bluuebunny/crossref_metadata_2025_split huggingface dataset:
SAT058 Diabetes Complicated By DKA, Mucormycosis And Covid-19
Authors: Gordon Ashley 
Published: October 2023

Abstract
Disclosure: A. Gordon: None.
Background: Mucormycosis is an invasive opportunistic fungal infection, that although rare, has an estimated all-cause mortality of 54%. Even with aggressive treatment, significant tissue necrosis and bone destruction can occur. Risk factors include diabetes, acidosis (as in diabetic ketoacidosis), excess iron, transplant, malignancy, kidney and liver disease, immunosuppressant and corticosteroid use - and in more recent times, the Covid-19 infection. We present a case of invasive rhino-orbital cerebral mucormycosis in the setting of DKA, uncontrolled diabetes and Covid-19. Case: A 60-year-old female with history of uncontrolled type 2 diabetes (HbA1c 14.3%) presented after being found unresponsive. Her exam was significant for encephalopathy, complete left eye ophthalmoplegia and right-sided hemiparesis. She was hemodynamically stable without fever. Initial labs showed significant hyperglycemia with an elevated anion gap and beta-hydroxybutyrate indicative of diabetic ketoacidosis (DKA). She had a lumbar puncture which showed leukocytosis with neutrophilia (146 WBC, 39% neutrophils), elevated glucose and normal protein; CSF culture was negative for growth. She also tested positive for Covid-19 although CXR was unremarkable and she did not require treatment. Initial CT/CTA head/neck showed complete occlusion of the left cervical internal carotid artery with associated acute/subacute infarcts in the left frontal, parietal, temporal and occipital lobes. CT sinuses showed severe frontal, maxillary and ethmoid sinus mucosal wall thickening with left premaxillary soft tissue swelling and leftward nasal septum deviation. Finally, orbital imaging showed left cavernous sinus thrombosis, left superior ophthalmic vein thrombosis, and necrosis vs devascularization of the extraocular muscles. She was initially treated with fluids, an insulin infusion and broad-spectrum antibiotics and was evaluated by neurology, ophthalmology, infectious disease and otolaryngology. She underwent surgical debridement with left maxillary antrostomy, left total ethmoidectomies and septoplasty. Sinus cultures were positive for MSSA (methicillin sensitive staph aureus) and tissue pathology showed angioinvasive fungal disease compatible with Mucor/Rhizomucor species. Antimicrobial coverage was changed to intravenous Nafcillin and Amphotericin B, and she underwent further surgical debridement twice more. Repeat brain imaging to evaluate for disease progression is pending. Discussion: Early detection and aggressive treatment of Mucormycosis with surgical debridement and Amphotericin B can improve patient outcomes, however the mortality rate is still significant. Therefore, tight glycemic control in high risk groups is imperative, particularly in those with uncontrolled diabetes complicated by DKA and Covid-19 infection (with or without steroid treatment).
Presentation: Saturday, June 17, 2023
SAT061 Deficiency Of ENaC Excites Pancreatic β Cells To Promote Insulin Secretion
Authors: Que Yanting , Ma Xiyang , Wu Yong , Chen Junjiang , Guo Jinghui , Chun Ruan Ye 
Published: October 2023

Abstract
Disclosure: Y. Que: None. X. Ma: None. Y. Wu: None. J. Chen: None. J. Guo: None. Y. Ruan: None.
