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Leukocyte telomere length (LTL) is a biomarker of biological aging, and has been associated with metabolic diseases such as type 2 diabetes (T2DM). Insufficient maternal vitamin D (25(OH)D) has also been associated with increased disease risk for many diseases and adverse later life outcomes. The association between maternal and cord blood total 25(OH)D and LTL was examined in mother-child pairs from the HAPO Hong Kong Field Centre, including 815 offspring at age (mean±SD) 6.96 ± 0.44 years. LTL was measured using real-time polymerase chain reaction and serum 25(OH)D2, 25(OH)D3 and 3-epi-25(OH)D3 were measured in maternal blood (at gestation 24-32 weeks) and cord blood by liquid chromatography-mass spectrometry. Children LTL at follow-up was significantly shorter in boys compared to girls (p Disclosure K. Wong: None. B. E. Metzger: None. W. Lowe: None. W. H. Tam: None. R. C. Ma: Other Relationship; Self; AstraZeneca, Medtronic, Research Support; Self; AstraZeneca, Bayer Healthcare Pharmaceuticals Inc., Novo Nordisk, Pfizer Inc., Sanofi-Aventis, Tricida, Inc. F. Cheng: None. D. Mao: None. C. K. P. Lim: Stock/Shareholder; Self; GemVCare Ltd. R. Wang: None. C. H. Tam: None. M. A. Joglekar: None. A. A. Hardikar: None. A. Jenkins: Advisory Panel; Self; Abbott Diabetes, Sanofi-Aventis, Other Relationship; Self; Amgen Inc., Research Support; Self; Abbott, International Diabetes Federation, JDRF, Medtronic, Mylan N. V.

Background: In the light of growing global epidemic of T2D and related microvascular complications, it is important to find early detection marker for the disease. Circulating miRNAs serve as a potential biomarker for monitering the progression to DR in diabetic individuals. Hypothesis and Aims: miRNAs play a substantial role in metabolic homeostasis through regulation of multiple genes, and could serve as a prognostic biomarker in DR. This study aims to assess a functional role of selected miRNAs in the Qatari population recruited for the Qatar Genome Project through Qatar BioBank. Methods: Plasma samples from 470 T2D subjects with and without DR and 500 matched-healthy controls has been included in this study. Total RNA was extracted and the expression profiling of 56 miRNA that previously reported as a player in the pathogenesis of DR was performed using custom open array panel. The miRNA expression data were normalized using ath-mir159a. Statistical analysis was performed to study the association between the phenotypes (HbA1c, C-peptide, age, BMI) and miRNA. Results: The patients and controls phenotypic and miRNA expression data were analyzed to interepret the differential expression of the miRNA in DR subjects. Initial analysis by applying the generalized additive model reveals that six miRNA (hsa-miR-27a, hsa-miR-376c, hsa-miR-92a, hsa-miR-1 hsa-miR-223, and mmu-miR-451) were found to be significantly associated with DR (p>0.05). Furthermore, two differentially expressed miRNAs, (hsa-miR-1274A and hsa-miR-24) found to be associated with higher level of C-peptide in DR patients, which suggest the correlation of these miRNAs with the glucose hemostasis and DR. Conclusion: The etiology of DR is unclear, and present treatments have limited effectiveness. The biomarkers and downstream functional studies are required to deepen the understanding of this chronic diseases and its complications. Disclosure A.S. Akil: None. S. Subash Padmajeya: None. L.A. Jerman: None. A. Al-Kurbi: None. A.M. Hussein: None. A. Hardikar: None. M. Joglekar: None. T. Habib: None. M. El Anbari: None. K. Fakhro: None.

Type 2 diabetes (T2D) is a major cause of morbidity and death worldwide. Women with gestational diabetes mellitus (GDM) have a seven-fold higher risk of developing T2D later in life. However, accurate methods for post-partum T2D risk stratification of GDM women, are lacking. MicroRNAs are now well recognised as circulating biomarkers or mediators of metabolic disease. We aimed to determine if post-partum circulating microRNAs can predict the devolvement of T2D in women with previous GDM. Participants were part of the GDM follow-up program at the Mercy Hospital for Women, Australia. Plasma samples were collected at 12-week post-partum from 103 women following an index GDM pregnancy. Women were followed for 10 years to assess development of T2D. In a discovery approach, we measured 754 miRNAs in plasma from T2D non-progressors (N=11) and in T2D progressors (N=10) using TaqMan-based real time PCR on an OpenArray platform. Data were analysed using penalised logistic regression followed by bootstrapping. Top 15 miRNAs with high frequencies in this discovery bootstrap analysis were selected for validation in the remaining samples (N=82; 71 non-progressors and 11 progressors). Validation data were normalized to spike-in control (ath-miR-172a). Four miRNAs (miR-543, miR-410, miR-491 and miR-369-3p) were significantly dysregulated (p Disclosure M. Joglekar: None. W.K. Wong: None. F. Ema: None. H.M. Georgiou: None. A. Hardikar: None. M. Lappas: None.

