bluuebunny/crossref_metadata_2025_split huggingface dataset:
115-OR: Microbiome Antigens as a Vaccination Strategy in Type 1 Diabetes
Authors: KOSTIC ALEKSANDAR 
Published: June 2021

Type 1 diabetes (T1D) is an autoimmune disorder that results from T cell-mediated destruction of the insulin-producing β cells of the pancreatic islets. Accumulating evidence from both human and animal studies points to the gut microbiome, the 100 trillion microbes inhabiting our intestinal tract, as a key component driving the inflammatory processes leading to T1D. However, the mechanisms therein are poorly understood. We propose that this inflammatory milieu occurs in the pancreas, where microbes or microbial fragments that translocate from their usual luminal, mucosal sites persist and induce the cytokine and chemokine signals needed for destructive, autoimmune cellular and humoral responses that destroy the insulin producing β cells. By controlling this inflammation via vaccination against a highly conserved microbial surface antigen, poly-N-acetyl glucosamine (PNAG), we can decrease the inflammatory response such that vaccination prevents diabetes. In analyses of human and murine pancreatic islets, we have found that PNAG is readily detectable within diabetic, but not healthy, pancreases and associated with both T cells and polymorphonuclear cells (PMNs). This finding suggests that inducing an effective immune response that can clear PNAG-containing microbes or microbial fragments may be critical in counteracting the microbiota-driven inflammation leading to autoimmunity. We report that NOD female mice vaccinated against PNAG exhibited a drastic reduction in diabetes incidence from in 70% of 21 controls to 17% of 18 PNAG-immune mice. Immunophenotyping on these mice indicates that PNAG vaccination dramatically increases Foxp3+ Tregs as well as CD4+ and CD4- IL-10-producing cells. Taken together, our work provides evidence that the translocation of microbial antigens to the pancreas may drive inflammation leading to T1D, and that vaccination against one particular microbial antigen, PNAG, may lead to protection against this disease.

Disclosure
A. Kostic: Other Relationship; Self; DeepBiome Therapeutics, Inc., FitBiomics, Inc.

Funding
American Diabetes Association/Pathway to Stop Diabetes (1-17-INI-13 to A.K.)
1152-P: Elucidating Protective Mechanisms of Targeting of hOGG1 to Mitochondria on the HFD-Induced Insulin Resistance in Mice Lacking OGG1
Authors: YUZEFOVYCH LARYSA , PASTUKH VIKTOR M. , RACHEK LYUDMILA 
Published: June 2021

Ogg1 (8-oxoguanine DNA glycosylase-1) is a DNA glycosylase mediating the first step in the base excision repair which removes 8-oxoguanine and repairs oxidized nuclear and mitochondrial (mtDNA). Obesity-induced mtDNA damage heightens mitochondrial oxidative stress, which leads to the development of insulin resistance. Thus, it is important to understand molecular mechanisms by which mtDNA damage induces insulin resistance and to develop therapies including preventative and treatment strategies. Previously we found that mitochondrial targeting of human OGG1 (mt-hOGG1) ameliorated high fat diet (HFD)-induced insulin resistance in Ogg1 deficient (KO) mice in both genetic model (KO/TG mice) and via TAT-protein delivery using intraperitoneal injections (Diabetes 68, Suppl 1, 2019). As we previously demonstrated that Ogg1 contributed to HFD-induced insulin resistance via hepatic insulin resistance (Diabetes 67, Suppl 1, 2018), the goal of the current study was to elucidate the underlying mechanisms for the protective effect of mt-hOGG1 delivery specifically in liver. In this study we found that targeting of mt-hOGG1 prevented hepatic mtDNA from oxidative damage in both genetic model (KO/TG) and via TAT-protein delivery (TAT-mt-hOGG1) to KO mice. Furthermore, delivery of mt-hOGG1 via TAT-protein transduction significantly increased copy number of mtDNA and expression of VDAC1, a mitochondrial marker in liver isolated from TAT-mt-hOGG1 compared to the vehicle treated mice. Studies are underway which will explore the mechanisms by which TAT-mt-hOGG1 increased mtDNA copy number, including evaluation of mitophagy and mitochondrial biogenesis. Collectively, our findings suggested that mt-hOGG1 is critical to preserve mtDNA integrity in liver and provides strong rationale to use TAT-mediated delivery of DNA repair enzymes into mitochondria as a new translational approach for treatment of obesity-induced insulin resistance.

