bluuebunny/crossref_metadata_2025 huggingface dataset:
1705-P: Expression of Selected MicroRNAs in Adipose Tissue-Derived Mesenchymal Stem Cells and Metabolic Syndrome
Authors: WYSTRYCHOWSKI GRZEGORZ , SIMKA-LAMPA KLAUDIA , WITKOWSKA AGNIESZKA , SOBECKO EWELINA , SKUBIS-SIKORA ALEKSANDRA , SIKORA BARTOSZ , WOJTYNA EWA , GWIZDEK KATARZYNA , GOLDA AGNIESZKA , WRÓBEL MARTA , KRUSZNIEWSKA-RAJS CELINA , GOLA JOANNA
Published: June 2023

Adipose tissue mesenchymal stem cells (AdMSC) secrete microRNAs (miR). Although miR are involved in pathogenesis of metabolic syndrome, the role of those secreted by AdMSC remains elusive.

We aimed to assess expression of selected miR by AdMSC in relation to basic metabolism parameters in nondiabetic adults.

For this preliminary study 10 obese, 8 overweight and 5 normal weight subjects with no inflammatory or cancerous disease were enrolled, including 19 female and 6 prediabetic ones. AdMSC were harvested from abdominal subcutaneous fat and cultured in standard conditions. Expressions of four miR reported in metabolic disturbances were estimated with RT-qPCR. Total and visceral fat were assessed with bioimpedance.

Expression of miR-21, miR-122, or miR-192 did not correlate with any clinical parameter. On the other hand, expression of miR-155 [2-ΔCt] was lowest in obese subjects (3.69±2.67 vs. 7.07±4.42 in overweight and 10.25±7.05 ×10-3 in normal weight ones, P =0.04, K-W ANOVA). It correlated inversely with BMI, total fat mass, visceral adiposity, and serum CRP, whereas positively (marginally significantly) with serum HDL-cholesterol (table 1).

Expression of miR-155 in AdMSC appears blunted in visceral obesity, which correlates with HDL-dyslipidemia and microinflammation. This calls for further study of AdMSC miR secretome in the pathogenesis of metabolic syndrome with focus on miR-155.

Disclosure

G. Wystrychowski: Other Relationship; Polfa Tarchomin. K. Simka-Lampa: None. A. Witkowska: None. E. Sobecko: None. A. Skubis-Sikora: None. B. Sikora: None. E. Wojtyna: None. K. Gwizdek: None. A. Golda: Other Relationship; Servier Laboratories. M. Wróbel: Other Relationship; Novo Nordisk, Lilly, Boehringer Ingelheim Inc. C. Kruszniewska-Rajs: None. J. Gola: None.

Funding

Medical University of Silesia, Katowice, Poland
1706-P: Obesity-Related Cancer Increased Sharply in Young Adults among Chinese Population from 2007 to 2021
Authors: YANG JIN-KUI , LIU CHANG , YUAN YINGCHAO
Published: June 2023

Objective: The burden of obesity-related cancer has rarely been investigated. This study aims to investigate the trends of incidence of obesity-related cancer in a Chinese population.

Method: During 2007-2021, a population-based registry of cancer covering 14.14 million people was performed in 207 medical sites by the Beijing Municipal Health Commission Information Center. We compared the trends of incidence of obesity-related cancers and non-obesity-related cancer.

Results: Overall, there were 651,482 incident cancer patients in this population from 2007 to 2021. Of these patients, 48.49% were obesity-related cancers. For all obesity-related cancers combined, the age-standardized incidence rates (ASR) showed a significant upward trend (3.6% per year, P< 0.001). Conversely, for non-obesity-related cancers, the ASR was stable during this period (Fig.1 Age-specific differences of all cancers: A,B: annual changes; C,D: incidence rate ratio; E,F: incidence). We predict that the ASR for obesity-related cancers will increase 2.62-fold in 2035 compared to 2021, and will increase mainly in people younger than 50 and decrease in those over 75 years.

Conclusion: Obesity-related cancer incidence significantly increased from 2007 to 2021. While most cases were diagnosed in old people, the obesity-related incidence increased sharply in young adults. These will increase the burden of cancer in the future.

Disclosure

J.Yang: None. C.Liu: None. Y.Yuan: None.

Funding

National Natural Science Foundation of China (81930019)
1708-P: Comparing 9- vs. 12-Month Implantation in the Worldwide EndoBarrier (EB) Registry
Authors: RYDER ROBERT E. , YADAGIRI MAHENDER , MUNRO LYNNE , FRYDENBERG HARRY , FISHMAN SIGAL , BYRNE JAMES P. , TEARE JULIAN P. , DE JONGE CHARLOTTE , WILLEM GREVE JAN , MCMASTER JESSICA J. , CHISHOLM JACOB , KOW LILIAN , MASON JOHN C. , COHEN RICARDO V. , SEN GUPTA PIYA
Published: June 2023

Uncertainty exists re risk: benefit of proximal intestinal exclusion with EB, a novel endoscopic duodenal jejunal liner device for obesity, both with and without diabetes. In view of this, during 2017, an independent, secure, on line registry was established under the auspices of the Association of British Clinical Diabetologists, for the collection of safety and efficacy data worldwide. As of December 2022, data had been entered on 1022 patients, of whom 195 (age 51.6±10.3 years, 53% male, 81% white ethnicity, BMI 39.9±6.9kg/m2) had both 9- and 12-month data entered. EB had considerable impact on weight and HbA1c (Table). There was no difference between the mean±SD reduction in HbA1c or weight at 9- vs 12-months (HbA1c: 1.39±1.67% vs 1.38±1.66% (p=0.98); weight: 11.7±8.3kg vs 12.0±8.3kg (p=0.44). The higher the HbA1c the greater the fall but again no difference between 9- and 12-months (Table). In the full registry, 43/1022 (4.2%) experienced serious adverse events (SAE). 15/43 (34.9%) SAE would have been avoided by removal at 9-months (9 liver abscess, 4 GI bleed, one cholecystitis). It was particularly noteworthy that 9/13 (69.2%) liver abscess SAEs would have been avoided by removal at 9-months. This international data from the EB registry suggests that the benefits of EB are achieved in 9-months and a reduction in the recommended implantation period from 12- to 9-months would reduce SAEs, especially the liver abscess SAE.

