bluuebunny/crossref_metadata_2025 huggingface dataset:
5156 Circulating MicroRNA-based Biomarker Combinations for Differentiating Uni- and Bilateral Primary Aldosteronism Using a Machine Learning Approach
Authors: Vekony Balint , Nyiro Gabor , Herold Zoltan , Fekete Janos , Ceccato Filippo , Gruber Sven , Kurzinger Lydia , Parasiliti-Caprino Mirko , Szeredas Balint , Nazri Siti K S M , Fell Vanessa , Bassiony Mohamed , Azizan Elena A , Bancos Irina , Beuschlein Felix , Igaz Peter
Published: October 2024

Disclosure: B. Vekony: None. G. Nyiro: None. Z. Herold: None. J. Fekete: None. F. Ceccato: None. S. Gruber: None. L. Kurzinger: None. M. Parasiliti-Caprino: None. B. Szeredas: None. S.K. Nazri: None. V. Fell: None. M. Bassiony: None. E.A. Azizan: None. I. Bancos: None. F. Beuschlein: None. P. Igaz: None.

Background: The differentiation of uni- and bilateral forms of primary aldosteronism (PA) is of pivotal clinical relevance. Whereas unilateral PA warrants surgical treatment, bilateral PA is treated by aldosterone antagonists. The most reliable way to differentiate these two entities is adrenal venous sampling (AVS) that is an invasive method with limited availability and requiring great expertise. Aim: To develop a circulating blood-borne microRNA-based approach using machine learning for the differentiation of uni- and bilateral PA. Methods: Circulating microRNA profiling on an Illumina MiSeq platform was performed on total RNA samples isolated from EDTA-anticoagulated blood specimens taken by AVS from 18 pairs of right and left suprarenal veins (10 uni-, 8 bilateral PA). Significantly differentially expressed microRNAs were analyzed by a neural network (90-10% learner-tester cross-validation simulation). From the top 29 significantly differentially expressed microRNAs, 9 were selected for validation by Taqman RT-qPCR on a separate cohort of 30 AVS-confirmed samples (15-15 uni- and bilateral forms) both on plasma samples from adrenal as well as from peripheral veins. The method was finally tested on another independent cohort of peripheral blood samples from 80 patients (40 uni- and 40 bilateral PA). Results: Regarding the 9 microRNAs selected for validation, microRNA abundance was non-significantly lower in peripheral samples compared to AVS samples. By analyzing peripheral plasma samples, ten combinations of 4-8 microRNAs from the pool of 9 microRNAs turned out to have maximal sensitivity and specificity values over 85 %. The best combination included 6 microRNAs with a sensitivity of 86.3 % and a specificity of 87.91 % (AUC = 0.871). The inclusion of further parameters such as imaging, potassium and renin levels did not improve the model’s performance. Conclusion: The analysis of circulating blood-borne microRNAs represents a minimally invasive and potentially reliable way of differentiating uni- and bilateral PA. If bilateral PA is confirmed by the model, no more localization efforts would be necessary thus simplifying the management of PA patients. This would be relevant both from the patient’s and a public health perspective.

Presentation: 6/3/2024
7386 GlycA as a Novel Biomarker of Systemic Inflammation: Determinants and Implications in South Asians
Authors: Deven A , Ramanathan A , Bukhari K , Rabel A , Muniyappa R
Published: October 2024

Disclosure: A. Deven: None. A. Ramanathan: None. K. Bukhari: None. A. Rabel: None. R. Muniyappa: None.

Background: South Asians are at an increased risk for developing diabetes and cardiovascular disease compared with their European counterparts. Increased systemic inflammation is a key contributor to cardiometabolic dysfunction. GlycA, a novel inflammatory biomarker, predicts incident type 2 diabetes mellitus (T2DM) in population-based studies of South Asians. However, the determinants of GlycA in this population are unknown. Objective: To investigate the relationships between GlycA, body composition, glycemia, lipoproteins, insulin sensitivity, and beta cell function in healthy individuals. Methods: This cross-sectional study involved 59 adults without T2DM (body mass index: 26.5 ± 5.2 kg/m2, age 36.5 ± 8.3 years, 44 % female) who underwent a frequently sampled intravenous glucose tolerance test and a mixed meal in the fasted state. Body composition was determined by dual-energy X-ray absorptiometry. Insulin sensitivity and beta cell function parameters were calculated by minimal model analysis. Nuclear magnetic resonance was used to measure GlycA and lipoprotein profiles. Results: GlycA was found to be positively associated with BMI, total fat mass, relative fat mass, total cholesterol, free fatty acids, LDL cholesterol, small LDL-particle number, triglycerides, triglyceride-rich lipoproteins particles (TRLP), fasting insulin, and insulin secretion. GlycA was also negatively associated with measures of insulin sensitivity, HDL, and large-HDL-particle number. The forward stepwise regression model, explaining 54% of the variance in GlycA levels (R2=0.54, P <.0001), identified relative fat mass, insulin sensitivity derived from mixed meal, and large HDL particle number as independent predictors. Conclusions: GlycA exhibited an independent association with relative fat mass, insulin resistance, and reduced levels of HDL particles (HDL-P) in South Asians, suggesting its potential as a predictive biomarker in this demographic. Future research should focus on exploring the underlying mechanisms driving these associations.

Presentation: 6/3/2024
8575 Hepatic Steatosis Index as a non-invasive marker for liver steatosis in patients with endogenous Cushing Syndrome, ERCUSYN Krakow database
Authors: Minasyan M , Gamrat A , Hubalewska-Dydejczyk A , Valassi E , Gilis-Januszewska A
Published: October 2024

Disclosure: M. Minasyan: None. A. Gamrat: None. A. Hubalewska-Dydejczyk: None. E. Valassi: None. A. Gilis-Januszewska: None.

