bluuebunny/crossref_metadata_2025_split huggingface dataset:
287-OR: An ABCC8 R1420H Loss-of-Function Variant Affects Insulin Secretory Function and Microvascular Disease in American Indians
Authors: VAZQUEZ ARREOLA ELSA , KNOWLER WILLIAM C. , BAIER LESLIE , HANSON ROBERT L. 
Published: June 2023

The ABCC8 gene regulates insulin secretion and plays a critical role in glucose homeostasis. An R1420H loss-of-function variant in ABCC8 was previously detected in ~3% of people in an American Indian Community with a high prevalence of type 2 diabetes (T2D); heterozygous carriers had a doubling in T2D risk and developed this disease at younger ages and lower BMIs. To further examine the phenotype of this variant, we analyzed the relationship between insulin secretion (2-h post-load corrected insulin response) and insulin sensitivity index [104/(fasting insulin*glucose)] in 5701 members of this population without diabetes aged ≥5 years using standard major axis regression, which adjusts insulin secretion for sensitivity. We used hierarchical logistic regression models in up to 7672 individuals to study cross-sectional associations with diabetes complications including increased albuminuria [albumin to creatinine ratio (ACR) ≥30 mg/g], severe albuminuria (ACR ≥ 300 mg/g), reduced eGFR (eGFR&lt;60 mL/min/1.73m2), and retinopathy assessed by fundoscopic examination. Variant carriers had worse beta-cell function than non-carriers both in children and adults (p=0.0003) with no difference in insulin sensitivity (p=0.50). This variant did not associate significantly with increased albuminuria, severe albuminuria or reduced eGFR after adjustment for age, sex, and presence and duration of diabetes. By contrast, carriers had significantly higher odds of retinopathy (OR=1.74, 95% CI: 1.19, 2.53). Increased odds of retinopathy were also observed among individuals with and without diabetes, and when adjusted for fasting plasma glucose (FPG) [carriers had higher FPG (b=6.1 mg/dl, 95% CI: 3.0, 9.2) adjusted for age, sex, and presence and duration of diabetes]. In sum, carriers of the R1420H ABCC8 variant have a higher risk of retinopathy but not of abnormal ACR or eGFR; this may be partially explained by higher glycemia levels and worse beta-cell function.

Disclosure
E.Vazquez arreola: None. W.C.Knowler: None. L.Baier: None. R.L.Hanson: None.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases
289-LB: Intestinal Stem Cells as a Source of Insulin-Positive Beta-Like Cells for Treating Diabetes
Authors: XIAO YU , XIAO YAO , JIANG YINAN 
Published: June 2023

Shortage of functional pancreatic beta-cells is the basic cause of type 1 and some late-stage type 2 diabetes, making generation of new beta-cells from stem cells a promising strategy for treating diabetes. Intestinal stem cells (ISCs) are adult stem cells found in the base of the crypts of the adult intestine, which is responsible for the continuous life-long renewal of intestinal epithelia cells and thus could also be used a potential source for autologous transplantation. Unlike the most extensively studied bone marrow derived stem cells, ISCs share the advantage of having a common endodermic origin with beta-cells, making their differentiation into beta-cells easier. However, purifying ISCs has been a challenge. Here, we reported a novel method that successfully purifies ISCs from wildtype mice by exploiting their specific binding to a lectin. The only other cell type in the intestine that also binds to this lectin was found to be Paneth cells, which could be excluded through a negative selection for CD24. The purified Lectin+CD24-ISCs, confirmed by gene profiling, were labeled with a fluorescent tag and shown to differentiate into insulin-producing beta-like cells when injected into day 11.5 mouse embryonic pancreatic explants and grown for a week. Further analysis of gene profile alterations and signaling pathways allowed us to develop a time-sensitive protocol with recombinant growth factors to optimize the differentiation of ISCs into beta-like cells in vitro, without the need for an embryonic environment. Our results not only demonstrate a new method for purifying ISCs, but also their potential to differentiate into insulin-producing beta-like cells. This study provides a promising strategy for beta-cell replacement therapy in diabetes.

