation are correlated with the severity of the diseases, which may serve as novel targets for antidiabetic or obese treatment. Higher numbers of blood Th1, Th17 and Th22 cells of T helper cells were found in disease patients, while Th2 and Treg cells were in reverse correlation of the severity of the disease. FATZO mouse is a new generation of T2D mouse model with intact leptin signaling. The model is featured with slow onset of diabetes and obesity. Here, we aim to characterize the peripheral immune cell responses in FATZO mice for the use of preclinical drug development targeting diabetes or obesity related inflammation.

Methods: Freshly collected blood from 12 FATZO mice at age 6, 12, 22 and 34 weeks were analyzed for the presence of monocytes, B, T, NK, NK-T, CD4+, CD8+ T cells and subsets of T helper cells (Th1, Th2, Th17, Th22, Treg) using FACS. Serum inflammatory cytokines such as IL1, IL2, IL6, IL17, TNFα and IFNγ were also measured.

Results: FATZO mice developed obesity and hyperglycemia from age of 12 weeks. Concomitantly, higher numbers of leukocytes were noticed with an increase and decrease in frequency of B and NK cells respectively. Within T cells, numbers of CD4+ T cells decreased by 20%, while CD8+ T cells increased by 25%. When subsets of T helper cells were assessed, percentages of Th1 and Th17 cells significantly elevated during the development of diabetes and obesity, while frequencies of Th2 and Treg cells declined. Serum inflammatory cytokine measurements showed a tendency in higher levels of serum IL1a when the animals became diabetic.

Conclusion: FATZO mice develop diabetes and obesity with changes in blood immune cell responses that closely mimic disease patients. The results suggest that FATZO might serve as an ideal model for preclinical research for the diabetes and obesity related inflammation.

Disclosure

G. Sun: None. W. Ye: Employee; Spouse/Partner; Crown Bioscience Inc.. X. An: None. Y. Xiao: None. Y. Wang: None.
Plasma Lipidomics and Gestational Diabetes—A Longitudinal Study in a Multiracial Cohort
Authors: RAHMAN MOHAMMAD L. , ANNE FENG YEN-CHEN , FIEHN OLIVER , TSAI MICHAEL Y. , TEKOLA-AYELE FASIL , LIANG LIMING , ZHU YEYI , ZHANG CUILIN
Published: June 2018

Dyslipidemia is implicated in glucose intolerance. Study of lipidomics in pregnancy may identify novel metabolites that may provide new insights to the etiology of gestational diabetes (GDM). Such studies, however, are sparse. We prospectively investigated lipidomics and GDM risk in a matched case-control study of 107 GDM and 214 non-GDM women nested in the NICHD Fetal Growth Studies. GDM diagnosis was based on Carpenter and Coustan Criteria. We measured plasma lipidome of 420 metabolites at gestational weeks (GW) 8-13, 16-22, 24-29 and 34-37 by gas chromatography mass spectrometry. Associations between individual lipids and GDM were assessed using linear mixed effect models at GW 8-13 and GW 16-22. At GW 8-13, 48 metabolites were significantly related to GDM risk (FDR <0.05). Specifically, the mid-to-long carbon-chain (C) glycerolipids (34-38C diglycerides [DGs]; 48-58C triglycerides [TGs]) were positively related to GDM, while long carbon-chain cholesteryl esters (18-22C CEs) were inversely related. At GW 16-22, 87 metabolites (34-36 C DGs, 48-58C TGs, 18-22C CEs, glycerophospholipids) were significantly related to GDM. Mean levels of top-identified metabolites between cases and controls are presented (Figure). We identified novel metabolites related to GDM with relations differed by lipid structure and the timing of pregnancy. Future studies are warranted to evaluate their roles in GDM early prediction.

