bluuebunny/crossref_metadata_2025_split huggingface dataset:
24-OR: Time-to-Event GWAS for Incident Cardiovascular Disease in Type 2 Diabetes Patients
Authors: KWAK SOO HEON , WU PEITAO , MEIGS JAMES B. ,   ON BEHALF OF THE CHARGE DIABETES WORKING GROUP
Published: June 2021

Patients with type 2 diabetes (T2D) are at two-fold increased risk of cardiovascular disease (CVD), which is the leading cause of T2D morbidity and mortality. Here, we conducted a time-to-event genome-wide association study (GWAS) to identify genetic variants associated with incident CVD among patients with T2D in the Cohorts for Heart and Aging Research in Genomic Epidemiology (CHARGE) consortium. Incident CVD was defined based on a composite of myocardial infarction, stroke, and cardiovascular death that occurred at least one year after the diagnosis of T2D. In each participating prospective cohort, we performed GWAS using a Cox proportional hazard survival model adjusting for age, and sex to identify genetic variants associated with incident CVD, and cohort-level estimated effect sizes were combined using inverse variance weighted fixed effects meta-analysis. A total of 37,327 participants were included in the analyses (26,928 European ancestry and 10,399 non-European ancestry) which comprised 6,012 primary event cases. We identified three distinct genetic loci associated with incident CVD among individuals with T2D that reached the threshold for genome-wide significance: rs231964 (intronic variant in NOX3, hazard ratio [HR] 1.4, P=8.3x10-9), rs139753656 (intronic variant in DCC, HR 2.0, P=2.4x10-8), and rs77202398 (intronic variant in TNS3, HR 1.4, P=4.4x10-8). Among 204 known coronary artery disease risk loci, 27 were associated with incident CVD with P&lt;0.05 and rs6842241 upstream of EDNRA was associated with increased risk of CVD after Bonferroni correction (HR 1.1, P=1.2x10-4). The data provide evidence of novel and known genomic regions associated with incident CVD among individuals with T2D that require validation but point to novel targets.

Disclosure
S. Kwak: None. P. Wu: None. J. B. Meigs: Consultant; Self; Quest Diagnostics. On behalf of the charge diabetes working group: n/a.

Funding
National Institutes of Health (1R01HL151855-01)
243-OR: Integrating Genetic Risk Score and Islet Autoantibody Characteristics in the Predictive Model for Type 1 Diabetes in the TrialNet Study
Authors: FERRAT LAURIC A. , STECK ANDREA , PARIKH HEMANG M. , YOU LU , ONENGUT-GUMUSCU SUNA , GOTTLIEB PETER , RICH STEPHEN S. , KRISCHER JEFFREY , ORAM RICHARD A. , REDONDO MARIA J. 
Published: June 2021

Background: Type 1 diabetes (T1D) genetic risk score (GRS) and islet autoantibody (AAb) specificity and titers influence T1D risk; however, these factors collected at baseline in cross sectional studies have not been effectively incorporated in the current models for prediction of progression along pre-clinical stages of T1D. Our aim was to develop an updated model of T1D prediction for research and clinical practice.
Methods: AAb-positive relatives of individuals with T1D (n=1,233, 52% female, 88% European ancestry, age mean 15 yrs [SD 12.2]) from the TrialNet Pathway to Prevention were followed for a median of 3.1 years (IQR 1.1 -7.1). T1D developed in 498/1,233 (40.4%). T1D GRS, AAb number, AAb combination, AAb titer, age and sex were evaluated on the combined T1D prediction model, estimating its performance using 3-fold cross-validation (10x repeated).
Results: Machine learning (survival random forest) and traditional methods (Cox models, extended Cox model) performed equivalently as measured by 5 yr horizon time-dependent AUC ROC (respectively 0.74, 0.74, 0.72), on calibration by integrated Brier score (respectively 0.15, 0.15, 0.16), and by calibration plots. AAb combinations with specific variables predicted T1D. In multivariate analysis, T1D GRS was a significant predictor of T1D in individuals positive for GADA, IAA or both. For any autoantibody combination including IA2A, the IA2A titer was predictive while the T1D GRS was not. Variable importance analyses showed that T1D was predicted by IA2A titer level, followed by T1D GRS and combinations of specific AAbs.
Conclusion: We modeled individual estimates of T1D risk with T1D GRS, AAb characteristics, age, and sex. T1D prediction improved by adding T1D GRS and AAb characteristics to other variables, and was similar using machine learning or traditional models. T1D GRS was a significant predictor only in the absence of IA2A. In the presence of IA2A, IA2A titer was the most influential factor.

