bluuebunny/crossref_metadata_2025 huggingface dataset:
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 <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 <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 <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)
297-OR: Triglyceride Reduction and Effect on Lipoprotein Metabolism in Patients with Type 2 Diabetes Mellitus Receiving Fixed-Dose Combination of Insulin Degludec and Liraglutide
Authors: PAPPA ELENI , KOSTARA CHRISTINA , DIMOU AIKATERINI , BAIRAKTARI ELENI T. , TSIMIHODIMOS VASILIS
Published: June 2021

Aim: To compare the effect of fixed-dose combination of Insulin Degludec and Liraglutide (IDegLira) on anthropometric, carbohydrate metabolism and lipid parameters, as well as on apoB-containing lipoprotein subclasses and HDL lipidome, in patients with type 2 diabetes (T2DM) regarding changes in triglyceride (TG) levels.

Methods: Seventy-two uncontrolled patients with T2DM on oral antidiabetics were randomized to receive IDegLira or Insulin Degludec (IDeg) for a 12-week treatment. In the present study, 42 patients receiving IDegLira were included. Twenty-one presented a decrease, while the remaining an increase in TG levels. ApoB-containing lipoprotein subclasses levels were measured by LDL-Lipoprint method, whereas HDL lipidomic analysis via 1H-NMR spectroscopy.

Results: In patients experiencing a decrease in TG levels after IDegLira administration, a significant decrease in body weight and BMI, by 4.4kg and 1.59kg/m2 respectively, was observed. Fasting glucose, HbA1c and HbA1c Glycation Index noted a decrease by 35 mg/dl, 1.4% and 1.1 respectively (p=0.018, p=0.004, p=0.004). IDegLira administration in this subgroup resulted in a significant reduction of total (TC) and LDL cholesterol (LDLC) by 22.5mg/dl and 13.2mg/dl, respectively. Small-dense LDLC (sdLDLC) levels significantly correlated with changes in TG levels (r=0.496, p=0.02). Furthermore, IDegLira administration resulted in significant enrichment of HDL particle core in esterified cholesterol, depletion of TG content and an atheroprotective shift in fatty acid pattern of HDLs. The aforementioned changes were significantly greater than those observed in patients receiving IDegLira and did not experience a TG reduction.

Conclusions: TG reduction as a result of IDegLira administration is accompanied by greater changes in anthropometric and carbohydrate metabolism parameters, TC, LDLC, sdLDLC and HDL lipidome.

Disclosure

E. Pappa: None. C. Kostara: None. A. Dimou: None. E. T. Bairaktari: None. V. Tsimihodimos: None.
3-OR: Racial/Ethnic Disparities in Diabetes Technology Use in Young Adults (YA) with Type 1 Diabetes (T1D): Multidisciplinary Stakeholder Perspectives and Solutions
Authors: AGARWAL SHIVANI , CRESPO-RAMOS GLADYS , LEUNG STEPHANIE , FINNAN MOLLY , PARK TINA , GONZALEZ JEFFREY S. , LONG JUDITH A.
Published: June 2021

Background: Black and Hispanic YA with T1D are less likely to use insulin pumps and continuous glucose monitors than White peers, which may be contributing to disparities in glycemic and long-term outcomes.

Methods: We conducted a series of interactive focus groups to elicit perspectives on racial/ethnic disparities in diabetes technology use, patient-centered barriers and facilitators, and solutions to enhance uptake. We invited multidisciplinary panels of stakeholders to participate, including YA T1D patients, physicians, nurses, psychologists, and community health workers. Sessions were recorded and analyzed for common themes.

Results: We convened 7 virtual 2-hour workshops and recruited 32 stakeholders from 11 states (5 Black and 7 Hispanic YA, 20 other stakeholders). Notably, many patients stated that they had never been offered technology until recent engagement in a specialized YA T1D clinic. Once offered technology, multiple barriers were present across a chronological journey consisting of three phases: 1) learning about and deciding to get technology; 2) initiating use; and 3) managing ongoing problems. Some barriers included a lack of knowledge about devices, mismanaged expectations for device wearability and use, and insurance coverage benefits. Facilitators included family support, T1D peer partnership, and a supportive healthcare team. Co-created solutions to address barriers were categorized into four main themes: 1) more equitable system of offering technology; 2) visual and hands-on education; 3) peer support; and 4) insurance navigation assistance.

Discussion: Engaging multiple stakeholders with lived experience of the problem is an important approach to diabetes disparities research. Inadvertent provider bias may be preventing equal access to technology for minority YA with T1D, while extra support for device use through the technology journey is needed to sustain use.

Disclosure

S. Agarwal: None. G. Crespo-ramos: None. S. Leung: None. M. Finnan: None. T. Park: None. J. S. Gonzalez: None. J. A. Long: None.

