bluuebunny/crossref_metadata_2025_split huggingface dataset:
385-P: Exosomal MiR-146a-5p from Macrophages Inhibits Diabetic Nephropathy Fibrosis by Targeting Smad4
Authors: ZHANG WENNING , JIANG HONGWEI , MA YUJIN , PENG HUIFANG , LIU JING , YUAN JINGYA , HU KEYAN , ZHAO BING , WU HONGLI 
Published: June 2021

Diabetic kidney disease(DKD) has become the leading cause of end-stage renal disease and the main cause of death of chronic kidney disease. Renal interstitial fibrosis is an important pathological change of DKD, and chronic microinflammation also plays a key role in the occurrence and development of Diabetic Nephropathy(DN). Macrophages are one of the main regulatory cells of inflammation and infiltrate in the kidney tissue of early DN. In addition to producing injury mediators, activated macrophages can also affect the kidney through other mechanisms. Interestingly, there are a large amount of exosomes generated by macrophage under hyperglycemia condition and it is not clear whether the exosomes has any effect on glomerular mesangial cell fibrosis in patients with diabetes. Here, we use a diabetic nephropathy db/db mouse model to determine the effect of macrophage-produced exosomes by miR-146a-5p overexpression or down-regulation on mesangial cell fibrosis. with high glucose, Macrophage proliferation is promoted in mice and miR-146a-5p expression is downregulated in both macrophages and glomerular mesangial cells. we confirmed that Smad4 is the target gene of miR-146a-5p through Luciferase reporter gene system. Fibrosis-associated col I, α-SMA, and Smad4 are upregulated in glomerular mesangial cells. In exosomes, we observed miR-146a-5p is downregulated in high glucose than low glucose. We use the exosomes from the macrophages that are transfected with miR-146a-5p mimics or inhibitor to culture the glomerular mesangial cells. Exosomes from miR-146a-5p mimics significantly downregulated glomerular Mesangial cell fibrosis-related factor α-SMA, col I protein, and Smad4. Exosomes from miR-146a-5p inhibition upregulated these factors. Finally, we found macrophage exosomes from miR-146a-5p overexpression can inhibit glomerular mesangial cell fibrosis, which could provide a potential treatment strategy for DKD.

Disclosure
W. Zhang: None. H. Jiang: None. Y. Ma: None. H. Peng: None. J. Liu: None. J. Yuan: None. K. Hu: None. B. Zhao: None. H. Wu: None.
386-P: Chronic Treatment with the Direct AMPK Activator PXL770 Improves Cardiac, Vascular, and Renal Function in Rodents with Diabetes-Related Cardiorenal Disease
Authors: STEPHAN YOHAN , SOULIÉ MATTHIEU , NICOL LIONEL , GLUAIS-DAGORN PASCALE , HALLAKOU-BOZEC SOPHIE , MULDER PAUL 
Published: June 2021