Disclosure
L. A. Ferrat: None. M. J. Redondo: Advisory Panel; Self; Provention Bio, Inc. A. Steck: None. H. M. Parikh: None. L. You: None. S. Onengut-gumuscu: None. P. Gottlieb: Advisory Panel; Self; Janssen Research & Development, LLC, Tolerion, Inc., Viacyte, Inc., Other Relationship; Self; ImmunoMolecular Therapeutics, Inc., Research Support; Self; Caladrius Biosciences, Inc., Immune Tolerance Network, Mercia Pharma Inc., National Institute of Diabetes and Digestive and Kidney Diseases, Precigen, Inc., Tolerion, Inc. S. S. Rich: None. J. Krischer: None. R. A. Oram: Consultant; Self; Janssen Research & Development, LLC.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (1R01DK121843-01); JDRF (3-SRA-2019-827-S-B)
245-OR: Comprehensive Genome-Wide Association Study (GWAS) Meta-analysis Using TOPMed Imputation in Latinos Identifies Rare Variation Associated with Type 2 Diabetes (T2D)
Authors: HUERTA ALICIA , COLE JOANNE B. , DORNBOS PETER , DICORPO DANIEL A. , LEONG AARON , FLOREZ JOSE C. , MERCADER JOSEP M. ,   CONSORTIUM SIGMA T2D,   BIOBANK MX,   BIOBANK MGB
Published: June 2021

The high prevalence of T2D among Latino individuals emphasizes the importance of studying both environmental and genetic factors that influence disease in this population. However, Latino is a recently admixed population with complex linkage disequilibrium patterns that impose methodological challenges, especially when using genotype imputation to test association with rare variation. The recent release of the multi-ethnic NHLBI Trans-Omics for Precision Medicine (TOPMed) reference panel, comprising more than 100,000 samples (~10% are Latinos), represents a unique opportunity to explore rare genetic variation in this population.
We performed the largest T2D GWAS meta-analysis in the Latino population imputed to the TOPMed panel. Our analysis includes 18,294 individuals (7,556 cases and 10,738 controls) with genotyping array and whole-exome sequencing data. We were able to impute 38M high-quality variants (rsq&gt;0.8) with TOPMed (5M with minor allele frequency [MAF] 0.001-0.0005) compared to 16M with 1000G Phase 3 panel (1.6M with [MAF] 0.001-0.0005). This allowed us to replicate previously reported T2D-associated common variants, and to identify a novel genome-wide significant variant (MAF=0.016, OR=2.06, P=1.7×10-9) near ORC5, a gene implicated in familial hyperinsulinemic hypoglycemia 2. Two additional suggestive novel signals merit further research: a variant in ROR2 (MAF=0.066, OR=1.3, P=8.8×10-8) and a Latino specific variant in HDAC2 only identified with the use of TOPMed (MAF=0.013, OR=1.82, P=3.2×10-7) and associated with insulin secretion in individuals without diabetes in this study.
Our results demonstrate the utility of the TOPMed imputation for the identification of novel and population-specific rare variation associated with T2D in understudied populations, which could lead to the identification of novel therapeutic targets and the improvement of polygenic risk scores.

Disclosure
A. Huerta: None. Mgb biobank: n/a. J. B. Cole: None. P. Dornbos: None. D. A. Dicorpo: None. A. Leong: None. J. C. Florez: Consultant; Self; Goldfinch Bio, Inc., Other Relationship; Self; Novo Nordisk. J. M. Mercader: None. Sigma t2d consortium: n/a. Mx biobank: n/a.

Funding
American Diabetes Association (1-19-ICTS-068 to J.M.M.)
25-OR: Role for TBC1D30 in Secretion of Mature Insulin
Authors: PARSONS VICTORIA A. , VADLAMUDI SWAROOPARANI , MOXLEY ANNE H. , MOHLKE KAREN L. 
Published: June 2021