Funding

National Institutes of Health (K23DK115896)
30-LB: Effect of Intense Lifestyle Therapy in People with Obesity and Type 2 Diabetes
Authors: YOSHINO MIHOKO , YOSHINO JUN , BITTEL ADAM J. , BITTEL DANIEL , REEDS DOMINIC , STEIN RICHARD , SINACORE DAVID R. , CADE WILLIAM T. , OKUNADE ADEWOLE L. , PATTERSON BRUCE W. , KLEIN SAMUEL
Published: June 2021

Weight loss and exercise are the cornerstones of therapy for patients with obesity and type 2 diabetes (T2D). However, successful compliance with therapy is difficult to achieve. We conducted an 8-month randomized, controlled trial in persons with obesity and T2D to determine the effect of intensive lifestyle therapy (ILT), delivered at a worksite setting, on body weight, cardiometabolic function, and cellular metabolic pathways. Subjects were assigned to Standard Care (SC; n=8, BMI:38±5, fasting blood glucose [FBG]:161±19 mg/dl, HbA1c:7.6±1.1%) or ILT (n=10, BMI:37±6, FBG:150±32 mg/dl, HbA1c:7.0±1.1%), which involved weekly diet-behavior therapy sessions and 4 d/wk of supervised resistance and endurance exercise training. Weight loss was greater in the ILT than SC group (17±7% vs. 1±2%, P<0.05). All cardiometabolic outcomes improved more in the ILT than SC group: i) insulin sensitivity (glucose disposal rate during a hyperinsulinemic-euglycemic clamp increased by 116±78% vs. 15±9%, P<0.05); ii) β-cell function (insulin secretion in response to an oral glucose load relative to insulin sensitivity increased by 194±205% vs. 12±20%, P<0.05); and iii) cardiorespiratory fitness (VO2 peak during cycle ergometer exercise increased by 28±22% vs. -2±15%, P<0.05). The most upregulated biological pathways enriched in differentially expressed genes (DEGs) in skeletal muscle, assessed by RNA-seq, after ILT involved mitochondrial function and sirtuin signaling pathways. Upstream regulators of these DEGs included NAD dependent deacetylase sirtuin-3 (SIRT3) and NAMPT (FDR<0.05). In addition, ILT increased muscle NAD content by 51±46% (P<0.05). FBG and HbA1c decreased to 108±31 mg/dl and to 6.1±0.9%, despite a 60% decrease in the use of diabetes medications after ILT, but did not change with SC. We conclude that marked diet-induced weight loss and exercise training can be achieved in a structured worksite setting and has profound therapeutic cardiometabolic effects in people with obesity and T2D.

Disclosure

M. Yoshino: None. B. W. Patterson: None. S. Klein: Advisory Panel; Self; Altimmune, Merck & Co., Inc., Novo Nordisk Inc., ProSciento, Research Support; Self; Janssen Research & Development, LLC. J. Yoshino: None. A. J. Bittel: None. D. Bittel: None. D. Reeds: None. R. Stein: None. D. R. Sinacore: None. W. T. Cade: None. A. L. Okunade: None.

Funding

American Egg Board; National Dairy Council (T32DK07296DK007120)
302-OR: Medical Prescriptions for Vegetables Are Associated with Reduced Consumption of Unhealthy Foods, Improved Food Security, and Lowered Cardiometabolic Risk among Adults with or at Risk of Type 2 Diabetes
Authors: GLANTZ NAMINO M. , CONNEELY CASEY , LAREZ ARIANNA J. , BEVIER WENDY C. , KERR DAVID
Published: June 2021

Previously, we have reported that improving access to fresh vegetables reduces cardio-metabolic risk for predominantly Mexican American adults with or at risk of type 2 diabetes (T2D). This analysis examined changes in self-reported consumption of less healthy foods (soda, sugary drinks, and tortillas) and the association with medical prescriptions for locally sourced fresh vegetables.

In this observational cohort study, 202 adults (152 female, 80% Hispanic/Latino, mean ±SD age of 53.8 ±12.6 years) with (n=62) or at risk (American Diabetes Association risk calculator) of T2D received 21 servings per week of locally sourced fresh vegetables prescribed by a physician for 10 weeks. No nutrition education was provided. For participants with paired data, there were reductions in waist circumference (-0.9 ±3.7 cm, p=0.003); systolic blood pressure for those with baseline ≥ 130 mmHg (-8.9±16.5 mmHg, p=0.001); and HbA1c values for participants with a baseline >7.0 % (-0.4 [-0.8, -0.1] %, p=0.016). Fewer participants had low or very low food security (USDA 6-item Food Security Module) after the intervention (25 versus 76, p<0.0001), associated with reductions in consumption of tortillas (p<0.00001); soda and other sugar-sweetened fruit drinks, sweet tea, and sports or energy drinks (p<0.001); and their weekly frequency of eating out (p=0.01). The improvement in food security was also associated with better sleep (ρ=0.22, p=0.007).

The use of medical prescriptions for vegetables is associated with greater food security and lower cardiometabolic risk in predominantly Mexican-American adults with or at risk of T2D, possibly related to less consumption of unhealthy foods.

Disclosure

N. M. Glantz: Research Support; Self; Abbott, Eli Lilly and Company. C. Conneely: Research Support; Self; Abbott, Eli Lilly and Company. A. J. Larez: Research Support; Self; Abbott, Eli Lilly and Company. W. C. Bevier: Research Support; Self; Abbott Diabetes, Eli Lilly and Company. D. Kerr: Advisory Panel; Self; Novo Nordisk, Sanofi-Aventis, Research Support; Self; Abbott Diabetes, Elsevier, Lilly Diabetes, Stock/Shareholder; Self; Glooko, Inc.