Introduction: PXL770 is a direct AMP kinase activator. In models of T2DM and NASH, PXL770 improves hyperglycemia and dyslipidemia and reduces: steatosis, inflammation, fibrogenesis. Whether these beneficial effects are associated with an improvement in diabetes-related cardiac, vascular and renal dysfunctions is unknown.
Methods: Diabetic ZSF-1 rats were treated for 90 days with PXL770 (150 mg/kg BID), then cardiac function (left ventricular (LV) hemodynamics), in vitro flow-mediated dilation (FMD; arteriograph) of mesenteric artery, renal histology and function (transcutaneous GFR) were assessed.
Results: Untreated ZSF-1 rats exhibited LV diastolic dysfunction, i.e., increases in LV End-Diastolic Pressure (LVEDP; 8.3±1.0 vs. 5.6±0.6 mm Hg in lean, p&lt;.05) and LV End-Diastolic Pressure Volume Relation (LVEDPVR; 5.4±0.6 vs. 1.1±0.2 mm Hg/RVU in lean, p&lt;.05). Myocardial tissue perfusion was reduced (4.0±0.2 vs. 7.4±0.9 ml/min/g in lean, p&lt;.05). Untreated rats exhibited impaired mesenteric FMD (% of preconstriction; 37±11 vs. 70±7% in lean, p&lt;.05) and renal function and structure as shown by a reduction in GFR (4.4±0.5 vs. 5.6±0.5 ml/min in lean, p&lt;.05) and increase in glomerular score (2.3±0.2 vs. 0.6±0.1 AU in lean, p&lt;.05). Compared to untreated ZSF-1 rats, 90-day PXL770 reduced glycaemia (-31 %, p&lt;.05) and plasma triglycerides (-44 %, p&lt;.05). PXL770 decreased LVEDP (-28 %) and LVEDPVR (2.7±0.2 mm Hg/RVU, p&lt;.05) and increased LV tissue perfusion (+30 %, p&lt;.05). PXL770 improved mesenteric FMD (64±3%, p&lt;.05) and increased GFR (6.0±0.1 ml/min, p&lt;.05) and reduced glomerular score (1.6±0.2, p&lt;.05).
Conclusion: In ZSF-1 rats, improvement of metabolic status induced by long-term treatment with the AMP kinase activator PXL770 is associated with an improved LV diastolic, vascular and renal function/structure. PXL770 could have therapeutic utility for the treatment of metabolic diseases and related co-morbid conditions.

Disclosure
Y. Stephan: None. M. Soulié: None. L. Nicol: None. P. Gluais-dagorn: Employee; Self; Poxel SA. S. Hallakou-bozec: None. P. Mulder: None.

Funding
Poxel SA
387-P: Protective Impact and Potential Mechanisms of Elabela on DKD via ß-Arrestins
Authors: SHI MIN , CHEN YANNA , ZHANG HONG 
Published: June 2021

Objective: ELABELA (ELA), an endogenous ligand for apelin receptor (APJ), is decreased in plasma of DKD patients. The effects of ELA on relieving diabetic renal lesion have not been reported. This study was performed to explore the protective impact and potential mechanisms of ELA on DKD via β-arrestins.
Methods: Sixteen eight-week-old male db/db mice were randomly divided into diabetic nephropathy group (db/db group) and intervention group (db/db+ELA group) with administration of ELA (5mg ∙ kg-1 ∙ day-1) for 8 weeks. 8 age-matched male db/m mice were recognized as normal control group (db/m group). Both db/db and db/m group received equivalent normal saline injection for 8 weeks. Body weights and blood glucose levels were measured once a week. After 8 weeks, blood and urine samples were collected. Serum creatinine and urinary albumin to creatinine ration (UACR) were examined. The kidneys were taken after sacrificing mice. The morphology change of kidney tissues were assessed by HE and PAS staining. The expression of β-Arrestin1/2 was detected by immunohistochemistry. The levels of fibronectin (FN), collagen type IV (Col-IV) and β-arrestin1/2 in kidney tissues were examined by western blotting.
Results: (1) With the intervention of ELA, UACR, serum creatinine and the ratio of kidney weight to body weight of db/db mice were significantly decreased (P&lt;0.05). (2) Compared with db/m mice, db/db mice exhibited renal tubular epithelial cells edema, decreased ratio of non-lumen area to total area, thickened basement membrane and increased glycogen accumulation, however, those pathological abnormality were mitigated by ELA. (3) The levels of FN, Col-IV and β-Arrestin-1/2 in db/db mice were higher than db/m mice, while ELA treatment suppressed the expression of FN, COL-IV and β-Arrestin-1/2 (P&lt;0.05).
Conclusions: ELA may play a role in protection from diabetic kidney damage via down-regulating the expression of β-Arrestin-1/2 and inhibiting the secretion of extracellular matrix.

Disclosure
M. Shi: None. Y. Chen: None. H. Zhang: None.

