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Background: Ectopic fat deposition resulting from impaired subcutaneous adipose tissue expandability may take center stage in the pathophysiological events linking low birth weight (LBW) with increased risk of developing Type 2 diabetes (T2D) . However, the extent to which adult LBW subjects exhibit increased hepatic fat accumulation is controversial. Methods and Results: Using 1H MR spectroscopy, we found a significantly increased hepatic fat content (P=0.014) in 26 early middle-aged and non-obese LBW males compared with 21 BMI- and age-matched normal birth weight (NBW) controls. Interestingly, 5 LBW (20%) versus no NBW subjects, had previously unrecognized overt nonalcoholic fatty liver disease (NAFLD) with a median hepatic fat content of 9.45%. The LBW w/NAFLD displayed widespread metabolic changes compared with both NBW and LBW w/o NAFLD subjects including hepatic insulin resistance (P=0.02) , increased fasting plasma triglycerides (TGs) (P=0.03) , cholesterol (P=0.05) , leptin, C-peptide and GLP-1 levels, as well as decreased adiponectin levels (P-range: 0.02-0.007) . Among 65 distinct metabolites and 279 lipids identified by untargeted serum metabolomic and lipidomic analyses, the levels of 7 metabolites including ornithine and citrulline, were different (P Conclusion: The data support a role for ectopic fat deposition in the pathophysiological events and trajectories linking LBW with increased risk of developing T2D and associated cardiometabolic traits including dyslipidemia. Disclosure L.Elingaard-larsen: None. T.Hansen: None. C.Brøns: Stock/Shareholder; Novo Nordisk. A.A.Vaag: Stock/Shareholder; AstraZeneca. L.Justesen: None. S.O.Villumsen: None. A.B.Thuesen: None. J.Størling: None. L.M.Sparks: None. M.Kim: None. C.Legido-quigley: None. E.R.Danielsen: None. Funding Novo Nordisk Foundation (NNF15OC0016692) , Aase and Ejnar Danielsens Fond, Augustinus Foundation, Simon Spies Fonden, TrygFonden.

Background: Fecal calprotectin, a marker of inflammation, is increased in persons with type 1 diabetes (T1D) and kidney disease. This may be explained by a reduction of butyrate-producing bacterial species in the gut. Butyrate supplementation has beneficial effects on intestinal inflammation in inflammatory bowel disease but has never been tested in persons with T1D. Objective: To assess the effect of sodium butyrate on fecal calprotectin and other inflammatory markers, kidney parameters, hba1c and gastrointestinal symptoms in persons with T1D and intestinal inflammation. Methods: Randomized placebo-controlled, double-blind, parallel clinical study including 53 participants with T1D, albuminuria and intestinal inflammation (elevated fecal calprotectin) . Participants were assigned to receive 3.6 g sodium butyrate daily (n=28) or placebo (n=25) for 12 weeks. Primary endpoint was fecal calprotectin changes, and additional outcomes were fecal short chain fatty acids, intestinal alkaline phosphatase activity and immunoglobulins; serum lipopolysaccharide, serum C-reactive protein, albuminuria, kidney function, hba1c and gastrointestinal symptoms. Results: Mean age of the participants was 54±13 years, 43% were women, diabetes duration was 30±15 years and median [Q1:Q3] urinary albumin excretion was 46 [14:121] mg/g. Median fecal calprotectin in the butyrate group was 48 [26:100] μg/g at baseline and change was -1.0 [-20:10] μg/g, the median in the placebo group was 61 [25:139] μg/g at baseline and change was -12 [-95:1] μg/g. Difference in change between groups was not significant (p=0.24) . Neither did we find an effect of butyrate compared to placebo on the other fecal and circulating inflammatory markers, kidney parameters, hba1c or gastrointestinal symptoms. Conclusion: Oral butyrate supplementation for 12 weeks did not reduce fecal calprotectin in persons with T1D and intestinal inflammation. Disclosure N.H.Tougaard: None. P.Groop: Advisory Panel; Novo Nordisk, Sanofi, Other Relationship; Astellas Pharma Inc., AstraZeneca, Boehringer Ingelheim International GmbH, Merck Sharp & Dohme Corp., Mundipharma, Speaker's Bureau; Medscape. M.Lehto: None. P.Rossing: Consultant; Astellas Pharma Inc., AstraZeneca, Bayer AG, Gilead Sciences, Inc., Merck Sharp & Dohme Corp., Novo Nordisk A/S, Sanofi, Speaker's Bureau; Eli Lilly and Company. M.Frimodt-moeller: None. H.Salmenkari: None. E.Buur stougaard: Other Relationship; Novo Nordisk. I.Mattila: None. T.W.Hansen: Stock/Shareholder; Novo Nordisk A/S. C.Legido-quigley: None. S.Hörkkö: None. C.Forsblom: None.