In response to glucose elevation or other physiological stimuli, pancreatic islet β cells are excited to mobilize intracellular Ca2+ leading to insulin secretion, which is a complex cellular event with underlying mechanisms not fully elucidated. The present study explored possible involvement of the epithelial Na+ channel (ENaC) in β cell excitability and insulin secretion. Analyzing human databases, primary rat/mouse pancreatic tissues as well as RINm5F, a rat β-cell line, we confirmed the expression of Scnn1a, Scnn1b and Scnn1g genes (encoding ENaC subunits, α, β and γ, respectively) in human and rodent β cells. To our surprise, inhibiting this Na+ channel by selective blockers, amiloride (1-10 µM) or benzamil (1 µM), did not retard insulin secretion, but instead triggered a slow membrane depolarization with electrical bursts (41.5 ± 5.9 mV, measured by patch-clamp, n = 5-6), elicited substantial Ca2+ oscillations (135.2 ± 2.5% of baseline, by Fura-2 imaging, n = 256-380) and promoted insulin secretion (158.2 ± 25.6% of control, by ELISA, t-test, p &lt; 0.05, n = 6) in RINm5F or isolated mouse β cells. siRNA-based knockdown of ENaCα, the rate-limiting subunit of ENaC, in RINm5F cells confirmed that deficiency of ENaC induced a significant increase in insulin secretion (230.2 ± 20.2% of control, t-test, p &lt; 0.001, n = 6). Proteomic analysis of RINm5F cells (n = 5) through mass spectrometry showed that signaling pathways key to glucose metabolism and insulin secretion were significantly activated in RINm5F cells with ENaC knockdown in comparison with control cells, consistently suggesting a role of ENaC deficiency in exciting β cells to release insulin. We next built a mouse model with β cell-speciﬁc knockout of ENaCα (Scnn1afl/fl, Ins1-Cre+), which exhibited disturbed responses in glucose tolerance test in comparison with the loxp-negative Cre control mice (Scnn1awt/wt, Ins1-Cre+). Taken together, these results have suggested an important role of ENaC in regulating the excitability of β cells and insulin secretion, which may contribute to the understanding of Na+ environment in relation to insulin homeostasis. This work was supported in part by National Natural Science Foundation of China (82071599), Areas of Excellence Scheme of Hong Kong (AoE/M-402/20) and Bai Cheng Bai Yuan Start-up Fund (I2022A008).
Presentation: Saturday, June 17, 2023
SAT063 Dopaminergic Agonist Cabergoline Inhibits Autophagic Flux In Beta Pancreatic Cells
Authors: Francisco Mendez Luis , Etcheverry Boneo Luz , Becu-Villalobos Damasia , Sorianello Eleonora 
Published: October 2023

Abstract
Disclosure: L.F. Mendez: None. L. Etcheverry Boneo: None. D. Becu-Villalobos: None. E. Sorianello: None.
Autophagy is a lysosomal degradation process necessary for the maintenance of cellular homeostasis, which is based on constantly removing ubiquitinated proteins and dysfunctional organelles that are eventually toxic to the cell. Since alteration of this process in pancreatic beta cells can lead to the development of diabetes, autophagy modulators arise as interesting therapeutic compounds for the treatment of this prevalent human pathology. Previous studies indicate that activation of the dopamine D2 receptor (D2R) alters autophagy in several cell types. In this context, we aimed at determining the effect of D2R activation by its agonist Cabergoline (CAB) on the autophagic process in pancreatic beta cells. To that end, we stimulated the murine pancreatic beta cell line MIN6B1 with 10-5 M CAB for 1, 6 and 24h to evaluate the kinetics of autophagic vesicle formation. Additionally, 10-5 M CAB in the presence or absence of Chloroquine (CQ), an inhibitor of late stages of the autophagy process, was used to study the autophagic flux. Using Western Blot and immunofluorescence and confocal microscopy, the autophagy markers LC3 (autophagic vesicle marker) and p62/SQSTM1 (autophagy cargo receptor and degradation substrate) were analyzed. CAB increased LC3 nucleation as a function of stimulation time, showing significant differences relative to the control after 6h (one-way repeated measures ANOVA p=0.0038; Tukey: Control vs CAB6h p&lt;0.05, vs CAB24h p&lt;0.01). Moreover, CAB significant increased p62/SQSTM1 levels after 24h of stimulation (one-way repeated measures ANOVA: p=0.0157; Tukey: Control vs CAB24h p&lt;0.05). When studying the effect of CAB on the autophagic flux, we observed that CQ significantly increased LC3 and p62/SQSTM1 levels at 24h of incubation (two-way repeated measures ANOVA (CQ and CAB): for LC3: interaction p&lt;0.05, Tukey: CQ vs Control p&lt;0.0001; for p62/SQSTM1: CQ pretreatment effect p&lt;0.05), as expected. Interestingly, CAB diminished the accumulation of LC3 (Test-T Delta (CQ-Control) vs Delta (CQCAB-CAB): p=0.04) and p62/SQSTM1 (Test-T Delta (CQ-Control) vs Delta (CQCAB-CAB): p=0.01) elicited by CQ at 24h. We conclude that CAB is able to increase autophagic vesicle formation and, in addition, to decrease autophagic flux after 24h of stimulation in MIN6B1 pancreatic beta cells. This work was funded by CONICET, ANPCyT, Fundación René Barón and Fundación Williams.
