bluuebunny/crossref_metadata_2025_split huggingface dataset:
280-OR: Metabolomic Signatures of Excess Fetal Growth in Pregnancies Complicated by Type 1 Diabetes (T1D)
Authors: ISGANAITIS ELVIRA M. , WOLFS DANIELLE , O'CONNELL JULIANNE C. , HANSBURY GEORGIA A. , WOLPOWITZ SHIRA , CLISH CLARY B. , JAMES-TODD TAMARRA , BROWN FLORENCE M. 
Published: June 2023

Background: In pregnancies complicated by T1D, glycemic control is only moderately associated with excess fetal growth, suggesting that nutrients other than glucose may be involved. Aims: We analyzed associations between metabolites in maternal and newborn plasma, and fetal growth patterns, in T1D pregnancies.
Methods: We obtained maternal plasma samples in the 2nd and 3rd trimester and at delivery, and cord blood samples immediately after delivery, in 22 mother/infant dyads. We used an untargeted metabolomic platform (Broad Institute) and calculated correlation coefficients between individual metabolites and newborn growth percentiles adjusted for gestational age, sex, parity, and ethnicity (GROW percentile). We used a false discovery rate threshold of FDR&lt;0.2. Large for gestational age (LGA) was defined as GROW percentile ≥90 and appropriate for gestational age (AGA) as between the 10th and 90th percentiles.
Results: Forty-five percent of newborns were classified as LGA. Maternal age, BMI, diabetes duration, insulin dose and pump use did not differ between LGA vs. AGA. We identified stronger associations between GROW percentiles and metabolites in cord blood than with metabolites in maternal plasma during the 2nd or 3rd trimester. In cord blood, 29 metabolites were associated with GROW percentile (FDR&lt;0.2). For 51 metabolites, the ratio between abundance in cord blood to abundance in maternal blood at delivery was associated with the GROW percentile (FDR &lt;0.2). Many of the top-ranking metabolites positively associated with infant growth were long-chain glycerophosphocholines (e.g., LPC(20:3) β 0.035, p=0.0002, FDR 0.01; LPC(22:5) β 0.033, p=0.0008, FDR 0.03).
Conclusion: Glycerophospholipids in cord blood were associated with excess fetal growth in T1D pregnancies, suggesting alterations in lipid metabolism. We also report associations between fetal growth and ratios of metabolites in cord blood to maternal blood, suggesting differences in placental transport.

Disclosure
E.M.Isganaitis: None. D.Wolfs: None. J.C.O'connell: None. G.A.Hansbury: None. S.Wolpowitz: None. C.B.Clish: None. T.James-todd: None. F.M.Brown: Other Relationship; Dexcom, Inc.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (P30DK036836, P30DK057521); Broad Institute
284-OR: An Alternative Downstream Translational Start Site in PPARG Produces a Novel Protein That Rescues Loss-of-Function Mutations in Humans
Authors: MENDEZ LARA KAREN , DU XIAOMI , GLASS KIMBERLY , HERNANDEZ RUBEN , DE ARRUDA SALDANHA CAMILA , HEINZ SVEN , MAJITHIA AMIT R. 
Published: June 2023

PPARG is the target of thiazolidinediones (TZD) and plays a central role in glucose and lipid metabolism. Loss-of-function (LOF) mutations in PPARG increase the risk of type 2 diabetes (T2D) and cause Mendelian lipodystrophy. In population-scale sequencing studies, we identified protein-truncating variants in PPARG expected to cause lipodystrophy, but surprisingly showing no burden of metabolic disease in their human carriers. To further understand this finding, we use CRISPR/CAS9 to systematically target deletions to every exon of PPARG revealing that mutations causing deleterious frameshifts in exons 1 and 2 resulted in cells with intact PPARG responses. We subsequently constructed a library of PPARG transgenes encoding termination sequences at every possible codon (1-505) in the PPARG2 cDNA and tested them for transactivation function. We found that PPARG transgenes encoding termination sequences prior to codon 135 (c.ATG402-404) corresponding to exon 2 in the genomic DNA (chr3:12381414 Hg38) retained transactivation activity. We hypothesized that c.ATG402-404 served as an alternative translational start site that generates a PPARG isoform lacking the N-terminus but retaining DNA-binding, ligand-binding, and transactivation properties. Indeed, cells engineered to disrupt PPARG prior to chr3:12381414 retained TZD responsiveness and produced a 40 kDa protein recognized by antibodies targeting the C-terminus but not the N-terminus of PPARG. Analysis of the global transcriptional activity of the PPARG p.M135-505 compared to full-length PPARG revealed that p.M135-505 produced a similar TZD-induced upregulation of known PPARG targets and gene sets. In summary, PPARG p.M135-505 is a truncated but fully functional protein that responds to PPARG agonists and may rescue disruptive mutations that would otherwise cause lipodystrophy.