Funding
National Natural Science Foundation of China (81700723)
40-OR: Effect of Naturally Occurring Genetic Variations in Incretin Receptors on Glucose Homeostasis
Authors: GAUR ANIRUDH 
Published: June 2021

Type 2 diabetes is characterized by disruption of glucose homeostasis. Insulin is the primary driver of glucose homeostasis and released by pancreatic β cells in response to nutritional and endocrine signals. Insulin release is also stimulated by incretin hormones including Glucagon-Like peptide 1 (GLP-1) which acts on the GLP-1 receptor (GLP1-R) in islet producing the “Incretin effect.” Common genetic variations in GLP-1R alter this incretin effect. Specifically, the R131Q variant increases while the G168S variant decreases the effect of GLP-1. We have found that these genetic variations do not affect the affinity of GLP-1 nor its efficacy for G protein signaling. Taken together, these findings present a “pharmacological mystery” as to how genetic variation at GLP-1R affects incretin function. In this study, we assessed whether genetic variation in GLP-1R alters receptor trafficking resulting in differential incretin response. HEK293 cells stably expressing wild type (WT), R131Q, and G168S variants were used to measure receptor endocytosis and post-endocytic sorting to the lysosome. We found that WT GLP-1R degrades after endocytosis via its interaction with the GPCR-associated sorting protein-1 (GASP1), a protein highly expressed in islets. Importantly, we found that GLP-1R variant R131Q recycled following endocytosis while variant G168S was more rapidly degraded compared to the WT GLP-1R. To assess whether GASP1 mediated GLP-1R degradation affects incretin function, we used CRISPR to generate rat INS-1 cells with a knockout (KO) of GASP1. Deletion of GASP1 had no effect on acute GLP-1R signaling. However, prolong agonist pretreatment produced a loss of the incretin response in WT INS-1 cells, while GASP1 KO cells showed preservation of the incretin effect. These results suggest GASP1 mediates the post-endocytic degradation of GLP-1R in β cells and that altered trafficking of the GLP-1R variants may underlie differential incretin response in humans who carry these variants.

Disclosure
A. Gaur: None.

Funding
University of California, Davis
408-P: Improving Knowledge and Confidence of Diabetologists and Endocrinologists Related to MRAs for CKD in T2D with Curriculum-Based Online Education
Authors: LARKIN AMY , HANLEY KELLY L. , LE ANNE 
Published: June 2021

The goal of continuing medical education (CME) is professional growth and improved patient care. We sought to determine if a curriculum of online continuing medical education (CME) activities could improve the clinical knowledge and confidence of diabetologists and endocrinologists (D/Es) related to use of mineralocorticoid receptor antagonists (MRAs) in the management of chronic kidney disease (CKD) in patients with type 2 diabetes (T2D). The online CME curriculum consisted of 5 online activities. Of these, 4 used repeated pairs pre-/post-assessment study design was used and McNemar’s test (P &lt;.05 is considered significant) to assess educational effect. The activities launched in 2020 and data were collected for up to 12 weeks. The education reached over 19,000 physicians, including 6,000 D/Es.
Overall, knowledge improved by 9% among D/Es (N=1,111, P&lt;.001) Specific improvements: • 8% relative increase in knowledge related to the role of MRAs in treatment of CKD in T2D (P&lt;.01) • 16% relative increase in knowledge related to the mechanism of action (MOA) of MRAs for CKD in T2D (P&lt;.01) • 9% relative increase in knowledge related to identifying CKD in patients with T2D (P&lt;.01)
Of the D/Es who were included, 34% had a measurable increase in confidence, and among those the average level of confidence increased 50%.
While this curriculum demonstrated success in educating on this topic, some gaps still remain after education. On average, 22% of the questions assessing knowledge on the MOA of MRAs in managing CKD in T2D and 29% of the questions assessing knowledge related to the role of MRAs in the management of CKD in T2D were still incorrectly answered after education, demonstrating a need for more education in these areas.

Disclosure
A. Larkin: None. K. L. Hanley: None. A. Le: None.