Introduction: Liver Steatosis(LS) can be one of the metabolic complications of Cushing Syndrome(CS). The data on the impact of cortisol on liver function are very limited, and there is only one study reporting a prevalence of LS of roughly 20%. Hepatic Steatosis Index (HSI) predicts LS based on ALT, AST, BMI, sex and co-existence of diabetes mellitus/impaired glucose intolerance. Objectives: To evaluate the prevalence of LS in patients with CS at the time of diagnosis by using HSI. Methods: We analyzed retrospectively adult patients from the ERCUSYN, Krakow database with complete HSI data available. 82 of 135 patients, aged 27-87 years, predominantly women (N=64), were eligible for the study. The HSI score was calculated using the following formula: 8×(ALT:AST)+BMI+2(if type 2 diabetes)+2(if female). Collected data were from the baseline CS diagnosis. Patients with score 36 or above were classified as having a high probability of LS. We compared HSI results with the liver steatosis diagnosed based on abdominal ultrasonography (USG) findings and clinical and demographic factors. Results: High HSI was observed in 81.7% patients (82.8% of females, 77.8% of males). HSI was elevated in 85%, 80% and 73% of patients with pituitary, adrenal and ectopic CS respectively. 41% of patients with elevated HSI were obese. HSI was elevated in among: 100% patients with confirmed liver steatosis on USG, 72% patients with normal USG and 78% patients who did not undergo USG. Conclusions: The prevalence of liver steatosis in active CS may be higher than previously reported. Further investigation should be undertaken to show if patients with high risk of LS based on HSI and normal liver image on USG, may benefit from liver MRI in order to confirm the diagnosis.

Presentation: 6/3/2024
5185 A Meta-Analysis Evaluating the Efficacy of Liraglutide in Reducing Cardiovascular Complications Among Patients With Type 2 Diabetes
Authors: Del Rosario Michelle Panol , Ong-Ramos Celeste
Published: October 2024

Disclosure: M.P. Del Rosario: None. C. Ong-Ramos: None.

Background: The objective of this study was to determine the efficacy of Liraglutide in reducing cardiovascular complications Methodology: Literature searches of electronic databases (PubMed and ScienceDirect) were performed to identify relevant studies. Relevant journals included were electronically searched for randomized controlled trials (RCTs) regarding the use of Liraglutide versus placebo in reducing the rate of cardiovascular complications. Data extraction and quality assessment were done from independent reviewers. Statistical analysis of the study was accomplished using Cochrane Systematic Review Software Review Manager. A p-value of less than 0.05 for the observed effect size was considered statistically significant. Results: A total of 2 RCTs with 16, 977 patients were included in this meta-analysis. The results showed that in the Liraglutide group there is a lower rate of the following: Major Adverse Cardiovascular Events (MACE) (risk ratio of 0.86 [0.78, 0.95]), cardiovascular events (risk ratio 0.75 [0.63, 0.89]), and all-cause mortality (risk ratio 0.81 [0.70, 0.93]), but is not statistically significant when compared to the placebo group in terms of non-fatal myocardial infarction and and non-fatal stroke. There is no significant difference in rates of severe adverse events in general (p > 0.05). However, Liraglutide caused significant reduction in rates of severe hypoglycemia as an independent secondary outcome (p < 0.05). Conclusions: Compared with placebo, Liraglutide provided significant reduction in MACE, cardiovascular events, all-cause mortality and severe hypoglycemia as an adverse event.

Presentation: 6/3/2024
12503 An Observational Study Of The Glycemic Variability In Type 1 Diabetes Mellitus Women Preconception And During Pregnancy And Its Associations With Adverse Maternal And Neonatal Outcomes
Authors: Avula Sreekant , Ankireddypalli Anvitha Reddy , Tessier Katelyn , Seaquist Elizabeth Rachel
Published: October 2024

Disclosure: S. Avula: None. A.R. Ankireddypalli: None. K. Tessier: None. E.R. Seaquist: None.

Background: Pregnancy in T1 diabetes mellitus (T1DM) women is associated with increased risk of maternal/fetal complications. Despite glycemic optimization during pregnancy in T1DM, offspring are at increased risk for neonatal complications. Aim: To examine glycemic variability by continuous glucose monitor (CGM) preconception and during pregnancy by trimester and assess the association with adverse maternal/fetal outcomes. Methods: This was a retrospective cohort study of pregnant patients with T1DM who used real-time CGM and delivered at a U.S. tertiary center (2012-2023). Multiple gestations, Cystic fibrosis-related diabetes (CFRD), twin or multiple births, and no valid CGM data for at least one trimester were excluded. 63 met the inclusion criteria. The primary outcome was composite neonatal morbidity (CNM), including large or small for gestational age (LGA/SGA), NICU admission, neonatal hypoglycemia, shoulder dystocia, respiratory distress syndrome, APGAR 1 or 5 minutes < 7, prematurity (<37 weeks), intrauterine growth restriction (IUGR), CPAP, and ventilator. We also collected data about maternal outcomes. Results:54 patients (86%) met at least one outcome included in the CNM. 20 patients (32%) were LGA, 3 (5%) were SGA, 32 (51%) premature, 29 (46%) required NICU admission, 44 (70%) had neonatal hypoglycemia, 3 (5%) had shoulder dystocia, 23 (37%) had respiratory distress, 22 (35%) needed CPAP, 3 (5%) were on ventilator, 11 (18%) had APGAR 1 minute <7. Among maternal outcomes, 18 (29%) developed pre-eclampsia, and 46 (73%) had a cesarean delivery. Overall, individuals with neonatal morbidity had a lower target in range (TIR) and higher target above range(TAR), average glucose, standard deviation(SD), and coefficient of variation (CV) compared with those without the primary composite, but statistical significance was seen for average glucose and TAR. For every 5 mg/dl increase in average glucose (mean of the 3-trimester values), there was a 25% increase in odds of neonatal morbidity (OR 1.25; 95% CI 1.04-1.56; p=0.028). There was a significant association between 1st trimester SD and neonatal morbidity (OR 1.11; 95% CI 1.02-1.24; p=0.027). There was a trend toward significance between the 3rd-trimester average blood glucose and neonatal morbidity (OR 1.26; 95% CI 1.02-1.63; p=0.051). Conclusion: In our single-center cohort, we found maternal average glucose and TAR, as well as first trimester SD, were significantly associated with CNM. This confirms that maternal hyperglycemia and demonstrates that first trimester SD are associated with poorer neonatal outcomes.