Background: We hypothesized that low birth weight (LBW) subjects with a known increased risk of type 2 diabetes exhibit increased sensitivity towards the deleterious metabolic effects of 4-weeks high carbohydrate overfeeding (HCOF) compared with normal birth weight (NBW) controls. Methods: Untargeted serum metabolomics and lipidomics were measured before and after 4-weeks HCOF (+25% energy) in 26 non-obese men with LBW (2789±174 g) and 21 normal birth weight (NBW) men/controls (3804±172 g) matched for BMI and age. Data were analyzed using a combination of advanced statistical and bioinformatical tools. Results: Among 65 identified metabolites, 8 metabolites were affected differentially by HCOF between the two groups (p Conclusion: The results document severe differential perturbations of lipid metabolism in LBW men exposed to HCOF. Public health initiatives promoting low carbohydrate diets in LBW subjects are warranted. Disclosure S.O.Villumsen: None. A.A.Vaag: Stock/Shareholder; AstraZeneca. L.Justesen: None. L.Elingaard-larsen: None. A.B.Thuesen: None. M.Kim: None. C.Legido-quigley: None. E.R.Danielsen: None. T.Hansen: None. C.Brøns: Stock/Shareholder; Novo Nordisk.

Mitochondrial glucose metabolism is essential for stimulated insulin release from pancreatic β-cells. Whether mitofusin gene expression, and hence, mitochondrial network integrity, is important for glucose or incretin signaling has not previously been explored. Here, we generated mice with β-cell-selective, adult-restricted deletion knock-out (dKO) of the mitofusin genes Mfn1 and Mfn2 (βMfn1/2 dKO). βMfn1/2-dKO mice displayed elevated fed and fasted glycemia and a more than fivefold decrease in plasma insulin. Mitochondrial length, glucose-induced polarization, ATP synthesis, and cytosolic and mitochondrial Ca2+ increases were all reduced in dKO islets. In contrast, oral glucose tolerance was more modestly affected in βMfn1/2-dKO mice, and glucagon-like peptide 1 or glucose-dependent insulinotropic peptide receptor agonists largely corrected defective glucose-stimulated insulin secretion through enhanced EPAC-dependent signaling. Correspondingly, cAMP increases in the cytosol, as measured with an Epac-camps-based sensor, were exaggerated in dKO mice. Mitochondrial fusion and fission cycles are thus essential in the β-cell to maintain normal glucose, but not incretin, sensing. These findings broaden our understanding of the roles of mitofusins in β-cells, the potential contributions of altered mitochondrial dynamics to diabetes development, and the impact of incretins on this process. G.A.R. was supported by a Wellcome Trust Senior Investigator Award (098424AIA) and Wellcome Trust Investigator Award (212625/Z/18/Z), Medical Research Council Programme grants (MR/R022259/1, MR/J0003042/ 1, MR/L020149/1), an Experimental Challenge Grant (DIVA, MR/L02036X/1), a Medical Research Council grant (MR/N00275X/1), and Diabetes UK grants (BDA/11/0004210, BDA/15/0005275, BDA16/0005485). I.L. was supported by a Diabetes UKD project grant (16/0005485). This project has received funding from the European Commission Innovative Medicines Initiative 2 Joint Undertaking, under grant agreement no. 115881 (RHAPSODY). This Joint Undertaking receives support from the European Union’s Horizon 2020 Research and Innovation Programme. This work is supported by the Swiss State Secretariat for Education, Research and Innovation (SERI), under contract no. 16.0097. A.T. was supported by Medical Research Council project grant MR/R010676/ 1. Intravital imaging was performed using resources and/or funding provided by National Institutes of Health grants R03 DK115990 (to A.K.L.), Human Islet Research Network UC4 DK104162 (to A.K.L., RRID:SCR_014393). BJ acknowledges support from the Academy of Medical Sciences, Society for Endocrinology, The British Society for Neuroendocrinology, the European Federation for the Study of Diabetes, an Engineering and Physical Sciences Research Council capital award, and the Medical Research Council (MR/R010676/1). S.A.S. was supported by the JDRF (CDA-2016-189, SRA-2018-539, COE-2019-861), the National Institutes of Health (R01 DK108921, U01 DK127747), and the U.S. Department of Veterans Affairs (I01 BX004444).

