bluuebunny/crossref_metadata_2025_split huggingface dataset:
1482-P: Neutrophil Inflammasome Is Activated in Diabetes and Contributes to the Exacerbated NETosis
Authors: DE WANG LIANG , DALAN RINKOO , LING WONG SIU 
Published: June 2023

Activation of the NOD-like receptor (NLR) family, pyrin domain-containing protein 3 (NLRP3) inflammasome has recently been implicated in driving the pathogenesis of diabetic complications. While diabetes is known to increase NETosis propensity, little is known whether neutrophil NLRP3 mediates NETosis in diabetes, and if organelles that govern metabolic homeostasis contribute to aberrant activation of NLRP3. In this study, we sought to examine if neutrophil NLRP3 is activated in diabetes, and if key metabolic organelles are involved in the activation of NLRP3, resulting in NETosis upregulation. We found that pre-treatment of blood neutrophils isolated from mice and humans with diabetes using MCC-950, a potent and specific NLRP3 inhibitor, normalized NET formation to levels similar to those of neutrophils isolated from healthy mice and humans. Similar to blood neutrophils, HL-60-derived neutrophils primed in high glucose also showed reduced NETosis when pre-treated with MCC-950. Notably, compared to their normoglycemic counterparts, neutrophils isolated from mice and humans with diabetes exhibited increased speck formation of the inflammasome adaptor protein, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) even at baseline, indicative of heightened NLRP3 activation in diabetes. In mouse neutrophils, confocal microscopy revealed that oxidized DNA release into the cytoplasm is a prerequisite for ASC speck formation, which co-localized with mitochondria. In addition, MCC-950 yielded no further reduction in NET formation when HL-60-derived neutrophils were pre-treated with tauroursodeoxycholic acid, an alleviator of endoplasmic reticulum (ER) stress. In summary, our findings suggest that mitochondria and ER stress may synergize in the activation of neutrophil NLRP3 that exacerbates NETosis in diabetes. Therapeutics that aim at intercepting the pathway may alleviate NET-mediated chronic inflammation in diabetes.

Disclosure
L.Wang: None. R.Dalan: None. S.Wong: None.

Funding
Nanyang Assistant Professorship; Singapore Ministry of Education Academic Research Fund Tier 1 (RG25/20); Nanyang President's Graduate Scholarship
1483-P: Distinct Immune Features in the Partial Remission Phase of Type 1 Diabetes Identified by Single-Cell RNA Sequencing and Functional Analysis
Authors: LI XIA , ZHONG TING , LI XINYU , KANG LEI , TANG RONG 
Published: June 2023

Type 1 diabetes (T1D) patients might experience a transient partial remission (PR) stage. However, the immunological mechanisms responsible for the PR stage still need to be elucidated. Here, we have applied single-cell RNA sequencing to delineate the immune underpinnings of peripheral immune cells among healthy individuals, new-onset T1D patients, and T1D patients in the PR phase. Combined with cohort validation and functional assays, the expansion of TIGIT+ CCR7− Tregs as well as the decrease of CD226+ CCR7− CD8+ T cells helps to maintain a window in which to transit the immune balance in the remission stage via the TGF-β signal. These findings, especially the identified cell subsets may provide novel mechanism for future therapeutic interventions.

Disclosure
X.Li: None. T.Zhong: None. X.Li: None. L.Kang: None. R.Tang: None.
1487-P: Inosine Monophosphate Preferentially Regulates Myeloid Progenitor Differentiation toward Neutrophil Production in Diabetes
Authors: DONG YUAN , YAN CEN , ZHANG YEUJIE , YANG XUANYI , FENG YINGMEI 
Published: June 2023