Presentation: 6/1/2024
9207 Anoctamin 4 in the Area Postrema Regulates Glucose and Energy Homeostasis
Authors: Tu Longlong , Xu Yong
Published: October 2024

Disclosure: L. Tu: None. Y. Xu: None.

Located on the floor of the 4th ventricle within the dorsal surface of the medulla oblongata, the area postrema (AP) is a circumventricular organ where a proper blood-brain barrier is missing. Numerous studies reveal that neurons in the AP are implicated into suppression of food intake (e.g., glucagon-like peptide 1 (GLP-1)-mediated anorexia), but the role of AP neurons in whole-body glucose homeostasis is less investigated. Here we identified and fully characterized the physiological functions of anoctamin-4 (Ano4, a calcium activated chloride channel)1 in the AP. CRISPR-mediated deletion of Ano4 in the AP caused a lower body weight and reduced blood glucose, concurrent with an improved glucose tolerance and increased insulin sensitivity. Chemogenetic activation of Ano4-expressing neurons in the AP (APAno4) increased food intake and blood glucose, and did not induce conditioned flavor avoidance. Both Ano4 neurons and non-Ano4 neurons (i.e., neurons do not express Ano4) sent projections to the lateral parabrachial nucleus (LPBN). Optogenetic stimulation of APAno4 ◊ LPBN circuit promoted feeding behavior and increased blood glucose, whereas stimulation of APnon-Ano4 ◊ LPBN circuit inhibited food intake and did not influence glucose homeostasis. Finally, RNAscope studies reveal that 45.7% of APAno4 neurons are glutamatergic neurons, and 41.8% of them are GABAergic neurons. Furthermore, APAno4 neurons showed only a 2.6% overlap with Glp-1r-expressing neurons in the AP, but had a 60.8% co-localization with protein tyrosine phosphatase receptor δ2, which serves as orexigenic asprosin receptor. Together, our results indicate that Ano4 neurons in the AP are the first reported orexigenic neurons, and represent potential therapeutic targets for human diseases with glucose imbalance and abnormal feeding behavior. Reference: (1) Tu, L. et al. Anoctamin 4 channel currents activate glucose-inhibited neurons in the mouse ventromedial hypothalamus during hypoglycemia. J Clin Invest 133,14. (2023). (2) Mishra, I. et al. Protein tyrosine phosphatase receptor δ serves as the orexigenic asprosin receptor. Cell Metab 34, 549-563. (2022).

Presentation: 6/2/2024
9300 Anti-PD1 induces T1D in an HLA-DQ8, HLA-A2.1 mouse model
Authors: Houeiss Pamela , luce Sandrine , Le-Gall Morgane , Onifarasoaniaina Rachel , Boitard Christian
Published: October 2024

Disclosure: P. Houeiss: None. S. luce: None. C. Boitard: None.

Anti-PD1 is an immune checkpoint inhibitor that has revolutionized cancer treatment. It blocks the interaction between PD1, a receptor on the surface of activated or exhausted T cells, and its ligands PDL1 and PDL2, to inhibit the immune response. However, anti-PD1 induces T1D in 0.4-2% of patients raising questions about the risk factors and the mechanisms behind T1D development. Since the pancreas of these patients is inaccessible, we focus on autoimmune diabetes induction in a transgenic mouse model modified to express the T1D susceptibility genes HLA-DQ8, HLA-A2.1 and the human preproinsulin and on identifying new targets to prevent or treat this disease. Thus, 8 to 12 weeks old mice were treated initially with 500 micrograms of anti-PD1 followed by 250 micrograms every other day for 10 days. Control mice received 6 injections of PBS. Mice were followed up for evidence of hyperglycemia for 60 days. Cells and tissues from the pancreas, and the spleens were isolated 15-, 30- or 60-days post-treatment initiation and analyzed by flow cytometry and immunohistochemistry. Although diabetes incidence in treated mice was 14% (n=14), insulitis incidence progressively increased over time reaching 86% on D60 (p=0.005). On Day 15, in the spleen, a significant 10% increase of CD4+Foxp3+CD25- T cells at the expense of the CD4+Foxp3- T cells were seen in the treatment group as compared to the control (37.1±4.25 vs 26±3.5, p<0.01, n=5 per group). In the treatment group, PD1 expression significantly increased on CD4+Foxp3+ T cells whereas PDL1 expression increased on CD4+Foxp3- and CD8+ T cells (p<0.01). On D30, in the pancreas, infiltrates show positive staining for F4/80+, CD4+ and CD8+ T cells with scarce foxp3+ T cells. While the absolute percentage of CD4+Foxp3+ T cells increased in the insulitis and diabetes group, their percentage among CD3+ T cells decreased in the insulitis (10.2%) and diabetes group (3.9%) as compared to the control group (13.5%). Infiltrates were concentrated around the islets and the efferent and afferent vessels, and extended to the exocrine pancreas, leading to secondary acinar destruction. In the pancreatic infiltrates from anti-PD1 treated mice, we noted a significant increase in CD4+ T cells recognizing Ins13-21 monomers as compared to the negative control. PD1 blockade induced an insulitis characterized by a Treg to effector T cells imbalance. Thus, it acted on the early phase of T1D. In conclusion, this model describes a new T1D endotype that shares similarities with ICI induced diabetes in patients.