Presentation: Saturday, June 17, 2023
SAT067 Bone Marrow-induced Epithelial-Mesenchymal Transition In Beta Cells
Authors: Lee Jihyun 
Published: October 2023

Abstract
Disclosure: J. Lee: None.
Normal cellular function is achieved via regulated cell differentiation cues. Deregulation of these intricately wired processes promotes cellular dysfunction. The pancreatic islet beta cell (β-cell) is a prime example of such integration between optimal cell differentiation and function. Fully differentiated adult β-cells synthesize and secrete insulin. But some express more than one single hormone, and cells containing insulin and glucagon (Ins+:Glu+ cells) are seen during prenatal pancreas development. Interestingly, Ins+:Glu+ cells have been observed in adult islets from diabetic mice or type 2 diabetes (T2D) human cadavers. It is still unclear the origins and relevance of these Ins+:Glu+ cells, and whether this phenomenon is a benefit in the context of severe β-cell stress. But, growing evidence suggests that Ins+:Glu+ cells represent attempts made by the mature β-cell to return to an immature stage with the goal of proliferating and reconstituting depleted β-cell reservoir when faced with a metabolic challenge. In this study, we thus investigated whether adult β-cells undergo epithelial to mesenchymal transition (EMT) that allows β-cells to retrace the developmental steps and attempt β-cell functional recovery. EMT can be induced by many growth factors and cytokines activated when epithelial cells encounter specific signals released by their immediate neighbors. The bone marrow microenvironment can activate EMT in adjacent cells through the secretion of chemokines, cytokines, and growth factors. We postulated bone marrow cells (BMCs) could induce EMT in β-cell. To accomplish this goal, we have developed islet cell and bone marrow cell co-culture conditions. Islets and bone marrow cells obtained from normal chow diet (NCD) and high-fat diet-fed (HFD) mice as well as specimens from non-diabetic and T2D humans are used in co-culture systems. RNA sequencing analysis showed that co-culture pancreatic islet cells with whole bone marrow cells induced the expression of genes relative to mesenchymal cells and suppressed beta cell marker genes like MafA, Ins. Results from immunofluorescence staining analysis also revealed that BMCs promoted mesenchymal phenotype in islet β-cells. We found that cytokines known to induce EMT were secreted in the co-culture media. Mice fed with HFD increased levels of that cytokines in serum compared to NCD. We further confirmed that cytokines secreted by BMCs activated EMT in mouse and human islet β-cells. Blocking of cytokines signaling decreased mesenchymal phenotypes in islet β-cells. Overall, our data suggest that EMT of β-cells is regulated by bone marrow cells via crosstalk between bone marrow cells and pancreatic islets using cytokines.