Disclosure
L. Yuzefovych: None. V. M. Pastukh: None. L. Rachek: None.
1158-P: Genetic Dissection of a Vagal Neurocircuit for Blood Glucose Regulation
Authors: CONLEY NICHOLAS J. , LI CHELSEA , CAMPBELL JOHN 
Published: June 2021

Neurons of the hindbrain’s dorsal motor nucleus of the vagus (DMV) in the hindbrain control key physiological functions related to diabetes, including pancreatic hormone secretion and food passage through the GI tract. Different subtypes of DMV neurons, or “functional units,” are thought to mediate distinct physiological roles. However, a lack of definition and access to these subtypes has greatly limited what is known about them. Thus, there is much to learn about the nature of the DMV neurons controlling insulin release and gastric motility, how they function, and how they can be targeted to treat diabetes. To gain traction on this issue, we began by classifying DMV neuron subtypes based on their gene expression, using high-throughput single-nuclei RNA-seq in a mouse model. Our analysis revealed seven, molecularly-distinct subtypes of DMV neurons, two of which were respectively marked by their expression of the genes Calb2 and Grp. Fluorescence in situ hybridization revealed that Calb2 DMV neurons and Grp DMV neurons occupy different subregions of the DMV, suggesting they may innervate different parts of the digestive system. To investigate this, we stained the axons of Calb2 DMV neurons and Grp DMV neurons and imaged them in optically cleared stomach and pancreas from mouse. These studies revealed that Calb2 DMV neurons innervate pancreas but not stomach, whereas Grp DMV neurons innervate neither organ. To then determine whether Calb2 DMV neurons control pancreas endocrine function, we used intersectional optogenetics to stimulate Calb2 DMV neurons while assessing blood glucose levels in vivo. Preliminary data indicate that activating Calb2 DMV neurons causes a rapid and substantial decrease in blood glucose levels. Together our results not only identify Calb2 DMV neurons as pancreas afferents that can lower blood glucose, but reveal a “labeled line” logic to the organization of vagal motor neurons, in which genetically-distinct neuron subtypes innervate different digestive organs to control their function.

Disclosure
N. J. Conley: None. C. Li: None. J. Campbell: None.

Funding
American Diabetes Association/Pathway to Stop Diabetes (1-18-INI-14 to J.C.)
1159-P: One-Hour Glucose during Oral Glucose Tolerance Test Predicts Hyperglycemia Relapse in Black Obese Patients with Hyperglycemic Crises
Authors: JAGANNATHAN RAM , SMILEY-BYRD DAWN , STEFANOVSKI DARKO , UMPIERREZ GUILLERMO E. , VELLANKI PRIYATHAMA 
Published: June 2021

Approximately 50% of Black individuals with unprovoked diabetic ketoacidosis (DKA) or severe hyperglycemia (SH, blood glucose [BG]&gt;400 mg/dl without ketosis) at new-onset of diabetes achieve near-normoglycemia remission (FBG &lt; 130 mg/dl, HbA1c &lt; 7%) with intensive insulin treatment. After remission, many patients experience hyperglycemia relapse. Previous studies showed that one-hour post glucose load (1-h PG) correlated with insulin sensitivity (Si) and secretion. We tested whether 1-h PG at remission predicts HR in obese Black individuals presenting with DKA or SH. Seventy-three Black individuals presenting with DKA (n=40) or SH (n=33) underwent a 2-h 75-gm OGTT after achieving remission and followed for a median of 58 weeks until hyperglycemia relapse (FBG&gt;130 mg/dl, random BG&gt;180 mg/dl, or HbA1c&gt; 7%). Si and was calculated from the oral minimal model. Disposition index (DI) was calculated as the product of Si and Area under the curve for insulin. Mean age was 47±10 years, with 36% (n=26) women, mean BMI 36.1±9.5 kg/m2 and 1-h PG 192±48 mg/dl. One-h PG was moderately correlated with Si (r=-0.26; P=0.0294) and DI (r=-0.28; P=0.023). Highest Youden index for 1-h PG was 0.38, corresponding to an optimal cut-point of 199mg/dl with 64% sensitivity and 71% specificity. Cox proportional hazards model adjusted for age, sex and BMI showed that 1-h PG level&gt;199 mg/dl was independently associated with increased hyperglycemia relapse (HR: 2.61 [95% CI:1.13-6.01]). In a multivariate model with fasting BG, adding 1-h PG level improved prediction of hyperglycemia relapse than 2-h PG level, with significant improvements in C index (Δ: +0.07; P=0.04), net reclassification index, and integrated discrimination index. A 1-h PG at the time of remission is a better predictor of hyperglycemia relapse than fasting or 2-h PG among obese Black individuals presenting with DKA or SH. 1-h PG can be used to identify individuals at high risk of developing hyperglycemia relapse.