Presentation: 6/2/2024
6560 Atypical Diabetes Mellitus in a Mother-Son Pair with Multiple Autoimmune Conditions & Pathogenic Variant in BCL11B Inherited from the Mother
Authors: Elrajabi Rawan , Gurtunca Nursen
Published: October 2024

Disclosure: R. Elrajabi: None. N. Gurtunca: None.

The etiology of diabetes can be challenging to determine at times and misdiagnosis is common in all age groups. To provide the right treatment and enhance glycemic control, which results in better outcomes and fewer comorbidities, an accurate diagnosis is crucial. We describe a mother-son pair who share a pathogenic variant in BCL11B gene with multiple autoimmune conditions including atypical autoimmune DM. A 13 year old young boy with a history of 33-week prematurity, developmental delay, autoimmune hypothyroidism, and vitiligo presented with concerns for DM given mother’s recent diagnosis of DM treated with Metformin and GLP-1 receptor agonist with poor control of her DM. His mother has a learning disability, hypothyroidism, alopecia totalis, vitiligo and hypertension. Interestingly, he and his mother share a genetic variant in BCL11B gene detected on whole exome sequencing sent as part of the workup of developmental delay. His weight and height were in the 13.5% and 13.9% centiles, respectively. He has vitiligo, but other than that, the physical examination was unremarkable. He is prepubertal. HbA1c was high, 6.4% despite the presence of anemia. An oral glucose tolerance test (OGTT) was consistent with DM as was his repeat HbA1c of 7.1%. He had elevated IA 2, zinc transporter 8, and GAD 65 antibodies, all consistent with autoimmune DM. He was admitted to the hospital and started on both long-acting and short acting insulin via multiple daily injections. BCL11B is a transcription factor that is expressed in all T cells and regulates important processes involved in T cell growth, survival, and function, including Treg cells and Foxp3 activity. Autoimmune diseases have been linked to abnormal Treg activity and Foxp3 deficiency. BCL11B functions as a checkpoint required for T cell lineage commitment while minimizing natural killer cell potential. Impaired BCL11B activity at a young age result in severe whole-body autoimmunity, defined by a fully stimulated immune system similar to animals lacking functioning Treg cells. DM associated with BCL11B variant has not previously been reported. If we follow the theory that autoimmunity affects beta cell function or mass, this patient's and his mother's DM and autoimmune disorders are most likely connected to the BCL11B variation they share.

Presentation: 6/2/2024
6519 BCG Clinical Trial Programs in Advanced Type 1 Diabetes - 2024 Update
Authors: Kühtreiber Willem M , Thach Bethany , Hayashi Hikaru , Adams Lindsey , Hullavarad Neeshi , Mounier Daniel A , Le Khoi , Bulczynski Emily , Dorsey Jack L , Kartsounis Nicholas , Braley Joan , Zheng Hui , Faustman Denise L
Published: October 2024

Disclosure: W.M. Kühtreiber: None. B. Thach: None. H. Hayashi: None. L. Adams: None. N. Hullavarad: None. D.A. Mounier: None. K. Le: None. E. Bulczynski: None. J.L. Dorsey: None. N. Kartsounis: None. J. Braley: None. H. Zheng: None. D.L. Faustman: None.

The bacillus Calmette-Guerin (BCG) vaccine, originally developed a century ago to prevent tuberculosis infection, has today advanced in multiple clinical trials for the treatment of the type 1 diabetes (T1D), other forms of autoimmunity and infectious disease. At our institution, over 500 individuals with type 1 diabetes have been enrolled in ongoing BCG clinical protocols to test BCG’s ability to lower HbA1c, reduce insulin requirements and stabilize day-to-day fluctuations in blood sugars; 300 have been treated with BCG. In a completed randomized, placebo-controlled Phase I trial in adults with established T1D, long-term follow up continues to show that HbA1c values are durably lowered for 8+ years after multi-dose BCG treatment. In two open-label clinical trials in adults with juvenile-onset diabetes, a 10-15% reduction of HbA1c was observed after multi-dose BCG treatment. A randomized, double-blinded Phase II trial in adults with longstanding T1D has followed all patients in the BCG-treated (n=100) and placebo groups (n=50) for five years and is pending read-out. The primary outcome of all the trials is HbA1c lowering. Two new, randomized, double-blinded pediatric trials are testing the potential benefits of BCG vaccination with active enrollment ongoing. One is testing BCG in children ages 12 to 18 with more than 2 years since T1D diagnosis. The other is enrolling children ages 8 to 18 with less than 1 year since T1D diagnosis. BCG vaccine therapy may provide a safe and affordable medical intervention in individuals with longstanding T1D and works decades after disease onset by changing glucose metabolism within lymphocytes, independently of the pancreas.

Presentation: 6/3/2024
7970 C3 and C4 Levels Decrease Post-prandial in Adolescents with type 1 Diabetes
Authors: Damilano Cecilia Paola , Yu Chack-Yung , Zhou Danlei , Mamilly Leena , Hoffman Robert Paul
Published: October 2024

Disclosure: C.P. Damilano: None. C. Yu: None. D. Zhou: None. R.P. Hoffman: None.