Funding
Bayer AG
409-P: Arteriolar Hyalinosis Is Associated with the Rate of eGFR Decline and Predicts the Onset of Macroalbuminuria and eGFR&lt;60 Ml/min/1.73 M2 in Patients with Type 2 Diabetes and Normo-microalbuminuria Regardless of Microalbuminuria Grade
Authors: MORIYA TATSUMI , HAYASHI AKINORI , SUZUKI AKIHIKO , MATSUBARA MADOKA , OUCHI MOTOSHI 
Published: June 2021

Microalbuminuria is thought to be a risk factor for progression to macroalbuminuria, an indicator of renal functional decline in patients with type 1 or 2 diabetes. However, it remains unknown which renal histological factors predict the onset of macroalbuminuria or eGFR&lt;60 mL/min/1.73 m2 (chronic kidney disease, CKD) during early diabetic nephropathy. Therefore, we sought to determine which histological factors, including index of arteriolar hyalinosis (IAH), are predictors for onset of macroalbuminuria and CKD. The study population consisted of 35 patients with type 2 diabetes (25 men; age, 47±9 years; eGFR, 92.7±18.0 mL/min/1.73 m2) with normoalbuminuria or microalbuminuria who underwent percutaneous renal biopsy. We followed these patients for at least 5 (18±6) years. The study endpoint was the onset of macroalbuminuria or CKD, or by March 2020. Light and electron microscopy-based morphometric analyses were performed to quantitatively evaluate glomerular and interstitial structural changes. During the observation period, 9 of 35 patients progressed to macroalbuminuria and 15 had CKD. The annual rate of eGFR decline was significantly related to IAH (r=-0.40, p=0.016), but not any other histological parameters. Kaplan-Meier analysis revealed that arteriolar hyalinosis and mesangial expansion are associated with a significantly higher risk of onset of macroalbuminuria and CKD, and microalbuminuria grade is associated with the onset of macroalbuminuria, but not the onset of CKD.
Conclusions: Aggravated arteriolar hyalinosis is a histological risk factor for renal functional decline and predicts the onset of macroalbuminuria and CKD in patients with type 2 diabetes, normo-microalbuminuria, and normal renal function regardless of microalbuminuria grade. This finding might reflect initial changes in early diabetic nephropathy.

Disclosure
T. Moriya: None. A. Hayashi: None. A. Suzuki: None. M. Matsubara: None. M. Ouchi: None.
41-LB: Changes in Circulating Metabolites, Including Butyrate, Points to Underlying Mechanism of a Probiotic Intervention That Improves Postprandial Hyperglycemia in Subjects with Type 2 Diabetes
Authors: MCMURDIE PAUL , STOEVA MAGDALENA K. , NEMCHEK MADELEINE , TYAGI SURABHI , GINES JESSICA , EID JOHN , KOLTERMAN ORVILLE G. 
Published: June 2021

Studies in type 2 diabetes (T2D) rodent models suggest the potential for individual microbial taxa in the human gut to improve or worsen metabolic disease via specific metabolites (signals) in hepatic or general circulation. We previously reported significant improvement in postprandial glucose control in T2D subjects following ingestion of a 5-strain novel probiotic formulation (INT) in a placebo (PBO) controlled setting. (DOI:10.1136/bmjdrc-2020-001319) Here we report measurements of fecal and fasting circulating plasma metabolites, implicated as mediators of microbiome effects, collected before and after the 12-week intervention. Consistent with the hypothesized benefits of increased colonic production of butyrate, plasma butyrate increased in INT but not PBO (within group, concentration increase = 0.15 µM, p = 0.007; between group, p = 0.11), but not acetate nor propionate, paralleling changes in subject stool and production by in vitro culture. We detected changes consistent with enhanced fatty acid oxidation in INT relative to placebo, especially a decrease in medium chain fatty acids, their carnitinylates, and a decrease in bilirubin degradation byproducts. We also detected increases in certain bile acids, especially ursodeoxycholic acid (UDCA). In vitro monoculture supernatant showed that the formulation’s C. butyricum strain specifically and efficiently synthesizes UDCA during butyrogenic growth in rich media amended with the primary bile acid chenodeoxycholic acid. Untargeted data also uncovered previously undocumented use of sulfonylurea (SFU), accounting for 6 out of 7 subjects with A1c non-response in the INT group. To our knowledge, this is the first description of an increase in circulating butyrate and UDCA due solely to a bacterial probiotic intervention in humans with T2D, and may help explain the observed improvement in postprandial glucose control.