Disclosure
R. Jagannathan: None. D. Smiley-byrd: Consultant; Self; LifeScan, Other Relationship; Self; Novo Nordisk, Speaker’s Bureau; Self; Merck & Co., Inc. D. Stefanovski: None. G. E. Umpierrez: Research Support; Self; AstraZeneca, Dexcom, Inc., Novo Nordisk. P. Vellanki: Consultant; Self; Boehringer Ingelheim Pharmaceuticals, Inc.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (K23DK11324-01A1, UL1TR002378, K08DK0830361)
1169-P: PNPLA3 148M Is More Susceptible to PA-Induced Endoplasmic Reticulum Stress-Associated Apoptosis in HepG2 Cells
Authors: LIANG HUA 
Published: June 2021

PNPLA3 is a major susceptibility gene for MAFLD and its polymorphic locus of rs738409 (I148M) is associated with MAFLD occurrence and progression. Endoplasmic reticulum (ER) stress and hepatocyte apoptosis induced by free fatty acids lipotoxicity in fatty liver cells play an important role in the progression of NAFLD [9]. PNPLA3 is a member of the non-calcium-dependent phospholipase A2 (iPLA2) family, iPLA2ε, and whether PNPLA3 148M has stronger phospholipase activity than 148I to promote apoptosis in hepatocyte is not clear. In this study, we constructed 148I/I knock-in HepG2 cell model by Cas9/sgRNA system on the basis of HepG2 carrying PNPLA3 148M/M. The PNPLA3 148MM and I/I cells were intervened with palmitic acid (PA) for 24 h, and cell lipid deposition was detected using oil red staining, apoptosis was observed by TUNEL, and western blot to detect PNPLA3 expression and ER stress-related CHOP apoptotic signaling pathway molecule expression. The results showed no difference in PNPLA3 expression and hepatocyte lipid deposition between PNPLA3 148M/M and 148I/I cells at baseline. However, after PA intervention, PNPLA3 148M/M cells exhibited more significant lipid deposition and apoptosis than 148I/I, as well as ER-related apoptotic molecules Bip, PERK, p-eIF2α, CHOP PUMA, BAX and cleaved caspase3 expression were more significantly increased.
In conclusion, PNPLA3 148M/M cells were more significantly susceptible to PA-induced lipid deposition and endoplasmic reticulum stress-related apoptosis than 148I/I cells.

Disclosure
H. Liang: None.

Funding
Guangdong Natural Science Foundation (2020A1515010226)
1178-P: Portal Vein Metabolites as Intermediate Regulators of the Gut Microbiome in Insulin Resistance
Authors: MOREAU FRANCOIS , BRANDAO BRUNA B. , AVILA JULIAN , PAN HUI , DREYFUSS JONATHAN , KOSTIC ALEKSANDAR , CLISH CLARY B. , KAHN C. RONALD 
Published: June 2021

Previous studies have shown that there is a strong interaction between the diet and the gut microbiome. This is often linked to circulating metabolites, however, in most studies these metabolites are measured in peripheral circulation, after hepatic clearance of intestinal venous drainage. In the present study, we have determined the effects of high fat diet (HFD) and the microbiome on changes in portal and peripheral metabolites and their link to physiology by performing global metabolomics using LC-MS/MS on portal versus systemic (cardiac) blood in C57BL/6 mice on chow diet, HFD or HFD supplemented with either vancomycin or metronidazole in the drinking water to change gut flora.
Among the 644 known metabolites that were identified, 77 were more abundant in cardiac blood whereas 160 were enriched in the portal blood (FC=1.5). In portal blood there was enrichment of secondary bile acids, short-chain fatty acids (butyrate), cinnamoylglycine, indoxylsulfate and hydrocinnamate that are strongly decreased by antibiotics, indicating a direct gut microbial origin. More surprisingly, we also identified several metabolites involved in the TCA cycle, including aconitate, mesaconate and the 2-hydroxyglutarate enriched in portal blood that were not affected by diet but significantly altered by both antibiotics, indicating origin in the gut microbiome. By contrast, some metabolites were enriched in cardiac blood, such as 6, 8 dihydroxypurine, 20-hydroxydocosahexaenoic acid and C46:0 triacylglyercide, were also significantly changed by antibiotics, suggesting they are peripherally produced but depend on precursors coming from gut bacteria.
Thus, the interaction between gut microbiota and circulating metabolites is complex and may affect their production from the small intestine but also from extra intestinal tissues. In addition, the gut microbiome is the source of important circulating metabolites which can enter and modify cellular metabolism at the level of the TCA cycle.