Introduction: Cardiovascular disease will be the primary cause of future death for adolescents with type 1 diabetes (T1D) and its origins are likely to occur in this age group. Adolescents with T1D have increased systemic inflammation which is likely to play a significant role in the pathogenesis of cardiovascular disease. The complement system, in particular, components C3 and C4 are part of the innate immune system that regulates inflammation. Increased C3 and C4 levels are associated with increased cardiometabolic risk. Objective: Most studies of inflammation are done while fasting but TNFα and IL6 increase postprandially in adolescent T1D following both high and low glycemic index (GI) meals. The goal of this study was to determine whether there are similar post-prandial changes in C3 and C4 levels. Methodology: Twelve, non-obese children and adolescents, 10-18 years old, with T1D were recruited from the Diabetes Clinic of Nationwide Children's Hospital. A randomized cross-over design for their two subsequent visits was used. After an overnight fast baseline blood samples were taken to measure plasma glucose, C3, total C4 protein, and total C4. Rapid acting insulin was given to cover meal carbohydrate without glucose correction, and blood samples were repeated (30, 60, 90, and 120 minutes) to measure glucose and C3 and C4 levels. Visits 1 and 2 were identical except for GI of the given meal. Visits were rescheduled if the fasting glucose was greater than 200 mg/dl. Repeated measures ANOVA and Spearman’s correlation were used. Results: Glucose response was significantly greater following high GI meal (Δglucose: high 52±12 vs low 6±5 mg/dl, mean±SE, p<0.001). For C3 levels, there was a significant post-prandial decrease (time effect p=0.049), but no meal effect or meal x time interaction. No specific time differences were found. C4 levels were also suppressed postprandially (p<0.001) and there was again no meal effect or meal by time interaction. Post hoc analysis found that C4 levels were lower at 60 min than at 0 min (p=0.007), 90 min (p=0.002) or 120 min (p<0.001). Conclusions: C3 and C4 levels decrease post-prandially in adolescents with T1D in contrast to increases in TNFα and IL6. Future research is clearly needed to clarify the postprandial interactions between the complement and inflammatory systems and potential pathogenic role in cardiovascular disease.

Presentation: 6/3/2024
8052 carRNA m6A methylation regulates transcription state and chromatin accessibility in human islets in normal physiology and type 2 diabetes
Authors: Shukla N , Ju C W , Li H , He C , Kulkarni R
Published: October 2024

Disclosure: N. Shukla: None. C. Ju: None. H. Li: None. C. He: None. R. Kulkarni: Advisory Board Member; Self; Novo Nordisk, Biomea Fusion Inc, REDD Pharma. Grant Recipient; Self; Inversago.

Chromatin-associated regulatory RNAs (carRNA), which include promoter- and enhancer-associated RNAs as well as several non-coding repeat elements have been established in the literature as important regulators of global gene transcription. However, the regulation of carRNAs themselves has been an enigma. Recent publications from the He Lab at UChicago have shown the role of RNA N6-methyladenosine (m6A) in regulating carRNA stability and function. m6A is among the most abundant mRNA modifications in mammals. Seminal work from our lab has reported that the “writers” of m6A (e.g. methyltransferase-like [METTL3] and METTL14) are significantly downregulated in islets of humans with type 2 diabetes (T2D). Consequently, the global m6A landscape in the islets segregates the T2D from the control group significantly better than the transcriptome (n=7 for Control and n=8 for T2D islets). Hypomethylation of key transcripts in the insulin/IGF-1-AKT-PDX1 pathway led to impaired insulin secretion and decreased β-cell proliferation. We now show this decreased m6A on RNA is not limited to the mRNA but also affects chromatin-associated regulatory RNAs (carRNA). Intriguingly, downregulation of METTL3 and METTL14 in human β-cell line EndoC-βH1 differentially regulates these regulatory versus protein-coding transcripts. We observe that while METTL14 KD causes global downregulation of transcripts belonging to pathways important for β-cell identity and function (including calcium signaling, PI3K-AKT/MAPK signaling pathway, pancreatic secretion etc.), it leads to an upregulation of pathways involved in RNA processing and global gene transcription (Transcription by RNA Pol II, metabolism and splicing of mRNA, chromatin organization and modification etc.) {false discovery rate (FDR) < 0.05}. Considering carRNAs have been previously reported to regulate global chromatin accessibility and transcription, our observations on their ability to get differentially regulated m6A writer proteins provide novel insights into the epigenetic regulation of human islet cells. These findings provide opportunities to develop therapeutic approaches to regulate β-cell identity and/or insulin secretion in T2D.

Presentation: 6/3/2024
7797 Characteristics of liver type 2 innate lymphocytes in blood-glucose lowering effect via IL13-IL-13R interaction through GATA3-dependent transcriptional factor network
Authors: Kozu Ryunosuke , Fujimoto Masanori , Taki Yuki , Kono Takashi , Onodera Atsushi , Tanaka Tomoaki
Published: October 2024

Disclosure: R. Kozu: None. M. Fujimoto: None. Y. Taki: None. T. Kono: None. A. Onodera: None. T. Tanaka: None.