Presentation: Saturday, June 17, 2023
SAT069 Targeting Human β-Cell G Protein-coupled Receptors To Alter Cytokine-mediated Caspase Activation
Authors: dos Santos Karina Rodrigues , Mukherjee Noyonika , Barella Luiz F , Armoo Fiona , Eizirik Decio L , Templin Andrew T , Kalwat Michael A 
Published: October 2023

Abstract
Disclosure: K. Rodrigues dos Santos: None. N. Mukherjee: None. L.F. Barella: None. F. Armoo: None. D.L. Eizirik: None. A.T. Templin: None. M.A. Kalwat: None.
In type 1 diabetes (T1D) autoimmune destruction of pancreatic islet β-cells causes a lack of insulin, leading to hyperglycemia. The development of therapies that can prevent β-cell death is critical. β-cell proliferation and survival pathways are affected by signaling through G protein-coupled receptors (GPCRs). Using high-depth RNA-seq, we found altered expression of 123 different GPCRs in human islets exposed to cytokines (IL1β 50U/ml + IFNγ 1000U/ml, 48 h) to model the T1D milieu. We hypothesized these candidates may be exploited for β-cell protection. Among the candidates, adenosine receptors (ADORA1 and ADORA2A), calcium-sensing receptor (CASR), and metabotropic glutamate receptor 4 (GRM4) stood out. Cytokines decreased the expression of ADORA1 and CASR, but increased ADORA2A and GRM4. To gauge the impact of agonizing/antagonizing these receptors, we pre-treated human EndoC-β H1 β-cells or islets with receptor ligands and co-treated with cytokines (IL1β 50U/ml + IFNγ or IL1β + 1000U/ml IFNγ + 1000U/ml TNFɑ, 48 h) and measured glucose-stimulated insulin secretion, cell viability, and gene expression. Each cytokine combination induced target gene expression (e.g. CXCL10), but only the combination of IL1β + IFNγ + TNFɑ reduced glucose-stimulated insulin secretion. Preliminary experiments indicated that adenosine receptor ligands had little impact in our assays. However, ligands for CASR and GRM4 exhibited significant suppression of cytokine-induced caspase 3/7 activation in EndoC-βH1 cells. Interestingly, suppressed caspase activation did not result in observable impacts on live/dead cell ratios or live-cell analysis using Incucyte in EndoC-βH1 cells. Consistently, we also observed cytokine-mediated cell death in the presence of the pan-caspase inhibitor ZVAD-fmk. We conclude that a variety of β-cell GPCR-mediated signaling pathways may converge to prevent cytokine-induced caspase activation, but additional measures are needed to enable protection from eventual cell death. Our results and this line of investigation is significant because understanding mechanisms of GPCR-cytokine crosstalk in β-cells will inform strategies to protect islets in other models of T1D.
Funding: JDRF 1-INO-2022-1113-A-N and Diabetes Research Connection #33.
Presentation: Saturday, June 17, 2023
SAT071 Inhibition Of 12 Lipoxygenase In A Humanized Mouse Model Of Type 1 Diabetes Delays Progression Of Hyperglycemia
Authors: Nargis Titli , Chakraborty Advaita , Figatner Kayla , Maloney David , Boxer Matthew , Tersey Sarah A , Mirmira Raghavendra G 
Published: October 2023

Abstract
Disclosure: T. Nargis: None. A. Chakraborty: None. K. Figatner: None. D. Maloney: None. M. Boxer: None. S.A. Tersey: None. R.G. Mirmira: None.