Disclosure

R.E.Ryder: Other Relationship; Novo Nordisk, Speaker's Bureau; GI Dynamics, BioQuest. J.J.Mcmaster: None. J.Chisholm: None. L.Kow: None. J.C.Mason: None. R.V.Cohen: Advisory Panel; GI Dynamics, Research Support; Medtronic, Johnson & Johnson Medical Devices Companies. P.Sen gupta: Other Relationship; Novo Nordisk. M.Yadagiri: None. L.Munro: None. H.Frydenberg: None. S.Fishman: None. J.P.Byrne: Advisory Panel; Oxford Medical Products, Join Embla, Stock/Shareholder; Join Embla. J.P.Teare: None. C.De jonge: None. J.Greve: Advisory Panel; GI Dynamics.

Funding

Association of British Clinical Diabetologists
1710-P: Uncovering Germline Copy Number Deletions Associated with Obesity and Lipid Traits by UK10K Whole-Genome Analysis
Authors: HUN OHN JUNG
Published: June 2023

We investigated the association of common high-confidence germline copy number deletions (CNDs) with metabolic traits by analyzing UK10K consortium data, where whole genome sequencing (WGS) was performed on 3,781 participants in two UK cohorts, TwinsUK (n=1,854) and ALSPAC (n=1,927). TwinsUK enrolled adult (mean age=59) females and ALSPAC followed children from birth to adolescence. BMI, total fat mass (TFM), truncal fat mass (TrFM), HDL, LDL, VLDL, glucose and insulin were measured. After excluding rare CNDs (MAF<5%) 1,222 (TwinsUK) and 1,211 (ALSPAC) common CNDs remained for association analyses. The genome-wide significance threshold was set at 4.09×10-5(TwinsUK) and 4.13×10-5(ALSPAC) after Bonferroni correction at level α=0.05 and five CNDs achieved genome-wide significance. At ALSPAC, a common (MAF=0.11) 7kb deletion on chromosome 11 (55031591-55038554;BMI at 15yrs,β=1.49, P=2.75×10-6;TFM at 9yrs,β=0.96, P=2.63×10-6;TrFM at 9yrs,β=0.45, P=2.15×10-6) with loss of exons from 2 to 6 in TRIM48 (Tripartite Motif Containing 48), E3 ubiquitin-protein ligase, was associated with childhood obesity. Two other loci at chromosome 8 (intergenic, 24972435-24990944;BMI,β=0.93, P=1.78×10-5;TFM,β=1.39, P=3.58×10-6; TrFM,β=0.64, P=4.56×10-6) and 11 (intron 2 deletion of lincRNA, LOC44040, 49759283-49760872;BMI,β=1.03, P=1.72×10-6; TFM,β=1.29, P=1.31×10-5; TrFM,β=0.60, P=1.66×10-5) were associated with childhood obesity. At TwinsUK, a common (MAF=0.29) 823bp deletion in intron 3 of lncRNA, LINC01923, on chromosome 18 (74347187-74348010,β=-0.22, P=4.15×10-7) was associated with low HDL. The genome-wide significant association of 3kb deletion in GUSBP1 (GUSB Pseudogene 1) enhancer on chromosome 5 (21611834-21614796) with childhood obesity (TFM at 9yrs,β=1.26, P=2.52×10-5) in ALSPAC was replicated in adult females of TwinsUK (TFM, P=3.79×10-2). Studies are warranted that investigate germline CNDs associated with metabolic traits.

Disclosure

J.Ohn: None.

Funding

Seoul National University Bundang Hospital Research Fund (18-2018-0006)
1714-P: Higher Initial Diet Quality Predicts Shorter Weight Loss Period prior to an Involuntary Weight Loss Plateau
Authors: MELHORN SUSAN J. , SEWAYBRICKER LETICIA E. , GAO HUI , ROSALYNN DE LEON MARY , WEBB MARY , LYLE MEGHAN , BEATTY SARAH J. , KRATZ MARIO , SCHUR ELLEN
Published: June 2023

It is currently unknown what factor(s) may promote entry into a weight loss plateau. Given intensive lifestyle interventions (ILI) for weight loss include changes in diet, we evaluated how diet quality impacts characteristics of a weight loss plateau. Daily weights were obtained remotely via electronic scale from 62 adults with obesity (73% female, mean age 42±11 y and BMI 37±5 kg/m2) undergoing a 24-week ILI. Periods (≥14 d) of active weight loss or plateau were identified by threshold regression modeling. Active weight loss was defined as a per day % weight change from baseline equivalent to ≥0.5 lb loss/wk and a weight loss plateau as ±0.25lbs/wk after a period of active weight loss (in which ≥3.5% weight loss was achieved). Three unannounced self-reported ASA24 dietary recalls were obtained at baseline and 3 mo. Diet quality was assessed by the healthy eating index (HEI)-2015. 53% reached a plateau after active weight loss (27% did not achieve >3.5% weight loss, 19% re-gained directly after loss). Weight loss (i.e., plateau depth) was associated with longer time to plateau (β=–10d, P<0.01). Higher baseline diet quality (total HEI) was associated with shorter time to plateau (β=–2.6d, P=0.03), but not overall depth of plateau (β=0.1%, P=0.28). Specifically, shorter time to plateau was related to lower baseline consumption of saturated fats (β=–15d, P=0.01) and greater plateau depth was related to lower baseline consumption of added sugars (β=1.3%, P=0.04). Mean diet quality improved minimally during ILI (ΔHEI 1.9±2.2). Lower baseline HEI correlated with greater improvement in diet quality at 3 mo (β=–0.5, P<0.001, N=46), but ΔHEI did not associate with any plateau characteristics (time: β=1.1d, P=0.15; depth: β=–0.1%, P=0.16, N=25). Higher diet quality upon entry into an ILI predicted a shorter duration of weight loss prior to reaching a plateau, suggesting that individuals with healthy eating patterns may derive less sustained weight loss benefit from participation in an ILI.

Disclosure

S.J.Melhorn: None. L.E.Sewaybricker: None. H.Gao: None. M.De leon: None. M.Webb: None. M.Lyle: None. S.J.Beatty: None. M.Kratz: Other Relationship; Nourished by Science LLC. E.Schur: None.