Introduction: The metabolic improving effects of type 2 innate lymphocytes (ILC2) have attracted attention in both mice and human; ILC2 was originally found as a lymphocyte population in mesenteric adipose tissue and is involved in many organs immune responses (Nature, 2010, 463,540). IL33-stimulated activated ILC2 in adipose tissue suppresses obesity (Nature, 2015, 519, 242), via IL13 production (Nature Commun., 2017, 8, 15781). In this context, we found that ILC2 in the liver exhibits a function of inhibiting gluconeogenesis via IL-13 and exerts a blood glucose-lowering effect (Nature Commun., 13, 5408, 2022). However, the regulatory mechanism of why liver ILC2 functions to improve glucose metabolism is not clarified. Here, we examined the molecular characteristics of liver ILC2 and its cell-cell interaction by single cell analysis of hepatocytes and liver ILC2 in mouse model. Method The subject was the liver and lung (for comparison) of a BALB/c mouse model in which ILC2 was activated by IL-33 administration. scRNA-seq followed by FACS sorting was performed. Cell–cell ligand–receptor interaction analysis was performed using CellPhoneDB (v.2.0.0) to document ligand–receptor analysis of cell populations in the liver, especially for assessment of the interactions between hepatocytes and ILC2. For TFs analysis, SCENIC (Single-Cell rEgulatory Network Inference and Clustering) was used. Results: IL-33 injection, an activator of ILC2, strongly stimulated liver ILC2, which in turn lead to reducing vivo blood glucose levels in accordance with gluconeogenesis inhibition. With the aim of clarifying the mechanism of action, single cell analysis revealed that ILC2 in the liver has strong IL-13 producing ability. In addition, cell-cell interaction analysis demonstrated that certain hepatocyte clusters with portal vein markers express IL-13R, and the interaction of liver ILC2 with IL-13R suppresses the expression of G6pc and Pck1 glycogenic enzymes, thereby lowering blood glucose. Finally, to address unique characteristics of liver ILC2, we performed SCENIC and found that Gata3, a key regulator of IL13, was significantly activated upon IL-33 treatment in liver ILC2 with transcriptional factor network including BATF (basic leucine zipper transcription factor, activating transcription factor-like) and AP1 family. Discussion Hepatocytes, particularly with ILC2 interaction, function differently in distinct areas due to the influx of high concentrations of nutrients from the portal vein. Thus, clarification of the relationship between GATA3 transcriptional complex mechanism and nutrients in liver ILC2 may lead to novel regulatory mechanisms and therapeutic targets for liver gluconeogenesis through immunometabolic linkage.

Presentation: 6/1/2024
8715 Cholinergic Signaling in the Mouse Embryonic Pancreas Targets Early Endocrine Cells
Authors: Duarte C , Panzer J K , Borowsky N , Caicedo A
Published: October 2024

Disclosure: C. Duarte: None. J.K. Panzer: None. N. Borowsky: None. A. Caicedo: None.

The parasympathetic vagus nerve affects pancreas function by activating local intrapancreatic neurons that release acetylcholine (ACh). These neurons are derived from progenitors of the vagal neural crest and colonize the pancreas during embryonic development. The specific mechanisms by which ACh is released from these neurons influence embryonic development of the islet and, specifically, beta cells remain largely unknown. The purpose of this study was to identify cholinergic signaling components and assess early physiological responses to ACh during murine development of endocrine lineage cells. We extracted pancreatic rudiments from mice at embryonic stages E15 to E18 and stained them for neuronal and endocrine markers, along with components associated with cholinergic signaling. We further performed Ca2+ imaging of the pancreatic rudiments from NGN3CRE-GCaMP3 mice to record responses to ACh in cells of the endocrine lineage. At E15, we observed a significantly higher concentration of glucagon+ cells compared to insulin+ cells, consistent with the earlier differentiation of alpha cells in the developing pancreas. Neuronal body clusters were identified in close proximity to endocrine lineage cells from E15 to E18, with the neuronal network becoming more dispersed and interconnected. At E15, wide distribution of the presynaptic vesicular acetylcholine transporter (VAChT) was observed later becoming localized around insulin+ and glucagon+ cells at E18. Postsynaptic acetylcholinesterase (AChE) colocalized with glucagon+ cells from E15 to E18 and with insulin+ cells by E18. ACh elicited Ca2+ responses in individual endocrine cells as early as E15. Our results show that cholinergic nerve terminals target early endocrine cells and elicit responses to ACh, indicating that local cholinergic neurons innervate and activate endocrine progenitors in the embryonic pancreas. Our current studies are aimed at determining how cholinergic signaling contributes to beta cell differentiation and function. We expect this knowledge to guide future protocols for deriving beta cells from stem cells.

Presentation: 6/2/2024
8757 Contingency Planning in the Event of Insulin Pump Failure or Malfunction - a Quality Improvement Initiative
Authors: Avula Sreekant , Hashmi Hiba , Ankireddypalli Anvitha Reddy , Inkollu Sindhura , Salim Michael , Ahmed Ammar , Kohlenberg Jacob , Harindhanavudhi Tasma
Published: October 2024

Disclosure: S. Avula: None. H. Hashmi: None. A.R. Ankireddypalli: None. S. Inkollu: None. M. Salim: None. A. Ahmed: None. J. Kohlenberg: None. T. Harindhanavudhi: None.

Background: People using insulin pump therapy are at risk of adverse outcomes, including hyperglycemia and diabetic ketoacidosis, in the event of pump failure or malfunction. A standing prescription for subcutaneous basal insulin, which can be filled as needed, has the potential to prevent these adverse outcomes. Aim: We aimed to increase the percentage of standing prescriptions for subcutaneous basal insulin by 20 % over 6-month duration for people using insulin pump therapy. Methods: Pre-intervention: We retrospectively obtained baseline electronic health record data for people with diabetes on insulin pump therapy in our health system (M Health Fairview, University of Minnesota) from January 1, 2022, to December 31, 2022. Intervention: Over a six-month duration, a multifaceted intervention was implemented. (1) During the rooming process for endocrinology appointments, the medical assistants inquired if the person using insulin pump therapy had a standing prescription for basal insulin in the event of insulin pump failure or malfunction. If not, the medical assistant encouraged the person with diabetes to review this with their Endocrinologist. (2) Informational brochures, placed in the exam rooms, encouraged people using insulin pump therapy to discuss a standing order for basal insulin. (3) Emails were sent to all endocrinology providers to encourage discussion of the provision of a standing prescription for basal insulin for people on insulin pump therapy. Post - intervention: We recorded the number of people with a standing prescription for subcutaneous basal insulin from 5/1/2023 to 5/14/2023. Results: The total number of patients with diabetes on insulin pumps prior to the intervention was 256. Of 256 patients with insulin pump therapy, 109 patients (42.6%) had standing basal insulin prescribed. A post-intervention chart review of patients with diabetes on an insulin pump was conducted for a total of 69 patients, of which 56 patients (81.2%) had backup basal insulin prescribed. The estimation of difference was - 0.3858 (p= <0.05; 95% CI for difference -0.4961, -0.2754). Conclusion: This multipronged approach increased the prescription availability and awareness among providers and patients for a standing order of basal insulin in the event of insulin pump failure or malfunction from 42% to 81%. Limitations of this study include the limited duration of intervention and the need to send repeated reminders to all providers, including clinic staff. The next plan is to create a best practice alert in EMR to alert providers to have a backup basal insulin plan for anyone on an insulin pump. Create a custom dot phrase of insulin backup plan in EMR.