Disclosure
Y. Xiao: None. Y. Xiao: None. Y. Jiang: None.

Funding
University of Pittsburgh; Children?s Hospital of Pittsburgh; Cochrane-Weber Endowed Fund in Diabetes Research
290-LB: Single-Cell Analysis of Endocrine Development Identifies Novel Roles for GLIS3 in Pancreatic ß-Cell Fate
Authors: SCOVILLE DAVID W. , GRIMM SARA , JETTEN ANTON M. 
Published: June 2023

The Krüppel-like zinc finger transcription factor GLIS3 has been identified as a critical regulator of pancreatic beta cell development and function. Mice lacking a functional Glis3 gene fail to produce enough insulin and suffer from severe hyperglycemia soon after birth. This phenotype matches what has been observed in humans with genetic mutations in GLIS3 and who have been diagnosed with permanent neonatal diabetes within the first month of life. Additional and ongoing work has begun to identify some of the transcriptional mechanisms utilized by GLIS3, yet many questions remain unanswered.
One such question is whether GLIS3 has roles in endocrine development prior to the formation of functional β-cells. To address this question, I have utilized single cell RNA-sequencing of mouse embryonic pancreas, at embryonic days (e)15.5 and e18.5, of both wild type (WT) and Glis3 knockout (KO) embryos. Analysis of this data has identified the expected developmental populations, from bipotent progenitor cells to pro-endocrine cells and ultimately hormone positive β-, α-, δ-, and ε-cells. Interestingly, there is little difference in gene expression between the WT and Glis3 KO pancreas in most of the populations observed (pro-endocrine, FEV+, α-/δ-/ε-cells). However, there are significant differences in gene expression between WT β-cells and KO β-cells. Surprisingly, most of the differences are in genes upregulated in the KO cells, with only a small number of genes downregulated. These upregulated genes are normally downregulated over the course of development from FEV+ to β-cells. These data indicate that GLIS3 plays a unique role in developing β-cells in that it primarily represses genes during the course of development but is also critical for the transcriptional activation of specific targets, particularly Ins2. Understanding how GLIS3 regulates β-cell generation, and how GLIS3 itself is regulated, could potentially provide new therapeutic avenues for the treatment of diabetes.

Disclosure
D. W. Scoville: None. S. Grimm: None. A. M. Jetten: None.

Funding
National Institutes of Health (Z01-ES-101585)
296-OR: Submicron Ultrasound Contrast Agents as Therapeutic Delivery Vehicles in Type 1 Diabetes
Authors: CICCAGLIONE MARK , MCDANIEL KRISTEN , MICHELS AARON W. , EXNER AGATA A. , BENNINGER RICHARD K. 
Published: June 2023

Type 1 diabetes (T1D) is characterized by immune-mediated destruction of insulin-producing beta cells resulting in severe hyperglycemia if untreated. Prior work has shown that islet microvasculature becomes hyperpermeable during presymptomatic T1D, allowing for ‘nanobubble (NB)’ submicron ultrasound contrast agents to accumulate in immune-infiltrated islets, which can be quantified using ultrasound contrast. We investigated the utility of these lipid-shelled NBs as peptide delivery vehicles to provide islet-targeted delivery, for acoustically driven therapeutic release and where therapeutic delivery can be visualized in real-time with ultrasound. We developed this platform to enhance efficacy of insulin peptide mimotopes, which can promote the expansion of anti-inflammatory regulatory T cells (Tregs) and prevent T1D in mice. We modified insulin B:9-23 peptides with a palmitoyl fatty acid tail to incorporate in the NB lipid shell. NBs accumulated specifically in NOD islets and not in other abdominal organs or NOD-SCID (control) islets (p &lt; 0.0001). We found that palmitoylation enhanced peptide retention in NBs (p &lt; 0.001) and is essential for delivery to NOD islets (p &lt; 0.0001), where the extent of peptide delivery correlates with extent of insulitis (p &lt; 0.05). Peptide incorporation did not affect NB echogenicity (in vitro and in vivo), ability to burst, and size distribution. Palmitoylation and incorporation in NBs does not affect the ability for peptide-pulsed antigen presenting cells to stimulate insulin-reactive T cells in vitro (p &lt; 0.0001). Finally, NOD mice treated with peptide-NBs had a higher proportion of Tregs amongst insulin-reactive CD4+ T cells in islets, as compared to mice treated with NBs or peptide alone (p &lt; 0.05). This serves as an important proof-of-principle for the usage of NBs for targeted peptide delivery and has the potential to improve therapeutic intervention in T1D.