Objective: Despite therapeutic advances, type 1 diabetes (T1D) is associated with a risk of development of micro- and macrovascular complications. Early treatment of complications is associated with a more favorable prognosis, hence biomarkers to enable early detection and to monitor treatments may be useful. Methods: Plasma (EDTA) samples were obtained from 30 adult (M/F = 20/10) individuals with T1D, age 42±16 years, T1D duration 24±11 years, HbA1c 7.5±1.2% and BMI 26.5±4.0 kg/m2 (15 with microvascular complications (retinopathy or nephropathy) - CX+, and 15 without complications - CX-) and compared with 15 nondiabetic controls (CON, matched age, gender, smoking and BMI). T1D CX+ and CX- individuals were matched for age, gender, diabetes duration, smoking, BMI, HbA1c and insulin therapy modality (CSII/MDI). In these discovery cohort samples, profiling of 754 miRs was done using a high throughput probe based quantitative PCR on nanofluidics Open Array platform . Results were analyzed using independent T test, Mann-Whitney test and classification trees. Statistical analysis was undertaken using R (ver. 3.5.1) and XLStat (ver. 2018.6). Results: Of 754 available miRs on the array, 186 miRs were detectable in plasma. Using an unbiased approach expression of 32 miRs differed significantly in CON vs. T1D and 16 miRs were differentially expressed between CX+ and CX- T1D subjects. Using a Random Forest wrapper algorithm 5 miRs were identified with importance for segregating CX+ vs. CX- status: miR-483-5p, miR-328, miR-1274B, miR-7 and miR-30a-5p. In logistic regression those 5 miRs provided a ROC curve with AUC=0.90 for the CX+ vs. CX- discrimination. Conclusion: In a discovery analysis of well- matched CX+ and CX- selected miRs provided good discrimination between T1D CX status. Validation studies in a larger cohort of cross-sectional and longitudinal samples are in progress. Disclosure A.S. Januszewski: None. E.S. Scott: None. M. Joglekar: None. L. Carroll: None. R. Farr: None. W. Wong: None. D.N. O’Neal: None. C. Karschimkus: None. G. Fulcher: Advisory Panel; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Janssen Research & Development, Merck Sharp & Dohme Corp., Novo Nordisk Inc. Consultant; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Janssen Research & Development, Merck Sharp & Dohme Corp., Novo Nordisk Inc. Research Support; Self; Novo Nordisk Inc. A. Hardikar: None. A.J. Jenkins: Advisory Panel; Self; Abbott, Australian Diabetes Society, Medtronic. Research Support; Self; Abbott, GlySens Incorporated, Medtronic, Mylan. Speaker’s Bureau; Self; Eli Lilly and Company, Novo Nordisk Inc. Funding Sydney Medical School Foundation; National Health and Medical Research Council of Australia

There is growing evidence that microRNAs, a class of non-coding RNA molecules, are associated with regulation of gene expression in general. Here we profiled microRNAs and key endocrine pancreatic gene transcripts in 698 human tissues, including ∼200 human islet preparations. Figure 1 shows the tissue types in our biobank and circos plots showing correlation of key pancreatic genes and microRNAs. Using novel approaches involving penalized regression and bootstrapping in our discovery set of 507 tissue samples, we identified microRNAs that are highly associated with (pro-)insulin expression. Then, using two different de-identified validation sets (N=91 and N=100) of human cells/tissue samples, we demonstrate that the levels of insulin-associated microRNAs can predict the level of insulin transcript with high accuracy. Finally, we show that forced expression of insulin gene does not change the levels of insulin-associated microRNAs, but forced expression of selected microRNAs promotes insulin gene expression. More generally, these data identify key microRNA genes associated with, predictive of and promoting for insulin gene transcription. Disclosure A. Hardikar: None. W. Wong: None. M. Joglekar: None. L.T. Dalgaard: Employee; Spouse/Partner; Novo Nordisk A/S. Stock/Shareholder; Spouse/Partner; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. A. Jenkins: Research Support; Self; Medtronic, Mylan, Sanofi-Aventis. R.C. Ma: Research Support; Self; AstraZeneca, Bayer AG, Merck Sharp & Dohme Corp., Pfizer Inc.. Advisory Panel; Self; Boehringer Ingelheim GmbH, Nippon Boehringer Ingelheim Co. Ltd..