Disclosure
F. Moreau: None. B. B. Brandao: None. J. Avila: None. H. Pan: None. J. Dreyfuss: None. A. Kostic: Other Relationship; Self; DeepBiome Therapeutics, Inc., FitBiomics, Inc. C. B. Clish: None. C. Kahn: Advisory Panel; Self; ERX Pharmaceuticals, Kaleido Biosciences, Inc., Consultant; Self; Flagship Pioneering, Sana/Cobalt.
1184-P: Return to Normal Glucose Control by Weight Loss in Nonobese People with Type 2 Diabetes: The ReTUNE Study
Authors: AL-MRABEH AHMAD , BARNES ALISON C. , IRVINE KEATON M. , KELLY TARA L. , HOLLINGSWORTH KIEREN G. , ROMERES DAVIDE , COBELLI CLAUDIO , TAYLOR ROY 
Published: June 2021

Type 2 diabetes (T2D) in the non-obese has been regarded as etiologically distinct from that in obese people. However, clinical observation suggests that loss of fat mass above a personal fat threshold brings about remission in the same way. ReTUNE aimed to determine what proportion of T2D people with BMI &lt;27kg/m2 could achieve remission, and to assess the fat threshold for this. 25 participants have been studied to date (T2D: 4M/10F, 58.3±2.1years, BMI 24.5±0.6kg/m2; matched nondiabetic controls: n=11). Other types of diabetes were excluded by anti-islet cell antibodies and MODY gene tests (2/14). From baseline, weight loss of 5% was achieved in 2-4 weeks using low calorie diet (800 kcal/day), then 6 weeks of weight maintenance. The cycle of weight loss plus maintenance was repeated 2-3 times until the clinical end point of remission was reached (HbA1c &lt; 6.5%). Only 2 cycles (~10% weight loss) were required in 8/12 participants. The planned decrease in weight (baseline to clinical endpoint: 69.0±3.5 to 60.8±2.9kg, p&lt;0.001) decreased total body fat from 33.1±1.9 to 27.4±2.0% (p&lt;0.001). Fasting plasma glucose decreased from 7.3±0.3 to 6.3±0.3 mmol/l, p&lt;0.05). HbA1c decreased from 7.1±0.2 to 6.5±0.1%, p&lt;0.05 with 8/12 in remission (&lt;6.5%). Fasting plasma insulin decreased (52.2±9.5 to 23.7±3.6 pmol/L, p=0.007) as did plasma triglyceride (1.6±0.2 to 1.0±0.1mmol/L, p=0.002). Liver fat fell from 4.4±0.8 to 1.4±0.1% (p=0.004) becoming similar to 11 matched healthy controls (1.9%). Pancreas fat content in T2D fell (5.1± 0.6 to 4.5±0.6%, p=0.026) towards control levels (3.3±0.3%, p=0.029 vs. T2D at baseline). Post meal insulin sensitivity (Si) and insulin secretion (disposition index and β-cell responsivity Φtot) improved after weight loss.
In conclusion, 2/14 people with BMI &lt;27kg/m2 were found to have other diagnoses, and weight loss brought about remission in 8/12 people accompanied by pathophysiological changes similar to those seen during remission in obesity-associated T2D.

Disclosure
A. Al-mrabeh: None. A. C. Barnes: None. K. M. Irvine: None. T. L. Kelly: Employee; Self; Diabetes Digital Media. K. G. Hollingsworth: None. D. Romeres: None. C. Cobelli: None. R. Taylor: Speaker's Bureau; Self; Novartis AG.

Funding
Diabetes UK (17/0005645)
1188-P: The New Myokine Myonectin Is Significantly Associated with Type 2 Diabetes in Elderly Patients
Authors: LEIHERER ANDREAS , MUENDLEIN AXEL , GEIGER KATHRIN , SAELY CHRISTOPH H. , LARCHER BARBARA , MADER ARTHUR , MAECHLER MAXIMILIAN , SPRENGER LUKAS , MUTSCHLECHNER BEATRIX , FRAUNBERGER PETER , DREXEL HEINZ 
Published: June 2021