Previous studies described hyperglycemia-induced myelopoiesis and atherosclerotic progression in mice with type I diabetes. By targeted metabolomics, we reported the increase of inosine monophosphate (IMP) in granulocyte/monocyte progenitors (GMP) in db/db mice. Moreover, IMP treatment stimulated GMP proliferation, resulting in skewed myelopoiesis and enhanced acute pancreatitis progression. However, how IMP modulates granulocyte production is not known. To investigate that, equal amount of FACS-sorted GMP of wild-type mice were cultivated with PBS or 15 mM IMP for 5 days in vitro. FACS analysis revealed that the absolute number of neutrophils was 1.7-fold higher but the number of CD11b+ monocytes was 3.2-fold lower in IMP group compared to control (p&lt;0.05 for both). Thereafter, FACS-sorted GMP treated with or without IMP for 3 days were harvested for single cell RNA sequencing. When classified by marker gene expression, 19 clusters representing 7 cell types were identified including neutrophils, monocytes, GMP, intermediate GMP/neutrophils, basophils, Megakaryocyte erythroid progenitors/megakaryocytes (MEP/Mgk), and platelets. The proportions of neutrophils and MEP/Mgk increased 1.3-, and 1.6-fold in IMP-treated GMP cells compared with controls (p=0.059 for neutrophils; p=0.032 for MEP/Mgk). Nevertheless, the percentage of other cell populations were comparable between two groups (p≥0.08 for all). Furthermore, cycling neutrophils in cluster 3 was 1.4-fold increase in IMP-treated GMP cells compared with controls (p=0.04). Accordingly, GO analysis and Gene set enrichment analysis illustrated that genes involved in cell cycles was enriched in cycling neutrophil clusters. In conclusion, our data demonstrated that IMP preferentially regulated GMP proliferation and differentiation toward neutrophil production, which provides new insights on the crosstalk between skewed metabolites and inflammation in diabetes.

Disclosure
Y.Dong: None. C.Yan: None. Y.Zhang: None. X.Yang: None. Y.Feng: None.

Funding
National Natural Science Foundation of China (81670765, 82070841)
1488-P: Reduced iPLA2b Expression in T-Lymphocytes Decreases Proinflammatory Eicosanoid Production and Cytokine Secretion Delaying Type 1 Diabetes Development in NOD Mice
Authors: WHITE TAYLEUR , RAMANADHAM SASANKA , DILORENZO TERESA P. , CHALFANT CHARLES E. 
Published: June 2023

Type 1 diabetes (T1D) is characterized by the destruction of insulin-producing pancreatic islet β-cells, but the causes of this process remain unclear. We find that Ca2+-independent phospholipase A2β (iPLA2β), which hydrolyzes membrane phospholipids at the sn-2 substituent to yield iPLA2β-derived lipids (iDLs), is induced under a diabetic milieu and mitigation of iPLA2β activity attenuates β-cell death. Several iDLs are pro-inflammatory and have been studied in different diseases but are not understood in diabetes. We investigated the role of immune-cell iDLs by utilizing T-cell adoptive transfer. Isolated CD4+/8+ T-cells from splenocytes prepared from donor female NOD and NOD. iPLA2β+/− (HET) mice, were transferred i.p. to NOD.scid recipient mice. We observed T1D onset for all T-cell recipient groups between 10-12 weeks of age. However, reduction of iPLA2β in CD4/8 T-cells delayed 50% incidence by 10 weeks in NOD4/HET8-recipient mice and HET4/NOD8 T-cell recipients compared to “matched” T-cell recipients. Furthermore, reduced insulitis and increased β-cell preservation was detected in HET4/NOD8-recipients compared to NOD4/8 recipients. The immune cell composition between our genotypes revealed no difference suggesting iDL signaling contributes to the observed incidence. Lipidomic analyses revealed a decrease in pro-inflammatory eicosanoids (PGs, DHETs and HETEs) in nondiabetic recipient mice, compared to diabetic counterparts. In addition, by targeting select eicosanoid signaling (PGE2 and DHET) utilizing chemical inhibitors (i.e., Grapiprant (EP4 receptor antagonist) and EC5026 (soluble epoxide hydrolase inhibitor)), we observed decreased IFN-γ production by Th1 helper cells in HET donors compared to NOD. These findings suggest that iPLA2β in T-cells is a critical contributor to the production of pro-inflammatory iDLs and could be a potential target in reducing T1D development.