Funding

U.S. Department of Agriculture (2018-33800-28404)
312-OR: Comparison of Processes of Care and Acute Care Utilization for Medicaid Enrollees with Type 2 Diabetes in Federally Qualified Health Centers vs. Other Sites of Primary Care
Authors: KNITTER ALEXANDRA C. , MURUGESAN MANORADHAN , SAULSBERRY LOREN , WAN WEN , NOCON ROBERT , HUANG ELBERT , BOLTON JOSH , CHIN MARSHALL , LAITEERAPONG NEDA
Published: June 2021

Background: Adults with type 2 diabetes and Medicaid insurance are a highly vulnerable population. Despite the vital role of primary care for diabetes management, few studies have examined how the type of primary care setting affects quality of care for this population, and existing studies have not examined data across the US. Since federally qualified health centers (FQHCs) are the primary care safety net for many Medicaid patients, we compared process of care outcomes and acute care utilization for non-elderly adult Medicaid enrollees with type 2 diabetes at FQHC vs. non-FQHC primary care settings.

Methods: We analyzed 2012 Medicaid fee-for-service and managed care claims data in all 50 states and DC in order to compare 8 processes of care and acute care utilization for patients receiving care at FQHCs (N=121,977) vs. non-FQHCs (N=700,401). We used propensity score overlap weighting to obtain groups balanced on covariates, then used generalized estimating equation models to compare the two groups.

Results: FQHC patients had 2.6-3.6% lower rates of each measure compared to non-FQHC patients, with the exception of a higher rate of diabetes education (relative risk, 1.09 [95% CI, 1.03-1.16]). Compared to non-FQHC patients, FQHC patients had more overall (incident rate ratio, 1.06 [1.05-1.07]) and diabetes-related emergency department (ED) visits (1.10 [1.08-1.13]), but fewer overall (0.87 [0.86-0.88]) and diabetes-related hospitalizations (0.79 [0.77-0.81]).

Conclusions: Our findings suggest that Medicaid enrollees with type 2 diabetes receiving primary care at FQHCs may have limited access to both preventative and urgent diabetes care. FQHC patients with diabetes may visit EDs more frequently as a result of limited appointment availability or access to specialists at FQHCs. These results underscore the need for increased support and funding for Medicaid patients with type 2 diabetes, especially in FQHCs.

Disclosure

A. C. Knitter: None. M. Murugesan: None. L. Saulsberry: None. W. Wan: None. R. Nocon: None. E. Huang: None. J. Bolton: None. M. Chin: Advisory Panel; Self; Bristol-Myers Squibb Company. N. Laiteerapong: Other Relationship; Self; American Diabetes Association.

Funding

Health Resources and Services Administration (HHSH250201300025I); National Institute of Diabetes and Digestive and Kidney Diseases (P30DK092949)
333-OR: Regulation of Adiponectin Receptor 2 Expression and Function in the Liver
Authors: HADLEY JASON , SCRUGGS DARAH A. , RYU JIYOON , DONG LILY Q.
Published: June 2021

Adiponectin, an adipokine, has been implicated in improving lipid and glucose metabolism and preventing diet-induced inflammation. Adiponectin transduces its effects by binding to its cognate receptors, AdipoR1 and AdipoR2. Compared to AdipoR1, AdipoR2 is specifically and heavily expressed in liver tissues, but its function in regulating Adiponectin-mediated signaling remains controversial. Our study indicates that AdipoR2 expression levels are reduced under feeding conditions as compared to fasting, suggesting that nutrition status regulates AdipoR2 expression. We also found that the levels of AdipoR2 in liver tissue are significantly less in mice fed with high fat diets compared to mice on chow diets, indicating down-regulation of AdipoR2 expression is associated with overnutrition-induced insulin resistance. Interestingly, we found that AdipoR2 undergoes protein palmitoylation, a diet-induced post-translational modification important in regulating protein trafficking and stability. By checking potential palmitoylation sites in AdipoR2, we identified several cysteine residues near the N-terminus that could be potential sites of palmitoylation. Site-direct mutagenesis studies indicated that C11A mutant, which mimics de-palmitoylation status at this residue, was more stable and had less plasma membrane located compared to the wild type receptor. Together, our results suggest that diet-induced palmitoylation of AdipoR2 is one of the mechanisms underlying high fat diet-triggered down-regulation of AdipoR2. This study provides critical information for understanding regulation of adiponectin signaling in liver and the potential theraputical values for treating insulin resistance and type 2 diabetes.