Disclosure
M.Ciccaglione: None. K.Mcdaniel: None. A.W.Michels: Stock/Shareholder; Immnomolecular Therapeutics. A.A.Exner: None. R.K.Benninger: None.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (R01DK102950, R01DK106412, P30DK116073, T32DK120520); National Center for Advancing Translational Sciences (TL1TR002533); JDRF (1-INO-2017-435-A-N); University of Colorado
299-OR: A Combinatorial Approach with Monoclonal Antibodies to SerpinB13 and CD3 Is Therapeutically Superior to Single Antibody Use in Autoimmune Diabetes
Authors: SZEPIETOWSKA BARBARA , AVDULOV SVETLANA V. , KRYVALAP YURY , CZYZYK JAN 
Published: June 2023

A successful approach to autoimmune diabetes therapy may require simultaneous insulin-producing cell regeneration and T-cell mediated autoimmunity resolution in pancreatic islets. While CD3 antibody has been proved to reduce islet inflammation, the monoclonal antibody (mAb) to serpinB13 protease inhibitor, developed in our laboratory, was shown to cause regenerative changes in beta cells. Whether a combination of both antibodies is superior to single antibody use in the treatment of autoimmune diabetes is unclear. To address this non-obese diabetic (NOD) female mice at late pre-diabetic stage were injected with serpinB13 mAb (clone B29, 10, 40 or 200 micrograms/animal) and CD3 mAb (clone 145-2C11; at a suboptimal dose of 10 micrograms). All experimental injections were controlled with nonspecific hamster IgG and mouse IgG (clone HB-158). After the last antibody injection, random blood glucose levels were assessed for 20 weeks, and mice were declared diabetic after two measurements of glucose exceeding 300 mg/dL. We found that the combination of 40 micrograms of serpinB13 mAb with CD3 mAb offered a significant level of protection from diabetes (p=0.0027, Kaplan-Meyer with log rank test), while the combination with 10 micrograms of serpinB13 mAb had no effect. The higher dose of 200 micrograms of serpinB13 mAb showed a trend in favor of diabetes inhibition. Interestingly, at 6 to 7 weeks after completing the treatment with 40 micrograms of serpinB13 mAb alone, NOD mice showed a strong tendency to remain healthy, and separated from the double-negative control, which continued to progress to diabetes. Our results indicate that a combinatorial approach with mAbs to serpinB13 and CD3 is beneficial in autoimmune diabetes. The observations also suggest that an intermediate dose of serpinB13 mAb, and a sufficient time following administration of this mAb, is critical for its positive impact on diabetes prevention.

Disclosure
B.Szepietowska: None. S.V.Avdulov: None. Y.Kryvalap: None. J.Czyzyk: None.