Disclosure
T. White: None. S. Ramanadham: None. T.P. DiLorenzo: None. C.E. Chalfant: None.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (R01DK069455); National Institute of Allergy and Infectious Diseases (R21AI146743, R21AI169214); JDRF (2-SRA-2022-1210-S-B); University of Alabama at Birmingham (5T32GM8111-34)
1496-P: Resatorvid, an Inhibitor of Toll-Like Receptor-4, Protects Allogenic Islets from Inflammatory and Immune Response
Authors: MATTKE JORDAN D. , NAZIRUDDIN BASHOO , KANE BOB 
Published: June 2023

Purpose: Loss of islet mass by instant blood mediated inflammatory reaction (IBMIR) and immunological rejection are major hurdles currently facing the field of islet transplantation. Toll-like receptor 4 (TLR4) plays a central role in inflammation and shaping adaptive immune response. We investigated the protective effects of resatorvid (TAK-242) using in vitro models of islet transplantation.
Methods: Soluble TAK-242 or TAK-242 conjugated to the surface of islets utilizing copper-free click chemistry with a cleavable chemical linker were used. Islets were mixed with allogenic blood for a period of 3 hours and plasma levels of miRNA-375 and miRNA-200c were analyzed by RT-qPCR. Allogenic peripheral blood mononuclear cells were cultured with islets. T cell and monocyte activation were monitored by flow cytometry. Cytokine and miRNA levels in culture supernatant were measured by Luminex assay and RT-qPCR respectively.
Results: The presence of TAK-242 in soluble form as well as released by cleavable linker from islet surface significantly reduced the release of miRNA-375 (p&lt;0.01) and miRNA-200c (p&lt;0.01) indicating less islet damage. In allogenic co-culture of islets and PBMCs, TAK-242 inhibited the expression of CD69 in T cells as well as decreased (p&lt;0.05) the expression of CD86 and CD80 in monocytes. The production of cytokines IL-1β (p&lt;0.001), IL-6 (p=0.01), IL-8 (p&lt;0.05), IP10 (p&lt;0.05), MCP-1 (p&lt;0.001) and TNFα (p=0.005) in culture supernatant was decreased in the presence of soluble TAK-242 as well as TAK-242 released from islet surfaces.
Conclusions: Inhibition of TLR4 using small molecule, TAK-242, protected islets from innate damage due to IBMIR as well as adaptive responses of activation of T cells and monocytes. These results validate the future study of inhibition of TLR4 using in vivo transplant model(s) to further investigate the efficacy of TLR4 inhibition to improve islet transplant survival and function.

Disclosure
J.D.Mattke: None. B.Naziruddin: None. B.Kane: None.
15-OR: The Impact of Insulin Out-of-Pocket Caps among Commercially Insured Diabetes Patients
Authors: GARABEDIAN LAURA F. , ROSS-DEGNAN DENNIS , COSTA REBECCA , ARGETSINGER STEPHANIE , ZHANG FANG , FRANK WHARAM J. 
Published: June 2023

In January 2021, 8 states capped insulin out-of-pocket (OOP) costs at $25-$100 per 30-day supply. We examined the impact of these state policies on insulin OOP costs and use among commercially insured members with diabetes. Given that members in high-deductible health plans with health savings accounts (HSA) pay the full price of insulin before reaching their annual deductible, we hypothesized that these members would be the most impacted by the policies. We took advantage of a natural experiment created by state variation in policy to employ a robust quasi-experimental design. We used a pre-post with control group study design, with unadjusted difference-in-differences analysis, to evaluate changes in outcomes from 2020 (pre-policy) to 2021 (post-policy) in the 8 intervention states compared to contemporaneous changes in outcomes in 40 control states. We measured (1) insulin OOP cost per insulin user per year and (2) 30-day insulin fills per diabetes patient per year, and stratified results by HSA vs. non-HSA plans. We included 450,351 commercially insured members diagnosed with diabetes enrolled in a large, national health plan (19% were in an HSA plan). For the OOP cost analyses, we included 70,904 members with non-zero insulin costs. Among members with diabetes in HSA plans, state insulin OOP caps were associated with a $178 (95% CI: -$239, -$116) absolute and 18% (95% CI: -24%, -12%) relative reduction in annual OOP insulin costs. However, insulin fills among members with diabetes in HSA plans were unchanged (0.01 fills [95%: -0.05, 0.08]). The state policies also had no impact on annual OOP insulin costs or insulin fills for members with diabetes in non-HSA plans. State policies that capped insulin OOP costs were effective in reducing financial burden among HSA members but did not increase insulin use. Furthermore, the policies had little impact among members in plan types with lower baseline cost-sharing. These findings shed light on the potential impact of Medicare’s insulin copayment cap policy.