Type 1 diabetes (T1D) is characterized by the loss of insulin-producing beta-cells. We currently lack diagnostic tools to measure T1D progression. Using systematic approaches involving unprejudiced discovery analyses in human tissues and clinical samples, we identify microRNAs that are: i) associated with insulin expression during embryonic development (N>60); ii) predictive of and necessary for insulin transcription (N=698); and iii) dysregulated in individuals with (vs. without) T1D (N=200). Insulin cfDNA (assessed by ddPCR) and microRNA (assessed by TaqMan qPCR) biomarkers were measured in around; i) N=400 Australian T1D individuals; ii) N=30 longitudinal samples from children at risk of T1D; iii) N=100 clinical trial samples from individuals receiving IFN-a or Exenatide vs. placebo; iv) N=400 Hong Kong Chinese individuals with T1D; v) N=700 Danish individuals at diagnosis of T1D and vi) N=400 Asian Indian T1D subjects. Among N=600 Australian samples, we identified miRNAs (N=5) that were significantly increased in the circulation of T1D individuals (vs. controls), decreased (at least 2 Fold Change/FC, P Disclosure M. Joglekar: None. R. Farr: None. W. Wong: None. K. Rother: None. A. Jenkins: Research Support; Self; Medtronic, Mylan, Sanofi-Aventis. C.S. Yajnik: None. R.C. Ma: Research Support; Self; AstraZeneca, Bayer AG, Merck Sharp & Dohme Corp., Pfizer Inc.. Advisory Panel; Self; Boehringer Ingelheim GmbH, Nippon Boehringer Ingelheim Co. Ltd.. M.E. Craig: None. T. Jones: None. A. Hardikar: None.

Transplantation of cadaveric islets is currently the only clinically recognized cell therapy for treatment of type 1 diabetes. Long-term success of this therapy is severely limited by multiple factors, including the immune-mediated destruction of transplanted islets. Development of molecular immunosuppressive strategies could offer much efficient graft survival thereby improving graft function and quality of life. Here we demonstrate that one of the novel mechanisms of immune regulation involves inter-cellular transfer of microRNAs via exosomes. We have established and extensively characterized human islet-derived progenitor cells (hIPCs), which are generated following epithelial-to-mesenchymal transition (EMT) and proliferation (expansion) of human cadaveric islet cells. Initial characterization of hIPCs indicates that they exhibit most of the characteristics of mesenchymal stem cells, including expression of CD73, CD90 and CD105. We observed that hIPCs can significantly inhibit (>90%) in vitro proliferation of different immune cell subsets (CD4+ T, CD8+ T, CD19+ B) of (PHA)-stimulated PBMCs in a co-culture system. RNA-sequencing and validation studies pointed to a potential role of microRNAs in inhibition of T cell proliferation. Recently, microRNAs have been demonstrated to shuttle between cells and are reported to be involved in inter-cellular gene regulation. When microRNAs were analysed in hIPCs alone and in hIPCs co-cultured with PBMCs, we found a small subset of significantly altered microRNAs. These microRNAs showed increase in abundance in PBMCs from co-culture setting and were higher in co-culture supernatant. When exosomal inhibitors were used to prevent the transfer of microRNAs from hIPCs in a co-culture setting, we found that T-cell proliferation was rescued back to normal. We believe that this study could help in discovering novel immunoregulatory molecules that may have a potential role for improving graft survival in islet transplantation for type 1 diabetes. Disclosure M. Joglekar: None. C. Maynard: None. A. Hardikar: None.

The incidence of diabetes is increasing globally, and in spite of modern treatments for glucose, blood pressure, and lipids, the vascular and neurologic complications of diabetes still occur at unacceptably high rates. The personal and economic burdens are enormous. Diabetes is a common cause for vision loss (diabetic retinopathy [DR]), renal impairment (nephropathy), peripheral and autonomic neuropathy and accelerated atherosclerosis leading to cardiovascular disease, cerebrovascular and peripheral vascular disease, and premature death (1). People with diabetes and with early renal damage are at particularly high risk of DR, renal failure, premature cardiovascular disease, and death. Moreover, people with DR are at increased risk of other micro- and macrovascular complications of diabetes (2). This may relate to common risk factors and mechanisms, such as damage to the vasculature, a chronic and clinically silent process. Vascular endothelial cells are the interface between blood and surrounding tissues, with key roles in regulation of inflammation, thrombosis, and angiogenesis and therefore a popular choice as a model for assessing endothelial damage and metabolic stress (3,4). Vision loss due to DR is often preventable. However, for many people with diabetes it is often difficult to predict the clinical course of disease. Furthermore, not all people take or respond well to proven primary and secondary treatments for DR, including risk factor control, renin-angiotensin system drugs, fenofibrate, and late-stage treatments, such as ocular laser, corticosteroids, and anti–vascular endothelial growth factor agents. This highlights the importance of improving our understanding of the pathogenesis of DR and developing …