Funding

National Institutes of Health (DK089036, DK1176223, K24HL144917); University of Washington (NORC, DK035816)
1717-P: Weight-Loss Differences with SGLT2 inhibitors, GLP-1 Receptor Agonists, and Repetitive Transcranial Magnetic Stimulation in Type 2 Diabetes and Obesity
Authors: FERRULLI ANNA , MASSARINI STEFANO , MACRÌ CONCETTA , SENESI PAMELA , TERRUZZI ILEANA , CANNAVARO DANIELE , LUZI LIVIO
Published: June 2023

Weight loss in individuals with obesity (O) and type 2 diabetes (T2D) may improve glycemic control, prevent obesity-related comorbidities and, in some cases, induce remission of T2D. Glucose-lowering drugs such as SGLT2i and GLP-1 RAs are able to control simultaneously body weight and glucose levels; their use is recommended in individuals with O and T2D. Recently, we have demonstrated the effectiveness of repetitive Transcranial Magnetic Stimulation (rTMS) in promoting weight loss up to 1 year in individuals with O, with and without T2D. The aim of this retrospective comparative study was to compare the efficacy of SGLT2i, GLP1-RA and rTMS in reducing body weight in the long term in patients with O and T2D. Data obtained from 31 patients with O and T2D were retrospectively analyzed: 11 were treated for O with high frequency rTMS for 5 weeks, 8 subjects were treated for T2D with SGLT2i for 1 year, and 12 individuals were treated for T2D with GLP1-RA for 1 year. Changes in body weight and HbA1c in the 3 groups were analyzed with ANOVA, after 6 months (FU1) and 1 year (FU2) from the beginning of the treatment. The change in body weight between the 3 groups was significant at both FU1 [−3.2±1.8% (SGLT2i) vs −3.5%±1.5% (GLP1-RA) vs −6.3±4.0% (rTMS);p=0.026] and FU2 [−2.1±2.1% (SGLT2i) vs −2.1±4.2% (GLP1-RA) vs −6.7±4.7% (rTMS); p=0.017]. Glycated hemoglobin variation was higher in the SGLT2i and GLP-1 RA groups than in rTMS at FU1 [−11.7±25.1% (SGLT2i) vs −11.9%±13.2% (GLP1-RA) vs −8.9±10.4% (rTMS)] and at FU2 [−15.9±24.2% (SGLT2i) vs −7.8±11.9% (GLP1-RA) vs −5.6±9.0% (rTMS)], although not statistically significant. TMS was found to be more effective than SGLT2i and GLP1-RAs in promoting long-term weight loss in a population with O and T2D, likely due to a prevalent effect of rTMS on appetite control at the meso-cortico-limbic system’s level. These findings set the stage for a potential combined use of rTMS with SGLT2i or GLP1-RAs in treatment of O and T2D.

Disclosure

A.Ferrulli: None. S.Massarini: None. C.Macrì: None. P.Senesi: None. I.Terruzzi: None. D.Cannavaro: None. L.Luzi: Advisory Panel; Eli Lilly and Company, Medtronic, Research Support; Gelesis, Speaker's Bureau; A. Menarini Diagnostics, Amgen Inc., Boehringer Ingelheim and Eli Lilly Alliance, Eli Lilly and Company, Novo Nordisk, Novartis.

Funding

IRCCS MultiMedica (Ricerca Corrente)
1719-P: Nomogram for Predicting Remission of Metabolic Syndrome One Year after Sleeve Gastrectomy Surgery in Chinese Patients with Obesity
Authors: PAN YUNHUI , HAN XIAODONG , TU YINFANG , ZHANG PIN , YU HAOYONG , BAO YUQIAN
Published: June 2023

Background: Sleeve gastrectomy (SG) is a widely used and effective treatment for patients with obesity and comorbid metabolic abnormalities. No specialized tool is available to predict metabolic syndrome (MS) remission after SG. We aimed to present a nomogram that evaluated the probability of MS remission 1 year after SG.

Methods: Patients who underwent SG were enrolled. Of these patients, those with baseline MS were analyzed at the end of follow-up. They were divided into a training set and a validation set. Multivariate logistic regression analysis was performed to identify independent predictors of MS remission 1 year after SG, and these predictors were employed to create a nomogram. Receiver operating characteristic (ROC) curve analysis was used to evaluate discrimination ability. Calibration was performed with the Hosmer-Lemeshow goodness-of-fit test. The net benefits of the nomogram were evaluated using decision curve analysis (DCA).

Results: Three hundred and eighteen patients (94 males and 224 females) with a median age of 34.0 years were analyzed at the end of follow-up. They were divided into a training set and a validation set with 159 individuals each. A combination of age, preoperative high-density lipoprotein cholesterol (HDL-c), presence of elevated triglycerides (TG) and the glycated hemoglobin (HbA1c) level independently and accurately predicted MS remission. The nomogram included all four factors. The model showed moderate discrimination in the training and validation sets (Area under curve 0.800 and 0.727, respectively). The Hosmer-Lemeshow X2 values of the nomogram were 8.477 (P=0.388) for the training set and 5.361 (P=0.718) for the validation set, both indicating good calibration. Moreover, DCA showed substantial clinical benefits of the nomogram in both datasets.

Conclusions: Our simple nomogram could assist in predicting MS remission in Chinese patients with obesity 1 year after SG.

Disclosure

Y.Pan: None. X.Han: None. Y.Tu: None. P.Zhang: None. H.Yu: None. Y.Bao: None.

Funding

Clinical Research Plan of SHDC (2020CR1017B)
1724-P: Caspase 3/7 Activation and RIPK3 Are Dispensable for Synthetic hIAPP-Induced ß-Cell Death
Authors: MUKHERJEE NOYONIKA , CONTRERAS CHRISTOPHER J. , LIN LI , CAI ERICA P. , KAHN STEVEN E. , TEMPLIN ANDREW T.
Published: June 2023