Presentation: 6/3/2024
8642 Continuous glucose monitor: reclaiming type 2 diabetes self-efficacy and mitigating disparities
Authors: Ni Kevin , Tampe Carolyn A , Sol Kayce , Cervantes Lilia , Pereira Rocio Ines
Published: October 2024

Disclosure: K. Ni: None. C.A. Tampe: None. K. Sol: Employee; Self; Patient accepted job with Dexcom Jan 2024. L. Cervantes: None. R.I. Pereira: None.

Context: The increase in continuous glucose monitor (CGM) use has been notable for widening disparities between the least and most socially marginalized. Given access barriers, there is limited CGM patient experience information that is inclusive of those with type 2 diabetes mellitus from socially marginalized backgrounds. Objective: To understand the CGM usage experience across a U.S. Medicaid population with type 2 diabetes at federally qualified health centers (FQHCs). Methods: This qualitative study used semi-structured phone interviews with 28 English or Spanish speaking participants prescribed CGM who were enrolled in a U.S. Medicaid program that subsidized CGM. Audio-recordings of interviews were transcribed and analyzed by inductive thematic analysis. Results: Participants had a median age 56 (interquartile range 48-60) with varying levels of formal schooling (55% with 9 years or less) and were from diverse racial or ethnic backgrounds (57% Hispanic, 18% non-Hispanic Black, 21% non-Hispanic White). Interviews were conducted in English (68%) or Spanish (32%). We identified six major themes: initial expectations and overcoming initiation barriers (subthemes: initial expectations, overcoming initiation barriers), convenience and ease promote daily use (subthemes: benefits over fingersticks), increased knowledge leads to improved self-management (subthemes: increased understanding and knowledge, glucose level self-awareness, motivating healthy behaviors), collaboration with provider and clinical team, improved self-reported outcomes (subthemes: improved glucose and non-glucose targets, positive emotional effect, essential for management), and barriers and burdens are generally tolerated (subthemes: barriers to monitoring, burden of self-monitoring). Conclusions: CGM was experienced as easy to understand and use and viewed as a tool for diabetes self-efficacy. Expanded CGM access for socially marginalized patients with type 2 diabetes can enhance diabetes self-management to help mitigate diabetes outcome disparities.

Presentation: 6/2/2024
12392 Decoding LADA: A Closer Look At A Suspected Case Influenced By Prior IVIG Therapy
Authors: Gondhi A , Antharam S , Bantu S S , Abdulhadi B
Published: October 2024

Disclosure: A. Gondhi: None. S. Antharam: None. S.S. Bantu: None. B. Abdulhadi: None.

Introduction: LADA(Latent Autoimmune Diabetes of Adults) is defined as a new diagnosis of diabetes after the age of 35, presenting with features of T2DM but with the presence of T1DM associated autoantibodies. Correct diagnosis of LADA is important since prognosis and therapy differs from T2DM. We report an interesting case of transient elevation of Glutamic Acid decarboxylase (GAD-65) antibodies due to IVIG therapy leading to a misdiagnosis of LADA. Case presentation:A 46-year-old women with hypothyroidism was referred for management of LADA. A few months prior to the referral, she was hospitalized for acute bacterial meningitis and developed flaccid quadriparesis concerning for Guillain Barre syndrome (GBS). She received multiple IVIG infusions, steroids with resultant hyperglycemia requiring insulin and was transferred to a rehabilitation facility. During her stay in rehab, her initial dose of glargine 20 units twice daily was titrated down rapidly once steroids are tapered. Blood work revealed HbA1C 6.1%, blood glucose 116 mg/dL, C-peptide levels 4.1 ng/ml (0.9 -7 ng/ml), and GAD-65 antibodies >250 (N<5 IU/ml). She was diagnosed with LADA and discharged on 2 units of glargine. Her primary care physician advised to stay on the 2 units of glargine since her GAD-65 antibodies were significantly elevated and then referred to the endocrine clinic. Repeat bloodwork in our clinic revealed HbA1c of 5.8%, GAD-65 antibodies were repeated twice and were found to be undetectable <5. She was taken off insulin and has been doing well since. Discussion:IVIG is derived from plasma pooled from donors, used to treat a variety of diseases. Autoantibodies contained in IVIG may result in false positive serum tests if levels are checked soon after the infusion. This phenomenon has been documented in previous cases involving treponemal, H. pylori, hepatitis C as well as ANA antibodies. Increases in anti-GAD levels post-IVIG therapy have been previously reported. We saw a similar scenario in our patient, who was initially diagnosed with LADA and started on insulin due to an observed elevation in GAD 65 levels. However, this elevation was transient with negative repeat levels a few months later. Conclusion: Our case highlights the importance of recognizing that IVIG infusions can lead to the transfer of and to transient elevations in autoantibodies including GAD-65, which may lead to misdiagnosis and unnecessary therapies. References: 1. de Beer, F. C., Schreurs, M. W. J., & Foncke, E. M. J. (2009). False positive autoantibodies to glutamic acid decarboxylase in opsoclonus-myoclonus-ataxia syndrome after intravenous treatment with immunoglobulin. Clinical neurology and neurosurgery, 111(7), 643-644.