Disclosure
P. Mcmurdie: Employee; Self; Pendulum Therapeutics. M. K. Stoeva: Employee; Self; Pendulum Therapeutics. M. Nemchek: Employee; Self; Pendulum Therapeutics. S. Tyagi: None. J. Gines: Employee; Self; Pendulum Therapeutics. J. Eid: Stock/Shareholder; Self; Pendulum Therapeutics. O. G. Kolterman: Board Member; Self; GlySens Incorporated, Consultant; Self; ADOCIA, Cirius Therapeutics, Nanoprecision Medical, Viacyte, Inc., Employee; Self; Pendulum Therapeutics.
41-OR: Roles of Transcription Factor EB (TFEB) in ß-Cell Function and Survival Under Stress
Authors: ZOU YUANJIE , PASULA DANIEL , VERCHERE BRUCE , LUCIANI DAN S. 
Published: June 2021

Autophagy can help protect stressed β-cells. We investigated if disruption of Transcription Factor EB (Tfeb), a transcriptional master-regulator of lysosomes and autophagy, contributes to β-cell failure and death under various ER stress-inducing conditions.
Short-term β-cell autophagy-deficiency (inducible Atg5 knockout) reduced β-cell survival under hypoxic stress in vitro and impaired the ability of syngeneic marginal mass islet grafts to maintain glucose homeostasis in streptozotocin (STZ)-induced diabetic mice, with &gt;80% of recipients of Atg5-deficient islets returning to diabetes within 10 days post-transplant (vs. 30% of controls, p=0.05). Hypoxic culture activated pro-apoptotic ER stress pathways, and confocal analysis of islet cells from LC3-RFP-EGFP autophagy reporter mice demonstrated that autophagic flux was interrupted. Hypoxia-induced impairment of autophagy was associated with reduced lysosomal cathepsin B activity and a significant loss of Tfeb, both of which could be partially rescued by Torin-1. Lipotoxicity and thapsigargin-induced ER-stress similarly reduced Tfeb protein and cell viability, suggesting ER stress-specific mechanisms may drive dysregulation of Tfeb and lysosomes. Conversely, overexpression of Tfeb:GFP reduced thapsigargin-induced death in MIN6 cells. To directly study the consequences of Tfeb loss we generated mice with Ins1Cre-driven, β-cell-specific, Tfeb deletion (TfebβKO). Notably, both male and female TfebβKO mice had normal glucose tolerance, but cultured Tfeb KO β-cells showed impaired lysosomal cathepsin B activity and TfebβKO mice had worsened glucose tolerance compared to Tfeb flox littermates following injection with multiple low-doses STZ.
In summary, autophagy protects β-cells under hypoxia but is impaired under prolonged hypoxic stress, possibly through Tfeb dysregulation. Our data further suggest loss of Tfeb is a common feature of ER stress and that this loss exacerbates stress-induced β-cell failure and death.

Disclosure
Y. Zou: None. D. Pasula: None. B. Verchere: Advisory Panel; Self; Integrated Nanotherapeutics Inc., Sirona Biochem, Stock/Shareholder; Self; Integrated Nanotherapeutics Inc. D. S. Luciani: None.