Type 1 diabetes (T1D) is an autoimmune disorder characterized by islet inflammation (insulitis). We have recently shown that a key mediator of inflammatory signaling in the islet beta cell is the enzyme 12-lipoxygenase (12-LOX), which generates proinflammatory eicosanoids that augment dysfunctional insulin secretion and beta-cell visibility to the immune system. Whereas inhibition of 12-LOX offers an opportunity to modify T1D progression by altering how the beta cell responds to insulitis, a barrier to testing inhibitors in preclinical models is the specificity of next generation inhibitors toward the human enzyme. To generate a preclinical platform for the study of human-specific 12-LOX inhibitors, we developed a mouse model in which the mouse-equivalent Alox15 coding region was replaced with the human ALOX12 gene, while retaining the mouse upstream control region. These mice (hALOX12) were developed on both the C57BL/6J and NOD genetic backgrounds, then subjected to treatment with the human 12-LOX inhibitor VLX-1005 to evaluate diabetes progression. C57BL/6J-wildtype and C57BL/6J-hALOX12 mice were confirmed to express exclusively mouse Alox15 and human ALOX12, respectively, using RNA isolated from islets. To test susceptibility to toxin-induced diabetes, C57BL/6J-wildtype and C57BL/6J-hALOX12 were subjected to multiple low-dose streptozotocin (STZ) to induce diabetes. Both wildtype and humanized mice exhibited gradual and equivalent hyperglycemia (within 25 days) following STZ injection. Following intraperitoneal VLX-1005 therapy (30 mg/kg), whereas C57BL/6J-wildtype mice developed diabetes (with a delay of 10 days) following STZ, C57BL/6J-hALOX12 mice showed complete protection against STZ-induced diabetes and exhibited significantly better glucose tolerance compared to vehicle- or VLX-1005-injected wildtype controls and vehicle-injected C57BL/6J-hALOX12 mice. Next, hALOX12 mice were backcrossed onto the NOD.ShiltJ background model of autoimmune T1D. NOD-hALOX12 mice received either vehicle or VLX-1005 (30 mg/kg) orally during the prediabetic phase (6-10 weeks of age) and were followed for diabetes incidence. Mice receiving VLX-1005 showed a significant delay with only 30% of female and 7% of male mice developing diabetes over 25 weeks of age. Pancreatic sections from the mice treated with VLX-1005 showed reduced insulitis and increased beta cell mass compared to vehicle controls. In summary, the hALOX12 mouse model serves as a preclinical translational platform to interrogate effects of next generation 12-LOX inhibitors and demonstrates the potential of VLX-1005 to modify T1D disease progression.
Presentation: Saturday, June 17, 2023
SAT092 A Case Of Mature Onset Diabetes Of The Young
Authors: Beshay Lauren , Thakkar Aditi 
Published: October 2023

Abstract
Disclosure: L. Beshay: None. A. Thakkar: None.
Background: Maturity onset diabetes of the young (MODY) is a disorder characterized by noninsulin-dependent diabetes diagnosed at a young age (&lt;25 years) with autosomal dominant inheritance and lack of autoantibodies. It accounts for 2 to 5 percent of diabetes cases. Several different genetic abnormalities have been identified causing disruption in pancreatic beta cells development and function leading to impaired insulin secretion. Mutations in hepatocyte nuclear factor-1-alpha (HNF1A) is the most commonly identified mutation occurring in 52 to 65 of MODY cases. Case Description: A 21 year old female with a history of diabetes and recurrent skin abscesses presented to the emergency room with right lower quadrant epidermal abscess. Endocrinology was consulted for evaluation of her diabetes. She was diagnosed at age 14 when she presented with weight loss, polyuria, polydipsia, BG 406 without diabetic ketoacidosis. She reported a history of type 2 diabetes in mother. Her medications included metformin, lantus and humalog. It was noted that her diabetes was never controlled with an A1c persistently &gt;10%. Patient reported skin swelling with insulin injections and working two jobs which limited her ability to take more than one injection per day. Her BMI is 23 kg/m2. Labs this admission showed HbA1c 13.7%, GAD 65 Ab and IA-2 Ab negative, c-peptide 1.1 ng/ml, with a glucose of 209 mg/dl. MODY was suspected due to her age, intact c-peptide, and lack of DM1 antibodies. The patient was discharged on Trulicity 0.75 mg weekly, Lantus 12U nightly, Glipizide 2.5mg daily. On follow up, a MODY genetic panel confirmed a HNF1A missense variant with AD inheritance. Since initiation of sulfonylurea, she has had improved control of diabetes. Glipizide dose is being titrated up with concomitant decrease in insulin dose with plan to discontinue basal insulin once her blood sugars are at goal. Conclusion: The HNF1A gene on chromosome 12 is a weak transactivator of the insulin gene in beta cells. Mutations of HNF1A can lead to abnormal insulin secretion and a low renal threshold for glucose. These patients are at risk for micro- and macrovascular complications of diabetes. In terms of treatment, they exhibit increased insulin sensitivity and marked sensitivity to the hypoglycemic effects of sulfonylureas compared with patients with type 2 diabetes. Thus, patients can be successfully treated with sulfonylurea monotherapy, and in one clinical study, approximately 70 percent of patients previously treated with insulin successfully switched to sulfonylureas. Thus, it is important to evaluate etiology of diabetes, as detection of MODY variants in which sulfonylureas are used for treatment simplifies management and improves outcomes as demonstrated in this case.