Disclosure
K.Mendez lara: None. X.Du: None. K.Glass: None. R.Hernandez: None. C.De arruda saldanha: None. S.Heinz: Research Support; Hologic Inc., Stock/Shareholder; Leash Laboratories. A.R.Majithia: None.

Funding
National Institutes of Health (RDK01DK123422)
288-LB: CHIP Haplodeficiency Exacerbates Experimental Type 1 Diabetes via the Regulation of DNA Damage Response and Apoptosis
Authors: HWANG HWANG AE-RANG 
Published: June 2023

Type 1 diabetes is a disease characterized by hyperglycemia resulting from defects in insulin secretion. Streptozotocin (STZ)-induced pancreatic β-cell damage is commonly used as causing hyperglycemia of type 1 diabetes model. It has been established that the carboxyl terminus of heat shock protein 70-interacting protein (CHIP) contributes to the pathogeneses of diabetic complications. However, the mechanisms linking type 1 diabetes and CHIP-dependent antidiabetic signaling pathway remain to be addressed. Here we utilized STZ-induced murine model of type 1 diabetes to examine DNA damage responses (DDR) during pancreatic β-cell apoptosis. This study was undertaken to explore the mechanism whereby CHIP instigates pancreatic β-cell apoptosis via DDR-dependent signaling pathway. Wild type and CHIP haplodeficient mice were i.p. injected with 50 or 100 mg/kg STZ in order to induce experimental mouse model of type I diabetes. STZ-induced hyperglycemia, glucose intolerance, and inflammatory responses were exacerbated in CHIP haplodeficient mice compared to Wild type. In addition, immunoblotting data revealed that depletion of CHIP with siRNA against rat stub1 enhanced STZ-induced apoptosis and DDR pathway in INS-1 cells, a rat insulinoma cell line. Notably, CHIP inhibition deteriorates STZ-induced cell cycle arrest and oxidative stress in INS-1 cells. In addition to in vitro study, immunohistochemical and immunoblotting study showed that CHIP haplodeficiency exacerbates DDR in pancreas from an experimental mouse model of type I diabetes. These results suggest that CHIP protects against STZ-induced pancreatic β-cell apoptosis and hyperglycemia by interrupting the DDR-mediated apoptotic pathway. Key words: diabetes, CHIP, DNA damage response, apoptosis, β-cell

Disclosure
A. Hwang: None.
292-OR: High-Fat Diets Containing Sucrose and Fructose, but Not Glucose, Induce Obesity and Hepatic Insulin Resistance via Accumulation of Diacylglycerols
Authors: PARK SE-HYUNG , FADHUL TAGHREED I. , RONALD KAHN C. , SOFTIC SAMIR 
Published: June 2023