Islet amyloid is a hallmark of type 2 diabetes that is associated with β-cell loss. Aggregation of the β-cell secretory product human islet amyloid polypeptide (hIAPP) to form islet amyloid promotes β-cell death 1) via direct effects on the β cell and 2) by stimulating islet immune cell cytokine production. We recently showed that β cells are susceptible to TNFα-induced necroptosis, a form of caspase-independent, receptor interacting protein kinase 3 (RIPK3)-mediated cell death. We hypothesized that RIPK3 also promotes β-cell death in response to islet amyloid formation. To test this hypothesis, we evaluated INS-1, NIT-1 and isolated mouse islet cells treated with synthetic hIAPP (20 μM) +/− the pan-caspase inhibitor zVAD (40 μM). We found synthetic hIAPP increased cell death and caspase activity in INS-1, NIT-1 and mouse islet cells over 48 h. Remarkably, inhibition of caspases with zVAD (as measured by caspase 3/7 activity) failed to reduce synthetic hIAPP-mediated cell death in INS-1 (n=3, p=0.99), NIT-1 (n=3, p=0.99) or mouse islet cells (n=4, p=0.98). To evaluate RIPK3 in hIAPP-induced β-cell death, we examined a small molecule RIPK3 inhibitor (GSK’872, 5 μM), gene-edited NIT-1 Ripk3Δ cells (gRNA targeting exon 6), and intact isolated Ripk3−/− mouse islets. We found GSK’872 failed to protect INS-1 (n=3, p=0.32) or NIT-1 (n=3, p=0.30) cells from hIAPP-induced cell death over 48 h. Similarly, neither NIT-1 Ripk3Δ cells (n=2, hIAPP: p=0.82, hIAPP+zVAD: p=0.97) nor Ripk3−/− islets (n=4, hIAPP: p=0.63, hIAPP+zVAD: p=0.86) were protected from synthetic hIAPP-induced cell death either when caspases were active or inhibited. Our data indicate that caspase 3/7 activity and RIPK3 are dispensable for synthetic hIAPP-induced β-cell death in vitro. However, we found that RIPK3 is upregulated in response to endogenous islet amyloid formation in vitro. Future studies will determine whether RIPK3 promotes β-cell loss in part via effects on endogenous amyloid-induced cytokine signaling.

Disclosure

N.Mukherjee: None. C.J.Contreras: None. L.Lin: None. E.P.Cai: None. S.E.Kahn: Advisory Panel; Anji Pharmaceuticals, Bayer Inc., Boehringer Ingelheim Inc., Eli Lilly and Company, Merck & Co., Inc., Other Relationship; Novo Nordisk. A.T.Templin: None.

Funding

U.S. Department of Veterans Affairs (IK2BX004659 to A.T.T.); National Institutes of Health (T32DK064466 to C.J.C.); Indiana University School of Medicine (to N.M.)
1727-P: Mitochondrial-Encoded Peptide MOTS-c Prevents Senescence-Induced Diabetes
Authors: KONG BYUNGSOO , HONG SERIN , MIN CHO YOUNG
Published: June 2023

A polymorphism MOTS-c, a mitochondrial-derived miniprotein, was reported to be associated with longevity in humans. Previous reports showed that MOTS-c regulates senescence in cells, enhances physical capacity in old mice and in humans. However, it is still unknown how MOTS-c is able to intervene the cellular senescence, what tissue or cell-type MOTS-c targets, and whether the regulation of aging by MOTS-c can lead to the prevention of aging- or senescence-induced disorders. Here, using S961 (or senescence)-induced mouse model, we show that MOTS-c is capable of targeting pancreas and is able to prevent senescence-induced diabetes. To test how MOTS-c is associated with β cell senescence, we firstly assess whether β cell senescence is associated with MOTS-c expression using pancreatic β cells isolated from 12-week-old mice. After isolation, pancreatic β cells were treated with H2O2 in concentration-dependent manner. Treatment of H2O2 lowered MOTS-c expression in concentration-dependent manner, but increased other senescence-related markers such as β-gal expression. Furthermore, MOTS-c treatment decreased H2O2-induced senescence-associated β-gal expression and SASPs (i.e., IL-1). To understand the inhibitory effect of MOTS-c against β cell senescence in vivo, we have transplanted S961-osmotic pump in 28-week-old mice with or without MOTS-c for 2 weeks. Mice treated with MOTS-c showed lowered random glucose level (310 mg/dL versus 180 mg/dL) and decreased level of pancreatic β-gal expression compared to S961-treated mice. Also, an i.p. glucose tolerance test (IPGTT) in 30-week-old mice revealed that MOTS-c treatment significantly improved glucose clearance. As a result, we showed that senescence/S961-induced diabetes were inhibited by MOTS-c treatment.

Disclosure

B.Kong: None. S.Hong: None. Y.Cho: Consultant; LG Chem.

Funding

Ministry of Health & Welfare, Republic of Korea (HI14C1277)
1733-P: Evaluating the Role of Chd3 Helicase Subunit on ß-Cell Function
Authors: KANOJIA SUKRATI , OSMULSKI MEREDITH E. , DAVIDSON REBECCA K. , SPAETH JASON
Published: June 2023

Evaluating the role of Chd3 helicase subunit on β-cell function

Pancreatic β-cells play a crucial role in maintaining whole-body glucose homeostasis by secreting insulin upon increased blood glucose levels. To develop effective therapies to combat hyperglycemia, it is of vital importance to understand how β-cells function normally and the mechanisms driving their dysfunction in settings of diabetes. Pdx1, an essential transcription factor involved in β-cell development and function, dynamically recruits coregulators to drive gene expression programs. Recently, Pdx1 was shown to interact with Chd4, a helicase of Nucleosome Remodeling and Deacetylase (NuRD) complex within pancreatic β-cells. Chd4 was found to regulate insulin secretion, modulate expression of β-cell functional genes by modifying chromatin accessibility. We discovered that Chd4 removal from mature β-cells (Chd4Δβ) increased Chd3 levels, an alternate Chd subunit of the NuRD complex. This led us to evaluate whether Chd3 alone plays a role in β-cell function and/or does it compensate in absence of Chd4. To investigate this possibility, we generated tamoxifen inducible, β-cell-specific Chd3 (Chd3Δβ) and Chd3/Chd4 double knockout (Chd3/4Δβ) mouse models. Whereas 4-weeks following Chd3 removal, Chd3Δβ mice did not display glucose intolerance, the Chd3/4Δβ mutants were profoundly glucose intolerant with elevated ad libitum fed blood glucose levels and near complete loss of insulin secretion in response to glucose, a phenotype much more severe than Chd4Δβ mutants previously characterized. This data suggests Chd3 plays a pertinent role in the absence of Chd4 within the β-cell. Current efforts are focused on determining the insulin secretion capacity of Chd3/4Δβ islets and evaluating gene expression and chromatin accessibility of Chd3/Chd4Δβ β-cells.

Disclosure

S.Kanojia: None. M.E.Osmulski: None. R.K.Davidson: Employee; Eli Lilly and Company. J.Spaeth: None.