Presentation: Saturday, June 17, 2023
SAT122 The Barriers Of Insulin Access In A Case Of Wolfram Syndrome
Authors: Sacoto Daniel Hernan , Villavicencio Camila Alejandra , Belokovskaya Regina , Franco-Akel Alberto A 
Published: October 2023

Abstract
Disclosure: D.H. Sacoto: None. C.A. Villavicencio: None. R. Belokovskaya: None. A.A. Franco-Akel, MD: None.
Wolfram syndrome (WS) is a rare and fatal condition with a median age of death of about 39 years and affecting around 1 of 100,000 people in North America. Insulin is the mainstay of treatment in WS though, access to it may affect adherence and glycemic control which is correlated with neurodegenerative progression. We present a patient with WS whose complications exacerbated due to insulin access. A 19-year-old African American male with a history of type 1 diabetes mellitus (T1DM), presented to the ED with a 2 day-history of general malaise and emesis. Diabetic ketoacidosis (DKA) (blood glucose 802 mg/dL, pH 7.2, positive serum ketones) was diagnosed upon arrival. Right optic nerve atrophy was diagnosed at age of seven. At age of eight, patient was diagnosed with bilateral sensory-neuronal hearing loss (SNHL) which led to school withdrawal. It was then when suspicion of WS arose. Single nucleotide polymorphism microarray confirmed homozygosity from the region of linkage on chromosome 4p16.1, where the WFS1 gene lies. At the age of eighteen, patient was diagnosed with neurogenic bladder for which he requires frequent self-catheterization. Since his T1DM diagnosis, patient has faced multiple barriers accessing insulin secondary to insurance issues and lack of social support resulting in multiple hospital admissions for DKA and urinary retention episodes. WS is an autosomal recessive disorder resulting from WFS1 or WFS2 gene mutation on chromosome 4p. WS commonly presents as childhood-onset diabetes mellitus, optic atrophy, SNHL, and neurogenic bladder. Insulin is the first line treatment for optimal glycemic control in WS. Despite optimal glycemic control, WS symptoms and neurodegenerative progression may be influenced by other factors. First, treatment access for rare diseases has shown to be impacted by medical unconscious bias, in addition to systemic, linguistic, and socioeconomic challenges patients may face. Furthermore, Medicaid insurance covering rare diseases still varies widely between 15-67%, and in those insured, there is still a 61% chance of delay or denial due to pre-approval requirements. In addition, poor glycemic control has been correlated with lower socioeconomic status, lack of access to devices such as insulin pumps, migratory status, social support and environment, food security, and ethnic background. Since diabetes control in WS patients is paramount to a better quality of life, additional studies are needed to understand the full scope of barriers that can contribute to policies that facilitate better treatment access for patient and families living with WS.
Presentation: Saturday, June 17, 2023