Hypoglycemia is a major limiting factor in achieving recommended glycemic target for patients in insulin treatment and can indirectly lead to diabetic complications, morbidity and mortality. While counterregulatory hormonal responses have been studied extensively in patients with type 1 diabetes, a more comprehensive assessment of the metabolic responses has not been done previously. This post-hoc study explored potential responses of the metabolome to hypoglycemia. Twenty-one patients with type 1 diabetes were examined using a hypoglycemic clamp at day one, and 24 hours later, at day two. Blood samples were taken during normoglycemia (5.0-6.0 mmol/L) and hypoglycemia (2.0-2.5 mmol/L). Non-targeted plasma metabolomic analyses was conducted using two-dimensional gas chromatography/time-of-flight mass spectroscopy. Metabolites were analyzed by a linear mixed effect model, adjusting for age and sex. P-values were adjusted for multiple testing using the Benjamini-Hochberg method. A total of 79 metabolites were identified. At day one, two amino acids decreased in response to hypoglycemia, leucine (β±SE: -0.78±0.18, p = 0.004) and isoleucine (β±SE: -0.72±0.16, p = 0.002). At day two, five amino acids including all three branched-chained amino acids decreased: Leucine (β±SE: -0.62±0.13, p = 0.002), isoleucine (β±SE: -0.59±0.19, p = 0.026), valine (β±SE: -0.52±0.12, p=0.002), methionine (β±SE: -0.62±0.19, p = 0.026) and phenylalanine (β±SE: -0.65±0.20, p = 0.026). Two fatty acids were elevated, oleic acid (β±SE: 0.50±0.14, p = 0.016) and tetradecanoic acid (β±SE: 0.63±0.17, p = 0.013). No significant changes were observed between the two days. In conclusion, this study found that the metabolome alternates in response to insulin-induced hypoglycemia resulting in decrement of several amino acids and increment of two fatty acids, contributing to the knowledge of human physiology in individuals with type 1 diabetes. Disclosure R. She: None. N. Al-sari: None. A. Sejling: Employee; Self; Novo Nordisk A/S. I. Mattila: None. P. Henriksen: None. J. Pedersen: None. C. Legido-quigley: None. U. Pedersen-bjergaard: Advisory Panel; Self; Novo Nordisk A/S, Sanofi-Aventis, Consultant; Self; Abbott, Speaker’s Bureau; Self; Novo Nordisk A/S.