Disclosure
L.F.Garabedian: None. D.Ross-degnan: None. R.Costa: None. S.Argetsinger: None. F.Zhang: Other Relationship; GlaxoSmithKline plc., Pfizer Inc. J.Wharam: None.

Funding
Centers for Disease Control and Prevention (U18DP006527)
1506-P: Effect of Beta-Agonist Treatment on Selenium Metabolism in Differentiated Brown Adipocytes
Authors: SWANSON SYDONIE 
Published: June 2023

Selenium (Se) is an essential trace element important to several body processes such as metabolism, thyroid function, and curbing of oxidative stress. Se is utilized as the amino acid known selenocysteine (Sec), decomposed by the enzyme selenocysteine lyase (Scly). Selenoproteins have a broad range of functions, including protecting against lipid peroxidation and apoptosis, metabolic dysregulation such as glucose intolerance, and other immune responses. In brown adipocytes, the selenoprotein glutathione peroxidase 4 (GPX4) plays a key role in lipid peroxidation which in turn allows for proper thermogenic responses triggered by adrenergic signals. GPX4 removes toxic lipid peroxides by converting them into non-toxic alcohol and water. When Se levels are insufficient in brown adipocytes, the selenoprotein GPX4 may be downregulated, leading cells to accumulate a lethal amount of lipid peroxidases and triggering ferroptosis, a type of cell death. Ferroptosis activation, in turn, negatively impact pathways dependent on adrenergic responses in brown adipocytes, such as thermogenesis activation. To determine how Se levels regulate the expression of Scly and GPX4 upon treatment with adrenergic agonist in differentiated PAZ6 brown adipocytes. Human PAZ6 pre-adipocytes were differentiated for 21 days, then treated with insufficient (20 nM), adequate (80 nM), and high sodium selenite (160 nM) levels for 48h. Differentiated brown adipocytes were also treated with 1 μM CL-316243, a β3-adrenergic receptor agonist, for 24h after selenite treatment. Cells were then harvested for qPCR and Western blot analysis. Differentiated PAZ6 cells had an increase in UCP1 expression, a marker of brown adipocytes. Upon Se treatment, both Scly and GPX4 expression were unchanged in PAZ6 cells. Treatment with β3-adrenergic receptor agonist also did not affect the expression levels of both proteins. In differentiated brown adipocytes, expression of Scly and GPX4 is not dependent on Se nor β-agonist signals.

Disclosure
S.Swanson: None.

Funding
National Institutes of Health (R01DK128390)
151-LB: Expansion of Medicaid Coverage of Continuous Glucose Monitor Reduces Health Disparity
Authors: MIYAZAKI BRIAN , ZEIER TROY , BARBER REBECCA O. , ESPINOZA JUAN C. , CHAO LILY C. 
Published: June 2023