Funding

National Institutes of Health (DK129287)
174-OR: Identification of Gestational Diabetes (GDM) Subtypes via Machine Learning and Associations with Perinatal Complications
Authors: ZHU YEYI , HARVILL RACHEL , GREENBERG MARA , NGO AMANDA , LIAO LAUREN D. , CHEHAB RANA F. , FERRARA ASSIAMIRA
Published: June 2023

There has been increasing recognition of GDM heterogeneity. We aimed to identify GDM subtypes using 45 variables from the electronic health records via Uniform Manifold Approximation and Projection and Density-Based Spatial Clustering of Applications with Noise, and assess associations with risk of perinatal complications via logistic regression. Since risk factors for GDM and complications were used to identify subtypes, association between subtypes and complications were not adjusted. In a cohort study of 37,561 racially and ethnically diverse individuals with GDM in 2011-2021, we split the sample into training/validation (70%/30%) sets. In the training set, 4 clusters (C) were identified. C1 (14.4%) and C2 (7.8%) were more likely to have early GDM diagnosed by Carpenter-Coustan criteria, pre-existing conditions (hypertension, prediabetes, polycystic ovary syndrome, and obesity), and to be older and multiparous, compared to C3 (65.7%) and C4 (12.1%; Table). Compared to C3 as the mildest and most common subtype, C2 and C4 had higher risk of preeclampsia/eclampsia, C2 had higher risk of placental abruption, and C1, C2, and C4 had higher risk of preterm birth, large for gestational age, and neonatal intensive care unit admission. Findings were confirmed in the validation set. Subtyping GDM may inform personalized risk assessment and management to improve perinatal outcomes.

Disclosure

Y.Zhu: None. R.Harvill: None. M.Greenberg: None. A.Ngo: None. L.D.Liao: None. R.F.Chehab: None. A.Ferrara: None.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases (K01DK120807); National Heart, Lung, and Blood Institute (R01HL157666)
1741-P: Placental Insulin Receptor and Insulin-Like Growth Factor 1 Receptor Regulate Offspring Glucose Homeostasis and Insulin Sensitivity
Authors: BEETCH MEGAN , ALEJANDRO EMILYN
Published: June 2023

Maternal hyperinsulinemia, often underlying pregnancies complicated by obesity and gestational diabetes, is associated with impaired placental insulin signaling, yet the consequences of altered placental insulin action on the metabolic health trajectory of the offspring remains unclear. Insulin binds to the insulin receptor (IR) and to the insulin-like growth factor receptor 1 (IGF1R). We reported that IR ablation in placental trophoblast cells mildly improved glucose homeostasis in adult male offspring when metabolically challenged by a high-fat diet. The current study examines metabolic phenotypes of offspring that experienced placental IGF1R knockout (IGF1RKO) or double IGF1R/IR knockout (DKO) in utero. We report that male IGF1RKO offspring have a decrease in body weight at embryonic day 17.5 but no differences in placenta weight. After weaning and throughout adulthood, no body weight differences were observed in male or female offspring. At 16 weeks of normal chow diet (NCD), random blood glucose was decreased in female IGF1RKO offspring but increased in male IGF1RKO offspring compared to respective littermate controls. Interestingly, these opposite changes in glycemia corresponded with reciprocal pancreas weights, prompting our investigation of beta-cell and alpha-cell mass and function. Glucose tolerance tests revealed improved glucose tolerance in IGF1RKO male offspring at 20 weeks NCD. Insulin tolerance tests have demonstrated comparable insulin sensitivity in IGF1RKO offspring of both sexes. In contrast, DKO offspring display reduced body weights at post-natal days 7 and 21. At 12 weeks of NCD, DKO females were significantly more insulin sensitive than littermate controls but had comparable glucose tolerance and body weights across adulthood. DKO males did not display glucose or insulin intolerance. Together, findings from these models suggest a need for further investigation of insulin action during pregnancy.

Disclosure

M.Beetch: None. E.Alejandro: None.

Funding

National Institutes of Health (T32DK007203, T32DK083250)
1745-P: Voltage-Gated K+ Channel Kv7.1 Is an Important Regulator of Insulin Secretion in Normal and Hyperinsulinemic Islets
Authors: JULIANA CHRISTINE , DE LEON DIVA
Published: June 2023

Fifty percent of infants with Beckwith-Wiedemann syndrome (BWS), an overgrowth syndrome caused by genetic / epigenetic abnormalities in imprinted genes on chromosome 11p15, have persistent hypoglycemia due to hyperinsulinism (BWS-HI). BWS-HI islets exhibit increased baseline and glucose-stimulated insulin secretion (GSIS) and are responsive to the KATP channel inhibitor glyburide, suggesting it is not caused by impaired KATP channels. KCNQ1 expression, encoding the pore-forming α subunit of voltage-gated K+ channel Kv7.1, is reduced in BWS-HI islets compared to normal islets (3-fold; p = 4.5×10−7) and KCNQ1 loss-of-function mutations in humans are associated with HI. We used genetic and pharmacologic methods to modify Kv7.1 activity and examined changes to insulin secretion. In response to glucose, Kcnq1−/− mice had lower plasma glucose and higher plasma insulin than wild-type controls. Perifusion of Kcnq1−/− pancreatic islets with increasing concentrations of glucose showed that GSIS in these islets was faster and greater than in wild-type islets. The perifusion GSIS responses in normal human, wild-type mouse and Sur1−/− mouse (KATP-HI mouse model) islets with two different Kv7.1 activators (hexachlorophene or ML277) were markedly slower and diminished. In contrast, wild-type mouse islet perifusion with the Kv7.1 inhibitor chromanol 293B strikingly fastened and increased the GSIS response compared to vehicle controls. Membrane potential (Vm) analysis of Kcnq1−/− islets demonstrated that depolarization occurs more quickly during the initiation of insulin secretion compared to wild-type controls. Treatment with Kv7.1 activators resulted in Vm changes in both wild-type mouse and normal human islets compared to controls. Our results reveal the capacity of Kv7.1 to regulate insulin secretion in pancreatic islets and that Kv7.1 is a viable target for development of therapeutics for HI.

Disclosure

C.Juliana: Research Support; Crinetics Pharmaceuticals, Inc., Twist Bioscience, AmideBio. D.De leon: Consultant; Zealand Pharma A/S, Crinetics Pharmaceuticals, Inc., Eiger BioPharmaceuticals, Hanmi Pharm. Co., Ltd., Research Support; Ultragenix, Crinetics Pharmaceuticals, Inc., Hanmi Pharm. Co., Ltd., Rezolute, Inc.