Diets high in sugar and fat promote the development of obesity and metabolic complications, while high-sugar, low-fat or high-fat, sugar-free (ketogenic) diets do not. We assessed the importance of sugar in a commonly used 60% high-fat diet (HFD), which contains 6.7% of calories from sucrose. The sucrose content was replaced either with an equal amount of fructose (HFD+RF) or glucose (HFD+RG) and all other components of the diet were identical. After 12 weeks on the diets, the mice on HFD and HFD+RF diets gained more weight, developed hepatic steatosis, glucose intolerance and insulin resistance, compared to the mice on normal chow or HFD+RG diets. A relatively small (6.7%) amount of sucrose or fructose, but not glucose, in a HFD was sufficient to increase the fructolysis (KHK-C, ALDO B and TKFC) and de novo lipogenesis (ME1, ACLY, ACC1, FASN, SCD1) pathways by WB. Metabolomics analysis revealed higher hepatic diacylglycerol (DAG) content in HFD and HFD+RF groups, while HFD+RG group had elevated hepatic triacylglycerol content. Proteomics analysis quantified 3058 unique proteins, 736 of which were significantly different between the mice on chow and HFD. Replacing sucrose with fructose altered only 2 proteins, while replacing sucrose with fructose significantly changed 196 proteins in the liver. Reactome pathway analysis showed that the most altered processes mediate lipid biosynthesis and metabolism. In conclusion, HFDs containing sucrose or fructose but not glucose increase the fructolysis pathway and lead to hepatic DAG accumulation and insulin resistance. Thus, a relatively small amount of fructose, regardless of its source, is sufficient to alter metabolic outcomes in mice consuming a HFD.

Disclosure
S.Park: None. T.I.Fadhul: None. C.Kahn: None. S.Softic: Research Support; Alnylam Pharmaceuticals, Inc.
293-OR: Macrophage-Derived Interleukin-12 Regulates Obesity-Mediated Insulin Resistance in the Liver
Authors: FRIEDLINE RANDALL H. , ALBUSHARIF MAHAA , KIM ALLISON M. , KIM LAUREN H. , BAEZ TORRES NATALIE M. , TAUER LAUREN A. , HU XIAODI , KIM JASON K. 
Published: June 2023

Inflammation and insulin resistance play a major role in nonalcoholic fatty liver disease (NAFLD), and IL-12, a pro-inflammatory cytokine primarily secreted by macrophages, is elevated in obesity. Here, we examined the effects of acute treatment with IL-12 in vivo on glucose metabolism in male C57BL/6J mice. Mouse recombinant IL-12 (0.25 μg/hour) or saline (control) was infused for 4 hours, followed by insulin clamps in mice (n=5/group). Glucose infusion rates during clamps were significantly reduced in IL-12-treated mice, resulting in a 34% decrease compared to controls (Fig. 1-2). Basal hepatic glucose production (HGP) did not differ, but clamp HGP was significantly increased in IL-12-treated mice (Fig. 3). As a result, IL-12 treatment caused a 43% decrease in hepatic insulin action compared to controls (Fig. 4). These IL-12 effects were associated with a 4-fold increase in STAT4 tyrosine-phosphorylation (0.071±0.026 vs. 0.018±0.004 in controls) and a 47% decrease in IRS1 protein levels (0.264±0.044 vs. 0.496±0.112 in controls) in the liver. Attenuated liver IL-12 level was further associated with improved hepatic insulin action in diet-induced obese mice with conditional loss of IFNγ signaling in myeloid cells. CONCLUSION: Our findings identify IL-12 as a novel cytokine regulating obesity-mediated insulin resistance and inflammation in the liver and a potential therapeutic target to treat NAFLD.

Disclosure
R.H.Friedline: None. M.Albusharif: None. A.M.Kim: None. L.H.Kim: None. N.M.Baez torres: None. L.A.Tauer: None. X.Hu: None. J.K.Kim: None.

Funding
National Institutes of Health (5U2CDK093000)
294-LB: Altered a-Cell Phenotype in Pancreata Is Associated with Increased Peri-islet Collagen Ceposition
Authors: KATTNER NICOLE , RICHARDSON SARAH , AL-SELWI YARA , TINIAKOS DINA , SHAW JAMES A. 
Published: June 2023