Background: Continuous glucose monitor (CGM) usage improves glycemia in people with type 1 diabetes (PWT1D) and is accepted as the standard of care. CGM utilization is lower in patients with public insurance and minorized ethnicities. In 2022, California Medicaid expanded CGM coverage to all PWT1D without requiring a minimum number of glucose tests per day. It is unknown if this policy change is sufficient to increase CGM usage. We hypothesize that Medicaid expansion narrowed but did not eliminate the disparity in CGM usage.
Methods: Data was extracted from electronic medical record of a large urban children's hospital in 2021 and 2022. CGM usage was determined based on clinician documentation or the presence of CGM downloads. We limited the data query to PWT1D who were seen at least once in the diabetes clinic each year. Chi-square and student T-tests were used to determine statistical significance (P&lt;0.05).
Results: We included 1471 and 1355 PWT1D in 2021 and 2022, respectively. Medicaid insured 66.8% and 62.2% of the patients in 2021 and 2022, respectively. CGM usage increased 1.4-fold in the entire patient population (2021: 52.2%, 2022: 71.1%). Between 2021 and 2022, CGM usage increased 1.6-fold (39% to 63%) and 1.1-fold (77% to 85%) for those with Medicaid and private insurances, respectively. CGM usage increased in all race/ethnicities, with the largest increase seen in patients who identify as Black and Latino (2021 vs 2022: White 69% vs 83%, Latino 38% vs 60%, Black 15% to 50%, P≤ 0.0001 for all comparisons). CGM usage increased 1.3-fold (58% to 74%) in English speakers and 2.1-fold (29% to 59%) in non-English speakers.
Conclusion: Our results demonstrate that Medicaid expansion of CGM coverage increases its utilization for pediatric PWT1D. Despite increased CGM usage in marginalized community, disparity persists among patients with public insurance, minority races/ethnicities, and non-English language. Future studies are needed to identify barriers that preclude equity in technology uptake.

Disclosure
B. Miyazaki: None. T. Zeier: None. R. O. Barber: None. J. C. Espinoza: Consultant; Sanofi, Research Support; NIH - National Institutes of Health, FDA, Speaker's Bureau; Glooko, Inc. L. C. Chao: None.

Funding
U.S. Food and Drug Administration (P50FD006425 to J.C.E.)
1513-P: IAPP25 in Beta Cells and DNSP11 in Alpha Cells Inhibit IAPP37 Fibrillation
Authors: LIU QING-RONG , ZHU MIN , EGAN JOSEPHINE M. 
Published: June 2023

Amyloidosis due to aggregation of mature islet amyloid polypeptide (IAPP37) in human islets causes β-cell dysfunction and there is a need to develop anti-amyloid therapeutics to protect β-cells. We used a bioinformatic approach to uncover novel human IAPP isoforms and we developed a quantitative selective reaction monitoring (SRM) proteomic assay to measure their translated peptide levels in human pancreatic islets. We used the Thioflavin T amyloid reporter assay to monitor the isoform-derived peptides for fibrillation propensity and for anti-amyloid potential to prevent aggregation of IAPP37. We uncovered hominid specific IAPP isoforms that are encoded by 4 exons instead of the conventional 3 exons: hIAPPβ, which encodes an elongated propeptide, and hIAPPγ, which is a frameshifted prepropeptide that is processed to mature IAPP25 instead of IAPP37. We found IAPP25 peptide levels to be significantly reduced in the islets of T2DM postmortem samples. We also found IAPP25, Cα-peptide (a novel short C-peptide of 19 AAs), and GDNF-derived DNSP11 present in α cells, to be nonaggregating peptides that inhibited the aggregation of IAPP37 in vitro. These novel peptides have potential to be developed as peptide based anti-amyloid therapies (This research was supported entirely by the Intramural Research Program of the NIH, National Institute on Aging).

Disclosure
Q.Liu: None. M.Zhu: None. J.M.Egan: None.

Funding
National Institute on Aging
152-LB: Implementing a Pilot Clinic for Steroid-Induced Hyperglycemia in Graft vs. Host Disease
Authors: PHAM DANIEL , GRDINOVAC KRISTINE , LEAL JORDAN , RODRIGUEZ LIZA , SALVA ALEXIS , WESSON WILLIAM , MCGUIRK JOSEPH 
Published: June 2023