Disclosure

M.L. Rahman: None. Y. Feng: None. O. Fiehn: None. M.Y. Tsai: None. F. Tekola-Ayele: None. L. Liang: None. Y. Zhu: None. C. Zhang: None.
Flash Glucose Monitoring Reflects Graft Function after Clinical Islet Transplantation—An Initial Report
Authors: LAM ANNA , FORBES SHAREEN , HAMMING CECILIA , MALCOLM ANDREW J. , JAMES SHAPIRO A.M. , SENIOR PETER A.
Published: June 2018

Metabolic assessment in clinical islet transplantation (CIT) has primarily been limited to the research setting. However, technological advances may now allow for this in routine practice. We describe here flash glucose monitoring (FGM) in islet transplant recipients with varying degrees of graft function as assessed by BETA-2 score (validated clinical index, range 0-42).

Method: Sensor data from 3 of 6 patients using FGM (Abbott FreeStyle Libre) at our center were available for this descriptive study. BETA-2 score was calculated from most recent A1c, insulin dose, fasting C-peptide and glucose.

Results: Patients (1,2,3) varied in age (68,49, 66 years), diabetes duration (59, 37, 48 years), total CITs (5, 2, 2), time from last CIT (28, 10, 1 months) and BETA-2 score (3.4, 10, 34). Patient 3 was insulin independent. FGM profile also varied and appeared to be related to BETA-2 score with improved mean glucose (7.8, 7.3, 5.2 mmol/L), time spent in target (4-8 mmol/L) (40,47, 97%) and decreased frequency/severity of hypoglycemia (Figure) with increasing BETA-2 score.

Conclusion: These observations are in line with previous studies showing an association between graft function (based on stimulated C-peptide) and continuous glucose monitoring and points to the potential of FGM and the BETA-2 score, which can be done in routine clinical care, to characterize graft function and glycemia post CIT.

Disclosure

A. Lam: None. S. Forbes: None. C. Hamming: None. A.J. Malcolm: None. A. Shapiro: Consultant; Self; Viacyte, Inc., Protokinetix. P.A. Senior: Consultant; Self; Abbott, AstraZeneca. Research Support; Self; AstraZeneca, Prometic Life Sciences Inc., Novo Nordisk Inc., Sanofi. Consultant; Self; Eli Lilly and Company. Speaker's Bureau; Self; Eli Lilly and Company, Novo Nordisk Inc.. Consultant; Self; Novo Nordisk Inc., Janssen Pharmaceuticals, Inc.. Speaker's Bureau; Self; Janssen Pharmaceuticals, Inc.. Consultant; Self; Boehringer Ingelheim Pharmaceuticals, Inc.. Speaker's Bureau; Self; AstraZeneca, Merck & Co., Inc., Abbott. Research Support; Self; Viacyte, Inc..
Impaired Insulin Resistance in IF1-Deficient Mice Subjected to a High-Fat Diet with Reduced-Fat Mass and Skeletal Muscle Lipid Accumulation
Authors: HUANG FENGYUAN , YANG KEVIN , SAJA KAMALAMMA , HUANG YICHENG , LONG QINGQIANG , YOSHIDA MASASUKE , YANG QINGLIN
Published: June 2018

Mitochondrial ATP synthase catalyzes the coupling step of oxidative phosphorylation. Under pathologic conditions, ATP synthase hydrolyzes ATP to replenish protons from the matrix into the intermembrane space, sustaining mitochondrial membrane potential. ATPase inhibitory factor 1 (IF1) is an ATP synthase-interacting protein that selectively inhibits the hydrolysis activity of ATP synthase to preserve ATP. The role of IF1 in patients with obesity and type 2 diabetes(T2D) has been reported recently but the in vivo role of IF1 in metabolic regulation in obesity and T2D remains unclear. To understand IF1 in the development of insulin resistance and T2D, we first assess IF1 expression in adipose and skeletal tissues in mice subjected to a high-fat diet (HFD). We found that IF1 transcript expression is upregulated in adipose, but reduced in skeletal muscle of the HFD-fed mice. IF1 knockout mice subjected to HFD showed a substantially slower increase of body weight in adult mice despite a similar food intake and body composition analysis using Magnetic resonance imaging(MRI) revealed that IF1 KO mice had less fat mass than WT mice littermates .Morphometric measurements showed that gonadal, subcutaneous and mesenteric fat mass were relatively lower than those of WT mice. Glucose and insulin tolerance assay revealed that IF1 KO mice impaired glucose and insulin tolerances after HFD feeding. Biochemical assays further revealed that the relatively lower fat mass in the IF1 KO mice was concomitant with increased circulating triglyceride and free fatty acids (FFA) levels. As a result, Gastrocnemius skeletal muscles showed increased triglyceride and FFA contents. Taken together, studies on the IF1 KO mice subjected to HFD uncover that IF1 is an important metabolic regulating factor that affects the development HFD-induced insulin resistance, probably due to an impairment in adipogenesis and skeletal muscle lipid accumulation.