Cystic fibrosis (CF)-related diabetes (CFRD) affects up to 50% of adults with CF, adding to treatment burden and impacting overall wellbeing. Major changes are observed in pancreas morphology with acinar tissue replaced by fibrosis and/or fat. Islets are still present even in advanced disease and may cluster together. It is believed CFRD is caused by impaired insulin secretion and loss of β-cell mass, however, detailed pathogenesis remains unknown. We investigated post-mortem pancreatic tissue from 19 donors with CF (age range: premature-27 years) and 10 donors without CF (6-65 years). CF pattern of injury (fibrotic/lipoatrophic) was assessed by routine histology. Sirius Red/Fast Green stain was used to assess fibrosis. Artificial intelligence analysis was performed to quantify intra-islet and peri-islet collagen distribution in 16 cases (7 CF, 9 control). Triple immunofluorescence staining for insulin, glucagon, and vimentin followed by confocal microscopy was used to image up to 50 islets per donor. Glucagon and vimentin-positive (glu/vim+) α-cells were observed in both CF and non-CF pancreata (mean (range) 32 (0-90)% of islets). The presence of this altered α-cell phenotype positively correlated with age in all donors (R=0.41, p&lt;0.05). Presence of glu/vim+ α-cells correlated with peri-islet collagen density in analysed CF and control cases (glu/vim+ islets 44 (0-82)%) (R=0.60, p&lt;0.05). In control pancreata alone glu/vim+ cells were seen in 1-82% (mean 41%) islets and correlated positively with intra-islet collagen density (R=0.76, p&lt;0.05). Interestingly, two CF cases with confirmed diabetes showed low prevalence of this altered phenotype (2% of islets) with 13 vs 40% fibrosis in the whole section. Our data provide further evidence for exocrine-endocrine signalling in the human pancreas and suggest that presence of vimentin-expressing α-cells may be driven by increased collagen deposition in the islet microenvironment during aging and in CFRD pathogenesis.

Disclosure
N. Kattner: None. S. Richardson: None. Y. Al-selwi: None. D. Tiniakos: None. J. A. Shaw: Advisory Panel; Betalin Therapeutics, Provention Bio, Inc., Consultant; Mogrify.

Funding
Cystic Fibrosis Trust (SRC019)
296-LB: Loss of SERCA2 Induces Mitochondrial Dysfunction, Increases ß-Cell Senescence, and Accelerates Type 1 Diabetes Development
Authors: WEAVER STACI A. , BONE ROBERT N. , ACRI DOMINIC J. , KIM JUNGSU , KONO TATSUYOSHI , DAHL RUSSELL , EIZIRIK DECIO L. , SYED FAROOQ , EVANS-MOLINA CARMELLA 
Published: June 2023

Type 1 diabetes (T1D) results from immune-mediated destruction of pancreatic β-cells. Senescence and activation of a senescence-associated secretory phenotype (SASP) in β-cells may contribute to T1D pathogenesis; however, the mechanisms responsible for this phenotype are not well understood. We hypothesized that loss of endoplasmic reticulum (ER) Ca2+ induces mitochondrial dysfunction and drives the development of β-cell senescence during T1D. To test this hypothesis, we generated sarco/ER Ca2+-ATPase 2 (SERCA2) haploinsufficient mice on the NOD background (NOD-S2+/-). NOD-S2+/- mice displayed accelerated T1D onset compared to NOD-WT mice (14 wks vs 18 wks; p≤ 0.0001 female and 21 wks vs 24.5 wks male; p&lt;0.05). As expected, basal ER Ca2+ levels were reduced in NOD-S2+/- islets, but basal mitochondrial Ca2+ levels were elevated compared to NOD-WT islets. Basal cytoplasmic Ca2+ levels remained unchanged. Single cell RNA sequencing showed an increase in immune cell markers in the NOD-S2+/- islets at 6 wks of age, with signatures of ER stress and mitochondrial dysfunction at both 6 and 10 wks of age. Islets from prediabetic female NOD-S2+/- mice showed decreased glucose-stimulated O2 consumption, reduced ATP production, and decreased mitochondrial membrane potential. Additionally, protein expression of ATP synthase and Complex III was decreased in female NOD-S2+/- islets, but mitochondrial content was unchanged. There was an increased expression of multiple markers of senescence and SASP in NOD-S2+/- islets between 6 and 12 wks of age compared to NOD-WT islets. Lastly, in vivo allosteric activation of SERCA decreased insulitis and delayed diabetes onset in WT NOD mice. Our results show that SERCA2 deficiency leads to mitochondrial dysfunction, β-cell senescence, and accelerated T1D development. In future studies, we will test whether senolytic agents can delay diabetes onset and rescue mitochondrial health in models of T1D and SERCA2 deficiency.