The novel myokine myonectin is predominantly expressed in skeletal muscle and is involved in the regulation of metabolic homeostasis. A putative association between myonectin and type 2 diabetes mellitus (T2DM) has been discussed controversially in current literature. The association between myonectin and T2DM at different ages is still obscure and thus is addressed in the present study. We measured myonectin in 410 vascular risk patients with a mean age of 66 years. Myonectin did not correlate with age (r=-0.19; p=0.697). Half of our patients (53%) were elderly (≥66 years; n=219) with a mean age of 74 years. The younger ones (≤65 years; n=191) had a mean age of 57 years. 40.6% (n=89) of elderly patients had T2DM and 42.4% (n=81) of the younger ones. Myonectin concentrations were significantly decreased in elderly patients with T2DM compared to nondiabetic ones (1.8 vs, 4.2 ng/ml; p=0.002), whereas no significant difference was seen in younger patients (2.6 vs. 2.3 ng/ml; p=0.183). Regression analysis revealed an unadjusted odds ratio (OR) of 0.24 [0.07-0.81] (p=0.021) for the association between myonectin and T2DM in elderly patients but not in younger patients (OR=1.08 [0.80-1.45]; p=0.609). This association remained significant after adjusting for sex, body mass index, LDL-cholesterol, HDL-cholesterol, current smoking, as well as statin intake in elderly but not in younger patients (OR=0.23 [0.07-0.81]; p=0.021 vs. OR=1.05 [0.76-1.46]; p=0.769). We conclude that plasma myonectin levels are significantly associated with T2DM particularly in elderly vascular risk patients.

Disclosure
A. Leiherer: None. P. Fraunberger: None. H. Drexel: None. A. Muendlein: None. K. Geiger: None. C. H. Saely: None. B. Larcher: None. A. Mader: None. M. Maechler: None. L. Sprenger: None. B. Mutschlechner: None.
1209-P: Profound Metabolic Benefits Observed in an Eight-Week GK Rat Study of a Novel Orally Administered Polymeric Duodenal Exclusion Therapy: Implications for Type 2 Diabetes (T2D) Therapy
Authors: NIMGAONKAR ASHISH , POLOMOSCANIK STEVEN , PASRICHA PANKAJ J. , COLBERT KEVIN , JOZEFIAK THOMAS H. , VORA JITEN , VIEIRA MARCELA , CARLSON TAYLOR , LIU LIANSHENG , KULKARNI SUBHASH , PETERSEN JOHN S. 
Published: June 2021

Background: Roux-en-Y gastric bypass surgery produces significant weight loss and improvement in T2D and insulin resistance, attributed predominantly to duodenal exclusion. To mimic this effect non-invasively, we developed an oral, gut-restricted, inert polymer (GLY-POL) that interacts with mucus in the proximal small intestine to form a dynamic barrier excluding the duodenal lining from luminal contents. We therefore hypothesized that such treatment would result in significant improvement in glycemia and weight loss in a rodent model of type 2 diabetes.
Methods: Goto-kakizaki (GK) rats were gavaged daily for 8 weeks. The treatment group (n = 7) received 1.5 ml of GLY-POL (8 wt %) solution and control group (n=7) received saline. Five metabolic responses were evaluated weekly: body weight, food intake, oral glucose (oGTT) and mixed meal tolerance (mMTT) tests, and HOMA-IR.
Results: oGTT performed after chronic GLY-POL therapy produced a profound reduction of post-prandial glycemia (PPG) (incremental area under curve (iAUC)  = 60 – 70%, p&lt;0.0005) compared to control; the PPG reduction amplified with duration of therapy through the study period. Similar robust reduction in PPG iAUC was evident during mMTT (iAUC  &gt; 55%, p &lt; 0.005). Fasting plasma glucose was lower in GLY-POL group (p&lt;0.05) with improvement in HOMA-IR (p&lt;0.005). GLY-POL therapy resulted in 6% weight loss compared to control (p&lt;0.005), without a difference in food intake between the groups.
Conclusion: Our results confirm a key role for chronic duodenal exclusion in improving the systemic metabolic milieu. Chronic once daily oral GLY-POL therapy results in a profound reduction in FPG, PPG, insulin resistance and weight loss. GLY-POL therapy offers the potential for a novel non-invasive approach for glycemic and weight control in T2D patients that mimics the effects of gastric bypass without the adverse effects associated with this surgery.

Disclosure
A. Nimgaonkar: Employee; Self; Glyscend Inc. S. Kulkarni: None. J. S. Petersen: Employee; Self; Glyscend Inc. S. Polomoscanik: Employee; Self; Glyscend Inc. P. J. Pasricha: Stock/Shareholder; Self; Glyscend. K. Colbert: Employee; Self; Glyscend Inc., LifeSprout, Inc. T. H. Jozefiak: Employee; Self; Glyscend Inc., Stock/Shareholder; Self; GLYCOLOGIX LLC. J. Vora: Advisory Panel; Self; Bayer Inc., Other Relationship; Self; AstraZeneca. M. Vieira: Employee; Self; AstraZeneca. T. Carlson: None. L. Liu: None.

Funding
National Science Foundation (1521347)