Disclosure

F. Huang: None. K. Yang: None. K. Saja: None. Y. Huang: None. Q. Long: None. M. Yoshida: None. Q. Yang: None.
Depletion of CD2M2-Like Macrophages Promotes the Browning of the White Adipose Tissue
Authors: NAWAZ ALLAH , FUJISAKA SHIHO , USUI ISAO , YAGI KUNIMASA , NAKAGAWA TAKASHI , KADO TOMONOBU , TOBE KAZUYUKI , IGARASHI YOSHIKO , OKABE KEISUKE , SAEKI KUMIKO
Published: June 2018

We have previously shown that CD2M2-like macrophages constitute a microenvironment for adipocyte progenitors (APs), in a Tgfβ-dependent manner, to retain systemic insulin sensitivity by tuning the quiescence/proliferation balance of APs to adapt to changes in nutritional status. Previously we generated CD206DTR transgenic mice and CD206-CreERT2/ Tgfβ1flox/flox (Tgfβ1 KO) mice to determine the role of CD2M2-like macrophages a role of CD206+ cell specific Tgfβ1 respectively, in the APs proliferation. Previous reports have shown that M2-like macrophages play a key role in the browning of WAT via activation of type 2 cytokines production during cold exposure. These findings revealed the relationship between the occurrence of beige adipocytes and M2-like macrophage recruitment. In order to investigate the role of CD2M2-like macrophages in the induction of browning into the WAT, we extended our study to see the browning phenomenon by using CD206DTR and Tgfβ1 KO mice. Here, we show that depletion of CD2M2-like macrophages promotes the browning of WAT. We report that cold stimulation resulted in the generation of smaller adipocytes, upregulation of UCP1 and CD137, a marker for beige progenitors, in the WAT of CD206-ablated mice. Flow cytometry analysis further confirmed enhanced beige progenitors in the WAT. Since blocking of Tgfβ is known to promote browning of the WAT. Mechanistically, we assume that enhanced browning into the WAT of CD206DTR mice might be due to the deletion of Tgfβ1 from CD206-M2-like macrophages. Taken together, we concluded that CD2M2-like macrophages induce the proliferation of beige progenitors in the WAT, which may serve as effective therapeutic tools for the prevention and treatment of obesity.

Disclosure

A. Nawaz: Research Support; Spouse/Partner; Kobayashi Foundation, Kobayashi Foundation, Kobayashi Foundation. S. Fujisaka: Research Support; Self; Mochida Pharmaceutical Co., Ltd., Eli Lilly and Company, MSD K.K., Toyama First Bank. I. Usui: None. K. Yagi: None. T. Nakagawa: None. T. Kado: None. K. Tobe: Research Support; Self; Bristol-Myers Squibb Company, Takeda Pharmaceutical Company, Teijin Pharma Limited, Japan Diabetes Society, Eli Lilly and Company, MSD K.K., Kyowa Hakko Kirin Co., Ltd., Mitsubishi Tanabe Pharma Corporation, Pfizer Inc., Astellas Parma Inc, Chugai Pharmaceutical Co., Ltd., Daiichi Sankyo Company, Limited, Novo Nordisk Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Tsumura & Co., Ono Pharmaceutical Co., Ltd., Novartis Pharma K.K., Sumitomo Dainippon Pharma Co. Y. Igarashi: None. K. Okabe: None. K. Saeki: None.
Adipose Tissue–Specific Knockout of AMPK alpha1/alpha2 Results in Normal AICAR Tolerance and Glucose Metabolism
Authors: CHOI RAN HEE , MCCONAHAY ABIGAIL , JOHNSON MACKENZIE B. , KOH HO-JIN
Published: June 2018