Funding

National Institutes of Health (5R01DK098517-08)
1749-P: Influence of the Biomechanical Environment on Islet Function and Maturity
Authors: VAZQUEZ KELLY , BENNINGER RICHARD K.
Published: June 2023

It is well-established that cells respond to mechanical cues via cytoskeletal remodeling. However, it is not known whether the mechanical environment influences β-cell maturity and function. To examine how mechanical cues influence the β-cell, we used biocompatible substrates with tunable mechanical properties synthesized with elastic moduli ranging from ~3kPa-33 kPa. The surface was coated with extracellular matrix and MIN6 β-like cells were seeded. To independently examine a link between β-cell function and maturity, we examined islets from mice in which β-cells express CaMPARI, a photoconvertible fluorescent protein that in the presence of high Ca2+ activity changes from green to red. For all conditions in MIN6 cells and CAMPARI-expressing islets, we imaged Ca2+ dynamics via Fluo4 at low (2mM) and high glucose (11, 20mM). qPCR was conducted on MIN6 cells (6, 22 kPa) and flow-sorted CaMPARI-expressing β-cells (green, red) to examine expression of genes linked to β-cell function and maturity. With increased substrate stiffness, MIN6 cell Ca2+ oscillations were more robust, including a significant increase in duty cycle (p<0.01) and were more coordinated, consistent with increased excitability. With increasing stiffness, there was significantly decreased expression of Sur1, Kir6.2, Ins2, Pdx1, Fltp, and Ecad (p<0.01). Thus, the dynamics and coordination of Ca2+ activity is mechanoresponsive and may result from altered β-cell maturity. In CaMPARI islets, Ca2+ oscillations were more robust and coordinated for photo-converted red cells which marks cells with higher Ca2+, compared to green cells. In these red cells that show higher Ca2+, qPCR showed significantly increased expression of cFos, but decreased expression of Pdx1 and Ins2, again showing more excitable cells showed altered maturity. In conclusion, our results suggest there may be an inverse relationship between β-cell maturity and function. Further, that the maturity and function of β-cells is responsive to the mechanical environment.

Disclosure

K.Vazquez: None. R.K.Benninger: None.

Funding

National Institutes of Health (R01DK102950, R01DK106412)
1750-P: Differential Impacts of Two Novel HNF1A Variants Associated with Familial Young-Onset Diabetes on Protein Function and Insulin Secretion In Vitro
Authors: CHERKAOUI INES , DU QIAN , DION CAMILLE , LEITCH HARRY , CHABOSSEAU PAULINE L. , EGLI DIETRICH M. , SACHEDINA DILSHAD , WASTIN JULES , MISRA SHIVANI , RUTTER GUY A.
Published: June 2023

Introduction: Mutations in the HNF1A are a cause of monogenic diabetes and are usually dominantly inherited. We report in vitro and clinical studies on two novel HNF1A variants to determine pathogenicity: a recessive (homozygous) HNF1A variant (p.A251T) and a dominant HNF1A variant (p.S19L). We also investigated the mechanisms by which the former variant affects insulin secretion using beta cells derived from induced pluripotent stem cells (iPSCs).

Methods: Nuclear localization, DNA binding, and transactivation activity were examined using standard techniques. iPSC lines from proband carrying the p.A251T variant were differentiated into beta-like cells according to Sui et al, CPHG, 2018. Ca2+ dynamics were monitored with Cal-520, and insulin secretion was measured by homogeneous time-resolved fluorescence assay compared to wild-type HNF1A (WT).

Results: HNF1A p.A251T showed a mild (10.4% ± 7.3, p= 0.16) decrease in transactivation activity compared with WT, and decreased (48% of WT, p=0.12) DNA binding. These reductions were significant in p.S19L (DNA binding 57% of WT, p=0.049). Significant impaired nuclear uptake (63% vs 90% cells for WT, p<0.0001) was only apparent for p.S19L. Glucose-stimulated insulin secretion was decreased in A251T iPSCs compared with WT (0.29% vs. 0.46% of total insulin). In contrast, glucose-induced Ca2+ increases were increased in mutant cells (arbitrary units: 7.39 ± 1.23, mean +/− SD, n=725 cells vs. 4.58 ± 0.77, n=724, p= < 0.001)

Conclusion: We demonstrate the value of combining multiple in vitro  functional studies to decipher novel HNF1A variant pathogenicity. The recessive p.A251T variant has relatively mild effects on protein function compared to a dominant p.S19L variant. However, when studying proband iPSC derived beta-cell A251T impairs glucose-stimulated insulin secretion despite preserved Ca2+ entry, suggesting a novel mechanism of action not captured by standard tests of HNF1A function.

Disclosure

I.Cherkaoui: None. G.A.Rutter: Consultant; Sun Pharmaceutical Industries Ltd. Q.Du: None. C.Dion: None. H.Leitch: None. P.L.Chabosseau: None. D.M.Egli: None. D.Sachedina: None. J.Wastin: None. S.Misra: Research Support; Dexcom, Inc.

Funding

Diabetes UK
1761-P: Verapamil Increases ß-Cells Proliferation in MIN6 Cells and ß-Cell Mass in Zebrafish Model
Authors: AREFANIAN HOSSEIN , AL MADHOUN ASHRAF , ALRASHED FATEMA , BAHMAN FATEMAH , ALSAEED HALEMAH , ABUKHALAF NERMEEN A. , SHENOUDA STEVE , WILLIAMS MICHAYLA R. , NIZAM RASHEEBA , JOHN SUMI E. , JACOB SINDHU , ALHUSAYAN MOHAMMED , KURIAN SHIBU J. , KOCHUMON SHIHAB P. , SINDHU SARDAR T.A.K. , ALZAID FAWAZ , ABU-FARHA MOHAMED , ABUBAKER JEHAD , THANGAVEL ALPHONSE T. , AHMAD RASHEED , ALMULLA FAHD
Published: June 2023