Funding
JDRF (2-SRA-2022-1215-S-B)
3-LB: Effects of Sustained Somatostatin Receptor 2 Antagonism (SSTR2a) on Glycemia in Type 2 Diabetic (T2D) Male Rats—A Pilot Study
Authors: ALEALI NADIA , D'SOUZA NINOSCHKA C. , SHAKERI DORSA , SIMONSON ERIC , LIGGINS RICHARD , CHAN OWEN , RIDDELL MICHAEL C. 
Published: June 2023

Introduction: Somatostatin may be elevated in diabetes, which can reduce glucagon secretion at mealtimes and during hypoglycemia. In this pilot study, we investigated the effect of a sustained low-dose delivery of a novel SSTR2a (ZT-01), using a mini-osmotic pump, on oral glucose tolerance (OGT) and response to an insulin-induced hypoglycemia challenge in a T2D rat model.
Methods: Male Sprague Dawley rats (n=4/5 per group) were placed on a high-fat diet (3 weeks) then injected with low-dose streptozotocin (35 mg/kg), inducing T2D. Basal bolus insulin was administered for 7 days (2-3 units/day) to maintain fed-state glycemia targeting 12-25 mmol/L, after which the rats were treated with ZT-01 (0.18 mg/day) or vehicle using implanted mini-osmotic pumps (Alzet). An OGT test on day 3 was followed by a hypoglycemic challenge (12U regular insulin) on day 5.
Results: Drug and vehicle groups displayed similar glycemic levels in the fasted state and post oral glucose ingestion. Glycemia was also similar prior to insulin challenge and there was no difference in basal glucagon between groups, but SSTR2a treatment attenuated the drop in blood glucose after bolus insulin.
Conclusion: Sustained exposure to low-dose SSTR2a does not appear to affect basal glucose or glucagon levels, nor affect the response to OGT but may improve the glucose response to insulin-induced hypoglycemia in this T2D model.

Disclosure
N. Aleali: None. N. C. D'souza: None. D. Shakeri: None. E. Simonson: None. R. Liggins: Employee; Zucara Therapeutics, Stock/Shareholder; Zucara Therapeutics. O. Chan: None. M. C. Riddell: Advisory Panel; Zealand Pharma A/S, Zucara Therapeutics, Indigo Diabetes, Consultant; Lilly Diabetes, Eli Lilly and Company, Jaeb Center for Health Research, Speaker's Bureau; Dexcom, Inc., Novo Nordisk, Sanofi, Stock/Shareholder; Supersapiens, Zucara Therapeutics.

Funding
GlycoNet (CD-85); Zucara Therapeutics Inc.
301-OR: Estrogen Signaling in the Ventromedial Hypothalamus Modulates Adipose Tissue Metabolic Adaptation
Authors: TORRES IRIZARRY VALERIA C. , YE HUI , LUO PEI , CARRILLO-SÁENZ LESLIE , YANG XIAOHUA , MUNOZ MARCOS D. , ANTONY NIMISHA , JIANG YUWEI , HE YANLIN , XU PINGWEN 
Published: June 2023

Brain estrogen receptor α (ERα) signaling plays a vital role in the regulation of energy homeostasis and adipose tissue metabolism. It has been demonstrated that ERα is abundantly expressed in the ventrolateral region of the ventromedial hypothalamus (vlVMH), a sex-dimorphic structure that directly modulates brown adipose tissue (BAT) thermogenesis. We previously showed that an ERαvlVMH-originated neural circuit responds to changes in ambient temperature and nutritional states, suggesting a potential role of this neural circuit in metabolic adaptation. Here, we found that chronic activation of ERαvlVMH neurons in male mice reduces body weight, decreases adiposity, and prevents diet-induced obesity (DIO), which is associated with increases in BAT thermogenesis and core temperature. Conversely, chronic inhibition of ERαvlVMH neurons increases body weight, promotes adiposity, and decreases baseline BAT temperature. Notably, ERαvlVMH neural inhibition also impairs cold-induced food consumption and BAT thermogenesis, resulting in lethal phenotypes during chronic cold exposure. Post hoc histology analysis further revealed that ERαvlVMH neural inhibition induces adipose tissue whitening. Finally, we explored the anatomical distribution of ERαvlVMH downstream neural circuitries. Anterograde tracing revealed ERαvlVMH-originated projections to the medial preoptic area (MPA), paraventricular nucleus (PVN), substantia nigra (SNR) and dorsal raphe nucleus (DRN). Interestingly, transsynaptic retrograde tracing further demonstrated that these brain regions send projections to inguinal white adipose tissue (iWAT), suggesting potential ERαvlVMH-originated circuits involved in adipose tissue adaptation. Together, these findings support a model that estrogens act through ERαvlVMH neurons to modulate fat-specific outputs and subsequently regulate adipose tissue adaptation to cold challenges.