AMP-activated protein kinase (AMPK) is a member of Ser/Thr kinases that has been shown to regulate energy balance. Although recent studies have demonstrated the function of AMPK in adipose tissue using different fat-specific AMPK knockout mouse models, the results were somewhat inconsistent. Our previous study has shown that exercise training increases AMPK activity in mouse adipose tissue, whereas high fat diet decreases its activity. For this study, we tested the hypothesis that AMPK in adipose tissue regulates whole body glucose metabolism. To determine the role of adipose tissue AMPK in vivo, we generated fat-specific AMPKα1/α2 knockout mice (AMPKFKO) using the Cre-loxP system. AMPK α1 and α2 protein expression in epididymal, inguinal, and brown adipose tissues was reduced by >90% in AMPKFKO mice compared to wild type littermates. Body weights of AMPKFKO mice were not different between 8-27 weeks of age. Furthermore, tissue weights (liver, kidney, muscle, heart and white and brown adipose tissue) were similar to wild type littermates and DEXA scan analysis revealed no differences in percentages of body fat and lean mass. Knockout of AMPKα1/α2 in adipose tissue abolished basal and AICAR-stimulated AMPK phosphorylation. Basal and AICAR-stimulated phosphorylation of Acetyl-CoA Carboxylase, a downstream of AMPK, was also reduced by 90% in AMPKFKO mice. Despite of the ablation of AICAR-stimulated AMPK phosphorylation, the blood glucose lowering effect of AICAR injection (i.p.) was normal in AMPKFKO mice. In addition, AMPKFKO displayed normal fasting blood glucose, glucose tolerance, and insulin tolerance, indicating normal whole body glucose metabolism. These data demonstrate that adipose tissue AMPK plays a minimum role in whole body glucose metabolism under normal condition.

Disclosure

R. Choi: None. A. McConahay: None. M.B. Johnson: None. H. Koh: None.
Role of Adenylyl Cyclase-Associated Protein 1 (CAP1) in Mediating Resistin Actions in Mouse Liver Cells
Authors: AVTANSKI DIMITER , GARCIA ANABEL , THANGESWARAN PRIYANTHAN , BORDON BEATRIZ CARABALLO , PORETSKY LEONID
Published: June 2018

Resistin is a pro-inflammatory adipokine produced by the white adipose tissue (WAT) adipocytes and macrophages. Obesity results in chronic inflammation of the WAT, marked by an increase in resistin and other inflammatory cytokines, and by infiltrating leukocytes. Elevated resistin levels are believed to play a major role in the development of insulin resistance in the peripheral tissues. Adenylyl cyclase-associated protein 1 (CAP1) was recently identified as a receptor for resistin. In the present study we aimed to investigate whether CAP1 mediates resistin actions which may affect insulin sensitivity in the liver. As a model we used BNL CL.2 mouse liver cell line. Concentration- and time-dependent experiments demonstrated that resistin upregulated TNFα, SOCS3, IL-1α, and IL-6 mRNA expression maximally when used in concentration of 12.5 ng/ml for 6 hours. In order to determine the CAP1 involvement in mediating resistin actions in the liver, we transfected BNL CL.2 cells with CAP1 siRNA and performed a real-time PCR array measuring the expression of 84 key genes involved in insulin signaling, adipokine signaling, and inflammation. Results demonstrated that resistin upregulated mRNA expression of IL-6; this effect was ameliorated when CAP1 was downregulated. Knock-down of CAP1 facilitated mRNA expression of genes involved in insulin signaling and adipokine signaling pathways, while it resulted in downregulation of infiltrating leukocyte markers expression. Taken together these results indicate that CAP1 is a mediator of resistin actions in the liver.