Background and Aims: Mitochondrial metabolism of glucose is vital for the initiation of insulin secretion from pancreatic β-cells. Whether mitochondrial ultra-structure, and the proteins controlling mitochondrial fission and fusion, are essential for glucose recognition is less clear. Here, β-cell-restricted deletion of Mfn1 and Mfn2 in adult mice was used to investigate the role of mitochondrial dynamics in insulin secretion and glucose homeostasis in vivo and in vitro. Methods: C57BL6 mice bearing Mfn1 and Mfn2 alleles with FloxP sites, were crossed to animals carrying an inducible Cre recombinase expressed under Pdx1 promoter control (PdxCreERT). Following tamoxifen induction, Mfn1 and Mfn2 knockdown was confirmed in KO islets. Live β-cell fluorescence imaging of Ca2+ accumulation and membrane potential were performed on isolated islets and in the living mouse. Mitochondrial ultra-structure was measured using super resolution fluorescence imaging and EM. Results: Mitochondrial length was strongly and selectively diminished in β-cells from βMfn1/2 KO mice. KO animals displayed higher fasting and fed glycaemia than control animals at 14 weeks and a considerable (>5-fold) decrease in plasma insulin post-IP glucose injection. Additionally, mitochondrial length, glucose-induced hyperpolarization, ATP synthesis, Ca2+ accumulation, O2 consumption and β-cell connectivity were all significantly reduced in βMfn1/2-KO mouse islets. Ca2+ dynamics and mitochondrial membrane potential changes were also suppressed in vivo. Remarkably, defective GSIS in islets isolated from βMfn1/2-KO mice was normalised by the addition of GLP-1 receptor agonists. Conclusions: Mitochondrial fusion and fission cycles are indispensable in the β-cell for normal glucose sensing in vitro and in vivo. GLP-1 receptor agonism may mitigate the effects of mitochondrial abnormalities in some forms of diabetes. Disclosure E. Georgiadou: None. M. Ibberson: None. I. Leclerc: Consultant; Spouse/Partner; Sun Pharmaceutical Industries Ltd. A. K. Linnemann: None. Y. Ali: None. T. Rodriguez: None. G. A. Rutter: Advisory Panel; Self; Sun Pharmaceutical Industries Ltd. C. Muralidharan: None. P. L. Chabosseau: None. A. Tomas: Other Relationship; Self; Sun Pharmaceutical Industries Ltd. T. Stylianides: None. C. Legido-quigley: None. N. Alsabeeh: None. C. Cruciani-guglielmacci: Consultant; Self; The Healthy Aging Company. C. Magnan: None. Funding UK Wellcome Trust (WT098424AIA, 212625/Z/18/Z); Medical Research Council (MR/R022259/1, MR/J0003042/1, MR/L020149/1, MR/N00275X/1); Experimental Challenge Grant (DIVA, MR/L02036X/1); Diabetes UK (BDA/11/0004210, BDA/15/0005275, BDA16/0005485)

Omics-based methods may provide new markers associated to diabetic retinopathy (DR). We investigated a wide omics panel of metabolites and lipids related to DR in type 1 diabetes. Metabolomic analyses were performed using two-dimensional gas chromatography with time-of-flight mass spectrometry and lipidomic analyses using an ultra-high-performance liquid chromatography quadruple time-of-flight mass spectrometry method in 648 individuals with type 1 diabetes. Subjects were subdivided into no DR, mild nonproliferative DR (NPDR), moderate NPDR, proliferative DR, and proliferative DR with fibrosis. End points were any progression of DR, onset of DR, and progression from mild to severe DR tracked from standard ambulatory care and investigated using Cox models. The cohort consisted of 648 participants aged a mean of 54.4 ± 12.8 years, 55.5% were men, and follow-up was 5.1–5.5 years. Cross-sectionally, 2,4-dihydroxybutyric acid (DHBA), 3,4-DHBA, ribonic acid, ribitol, and the triglycerides 50:1 and 50:2 significantly correlated (P < 0.042) to DR stage. Longitudinally, higher 3,4-DHBA was a risk marker for progression of DR (n = 133) after adjustment (P = 0.033). We demonstrated multiple metabolites being positively correlated to a higher grade of DR in type 1 diabetes and several triglycerides being negatively correlated. Furthermore, higher 3,4-DHBA was an independent risk marker for progression of DR; however, confirmation is required.