Genome-wide association studies (GWAS) have identified loci associated with the processing and secretion of mature insulin. Increased proinsulin (PI) secretion due to incomplete processing of insulin precursors has been linked to pancreatic beta-cell stress. At one PI GWAS locus, rs150781447-T is associated with increased PI secretion in Finns (p=5.5X10-11) and causes an amino acid (AA) change (R279C) at a conserved position in exon 4 of TBC1D30, a putative Rab GTPase Activating Protein (GAP), while no GWAS proxy variants (r2&gt;0.8, 1000G Finns) are located in transcribed or enhancer regions. We hypothesized that TBC1D30 is involved in PI processing and that missense variant rs150781447, predicted to be damaging, has a negative impact on gene function. We used genome editing to generate four sets of TBC1D30 mutants in 832/13-INS1 cells. Compared to mock-edited control cells, apparent loss-of-function in frame insertion-deletion cells (106% expression) secreted 2.1-fold more PI (p=0.0001), and heterozygous truncation knockout cells (44% expression) secreted 1.9-fold more PI (p=0.005). Cells heterozygous for rs150781447, generated by prime editing, secreted 1.2-fold more PI (p=0.89), suggesting that the heterozygous variant has subtle or no effect on gene function. Unexpectedly, when exon 4 (42 AA) was deleted in frame, the cells secreted 3.2-fold less PI (p=0.02), consistent with a gain-of-function effect and suggesting that exon 4, part of the conserved Rab GAP protein domain, has a critical function. Preliminary insulin secretion data, while variable, demonstrate trends similar to proinsulin secretion, consistent with a stronger effect of TBC1D30 on processing than secretion. Together, these data demonstrate TBC1D30 as the likely target of the nearby PI GWAS locus and suggest that Rab GAP TBC1D30 plays a critical role in the processing of mature insulin.

Disclosure
V. A. Parsons: None. S. Vadlamudi: None. A. H. Moxley: None. K. L. Mohlke: None.

Funding
National Institutes of Health (1T32GM135128-01)
260-OR: The L-Cell Clock Is Required for Circadian GLP-1 Secretion and Maintenance of Intestinal Homeostasis
Authors: MARTCHENKO SARAH E. , MARTCHENKO ALEXANDRE , BIANCOLIN ANDREW , BRUBAKER PATRICIA L. 
Published: June 2021

Glucagon-like peptide-1 (GLP-1) is an incretin hormone released by the intestinal L cell in response to nutrient intake. Recent studies identified a circadian rhythm in GLP-1 secretion that is disrupted in whole-body Bmal1 KO mice. However, the extent to which the L cell clock regulates circadian GLP-1 secretion and overall diurnal metabolic and intestinal homeostasis is unknown. Gcg-creERT2/+;Bmal1flox/flox KO mice with a disrupted clock in L cells, alpha cells and some CNS cells were thus generated. M/F- and age-matched KO mice had normal body weight and 24hr food intake patterns vs. all genetic and chemical controls. Upon oral glucose gavage at the known peak and trough timepoints of GLP-1 release, KO animals lost the rhythmic GLP-1 secretory pattern in association with impaired secretion (p&lt;0.001) at the peak timepoint. Interestingly, the reduced GLP-1 secretion did not translate to decreased insulin or hyperglycemia, perhaps due to decreased glucagon levels (p&lt;0.05). However, L cell Bmal1 KO caused increases in colonic weight and interleukin-6 (p&lt;0.05) and interferon gamma (p&lt;0.05) expression, suggesting a pro-inflammatory state. In line with findings that altered intestinal epithelial cell function decreases the frequency of intraepithelial CD4+ T cells, KO animals had reduced proportions of these immune cells (p&lt;0.05). Furthermore, 16S rRNA gene analysis revealed increased actinobacteria in the colon, consistent with data from GLP-1 receptor KO mice. KO mice also had decreased levels of cecal bile acids (p&lt;0.05-0.001) and short-chain fatty acids (p&lt;0.05) suggestive of a functional impairment in the KO microbiome. Finally, siRNA-mediated knockdown of Bmal1 in the murine GLUTag L cell line also resulted in impaired rhythmic GLP-1 secretion (p&lt;0.001). Together, these data establish the L cell clock as an essential regulator of circadian GLP-1 secretion which is, then, critical to the maintenance of overall intestinal homeostasis.

Disclosure
S. E. Martchenko: None. A. Martchenko: None. A. Biancolin: None. P. L. Brubaker: None.

Funding
Canadian Institutes of Health Research (PJT-15308)
271-OR: ADA Presidents’ Select Abstract: MyD88 Signaling in Trophoblasts Is Necessary for Maternal High-Fat Diet-Associated Developmental Programming of Obesity
Authors: RUEBEL MEGHAN L. , THAKALI KESHARI M. , CHINTAPALLI SREE V. , SHANKAR KARTIK 
Published: June 2021