Disclosure

D. Avtanski: None. A. Garcia: None. P. Thangeswaran: None. B. Caraballo Bordon: None. L. Poretsky: None.
Mitochondrial DNA DAMPs Induce Inflammation and Insulin Resistance
Authors: YUZEFOVYCH LARYSA , PASTUKH VIKTOR M. , RACHEK LYUDMILA
Published: June 2018

Increasing evidence showed that mitochondrial DNA (mtDNA), as a part of mitochondrial damage-associated molecular patterns (DAMPs), induces an immune response and inflammation in immune, as well as in cells of nonnimmune origin. As insulin resistance and type 2 diabetes are characterized by a persistent chronic inflammation, the aim if this study was to evaluate if mtDNA DAMPs induce an inflammatory responses in skeletal muscle cells and induce insulin resistance both in vitro and vivo. In vitro study showed that exogenous mtDNA fragments induced TLR9-mediated NF-κB activation in primary muscle cells. Also, we found that mtDNA DAMPs and palmitate increased TLR9 content in muscle cells. Furthermore, mtDNA DAMPs reduced insulin signaling and insulin stimulated glucose uptake in primary muscle cells. Importantly, administration of exogenous mtDNA DAMPs induced inflammatory signaling, hyperglycemia and insulin resistance in vivo, providing a direct causative role for mtDNA DAMPs in the development of insulin resistance. Parallel studies in vivo showed that high fat diet-induced insulin resistant mice (Yuzefovych et al, 2013, PLOS One) have elevated circulating mtDNA DAMPs contents which were associated with increased mtDNA damage in skeletal muscle and liver, increased oxidative stress and content of TLR9 in skeletal muscle. Collectively, this study identifies a novel role for mtDNA as a pro-inflammatory molecule which promotes insulin resistance. Ultimately, this can provide new clinical implications both to design novel therapeutic strategies and cost-effective biomarkers of insulin resistance progression.

Disclosure

L. Yuzefovych: None. V.M. Pastukh: None. L. Rachek: None.
Hierarchical Brain and Physiological Responses to Hypoglycemia and HAAF
Authors: LEE JOHN J. , TANENBAUM AARON , ARBELAEZ ANA MARIA ,   HYPOGLYCEMIA AND HAAF IN T1DM STUDY GROUP HIERARCHICAL BRAIN AND PHYSIOLOGICAL RESPONSES TO
Published: June 2018

Glycemic control is essential for the management of diabetes. However, intensive glycemic control brings increased morbidity and mortality associated with hypoglycemia. This work tested the hypothesis that human brain networks have functional patterns of hierarchical recruitment in physiologic responses to induced hypoglycemia.

Design and Methods: Glycemic levels were clamped to 90, 65, 55 and 45 mg/dL in seven normal subjects and six persons with T1DM. Cerebral blood flow (CBF) was measured by arterial spin labelling with MRI.

Results: The thalamus, medial prefrontal (MPFC) and orbital prefrontal cortices (OPFC), and striatum have differential changes in CBF with progressive hypoglycemia. Significances were verified using threshold-free cluster enhancement methods.

Discussion: With progressively induced hypoglycemia, specific cortical and subcortical regions demonstrate characteristic alterations of CBF. These changes confirm existing data regarding the roles of the thalamus, MPFC, OPFC and striatum in responses to hypoglycemia. Furthermore, regional CBF reactivity to progressive, induced hypoglycemia has distinctive features for acute/recurrent hypoglycemia and hypoglycemia in T1DM. These characteristic features suggest mechanistic roles for human brain networks in physiologic responses to severe hypoglycemia and hypoglycemia-associated autonomic failure (HAAF).

Disclosure

J.J. Lee: None. A. Tanenbaum: None. A. Arbelaez: None.
Impact of Hepatic Mig6 on Glucose Homeostasis and Oxidative Stress
Authors: IRIMIA-DOMINGUEZ JOSE M. , LUTKEWITTE ANDREW J. , FUEGER PATRICK T.
Published: June 2018