In Europid populations, low birth weight of offspring predicts insulin resistance in the mother and cardiovascular disease in both parents. We investigated the association between birth weight of offspring and obesity and cardiovascular risk in the parents of 477 8-year-old children born at the King Edward Memorial Hospital, Pune, India. Eight years after the birth of the child, mothers (33 years of age, n = 459) of heavier babies were taller and more obese (BMI, fat mass, and waist circumference, all P < 0.001) than mothers of lighter babies. Increasing offspring birth weight predicted higher homeostasis model assessment for insulin resistance (P < 0.01) and metabolic syndrome in mothers (P < 0.001) (adjusted for offspring sex and birth order, maternal age, and socioeconomic status) but not hyperglycemia. Fathers (39 years of age, n = 398) of heavier babies were taller and heavier, independent of maternal size (P < 0.01, both), but were not more insulin resistant. Unlike other reports, lower offspring birth weight did not predict insulin resistance in fathers. Thus, urban Indian parents have a higher risk of being obese 8 years after delivery of a heavier child. Mothers but not fathers of heavier babies also have a higher risk of being insulin resistant and developing the metabolic syndrome. Our findings highlight the need for a better understanding of the relation between fetal growth and future health before contemplating public health interventions to improve fetal growth.

We have studied 477 8-year-old Indian children to define the relationship between birth weight and cardiovascular risk factors, including insulin resistance syndrome (IRS) variables and plasma total and LDL cholesterol concentrations. All risk factors were strongly related to current weight. After adjustment for current weight, age, and sex, lower birth weight was associated with higher systolic blood pressure (P = 0.008), fasting plasma insulin and 32-33 split proinsulin concentrations (P = 0.08 and 0.02), glucose and insulin concentrations 30 min postglucose (P = 0.06 and 0.04), subscapular/triceps skinfold ratio (P = 0.003), and plasma total and LDL cholesterol concentrations (P = 0.002 and 0.001). Lower birth weight was associated with increased calculated insulin resistance (homeostasis model assessment [HOMA], P = 0.03), but was not related to the HOMA index of beta-cell function. The highest levels of IRS variables and total and LDL cholesterol were in children of low birth weight but high fat mass at 8 years. Taller height at 8 years predicted higher fasting plasma insulin concentrations, insulin resistance, and plasma total and LDL cholesterol concentrations. The most insulin-resistant children were those who had short parents but had themselves grown tall. Although the implications of our findings in relation to height are unclear, interventions to improve fetal growth and to control obesity in childhood are likely to be important factors in the prevention of cardiovascular disease and IRS in India.

In Europid populations, low birth weight of offspring predicts insulin resistance in the mother and cardiovascular disease in both parents. We investigated the association between birth weight of offspring and obesity and cardiovascular risk in the parents of 477 8-year-old children born at the King Edward Memorial Hospital, Pune, India. Eight years after the birth of the child, mothers (33 years of age, n = 459) of heavier babies were taller and more obese (BMI, fat mass, and waist circumference, all P0.001) than mothers of lighter babies. Increasing offspring birth weight predicted higher homeostasis model assessment for insulin resistance (P0.01) and metabolic syndrome in mothers (P0.001) (adjusted for offspring sex and birth order, maternal age, and socioeconomic status) but not hyperglycemia. Fathers (39 years of age, n = 398) of heavier babies were taller and heavier, independent of maternal size (P0.01, both), but were not more insulin resistant. Unlike other reports, lower offspring birth weight did not predict insulin resistance in fathers. Thus, urban Indian parents have a higher risk of being obese 8 years after delivery of a heavier child. Mothers but not fathers of heavier babies also have a higher risk of being insulin resistant and developing the metabolic syndrome. Our findings highlight the need for a better understanding of the relation between fetal growth and future health before contemplating public health interventions to improve fetal growth.