Disclosure
V.C.Torres irizarry: None. P.Xu: None. H.Ye: None. P.Luo: None. L.Carrillo-sáenz: None. X.Yang: None. M.D.Munoz: None. N.Antony: None. Y.Jiang: None. Y.He: None.

Funding
National Institutes of Health (R01DK123098, T32AA026577); U.S. Department of Defense (W81XWH-19-PRMRP-DA); National Institute of Diabetes and Digestive and Kidney Diseases (DK020595); American Heart Association (915789)
304-OR: Reduced VDAC1 Limits ROS Release to Restrain Oxidative Damage in Muscle of 3-Year-Old Nonhuman Primates with Early-Life Exposure to Maternal Western Diet or Obesity
Authors: GREYSLAK KEENAN T. , MCCURDY CARRIE E. 
Published: June 2023

Offspring from obese pregnancies have an elevated risk for childhood obesity and chronic metabolic diseases. Using a Japanese macaque model of early-life maternal Western-style diet (mWD) exposure, we previously observed decreased oxidative capacity and elevated markers of mitophagy in skeletal muscle of 3-yr-old offspring. We also found substantial reductions in VDAC1 content, the loss of which significantly correlated with decreased whole-muscle lipid peroxidation in mWD offspring. For this study, we isolated the impact of mWD from mWD-induced obesity by studying offspring from both lean (Ln, body fat &lt;25%) or obese (Ob, body fat &gt;30%) dams fed a WD or lean dams fed a control diet (CD) prior to and throughout pregnancy and lactation. All offspring were weaned to CD and offspring gastrocnemius was collected at 40 months of age. This design created three offspring groups: LnCD, LnWD and ObWD. In offspring muscle, VDAC1, but not VDAC2, was significantly decreased in LnWD, and was further reduced in ObWD. In contrast to whole-tissue, lipid peroxidation in a mitochondrial-enriched fraction was increased in offspring muscle from LnWD and ObWD groups. Markers of mtDNA damage were also elevated in ObWD muscle. Lastly, Sirt3 abundance, a master regulator of mitochondrial antioxidant defense, was reduced in muscle from both LnWD and ObWD. These data suggest that ROS are retained within the mitochondrial compartment due, in part, to VDAC1 suppression, resulting in an overall reduction in cellular oxidant burden. These findings elucidate a long-term adaptation to early life stress caused by mWD and obesity which limits cellular damage at the expense of mitochondrial function.

Disclosure
K.T.Greyslak: None. C.E.Mccurdy: None.

Funding
National Institutes of Health (R24DK090964); Eugene Evonuk Memorial Graduate Fellowship
306-OR: PEA15 Regulates Metabolism and Adiposity in a Sex-Dependent Manner through Alterations in Metabolic Flexibility
Authors: J. TOWNS T. , BRINKER EMILY , WATANABE RIE , ODENIYI IFEOLUWA A. , MCCAFFERTY KAYLEEN J. , GRABAU NATASHA , KROEGER DANIEL , GREENE MICHAEL W. , JUDD ROBERT L. , GRAFF EMILY 
Published: June 2023