Introduction: Malignant hematologic disorders are often treated with hematopoietic stem cell transplantation. This can be complicated by graft versus host disease (GVHD). Treatment is high dose corticosteroids, which often leads to steroid induced hyperglycemia (SIH). Uncontrolled hyperglycemia is associated with increased morbidity and mortality. Despite evidence favoring glycemic control in this population, there are no formal protocols or clinics in place to achieve this.
Methods: The University of Kansas Blood and Marrow Transplant Program collaborated with the Department of Endocrinology to establish a pilot clinic for patients with GVHD who have SIH. Over 12 months, patients were referred to a physician assistant specializing in diabetes. They were seen at 1-4 week intervals based on acuity and glycemic control. Data was collected on blood glucose, hemoglobin A1c, fructosamine, steroid doses, insulin doses, hospitalizations, infections, and death.
Results: 64 patients were referred to this clinic. 24 were excluded due to lack of hyperglycemia or declined being seen. Median time to index visit from referral was 20.5 days (IQR 8.25-29). Median A1c at index visit was 5.9 (IQR 5.4-7.2) and at 3 months was 5.3 (IQR 4.6-6.3). Median highest glucose within the first 3 months was 230 mg/dL (IQR 183-362) and at 3-6 months was 161 mg/dL (IQR 119.8-247, p-value = 0.0025). Median lowest glucose at 3 months was 86 mg/dL (IQR 78.5-103) and at 6 months was 87 mg/dL (IQR 75-115.8). There were 20 patients who were hospitalized and 9 patient deaths.
Conclusions: Managing SIH is complex and requires a multidisciplinary approach. Patients who were seen had reduced episodes of hyperglycemia without increased episodes of hypoglycemia. A1c is an unreliable measurement in this patient population due to high cell turnover and frequent transfusions. Future studies using continuous glucose monitors could be investigated to assess the efficacy and feasibility of this pilot clinic.

Disclosure
D. Pham: None. K. Grdinovac: None. J. Leal: None. L. Rodriguez: None. A. Salva: None. W. Wesson: None. J. McGuirk: None.
1521-P: Healthy, Nonobese South Asian Men Have Both Greater Peripheral and Hepatic Insulin Resistance than Chinese Men
Authors: LIM AMANDA Y.L. , CHANG TAN HONG , MENG KHOO CHIN 
Published: June 2023

Introduction: South Asian individuals, despite a healthy weight, are at a greater risk of type 2 diabetes mellitus than Chinese individuals. This may be secondary to higher peripheral and/or hepatic insulin resistance (IR). Using the 2-stepped hyperinsulinemic-euglycemic clamp, we examined the differences in peripheral and hepatic insulin resistance between healthy and non-obese South Asian and Chinese men. Bioimpedance analysis was performed to assess body composition.
Methods: A 2-stepped hyperinsulinemic-euglycemic clamp was performed together with a “pancreatic” clamp by infusing somatostatin. A low-dose insulin (10 mU/m2/min) was infused for 2 hours, followed by high-dose insulin infusion (40 mU/m2/min) for 2 hours. Intravenous 20% glucose was infused to maintain euglycemia (blood glucose = 95-100 mg/dl) for the clamp duration. A higher M value (mg glucose per kg bodyweight per minute) during low-dose insulin infusion reflects lower hepatic IR while a higher M value during high-dose insulin infusion reflects lower peripheral IR.
Results: The 2 groups (26 South Asian and 26 Chinese men) had similar age (27.3 vs 26.2 years, p = 0.338), but the South Asian group had slightly higher BMI (24.6 vs 22.9 kg/m2, p = 0.027) and body fat % (23.9 vs. 20.6%, p &lt; 0.05). During low-dose step, the M-value was lower among South Asians compared to Chinese (0.8 ± 0.58 vs. 1.1 ± 0.55 mg/kg/min, p = 0.027). During high-dose step, the M-value was also lower among South Asians compared to Chinese (3.2 ± 2.6 vs. 5.2 ± 3.3 mg/kg/min, p = 0.015). The differences in the M-values between South Asian and Chinese individuals remained significant after correcting for body weight and fat-free mass.
Conclusion: Despite a healthy weight, young South Asians have both greater hepatic and peripheral muscle IR than Chinese.

Disclosure
A.Y.L.Lim: None. H.Tan: None. C.Khoo: None.

Funding
National Medical Research Council of Singapore (MOH-CNIG19NOV-0002)