Disclosure

J. Hadley: None. D. A. Scruggs: None. J. Ryu: None. L. Q. Dong: None.
35-OR: Epigenetic Regulation of White Adipose Tissue Browning by HMGN Proteins
Authors: NANDURI RAVIIKANTH , FURUSAWA TAKASHI , LOBANOV ALEXEI , GAVRILOVA OKSANA , BUSTIN MICHAEL
Published: June 2021

Chromatin architecture proteins HMGNs (High Mobility Group Nucleosome-binding) affect local and global chromatin structure and colocalize with cell-specific regulatory sites and regulate cell identity. The role of HMGN proteins in adipocyte function and energy metabolism is yet to be investigated. Here we report that HMGN1 and HMGN2 double knock-out (DKO) mice on chow diet exhibited decreased weight gain (10%) and total activity with increased food intake and energy expenditure compared to control mice. DKO mice on a high-fat diet also displayed increased food intake and decreased weight gain. We have observed that DKO mice possess less fat mass, which is correlated with decreased WAT size, but not BAT. Histological analysis of WAT from mice revealed smaller adipocytes and increased numbers of cells in DKO, while immunofluorescence analysis indicated more UCP1 in DKO, suggestive of increased WAT browning in DKO mice. Investigations on white preadipocytes or MEF cells differentiation by adipogenic cocktail revealed faster differentiation and elevated UCP1 in DKO cells. Time course RNA seq analysis on in-vitro differentiated adipocytes also confirmed enhanced browning in DKO cells. We further investigated the distribution of active histone marks such as H3K9 acetylation and H3K27 acetylation in adipocytes differentiated from preadipocytes. Our time-course ChIP seq analyses indicated that HMGN’s absence altered both H3K9 and H3K27 acetylation patterns in DKO cells compared to WT. Interestingly, correlation analysis of ChIP seq and RNA seq suggests that DKO preadipocytes and adipocytes lost active chromatin marks at certain white adipocyte-specific enhancers.

In summary, loss of HMGN attributes thermogenic character to WAT in DKO mice, thereby contributing to increased energy expenditure, which might protect mice from metabolic disorders—increased white adipocyte browning in DKO is partly by reducing the active histone marks at white specific enhancers, thereby altering white adipocyte identity.

Disclosure

R. Nanduri: None. T. Furusawa: None. A. Lobanov: None. O. Gavrilova: None. M. Bustin: None.

Funding

National Cancer Institute
353-P: Safety and Tolerability of Sodium-Glucose Cotransporter 2 Inhibitors during Ramadan Fasting
Authors: EID ELSAYED M.A. , HUSSEIN ABDELAZIZ , GHAZI HOSSAM A.
Published: June 2021

Background: Diabetic management during Ramadan fasting is a significant clinical challenge. Sodium glucose co-transporter -2 (SGLT-2) inhibitors are new class of antidiabetic medications known for low frequency of associated hypoglycemia. The present study aimed to evaluate efficacy, safety and tolerability of SGLT-2 inhibitors in diabetic patients practicing Ramadan fasting.

Patients and Methods: The study included 94 patients. They comprised 51 patients who received metformin and sulfonylureas: Glimepiride (1mg-6mg/d) or Gliclazide MR (60-120mg/d) and 43 patients who received SGLT2 inhibitors: Empagliflozin (25 mg), Dapagliflozin (10 mg) or Canagliflozin (300 mg). The study outcome parameters were frequency of hypoglycemia episodes, volume depletion episodes, number of days with early breaking of fasting and missed fasting days.

Results: It was found that patients in SGLTI group experiences significantly fewer symptomatic (9.3 % versus 35.3 %, p=0.003) and documented (7.0 % versus 25.5 %, p=0.017) hypoglycemic episodes as compared to the SU group. However, there were no significant differences between the studied groups regarding the frequency of patients with volume depletion episodes (5.9 % versus 16.3 %, p= 0.1). Moreover, there were no significant differences between groups regarding the frequency of patients with early breaking of fasting (11.8 % versus 9.3 %, p=0.7) or missed fasting (3.9 % versus 2.3 %, p=0.66). None of the studied patients discontinued the prescribed medications.

Conclusions: SGLT-2 inhibitors combined with metformin for diabetic patients during Ramadan fasting are effective, safe and well-tolerated with the advantage of reduced hypoglycemic events.

Disclosure

E. M. A. Eid: None. A. Hussein: None. H. A. Ghazi: None.
355-P: Decreased BCAAs and Elevated Fatty Acids during Repeated Episodes of Hypoglycemia in Individuals with Type 1 Diabetes
Authors: SHE RUI , AL-SARI NABA , SEJLING ANNE-SOPHIE , MATTILA ISMO , HENRIKSEN PETER , PEDERSEN JENS , LEGIDO-QUIGLEY CRISTINA , PEDERSEN-BJERGAARD ULRIK
Published: June 2021

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.
382-P: Thyroid Hormone Is Associated with Arterial Stiffness in Chinese Patients with Type 2 Diabetes Mellitus
Authors: CHEN YAO , HONG ZHAI , MA BINGWEI , WANG XINGCHUN , QU SHEN
Published: June 2021

Objectives: Thyroid hormone has effects on cardiovascular system other than metabolism. This study aimed to investigate the association between thyroid hormone and Arterial stiffness (AS) in type 2 diabetes mellitus (T2DM).

Methods: 1064 T2DM patients were included in the cross-sectional study. They were divided into two groups: AS group (baPWV≥1550cm/s) and Non-AS group (baPWV<1550cm/s) or PAD group (ABI≤0.9)or Non-PAD group(ABI>0.9).