Disclosure
S. A. Weaver: Employee; Eli Lilly and Company. R. N. Bone: None. D. J. Acri: None. J. Kim: None. T. Kono: None. R. Dahl: None. D. L. Eizirik: None. F. Syed: None. C. Evans-molina: Advisory Panel; Provention Bio, Inc., DiogenX, Avotres Inc., Neurodon, MaiCell Therapeutics, Other Relationship; Isla Technology, Bristol-Myers Squibb Company, Nimbus Therapeutics, Research Support; Lilly, Astellas Pharma Inc.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (F31DK134168, R01DK093954, R01DK127308, R01DK127236, UC4DK104166, U01DK127786); U.S. Department of Veterans Affairs (I01BX001733, 2I01BX001733-05); JDRF (3-PDF-2017-385-A-N, 3-IND-2022-1235-I-X); National Center for Advancing Translational Sciences (UL1TR002529)
297-LB: Dorzagliatin, a Glucokinase Activator, Repairs Defective Islet Function in High-Fat Diet-Induced Diabetic Mice
Authors: HAN DONGNA , MENG SHI , ZHOU XUE , LI RUYUE , LI CHANGHONG 
Published: June 2023

The DREAM study showed long-term benefits for glucose control in newly diagnosed type 2 diabetes (T2D) patients even after withdrawal of dorzagliatin treatment, a newly approved glucokinase activator for T2D treatment, who were considered to be in the remission for diabetes. The purpose of the current study is to examine the mechanisms of dorzagliatin-induced diabetes remission by evaluating the changes in islet function over time after a period of administering dorzagliatin in high-fat diet (HFD) induced obesity and diabetes mouse models. Diabetes mice were generated after 7 months of HFD, then received dorzagliatin or vehicle controls via oral gavage once a day for 19 days (8 days of 30 mg/Kg and 11 days of 15 mg/Kg). After drug withdrawal, one group of mice remained in HFD and another group changed to a normal diet. Islets were then isolated at different time points after drug withdrawal, and the serum and liver were collected for evaluation. The glucose ramp-stimulated insulin secretion (GSIS) was evaluated in islet perifusion. Gene expression was measured by qPCR in islets and by RNA-seq in the liver. Compared to normal mouse islets, HFD-induced diabetic islets in vehicle groups have impaired GSIS with about 40% reduction in maximum insulin secretion. In contrast, GSIS was fully recovered to the levels of normal islets in 10, 18 and 38 days after dorzagliatin withdrawal, but returned to defective GSIS levels after 45 days of drug discontinuation. In addition, islet GSIS can be fully recovered after 80 days of diet change.
In conclusion, the defective GSIS in HFD-induced diabetes mouse islets is reversible and can be fully restored by dorzagliatin even after a long period of drug withdrawal while still remaining in HFD. The mechanisms are likely due to increased glucokinase gene expression caused by a short treatment of dorzagliatin. Diet change alone can recover islet function but requires weight reduction and a longer duration.

Disclosure
D. Han: Employee; Nanjing AscendRare Pharmaceutical Technology. S. Meng: Employee; Nanjing AscendRare Pharmaceutical Technology. X. Zhou: Employee; Nanjing AscendRare Pharmaceutical Technology. R. Li: Employee; Nanjing AscendRare Pharmaceutical Technology. C. Li: Employee; Hua Medicine, Nanjing AscendRare Pharmaceutical Technology.
323-OR: Association of Silent Myocardial Infarction with Cardiac Autonomic Neuropathy among Asymptomatic Patients with Type 2 Diabetes
Authors: FU YIMAN , WU LIN , YANG YI , QIN BAODING , LUO QINWEN , CHEN YANMING , ZHU YANHUA 
Published: June 2023