Maternal obesity (OB) contributes to programming of offspring obesity. A causal role for placental inflammatory mechanisms in developmental programming remains to be established. We examined the role of the toll-like receptor (TLR) pathway in mediating maternal obesity associated programming by trophoblast deletion of the TLR adapter protein MyD88. We generated MyD88fl/fl:Cyp19 Cre+ (conditional-KO, MyD88-CKO) and MyD88fl/fl:Cyp19 Cre- (flox-control, f/f) mice. Beginning at 5 wk of age, female MyD88-CKO mice were fed ad libitum control (17% fat calories, TD95092) or high-fat diets (HFD, 45% fat calories, TD08811) for 12 wk and then bred with lean MyD88fl/fl:Cyp19 Cre-(flox-control) males (fed control diet). This breeding strategy generated 50% of placenta/offspring with the Myd88-CKO genotype and genetically matched f/f littermate controls. At weaning, offspring (n = 10-12 from distinct litters) were randomized to control or HFD for 16 wk. All offspring have normal TLR signaling, since Cyp19 activity is only present in the placenta. Deletion of placental MyD88 had no effect on offspring growth on control diet. Male f/f offspring from HFD-dams fed HFD showed greater weight gain relative to f/f offspring from control-fed dams fed HFD, indicating an effect of maternal HFD. However, this effect of maternal HFD was not present in MyD88-CKO offspring from HFD-dams fed HFD, revealing a pivotal role for placental TLR signaling in the maternal HFD programming of obesity. Transcriptomic analyses of sex-identified placenta at dpc 17.5 showed that HFD altered more genes in male placenta (864 vs. 580, ±1.5 fold, p&lt;0.05) in a MyD88-dependent manner. Gene ontology analyses showed that alterations in genes related to inflammation, extracellular matrix, and mitochondrial function were absent in placenta lacking MyD88. These findings provide the first unequivocal evidence that placental changes due to maternal HFD are causally related to offspring obesity risk.

Disclosure
M. L. Ruebel: None. K. M. Thakali: None. S. V. Chintapalli: None. K. Shankar: None.

Funding
U.S. Department of Agriculture (ARS-CRIS 6026-51000-012-00-D)
275-OR: Maternal Vitamin D Levels during Pregnancy Is Associated with Childhood Telomere Length: Analysis from a Longitudinal Mother-Child Cohort
Authors: WONG KWUN KIU , CHENG FEIFEI , MAO DI , LIM CADMON K.P. , WANG RONALD , TAM CLAUDIA H. , JOGLEKAR MUGDHA A. , HARDIKAR ANANDWARDHAN A. , JENKINS ALICIA , METZGER BOYD E. , LOWE WILLIAM , TAM WING H. , MA RONALD C. 
Published: June 2021

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&lt;0.001) at age 7. Childhood LTL was positively associated with insulin sensitivity according to the Matsuda index after sex and age adjustment (β±SE=0.038±0.017, p=0.022). Multiple linear regression was used to evaluate the relationship between 25(OH)D levels and LTL, with covariate adjustments for child age, sex, body mass index (BMI), systolic blood pressure (SBP), exercise, gestational age, mode of delivery, and maternal age, SBP, parity, smoking, current parental DM status and pre-pregnancy BMI. Childhood LTL positively correlated with total maternal 25(OH)D concentrations (β±SE=0.003±0.001, p=0.001), maternal D3 (β±SE=0.003±0.001, p=0.002) and maternal D3 epimer (β±SE=0.138±0.042, p&lt;0.001), and similar associations were noted for cord D3 epimer (β±SE=0.101±0.045, p=0.024) and cord total 25(OH)D (β±SE=0.1±0.05, p=0.048), even after covariates adjustments. Our findings suggest 25(OH)D, D3 and D3 epimer both in utero and at birth may impact on childhood LTLs, highlighting a potential link between maternal vitamin D and biological aging.

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.
276-OR: Predictive Primary Metabolomics Signatures in Early to Mid-pregnancy for Risk of Gestational Diabetes
Authors: ZHU YEYI , NGO AMANDA , FAN SILI , BARUPAL DINESH , QUESENBERRY CHARLES , FIEHN OLIVER , FERRARA ASSIAMIRA 
Published: June 2021