Hepatic epidermal growth factor receptor (EGFR) expression and activation are decreased during steatosis in humans and several animal models of obesity. Restoring EGFR activation in obesity-induced endoplasmic reticulum (ER) stress and diabetes is a potential therapy to improve liver function in type 2 diabetes (T2D). Mitogen-inducible gene 6 (Mig6) is an inducible feedback inhibitor of EGFR activity and, therefore, a target for enhancing EGFR signaling during diet-induced obesity and T2D. We previously observed that liver Mig6 expression was increased during obesity-induced insulin resistance in C57Bl/6J mice fed a high fat diet and by both pharmacological- and fatty acid-driven ER stress in hepatocytes, leading to decreased EGF-mediated EGFR activation. To test the hypothesis of Mig6 as a target to treat insulin resistance in T2D, we examined liver-specific Mig6 knockout mice (LKO) and their littermate controls (CON) during diet-induced obesity. We identified enhanced whole-body glucose tolerance and hepatic insulin action in LKO compared to CON mice. Moreover, liver ballooning, an oxidative stress marker, was also improved in LKO mice. Using whole transcriptome sequencing, we sought to determine the molecular mechanisms leading to liver function improvement in obesity-induced insulin resistance in LKO mice. We identified that gene expression and protein accumulation of key enzymes of both peroxisomal (e.g., Acox1, Ehhadh) and mitochondrial (e.g., Cpt2, Acadl) beta-oxidation were decreased by Mig6 ablation. This observation correlated with a compromised gene expression of liver gluconeogenic enzymes.

In conclusion, we posit that hepatic Mig6 deficiency partially reverts the effects of obesity-induced diabetes by decreasing hepatic glucose output and reducing cellular oxidative stress, thereby establishing Mig6 as a therapeutic candidate for treating insulin resistance in obesity and T2D.

Disclosure

J.M. Irimia-Dominguez: None. A.J. Lutkewitte: None. P.T. Fueger: None.
Evaluating the Sub-chronic Effects of MEDI0382, a GLP-1/Glucagon Receptor Dual Agonist, on Weight Loss in Male ob/ob Mice
Authors: OLDHAM STEPHANIE , CHURCH CHRISTOPHER D. , WILL SARAH , KONKAR ANISH , TREVASKIS JAMES , DAVIES GRAEME R.
Published: June 2018

MEDI0382, a balanced GLP-1/glucagon dual receptor agonist, is under development for the treatment of type 2 diabetes (T2DM). MEDI0382 reduces food intake and improves glucose control in lean and obese mice. Here, we determined the effects of MEDI0382 on body weight in leptin-deficient ob/ob mice. MEDI0382 (10, 20 or 30 nmol/kg), liraglutide (30 nmol/kg; a GLP-1 analog), or G1437 (30 nmol/kg; a lipidated glucagon receptor agonist) was administered to ob/ob mice for 21 days. Body weight was reduced by MEDI0382 treatment compared to vehicle controls, with 15% weight loss observed in the highest dose group. In comparison, liraglutide reduced body weight by 8% and G1437 by 29% (both p<0.0001 vs. vehicle and high-dose MEDI0382). Glucose tolerance was improved by MEDI0382 (30 nmol/kg), similar to liraglutide, whereas G1437 worsened glucose tolerance. Additionally, 4-h fasted glucose on day 7 and 14 in high-dose MEDI0382- and liraglutide-treated groups was lower than vehicle, whereas G1437-treated mice exhibited higher glucose levels. Overnight fasting glucose was also reduced in all groups relative to vehicle except G1437. Liver fat was reduced in a dose-dependent manner (11%, 14% and 22%, respectively) by MEDI0382 compared to vehicle. Both liraglutide (12%) and G1437 (10%) also reduced liver fat (both p<0.vs. vehicle and MEDI0382 30 nmol/kg dose). Serum ALT levels were also dose-dependently reduced by MEDI0382 and liraglutide, but not G1437. Expression of thermogenic genes (Ucp1, Ppargc1a and/or Adrb3) tended to be increased in brown and white adipose tissue by MEDI0382 administration, whereas negligible effects were observed following liraglutide or G1437 administration.

In summary, these data indicate that high-dose MEDI0382 produced superior body weight loss and improved liver health compared to liraglutide but the improved glucose homeostasis was similar to liraglutide.

Disclosure

S. Oldham: Stock/Shareholder; Self; AstraZeneca. Employee; Self; MedImmune. C.D. Church: Employee; Self; MedImmune. S. Will: Employee; Self; MedImmune. Stock/Shareholder; Self; AstraZeneca. A. Konkar: Employee; Self; Sanofi. Other Relationship; Self; MedImmune. J. Trevaskis: Employee; Self; MedImmune. Stock/Shareholder; Self; AstraZeneca. G.R. Davies: Employee; Self; MedImmune.