Background: Silent myocardial infarction (SMI) is a severe cardiovascular disease among patients with DM. Nowadays, the mechanism of SMI is not clear, but diabetic cardiac autonomic neuropathy (CAN) is assumed to be the most probable reason. However, whether SMI is associated with CAN remains controversial. Therefore, we aimed to evaluate the relationship between SMI and CAN in asymptomatic patients with T2DM.
Methods: This is a hospital-based observational study. 4055 T2DM patients who were screened for CAN by Holter monitoring were investigated. Q-waves SMI was detected by ECG. The sensitivity and specificity of ECG-detected SMI by computed tomography-coronary angiography (CTA).
Results: A total of 4055 patients were screened. At last, we included 1015 patients with T2DM. In the included population, 508 cases (50%) had CAN. Patients with CAN were more likely to be older and have higher levels of TC, LDL-c, Cr, UA, and Hcy (p&lt;0.05). A total of 466 cases of SMI were detected by ECG, and the prevalence of SMI was 45.9% in 1015 patients. Compared with T2DM patients without CAN, patients with CAN had a higher prevalence of SMI (73.4% vs 26.6%, p&lt;0.001). Multivariate logistic regression analysis confirmed that concomitant CAN was an independent predictor of the risk of SMI in T2DM patients (OR=7.53, 95%CI:2.98-19.02, p&lt;0.001). Among the 133 patients who underwent coronary CTA, 55 (41.0%) cases had SMI, and 36 of them had Q-waves on ECG. The sensitivity and specificity of SMI detected by ECG were 65.5% and 91.0%, respectively. In 133 T2DM patients, patients with CAN also had a higher risk of occurring SMI than those without CAN (65.5% vs 34.5%, p&lt;0.001).
Conclusion: In asymptomatic T2DM patients, the occurrence of SMI was independently associated with CAN. Therefore, we should pay attention to the possibility of SMI in asymptomatic T2DM patients clinically. In particular, the monitoring should be strengthened when patients combined with diabetic CAN.

Disclosure
Y.Fu: None. L.Wu: None. Y.Yang: None. B.Qin: None. Q.Luo: None. Y.Chen: None. Y.Zhu: None.
325-OR: Glucagon-Like Peptide 1 Receptor Agonists Preserved Vascular Endothelial Junction through Inhibition of STING Signaling–Elicited Inflammation in Diabetic Retinopathy
Authors: HE XUEMIN , WEN SIYING , TANG XIXIANG , WEN ZHEYAO , ZHANG RUI , LI SHASHA , GAO RONG , ZHU YANHUA , LU YAN , SHI GUOJUN , CHEN YANMING 
Published: June 2023

Background: Glucagon like-peptide 1 receptor agonists (GLP-1 RAs) are protective to macrovascular diseases and nephropathy in diabetic patients; however, their role in diabetic retinopathy (DR) are still debatable. This study aimed to address the effect of GLP-1 RAs on endothelial junction and inflammation in DR.
Methods: GLP-1 RAs were injected to db/db mice, then retinal inflammation and vascular changes were determined by RNA-sequencing analysis, immunostaining, fluorescein fundus imaging and Optical coherence tomography. Cross-sectional study of 1626 type 2 diabetic patients was performed.
Findings: GLP-1R was majorly detected in retinal vessels, which was down-regulated by diabetes. GLP-1 RAs significantly rescued diabetes-induced retinal thinning and vascular tortuosity, likely through altered the expression profiles of junction- and mitochondria-related genes, which was confirmed by exedin-4 treatment in vitro. Stimulator of interferon genes (STING) was most abundant in the endothelial cells from fibrovascular membranes, which was positively correlated with the levels of angiogenic factors and inflammatory factors in endothelial cells from retinas with retinopathy and fibrovascular membranes. Further analysis showed prominent inhibition by GLP-1 RAs on STING activation in db/db retinas. Cross-sectional study further showed decreased DR incidence by GLP-1 RAs.
Conclusions: Activation of the STING pathway induced inflammatory factors while suppressed junctional proteins in endothelial cells of DR. GLP-1 RAs ameliorated diabetes-elicited vascular damages through inhibition of the STING pathway.

Disclosure
X.He: None. G.Shi: None. Y.Chen: None. S.Wen: None. X.Tang: None. Z.Wen: None. R.Zhang: None. S.Li: None. R.Gao: None. Y.Zhu: None. Y.Lu: None.

Funding
National Natural Science Foundation of China (82000782, 82270886, 82070811); Third Affiliated Hospital of Sun Yat-sen University (2020G2RPYQN11); China International Medical Foundation (2018-N-01); Science and Technology Plan Project of Guangzhou City (202007040003)