Recent clinical studies highlighted a significant clinical utility for verapamil in both type 1 (T1D) and type 2 (T2D) diabetes. Verapamil treatment in T1D lowered dependency on exogenous insulin. In T2D, addition of verapamil to metformin therapy improved glycemic control. These findings led us to further study the molecular and cellular mechanisms of verapamil in the treatment of T1D and T2D using an in vitro (MIN6 mouse β cell-line) and in vivo zebrafish (Danio rerio) model. In this study, MIN6 cells were cultured in growth media containing 5.6 mM glucose (mimicking normoglycemia) and supplemented with 2% BSA to eliminate any proliferative effect of high glucose in the media, and growth factors in FBS. The cells were treated with different concentrations of verapamil. The levels of cell proliferation (MTT assay), cell growth (cell counts), Ki67 expression (IF), and cell cycle (flow cytometry) were studied in MIN6 cells treated with the optimal dose detected (50 μM) of verapamil. Larvae (24hpf) of the Ins:NfsB-mCherry transgenic zebrafish (expressing mCherry fluorescence reporter under the control of insulin proximal promoter) were treated with 10 uM of verapamil for 3 days. Full larvae images were acquired using a stereo microscope. Our results show that verapamil induced significantly increased proliferation of MIN6 cells. Cell cycle analysis shows initiation of the proliferative effect during an early time point (1-4 hours) after verapamil treatment. Higher Ki67 expression indicated enhanced cellular proliferation at 24 hours and growth curve data confirmed that the proliferative effect of verapamil lasted for 7 days. As evident from increased mCherry fluorescence intensity in zebrafish larvae, verapamil treatment increased the β cell mass during the exposure period of three days. Taken together, our findings support the positive impact of verapamil on mouse β cell proliferation in vitro as well as on increased β cell mass in Zebrafish model.

Disclosure

H.Arefanian: None. S.E.John: None. S.Jacob: None. M.Alhusayan: None. S.J.Kurian: None. S.P.Kochumon: None. S.T.K.Sindhu: None. F.Alzaid: None. M.Abu-farha: None. J.Abubaker: None. A.T.Thangavel: None. A.Al madhoun: None. R.Ahmad: None. F.Almulla: None. F.Alrashed: None. F.Bahman: None. H.Alsaeed: None. N.A.Abukhalaf: None. S.Shenouda: None. M.R.Williams: None. R.Nizam: None.

Funding

Kuwait Foundation for the Advancement of Sciences (RACB-2019-001)
1763-P: Pretreatment with Verapamil Increases MIN6 Mouse ß-Cell Survival after Exposure to Cytotoxic Stressors
Authors: AREFANIAN HOSSEIN , BAHMAN FATEMAH , AL MADHOUN ASHRAF , SINDHU SARDAR T. , ALRASHED FATEMA , KOCHUMON SHIHAB P. , NIZAM RASHEEBA , JOHN SUMI E. , JACOB SINDHU , ALHUSAYAN MOHAMMED , ABUKHALAF NERMEEN A. , SHENOUDA STEVE , ALSAEED HALEMAH , KURIAN SHIBU J. , WILLIAMS MICHAYLA R. , ALZAID FAWAZ , ABU-FARHA MOHAMED , ABUBAKER JEHAD , THANGAVEL ALPHONSE T. , AHMAD RASHEED , ALMULLA FAHD
Published: June 2023

Verapamil was found to lower the risk of developing new-onset diabetes and to lower fasting glucose levels in persons with diabetes. While previous studies have focused on treatment with verapamil, our objective in this study was to determine the effect of verapamil pre-treatment on MIN6 mouse β cell-line viability in response to different stressors mimicking conditions of in vitro models of type 1 (T1D) and type 2 diabetes (T2D). MIN6 cells were treated with either streptozotocin (STZ), T1D-cytomix containing IL-1β, TNF-α, and INF-γ, or a T2D-cytomix containing IL-1β, TNF-α, and palmitic acid. MTT assay was performed to determine viability of the cells pre-treated with verapamil (50 µM) for 24 hours, followed by each stressor treatment for another 24 hours, under normal (5.6 mM) glucose conditions. For all stressors, pre-treatment with verapamil significantly increased the cell viability, compared to verapamil-untreated controls. To study the synergistic effect of verapamil pre-treatment on the combined treatment of verapamil plus stressors, MIN6 cells were pre-treated with verapamil, then exposed to co-treatment conditions. We found that co-treatment of verapamil with each stressor significantly improved the cell survival, compared to cells treated with the stressor only. In case of STZ and T2D-cytomix, pre-treatment of verapamil significantly increased the cell viability, compared to cells that only received co-treatment alone. No significant synergistic effect of verapamil was observed as pre-treatment on co-treatment of verapamil and T1D-cytomix. In all cases showing protective effect of verapamil pre-treatment, verapamil improved cell viability in a dose-dependent manner. These results suggest that verapamil may have cytoprotective effect in MIN6 β cells under conditions mimicking T1D/T2D insults, and pre-treatment with verapamil, in particular, offers higher protection.

Disclosure

H.Arefanian: None. M.Alhusayan: None. N.A.Abukhalaf: None. S.Shenouda: None. H.Alsaeed: None. S.J.Kurian: None. M.R.Williams: None. F.Alzaid: None. M.Abu-farha: None. J.Abubaker: None. A.T.Thangavel: None. F.Bahman: None. R.Ahmad: None. F.Almulla: None. A.Al madhoun: None. S.T.Sindhu: None. F.Alrashed: None. S.P.Kochumon: None. R.Nizam: None. S.E.John: None. S.Jacob: None.

Funding

Kuwait Foundation for the Advancement of Sciences (RACB-2019-001)
1768-P: Sel1L-Hrd1 Endoplasmic Reticulum-Associated Degradation Regulates PC2 Function and Proglucagon Processing in Islet α-Cells
Authors: REINERT RACHEL B. , ZHU WENZHEN , PAN LINXIU , CHIARA RUSSO ANNA , RAY ROHIT , SHRESTHA NEHA , LINDBERG IRIS , QI LING
Published: June 2023