Results: 1) FT3, FT4, TT3 and TT4 were comparable (all P>0.05) while TSH were significantly higher in PAD group than Non-PAD group(2.19±1.86μIU/mL vs. 1.85±1.95μIU/mL, P=0.031). FT3 was significantly lower while TSH was significantly higher in AS group than Non-AS group (4.52±0.74 pmol/L vs.4.72±1.45pmol/L, P=0.007; 2.28±2.03μIU/mL vs.1.95±1.38μIU/mL, P=0.009). FT4, TT3 and TT4 had no difference between the two groups. 2) ABI was significantly positively associated with TT3(r=0.091, P=0.006) while baPWV was significantly negatively associated with TT3 and FT3(r=-0.065, P=0.049; r=-0.149, P<0.001). Multiple linear regression showed that FT3 were the negatively influencing factor of baPWV (β=-75.24, P<0.001). 3) baPWV was significantly higher in patients with subclinical hypothyroidism than without subclinical hypothyroidism (1831.11±347.39 cm/s vs.1729.32±377.12, P=0.027). 4) Logistic regression showed that FT3 was protective factor of AS assessed by baPWV (OR:0.708, 95% CI: 0.506-0.990, P=0.044).

Conclusions: T2DM patients with Arterial stiffness defined by ABI or baPWV have higher TSH and lower FT3 while baPWV was higher in patients with subclinical hypothyroidism. FT3 was negatively influencing factor of baPWV and protective factor of AS assessed by baPWV.

Disclosure

Y. Chen: None. Z. Hong: None. B. Ma: None. X. Wang: None. S. Qu: None.

Funding

National Natural Science Foundation of China (81970677)
385-P: Exosomal MiR-146a-5p from Macrophages Inhibits Diabetic Nephropathy Fibrosis by Targeting Smad4
Authors: ZHANG WENNING , JIANG HONGWEI , MA YUJIN , PENG HUIFANG , LIU JING , YUAN JINGYA , HU KEYAN , ZHAO BING , WU HONGLI
Published: June 2021

Diabetic kidney disease(DKD) has become the leading cause of end-stage renal disease and the main cause of death of chronic kidney disease. Renal interstitial fibrosis is an important pathological change of DKD, and chronic microinflammation also plays a key role in the occurrence and development of Diabetic Nephropathy(DN). Macrophages are one of the main regulatory cells of inflammation and infiltrate in the kidney tissue of early DN. In addition to producing injury mediators, activated macrophages can also affect the kidney through other mechanisms. Interestingly, there are a large amount of exosomes generated by macrophage under hyperglycemia condition and it is not clear whether the exosomes has any effect on glomerular mesangial cell fibrosis in patients with diabetes. Here, we use a diabetic nephropathy db/db mouse model to determine the effect of macrophage-produced exosomes by miR-146a-5p overexpression or down-regulation on mesangial cell fibrosis. with high glucose, Macrophage proliferation is promoted in mice and miR-146a-5p expression is downregulated in both macrophages and glomerular mesangial cells. we confirmed that Smad4 is the target gene of miR-146a-5p through Luciferase reporter gene system. Fibrosis-associated col I, α-SMA, and Smad4 are upregulated in glomerular mesangial cells. In exosomes, we observed miR-146a-5p is downregulated in high glucose than low glucose. We use the exosomes from the macrophages that are transfected with miR-146a-5p mimics or inhibitor to culture the glomerular mesangial cells. Exosomes from miR-146a-5p mimics significantly downregulated glomerular Mesangial cell fibrosis-related factor α-SMA, col I protein, and Smad4. Exosomes from miR-146a-5p inhibition upregulated these factors. Finally, we found macrophage exosomes from miR-146a-5p overexpression can inhibit glomerular mesangial cell fibrosis, which could provide a potential treatment strategy for DKD.

Disclosure

W. Zhang: None. H. Jiang: None. Y. Ma: None. H. Peng: None. J. Liu: None. J. Yuan: None. K. Hu: None. B. Zhao: None. H. Wu: None.
386-P: Chronic Treatment with the Direct AMPK Activator PXL770 Improves Cardiac, Vascular, and Renal Function in Rodents with Diabetes-Related Cardiorenal Disease
Authors: STEPHAN YOHAN , SOULIÉ MATTHIEU , NICOL LIONEL , GLUAIS-DAGORN PASCALE , HALLAKOU-BOZEC SOPHIE , MULDER PAUL
Published: June 2021

Introduction: PXL770 is a direct AMP kinase activator. In models of T2DM and NASH, PXL770 improves hyperglycemia and dyslipidemia and reduces: steatosis, inflammation, fibrogenesis. Whether these beneficial effects are associated with an improvement in diabetes-related cardiac, vascular and renal dysfunctions is unknown.

Methods: Diabetic ZSF-1 rats were treated for 90 days with PXL770 (150 mg/kg BID), then cardiac function (left ventricular (LV) hemodynamics), in vitro flow-mediated dilation (FMD; arteriograph) of mesenteric artery, renal histology and function (transcutaneous GFR) were assessed.