Gestational diabetes (GDM) predisposes 7-10% of pregnant women in the U.S. to a myriad of adverse health sequelae. Early prediction is critical. By leveraging metabolomics and clinical data, we aimed to develop and validate predictive metabolic signatures in early to mid-pregnancy for GDM. In a nested matched case-controls study of 91 women with GDM and 180 non-GDM controls in the Pregnancy Environment and Lifestyle Study (PETALS) cohort (discovery set), 157 annotated primary metabolites were measured by gas chromatography/time-of-flight mass spectrometry using fasting serum at gestational weeks (GW) 10-13 and 16-19. Machine learning algorithm using 10-fold cross validated Lasso regression identified predictive metabolomics signatures for GDM risk in the discovery set. We validated the signature at GW 10-13 in a random PETALS subsample (42 GDM, 372 non-GDM; validation set 1) and a matched case-control study within the GLOW trial of overweight/obese women (35 GDM, 70 non-GDM; validation set 2) and the signature at GW 16-19 in validation set 1. Upregulated aromatic and acidic amino acids and purinones at both GW 10-13 and 16-19 and additionally hexoses at GW 16-19 were significantly associated with GDM risk (FDR-adjusted P &lt;0.05). A 33-metabolite signature of organic acids, lipids, and organooxygen compounds at GW 10-13 revealed incremental predictive ability beyond conventional risk factors (CRF) including age, pre-pregnancy BMI, preexisting hypertension, previous GDM, diabetes family history, and fasting glucose (discovery set AUC: 0.871 vs. 0.742; validation set 1: 0.869 vs. 0.731; validation set 2: 0.972 vs. 0.742; all P &lt;0.01). Similar predictive ability was found for 13 metabolites at GW 17-19 beyond CRF (discovery AUC: 0.838 vs. 0.732; validation 1: 0.830 vs. 0.774; both P &lt;0.05). Our findings suggest potentially important roles of increased amino acids, purinones, and hexoses in the pathophysiology of GDM and a predictive ability of selected primary metabolites beyond CRF.

Disclosure
Y. Zhu: None. A. Ngo: None. S. Fan: None. D. Barupal: None. C. Quesenberry: None. O. Fiehn: None. A. Ferrara: None.

Funding
National Institutes of Health (K12HD052163); National Institute of Diabetes and Digestive and Kidney Diseases (K01DK120807)
279-OR: Hydroxylation Regulation of Hepatic Gluconeogenesis via Prolyl-Hydroxylase 3 in Mice
Authors: XUE YAQIAN , CAI GENXIANG , MA FENGGUANG , LIU ZHENGSHUAI , LIU YUXIAO , SU WEITONG , ZHENG ZENGPENG , DING DONG , JIANG YANG , GAO JING , DAI XIAOZHEN , CUI AOYUAN , LI YU 
Published: June 2021

Hepatic gluconeogenesis is critical for metabolic adaption in response to fasting, and its abnormal elevation contributes to hyperglycemia and type 2 diabetes. However, whether and how hydroxylation regulates hepatic gluconeogenesis remains largely unknown. Here we identified prolyl hydroxylase 3 (PHD3) of the hydroxylase family as a key regulator for hepatic gluconeogenesis. Liver-specific knockout of PHD3 (PHD3 LKO) mice show attenuated fasting gluconeogenic genes, glycaemia and hepatic capacity to produce glucose as evidenced by a pyruvate tolerance test. The efficacies of glucagon or adenylyl cyclase activator forskolin on stimulating the expression of gluconeogenic enzymes, such as PEPCK and G6Pase, and on glucose output are abolished in PHD3-/- mouse primary hepatocytes. To determine whether the prolyl hydroxylase activity is required for PHD3’s effects on promoting gluconeogenesis, liver-specific prolyl hydroxylase-deficient knockin mice with alanine mutation in PHD3 at Pro196 (PHD3 KI) were generated. Similar to PHD3 LKO mice, PHD3 KI mice exhibit attenuated fasting gluconeogenic genes, glycaemia and hepatic capacity to produce glucose, suggesting that PHD3 regulates hepatic gluconeogenesis in a proline hydroxylase-dependent manner. Moreover, several cellular proteins that associate with PHD3 were identified by comprehensive protein analysis using LC-MS/MS followed by bioinformatic analysis including GSK3β, IRS2 and OGT, which may act as PHD3 substrates and play roles in mediating PHD3’s effects on regulating glucose metabolism. Intriguingly, expression levels of PHD3 are increased in the liver diet-induced insulin resistant mice. Together, these results demonstrate that hydroxylation plays a critical role in the regulation of hepatic gluconeogenesis under physiological and insulin resistant conditions. Hyperactivation of PHD3 may underscore the potential role of PHD3 in the development of sustained gluconeogenesis, hyperglycemia and type 2 diabetes.

Disclosure
Y. Xue: None. J. Gao: None. X. Dai: None. A. Cui: None. Y. Li: None. G. Cai: None. F. Ma: None. Z. Liu: None. Y. Liu: None. W. Su: None. Z. Zheng: None. D. Ding: None. Y. Jiang: None.

Funding
National Key Research and Development Project of China (2019YFA0802502); National Natural Science Foundation of China (81925008); Shanghai Science and Technology Commission (19140903300)