Pancreatic islets rely on cellular protein quality control to maintain glucoregulatory hormone production. Proglucagon, precursor of glucagon and glucagon-like peptide 1 (GLP1), is expressed in islet α cells, intestinal L-cells, and some neurons. Islet α cells predominantly convert proglucagon to glucagon through prohormone convertase 2 (PC2). Islet-derived GLP1 is increasingly recognized as an important contributor to islet function, but the factors controlling its production remain unclear. To understand how protein quality control affects proglucagon processing in α cells, we inactivated the highly conserved Sel1L-Hrd1 endoplasmic reticulum-associated degradation (ERAD) system using Cre-loxP recombination in mice. Compared to littermate controls, Sel1LΔGcg mice showed an age-related decline in glucagon secretion in vivo, associated with a 43% decrease in pancreatic glucagon content (76.1 +/- 14.1 vs. 134.4 +/- 14.9 ng/g). Pancreatic GLP1 content was unchanged in Sel1LΔGcg mice (779.4 +/- 60.7 vs. 773.4 +/- 128.6 pg/g), despite a 37% reduction in α cell mass (0.19 +/- 0.04 vs. 0.12 +/- 0.04 mg/pancreas). Expression of the precursor form of PC2 (proPC2) was increased in Sel1LΔGcg α cells, suggesting accumulation of misfolded protein and/or impaired maturation. To investigate the mechanism underlying these changes, we used CRISPR to delete Sel1L in αTC cells. ERAD-deficient αTC-ΔSel1L cells showed 1.75-fold higher expression of proPC2, in addition to appearance of a novel truncated (~56 kD) form of proPC2. Though expression of the mature (64 kD) PC2 form was unchanged in αTC-ΔSel1L cells, PC2 activity was reduced by half in a specific enzyme assay. This was accompanied by decreased expression of 7B2, a chaperone required for proPC2 transit and activation in the secretory pathway. These data show that Sel1L-Hrd1 ERAD is required to ensure proper proPC2 maturation and glucagon production in α cells, thus maintaining classic α cell identity.

Disclosure

R.B.Reinert: None. W.Zhu: None. L.Pan: None. A.Russo: None. R.Ray: None. N.Shrestha: None. I.Lindberg: None. L.Qi: None.

Funding

National Institutes of Health (K08DK129719, T32DK007245)
1769-P: Single-Nucleus Multiomics Reveals Unique Molecular Signatures of ß-Cell Heterogeneity and Compensation in Human Obesity
Authors: KIM HYUNKI , LI YAN , ZHU HOUXIANG , REN ZIYOU , EL JELLAS KHADIJA , SHUKLA NAMRATA , CHEN MENGJIE , KULKARNI ROHIT
Published: June 2023

The increasing prevalence of obesity and type 2 diabetes (T2D) continues to be a major economic burden worldwide. Healthy humans adapt to metabolic stress by increasing their functional β-cell mass. Failure of this process, called de-compensation, potentially triggers the development of T2D. However, molecular mechanism(s) underlying the ability of heterogenous human β-cells to compensate remain elusive. To interrogate the heterogeneity of gene expression regulation during successful β-cell compensation, we performed single-nucleus multiome (RNA/ATAC) sequencing in human islet cells. In this study, we analyzed a total of 160,954 islet nuclei isolated from nondiabetic lean (LN-ND, n=7), overweight (OW-ND, n=5), obese (OB-ND, n=7), or T2D (n=10) cadaveric donors. Following total cell integration and subclustering, we identified 16 islet/pancreatic cell types including 6 human β-cell subtypes (β1-6). Single-nucleus multiomics revealed unique molecular signatures of the β-cell subtypes that compensate for obesity. We identified unique sets of genes and biological pathways for each β-cell subtype that correlated with the body mass index among the ND individuals. We also identified β-cell subtype-specific transcription factor motifs and intra-islet interactions that are enriched during compensation. Finally, the intersection of publicly available human islet/β-cell-specific chromatin looping data with our datasets revealed compensatory β-cell subtype-specific cis-regulatory elements that overlap with T2D genomic variants. In summary, we report the discovery of unique signatures of chromatin landscape and gene expression regulation in heterogenous β-cells secondary to potential cell-cell interactions during islet compensation in human obesity. These results provide novel insights into the epigenetic regulation in human islet cells that can be harnessed to develop new therapeutic approaches in precision medicine to counter T2D.

Disclosure

H. Kim: None. Y. Li: None. H. Zhu: None. Z. Ren: None. K. El Jellas: None. N. Shukla: None. M. Chen: None. R. Kulkarni: Advisory Panel; Novo Nordisk, Inversago. Research Support; Inversago. Advisory Panel; Biomea Fusion, Inc., REDD Pharma.

Funding

National Institutes of Health (R01067536)
1773-P: Glucocorticoid Receptor Signaling Regulates Pancreatic Beta-Cell Circadian Transcriptome and Function
Authors: SEN SATISH K. , WANG LIU , BROWN MATTHEW , WU JIE , WEI ZONG , MATVEYENKO ALEKSEY
Published: June 2023

Glucocorticoid (GC) signaling is an important regulator of glucose homeostasis. GC secretion also serves as an entrainment signal that coordinates circadian gene expression of peripheral circadian clocks. Pancreatic beta cells are key biological controllers of glucose homeostasis through the coordinated circadian release of insulin. However, the role of GC signaling in the regulation of beta cell circadian clocks and insulin secretion remains unknown. To address this, we first performed chromatin immunoprecipitation/sequencing (ChIP-seq) in beta cell lines (MIN6 and INS1) and identified >1,000 glucocorticoid receptor (GR)-bound regions (FDR<0.01) in response to the synthetic GC dexamethasone (DEX, 100nM). Annotation of GR-bound regions showed enrichment for pathways regulating insulin secretion and circadian rhythms. To further explore the effects of GC on circadian beta cell gene expression and function in vivo, we used C57BL/6 mice housed under either 12h/12h light-dark cycles (LD) or constant light-mediated circadian disruption (CD). Mice were subsequently administered daily intraperitoneal injections of DEX (2 mg/kg) or vehicle, designed to restore circadian secretion of GC disrupted in CD group. DEX treatment normalized circadian regulation of glucose tolerance (p<0.05 vs. vehicle) and glucose-stimulated insulin secretion (p<0.05 vs vehicle) in CD group. We next utilized RNA sequencing to assess circadian transcriptional profiles in islets isolated from LD, CD, and CD + DEX mice. Consistent with previous studies, CD resulted in abrogation of circadian islet mRNA expression, whereas DEX restored circadian rhythmicity and enhanced expression of core clock genes (e.g., Bmal1, Dbp, Nr1d1) and genes regulating beta cell identity (e.g., Ins1, Nkx6.1) and insulin secretion (e.g., Gck, Irs2). These data highlight the GC/GR axis as a direct regulator of beta cell circadian gene expression, insulin secretion and glucose tolerance.

Disclosure

S.K.Sen: None. L.Wang: None. M.Brown: None. J.Wu: None. Z.Wei: None. A.Matveyenko: None.

Funding

National Institutes of Health (DK098468)