Results: Untreated ZSF-1 rats exhibited LV diastolic dysfunction, i.e., increases in LV End-Diastolic Pressure (LVEDP; 8.3±1.0 vs. 5.6±0.6 mm Hg in lean, p<.05) and LV End-Diastolic Pressure Volume Relation (LVEDPVR; 5.4±0.6 vs. 1.1±0.2 mm Hg/RVU in lean, p<.05). Myocardial tissue perfusion was reduced (4.0±0.2 vs. 7.4±0.9 ml/min/g in lean, p<.05). Untreated rats exhibited impaired mesenteric FMD (% of preconstriction; 37±11 vs. 70±7% in lean, p<.05) and renal function and structure as shown by a reduction in GFR (4.4±0.5 vs. 5.6±0.5 ml/min in lean, p<.05) and increase in glomerular score (2.3±0.2 vs. 0.6±0.1 AU in lean, p<.05). Compared to untreated ZSF-1 rats, 90-day PXL770 reduced glycaemia (-31 %, p<.05) and plasma triglycerides (-44 %, p<.05). PXL770 decreased LVEDP (-28 %) and LVEDPVR (2.7±0.2 mm Hg/RVU, p<.05) and increased LV tissue perfusion (+30 %, p<.05). PXL770 improved mesenteric FMD (64±3%, p<.05) and increased GFR (6.0±0.1 ml/min, p<.05) and reduced glomerular score (1.6±0.2, p<.05).

Conclusion: In ZSF-1 rats, improvement of metabolic status induced by long-term treatment with the AMP kinase activator PXL770 is associated with an improved LV diastolic, vascular and renal function/structure. PXL770 could have therapeutic utility for the treatment of metabolic diseases and related co-morbid conditions.

Disclosure

Y. Stephan: None. M. Soulié: None. L. Nicol: None. P. Gluais-dagorn: Employee; Self; Poxel SA. S. Hallakou-bozec: None. P. Mulder: None.

Funding

Poxel SA
387-P: Protective Impact and Potential Mechanisms of Elabela on DKD via ß-Arrestins
Authors: SHI MIN , CHEN YANNA , ZHANG HONG
Published: June 2021

Objective: ELABELA (ELA), an endogenous ligand for apelin receptor (APJ), is decreased in plasma of DKD patients. The effects of ELA on relieving diabetic renal lesion have not been reported. This study was performed to explore the protective impact and potential mechanisms of ELA on DKD via β-arrestins.

Methods: Sixteen eight-week-old male db/db mice were randomly divided into diabetic nephropathy group (db/db group) and intervention group (db/db+ELA group) with administration of ELA (5mg ∙ kg-1 ∙ day-1) for 8 weeks. 8 age-matched male db/m mice were recognized as normal control group (db/m group). Both db/db and db/m group received equivalent normal saline injection for 8 weeks. Body weights and blood glucose levels were measured once a week. After 8 weeks, blood and urine samples were collected. Serum creatinine and urinary albumin to creatinine ration (UACR) were examined. The kidneys were taken after sacrificing mice. The morphology change of kidney tissues were assessed by HE and PAS staining. The expression of β-Arrestin1/2 was detected by immunohistochemistry. The levels of fibronectin (FN), collagen type IV (Col-IV) and β-arrestin1/2 in kidney tissues were examined by western blotting.

Results: (1) With the intervention of ELA, UACR, serum creatinine and the ratio of kidney weight to body weight of db/db mice were significantly decreased (P<0.05). (2) Compared with db/m mice, db/db mice exhibited renal tubular epithelial cells edema, decreased ratio of non-lumen area to total area, thickened basement membrane and increased glycogen accumulation, however, those pathological abnormality were mitigated by ELA. (3) The levels of FN, Col-IV and β-Arrestin-1/2 in db/db mice were higher than db/m mice, while ELA treatment suppressed the expression of FN, COL-IV and β-Arrestin-1/2 (P<0.05).

Conclusions: ELA may play a role in protection from diabetic kidney damage via down-regulating the expression of β-Arrestin-1/2 and inhibiting the secretion of extracellular matrix.

Disclosure

M. Shi: None. Y. Chen: None. H. Zhang: None.

Funding

National Natural Science Foundation of China (81700723)
40-OR: Effect of Naturally Occurring Genetic Variations in Incretin Receptors on Glucose Homeostasis
Authors: GAUR ANIRUDH
Published: June 2021

Type 2 diabetes is characterized by disruption of glucose homeostasis. Insulin is the primary driver of glucose homeostasis and released by pancreatic β cells in response to nutritional and endocrine signals. Insulin release is also stimulated by incretin hormones including Glucagon-Like peptide 1 (GLP-1) which acts on the GLP-1 receptor (GLP1-R) in islet producing the “Incretin effect.” Common genetic variations in GLP-1R alter this incretin effect. Specifically, the R131Q variant increases while the G168S variant decreases the effect of GLP-1. We have found that these genetic variations do not affect the affinity of GLP-1 nor its efficacy for G protein signaling. Taken together, these findings present a “pharmacological mystery” as to how genetic variation at GLP-1R affects incretin function. In this study, we assessed whether genetic variation in GLP-1R alters receptor trafficking resulting in differential incretin response. HEK293 cells stably expressing wild type (WT), R131Q, and G168S variants were used to measure receptor endocytosis and post-endocytic sorting to the lysosome. We found that WT GLP-1R degrades after endocytosis via its interaction with the GPCR-associated sorting protein-1 (GASP1), a protein highly expressed in islets. Importantly, we found that GLP-1R variant R131Q recycled following endocytosis while variant G168S was more rapidly degraded compared to the WT GLP-1R. To assess whether GASP1 mediated GLP-1R degradation affects incretin function, we used CRISPR to generate rat INS-1 cells with a knockout (KO) of GASP1. Deletion of GASP1 had no effect on acute GLP-1R signaling. However, prolong agonist pretreatment produced a loss of the incretin response in WT INS-1 cells, while GASP1 KO cells showed preservation of the incretin effect. These results suggest GASP1 mediates the post-endocytic degradation of GLP-1R in β cells and that altered trafficking of the GLP-1R variants may underlie differential incretin response in humans who carry these variants.