Funding
JDRF (2-2013-50)
413-P: Renal Tubular Damage Marker, Urinary N-acetyl-b-d-glucosaminidase, as a Predictive Marker of Hepatic Fibrosis in Type 2 Diabetes
Authors: KIM HAE KYUNG , KIM YOUNG-EUN , LEE MINYOUNG , LEE YONG-HO , KANG EUN SEOK , CHA BONG-SOO , LEE BYUNG-WAN 
Published: June 2021

Background: Nonalcoholic steatohepatitis is closely associated with the progression of diabetic kidney disease (DKD) in type 2 diabetes (T2D). We investigated whether urinary N-acetyl-β-D- glucosaminidase(u-NAG), an early renal tubular damage biomarker in DKD, could be related to the degree of hepatic fibrosis in patients with T2D.
Methods: A total of 300 patients with T2D were enrolled in the study. Hepatic steatosis and fibrosis were determined using transient elastography. The levels of urinary biomarkers, including u-NAG, albumin, protein, and creatinine, and glucometabolic parameters were measured.
Results: Based on the median value of u-NAG to creatinine (u-NCR), subjects were divided into normal and high u-NCR groups. The high u-NCR group showed a significantly longer duration of diabetes, worsened hyperglycemia, and a more enhanced hepatic fibrosis index. A higher u-NCR was associated with a greater odds ratio for the risk of higher hepatic fibrosis stage (F2 OR 1.99; 95% CI [1.04-3.82]; F3 and F4 OR 2.40, 95% CI [1.52-3.80]). In addition, u-NCR was an independent predictive marker for more advanced hepatic fibrosis even after adjusting for several confounding factors (β = 1.58, P &lt;0.01).
Conclusions: The elevation of u-NAG was independently associated with a higher degree of hepatic fibrosis in patients with T2D. Considering the common metabolic milieu of renal and hepatic fibrosis in T2D, the potential use of u-NAG as an effective urinary biomarker reflecting hepatic fibrosis in T2D needs to be validated in the future.

Disclosure
H. Kim: None. Y. Kim: None. M. Lee: None. Y. Lee: None. E. Kang: None. B. Cha: None. B. Lee: None.
415-P: Prevalence of Gastroparesis Symptoms in Diabetic Patients in the Western Region of Saudi Arabia, 2019
Authors: BUKHARI ABDULRAHMAN A. , ALSAYARI MAYADA M. 
Published: June 2021

Gastroparesis is a well-established diabetes complication with conflicting data regarding its prevalence. Gastroparesis impacts quality of life, increases direct health-care costs and is associated with morbidity and mortality. Other possible consequences include erratic glycemic control, alteration in drug absorption and hypoglycemia.
Objectives: To estimate the prevalence of diabetic gastroparesis symptoms in T2DM patients attending PHC centers and to determine the associated factors
Methods: A cross sectional study which included a representative sample of patients attending PHC centers of the Ministry of National Guards Health Affairs, Western Region-Saudi Arabia. Simple random sampling technique was adopted to select the study population. Patient interviewing technique was adopted to fill the Arabic version of GSCI. Information on demographics and personal characteristics, diabetes duration, type of treatment and other variables were obtained.
Results: The study involved 365 participants, 66.3% were females, 52.9% aged 40-59 years and 31.8% with disease duration 10-20 years. A GCSI with a score of ≥ 1.9 was used to diagnose gastroparesis symptoms. Prevalence of gastroparesis symptoms was 7.9%. The most frequent symptoms were bloating (34.8%) then stomach fullness (26.6%) and the least frequent was vomiting (4.1%). The most frequently used agent was metformin (84.9%). 47.1% of participants had HbA1C between (7-9%) and 37.9% of them have gastroparesis symptoms (p 0.318). 69% of participants with gastroparesis symptoms were using metformin (p 0.012). 10.3% of participants with gastroparesis symptoms were not using any diabetes medications (p 0.037). 25% of participants with CHD had gastroparesis symptoms (p 0.001).
Conclusion: Up to our knowledge, this is the 1st local multi-city and primary care-based study. The prevalence of gastroparesis symptoms was 7.9%. Gastroparesis symptoms were significantly associated with the use of metformin, CHD and participants not on any medication.

Disclosure
A. A. Bukhari: None. M. M. Alsayari: None.