Presentation: 6/3/2024
8569 Defining Cell-Intrinsic Defects In Hepatic Insulin Resistance In Type 2 Diabetes Using A Human iPS Cell-Derived Disease-In-A-Dish Model
Authors: Gattu A , Tanzer M , Yaron T , Cantley L , Kahn C R
Published: October 2024

Disclosure: A. Gattu: None. M. Tanzer: None. T. Yaron: None. L. Cantley: None. C.R. Kahn: None.

Hepatic insulin resistance is central to type 2 diabetes (T2D). Understanding the cellular drivers underlying insulin resistance in the liver is challenging since, in vivo, this can be due to cell-intrinsic defects in insulin signaling, as well as the effects of multiple circulating factors that can modify insulin signaling. Here, we have utilized inducible pluripotent stem cells (iPS) from 8 control and 8 T2D patients differentiated in vitro into hepatocytes (iHeps) to dissect the cell-intrinsic defects in hepatic insulin resistance. We find that following stimulation with insulin, T2D iHeps displayed “pathway-selective insulin resistance,” in which insulin failed to suppress gene expression of critical gluconeogenic enzymes (PCK1, G6PC) but continued to stimulate expression of the lipogenic enzymes (FASN, ACACA). This was associated with a 1.5-fold higher level of insulin stimulation of C[1]4 acetate into lipids, i.e., de novo lipogenesis. Immunoblotting analysis showed T2D iHeps exhibited a 50% reduction in phosphorylation of the insulin receptor IRY[1][1]35 and ∼ 30% reductions in AKTT308, GSK3αS2[1]/GSK3βS9, and FOXO1T24/FOXO3aT32 in T2D following insulin stimulation for 10 minutes. To define the full spectrum of alterations in signaling in iHeps, we used unbiased LC-MS/MS-based phosphoproteomics. This revealed alterations in 987 insulin-regulated phosphosphorylations that had increased phosphorylation after insulin treatment and alterations in 723 sites for which insulin decreased phosphorylation (with a |fold-change| >1.5 and adj. p < 0.05). These alterations were of two types: 43% of the altered insulin-regulated phosphosites showed reduced or “impaired” insulin regulation, but 57% of the phosphosites displayed new or significantly increased insulin-regulated phosphorylations in T2D, i.e., “emergent” signaling. The impaired signaling in T2D included losses in the insulin receptor signaling cascade, Rho-GTPase pathway, Notch-HLH, and anti-viral mechanisms of IFN-stimulated genes. In contrast, emergent signaling included other components of the Rho-GTPase pathway, metabolism of RNA, membrane trafficking, and chromatin-modifying enzymes. Using an AI-based kinase-substrate association analysis, we found that the deficiencies in signaling represented reduced actions of kinases AKT2, AKT3, PKCθ, CHK2, PHKG2, and/or STK32C kinases, while the emergent changes represented increases in ROCK1, ROCK2, MST4 and/or BCKDK kinase activity. In summary, phosphoproteomic analyses of iHeps revealed a comprehensive insulin signaling network disrupted in insulin resistance in the liver in T2D, which underlies the etiology of insulin resistance. These alterations can be ascribed to changes in the activity of two classes of upstream kinases, representing potential new targets for treating T2D.

Presentation: 6/1/2024
12474 Demographic Characteristics Of Patients With Prediabetes Who Reverted To Normoglycemia Or Progressed To Diabetes At 1 Year Follow Up
Authors: Niedzialkowska Ewelina Marta , Sayed Farah El , Townsend Mary , Bhatia Unnati , Roy Sujoy , Homayouni Ramin , Halalau Alexandra
Published: October 2024

Disclosure: E.M. Niedzialkowska: None. F. El Sayed: None. M. Townsend: None. U. Bhatia: None. S. Roy: None. R. Homayouni: None. A. Halalau: None.

Background: Pre-diabetes (PD), characterized by hyperglycemia influenced by lifestyle factors, genetics, age, and ethnicity, is considered a risk factor for development of diabetes. However, only a small proportion of PD individuals progress to type-2 diabetes (T2D) each year. More refined diabetes risk prediction tools are needed to direct proactive health management strategies for at-risk populations. Question and Significance: Our study explored the associations between age, race, gender and BMI in PD individuals who either progressed to T2D or reverted to normoglycemia (NG) over a one-year follow-up period. Methods and Design: This is a retrospective observational study of 38,228 patients with PD, based on having two consecutive years of hemoglobin A1c (HbA1c) between 5.7% and 6.5%. We compared the demographic characteristics of individuals who progressed to T2D or regressed to NG within one year using bivariate (Kruskal-Wallis or Chi-squared) or multivariate logistic regression analysis. Results: The study cohort included 57.36% females, 66.13% Whites, 19.41% Blacks and 4.36% Asians with a median age of 69. Bivariate analysis revealed significant age differences between PD patients who reverted to NG with median age of 67 in those who reverted vs 70 in those who did not revert (p< 0.0001). In addition, the proportion of males who progressed to T2D was significantly higher and the proportion of Blacks who reverted to NG was significantly lower.Using multivariate regression (adjusting for age, gender, race and BMI), we found that each year in age significantly lowered the odds of reversion from PD to NG (OR 0.98, p<0.0001) but was not associated with progression to T2D. PD males had significantly higher odds of progression to T2D (OR 1.28, p<0.0001). Both Asians (OR 0.62, p<0.0007) and Blacks (OR 0.68, p<0.0004) had significantly lower odds of reversion to NG from PD compared to Whites. Surprisingly, BMI was not associated with neither reversion to NG nor progression to T2D, after adjusting for age, gender, and race. Discussion: Our findings underscore significant racial disparities and gender differences in regression to normoglycemia and progression to diabetes. This emphasizes the need for personalized interventions and targeted strategies for prediabetic populations to mitigate progression to T2D and to achieve normoglycemia.

Presentation: 6/1/2024