Disclosure

A. Gaur: None.

Funding

University of California, Davis
408-P: Improving Knowledge and Confidence of Diabetologists and Endocrinologists Related to MRAs for CKD in T2D with Curriculum-Based Online Education
Authors: LARKIN AMY , HANLEY KELLY L. , LE ANNE
Published: June 2021

The goal of continuing medical education (CME) is professional growth and improved patient care. We sought to determine if a curriculum of online continuing medical education (CME) activities could improve the clinical knowledge and confidence of diabetologists and endocrinologists (D/Es) related to use of mineralocorticoid receptor antagonists (MRAs) in the management of chronic kidney disease (CKD) in patients with type 2 diabetes (T2D). The online CME curriculum consisted of 5 online activities. Of these, 4 used repeated pairs pre-/post-assessment study design was used and McNemar’s test (P <.05 is considered significant) to assess educational effect. The activities launched in 2020 and data were collected for up to 12 weeks. The education reached over 19,000 physicians, including 6,000 D/Es.

Overall, knowledge improved by 9% among D/Es (N=1,111, P<.001) Specific improvements: • 8% relative increase in knowledge related to the role of MRAs in treatment of CKD in T2D (P<.01) • 16% relative increase in knowledge related to the mechanism of action (MOA) of MRAs for CKD in T2D (P<.01) • 9% relative increase in knowledge related to identifying CKD in patients with T2D (P<.01)

Of the D/Es who were included, 34% had a measurable increase in confidence, and among those the average level of confidence increased 50%.

While this curriculum demonstrated success in educating on this topic, some gaps still remain after education. On average, 22% of the questions assessing knowledge on the MOA of MRAs in managing CKD in T2D and 29% of the questions assessing knowledge related to the role of MRAs in the management of CKD in T2D were still incorrectly answered after education, demonstrating a need for more education in these areas.

Disclosure

A. Larkin: None. K. L. Hanley: None. A. Le: None.

Funding

Bayer AG
409-P: Arteriolar Hyalinosis Is Associated with the Rate of eGFR Decline and Predicts the Onset of Macroalbuminuria and eGFR<60 Ml/min/1.73 M2 in Patients with Type 2 Diabetes and Normo-microalbuminuria Regardless of Microalbuminuria Grade
Authors: MORIYA TATSUMI , HAYASHI AKINORI , SUZUKI AKIHIKO , MATSUBARA MADOKA , OUCHI MOTOSHI
Published: June 2021

Microalbuminuria is thought to be a risk factor for progression to macroalbuminuria, an indicator of renal functional decline in patients with type 1 or 2 diabetes. However, it remains unknown which renal histological factors predict the onset of macroalbuminuria or eGFR<60 mL/min/1.73 m2 (chronic kidney disease, CKD) during early diabetic nephropathy. Therefore, we sought to determine which histological factors, including index of arteriolar hyalinosis (IAH), are predictors for onset of macroalbuminuria and CKD. The study population consisted of 35 patients with type 2 diabetes (25 men; age, 47±9 years; eGFR, 92.7±18.0 mL/min/1.73 m2) with normoalbuminuria or microalbuminuria who underwent percutaneous renal biopsy. We followed these patients for at least 5 (18±6) years. The study endpoint was the onset of macroalbuminuria or CKD, or by March 2020. Light and electron microscopy-based morphometric analyses were performed to quantitatively evaluate glomerular and interstitial structural changes. During the observation period, 9 of 35 patients progressed to macroalbuminuria and 15 had CKD. The annual rate of eGFR decline was significantly related to IAH (r=-0.40, p=0.016), but not any other histological parameters. Kaplan-Meier analysis revealed that arteriolar hyalinosis and mesangial expansion are associated with a significantly higher risk of onset of macroalbuminuria and CKD, and microalbuminuria grade is associated with the onset of macroalbuminuria, but not the onset of CKD.

Conclusions: Aggravated arteriolar hyalinosis is a histological risk factor for renal functional decline and predicts the onset of macroalbuminuria and CKD in patients with type 2 diabetes, normo-microalbuminuria, and normal renal function regardless of microalbuminuria grade. This finding might reflect initial changes in early diabetic nephropathy.