Phosphoprotein enriched in astrocytes 15kDa (PEA15) is a ubiquitously expressed cytosolic protein with high expression in the brain, and enhanced expression in the hypothalamus. Humans with type 2 diabetes mellitus have increased expression of PEA15 in adipose tissue and skeletal muscle, and overexpression of PEA15 in transgenic mice and pigs results in peripheral insulin resistance. Despite these findings, studies of PEA15 in high fat diet (HF)-mediated obesity are limited. Male and female Pea15-/- mice (KO) and littermate controls (WT) were fed a control diet (CD) or HF for 20 weeks, with a GTT at 19 weeks and metabolic cage data at 20 weeks, just prior to serum and tissue collection. The effects of PEA15 on metabolic parameters varied based on sex. Female KO mice fed HF had a significant increase in body weight due to increased inguinal adipose and liver weight, and impaired insulin sensitivity compared to HF-fed WT females. The KO mice had similar food intake but decreased activity during the light cycle compared to HF-fed WT female mice. In contrast, a genotypic effect in male mice was noted in CD-fed mice with KO mice having an increased body weight and corresponding increases in inguinal and gonadal adipose compared to WT with no significant change in insulin sensitivity, food intake or energy expenditure. KO males on CD were significantly more active during the light phase than WT controls despite increased body weight. Substrate utilization and its associated diurnal switch were altered in both sexes of KO mice compared to WT, with KO male mice having increased preference towards carbohydrate utilization while female KO mice had increased lipid utilization. These findings suggest that PEA15 regulates metabolism in a sex-dependent manner through alterations in metabolic flexibility, energy substrate utilization, and diurnal rhythms. Further work will investigate the sex-dependent role of PEA15 on central and peripheral regulators of diurnal rhythm and metabolism.

Disclosure
T.J.Towns: None. E.Graff: None. E.Brinker: None. R.Watanabe: None. I.A.Odeniyi: None. K.J.Mccafferty: None. N.Grabau: None. D.Kroeger: None. M.W.Greene: None. R.L.Judd: None.

Funding
Auburn University
307-OR: Delineating the Origin of Supraclavicular Brown Adipocytes
Authors: RAN YALI , WANG ZIYI , WATERSTRAAT MARK G. , CABALLERO-JUAREZ ROMMEL , CHEN MIAO-HSUEH 
Published: June 2023

Brown adipose tissue (BAT) can use nutrients through non-shivering thermogenesis to generate heat. Thus, activation of BAT can be utilized to maintain body temperature and regulate metabolism. Healthy adult humans possess small amounts of BAT, with the most visible depot, the supraclavicular BAT (scBAT), located in the supraclavicular region of the neck. To understand how human scBAT functions, we previously identified BAT depots in the supraclavicular region of the mouse neck that anatomically resemble human scBAT. Similar to human scBAT, mouse scBAT contains a high density of mitochondria and expresses high levels of uncoupling protein 1. Mouse scBAT first appears in the late fetal stage in the ventral neck and continues to expand toward the lateral neck after birth. To identify the stem/progenitor cells that give rise to supraclavicular brown adipocytes, we performed a series of lineage tracing analyses. Unlike the well-studied mouse interscapular BAT depot, scBAT did not originate from the Myf5, Pax3, or Pax7-labeled progenitor cells that also give rise to skeletal muscle. Rather, a large portion of the supraclavicular brown adipocytes were labeled by Cre recombinase driven by the Mef2c anterior heart field promoter (AHF), which marks cells that give rise to the outflow tract and right ventricle of the heart. Mef2c-AHF+ cells isolated from the scBAT can spontaneously differentiate into highly thermogenic brown adipocytes in vitro, and the differentiation efficiency is further enhanced by addition of Bmp7 and PPARγ agonist, rosiglitazone. Together, our lineage tracing studies revealed that the scBAT is composed of heterogeneous brown adipocyte progenitor cells and that Mef2c-AHF-marked cardiac progenitor cells are one of the ancestors of these brown adipocytes. Understanding the trajectories of and molecular mechanisms by which these progenitor cells give rise to scBAT will provide new insights for developing a novel cell-based therapeutic tool for treating obesity and related metabolic complications.

Disclosure
Y.Ran: None. Z.Wang: None. M.G.Waterstraat: None. R.Caballero-juarez: None. M.Chen: None.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (R01DK116899)