Disclosure

T. Moriya: None. A. Hayashi: None. A. Suzuki: None. M. Matsubara: None. M. Ouchi: None.
41-LB: Changes in Circulating Metabolites, Including Butyrate, Points to Underlying Mechanism of a Probiotic Intervention That Improves Postprandial Hyperglycemia in Subjects with Type 2 Diabetes
Authors: MCMURDIE PAUL , STOEVA MAGDALENA K. , NEMCHEK MADELEINE , TYAGI SURABHI , GINES JESSICA , EID JOHN , KOLTERMAN ORVILLE G.
Published: June 2021

Studies in type 2 diabetes (T2D) rodent models suggest the potential for individual microbial taxa in the human gut to improve or worsen metabolic disease via specific metabolites (signals) in hepatic or general circulation. We previously reported significant improvement in postprandial glucose control in T2D subjects following ingestion of a 5-strain novel probiotic formulation (INT) in a placebo (PBO) controlled setting. (DOI:10.1136/bmjdrc-2020-001319) Here we report measurements of fecal and fasting circulating plasma metabolites, implicated as mediators of microbiome effects, collected before and after the 12-week intervention. Consistent with the hypothesized benefits of increased colonic production of butyrate, plasma butyrate increased in INT but not PBO (within group, concentration increase = 0.15 µM, p = 0.007; between group, p = 0.11), but not acetate nor propionate, paralleling changes in subject stool and production by in vitro culture. We detected changes consistent with enhanced fatty acid oxidation in INT relative to placebo, especially a decrease in medium chain fatty acids, their carnitinylates, and a decrease in bilirubin degradation byproducts. We also detected increases in certain bile acids, especially ursodeoxycholic acid (UDCA). In vitro monoculture supernatant showed that the formulation’s C. butyricum strain specifically and efficiently synthesizes UDCA during butyrogenic growth in rich media amended with the primary bile acid chenodeoxycholic acid. Untargeted data also uncovered previously undocumented use of sulfonylurea (SFU), accounting for 6 out of 7 subjects with A1c non-response in the INT group. To our knowledge, this is the first description of an increase in circulating butyrate and UDCA due solely to a bacterial probiotic intervention in humans with T2D, and may help explain the observed improvement in postprandial glucose control.

Disclosure

P. Mcmurdie: Employee; Self; Pendulum Therapeutics. M. K. Stoeva: Employee; Self; Pendulum Therapeutics. M. Nemchek: Employee; Self; Pendulum Therapeutics. S. Tyagi: None. J. Gines: Employee; Self; Pendulum Therapeutics. J. Eid: Stock/Shareholder; Self; Pendulum Therapeutics. O. G. Kolterman: Board Member; Self; GlySens Incorporated, Consultant; Self; ADOCIA, Cirius Therapeutics, Nanoprecision Medical, Viacyte, Inc., Employee; Self; Pendulum Therapeutics.
41-OR: Roles of Transcription Factor EB (TFEB) in ß-Cell Function and Survival Under Stress
Authors: ZOU YUANJIE , PASULA DANIEL , VERCHERE BRUCE , LUCIANI DAN S.
Published: June 2021

Autophagy can help protect stressed β-cells. We investigated if disruption of Transcription Factor EB (Tfeb), a transcriptional master-regulator of lysosomes and autophagy, contributes to β-cell failure and death under various ER stress-inducing conditions.

Short-term β-cell autophagy-deficiency (inducible Atg5 knockout) reduced β-cell survival under hypoxic stress in vitro and impaired the ability of syngeneic marginal mass islet grafts to maintain glucose homeostasis in streptozotocin (STZ)-induced diabetic mice, with >80% of recipients of Atg5-deficient islets returning to diabetes within 10 days post-transplant (vs. 30% of controls, p=0.05). Hypoxic culture activated pro-apoptotic ER stress pathways, and confocal analysis of islet cells from LC3-RFP-EGFP autophagy reporter mice demonstrated that autophagic flux was interrupted. Hypoxia-induced impairment of autophagy was associated with reduced lysosomal cathepsin B activity and a significant loss of Tfeb, both of which could be partially rescued by Torin-1. Lipotoxicity and thapsigargin-induced ER-stress similarly reduced Tfeb protein and cell viability, suggesting ER stress-specific mechanisms may drive dysregulation of Tfeb and lysosomes. Conversely, overexpression of Tfeb:GFP reduced thapsigargin-induced death in MIN6 cells. To directly study the consequences of Tfeb loss we generated mice with Ins1Cre-driven, β-cell-specific, Tfeb deletion (TfebβKO). Notably, both male and female TfebβKO mice had normal glucose tolerance, but cultured Tfeb KO β-cells showed impaired lysosomal cathepsin B activity and TfebβKO mice had worsened glucose tolerance compared to Tfeb flox littermates following injection with multiple low-doses STZ.

In summary, autophagy protects β-cells under hypoxia but is impaired under prolonged hypoxic stress, possibly through Tfeb dysregulation. Our data further suggest loss of Tfeb is a common feature of ER stress and that this loss exacerbates stress-induced β-cell failure and death.

Disclosure

Y. Zou: None. D. Pasula: None. B. Verchere: Advisory Panel; Self; Integrated Nanotherapeutics Inc., Sirona Biochem, Stock/Shareholder; Self; Integrated Nanotherapeutics Inc. D. S. Luciani: None.

Funding

JDRF (2-2013-50)