Boehringer Ingelheim Japan Research Articles

748-P: GLP-1 Receptor Agonists Dulaglutide and Semaglutide in Japanese Patients with Type 2 Diabetes—Randomized, Parallel-Group, Multicenter, Open-Label Trial

The clinical usefulness of once weekly GLP-1 receptor agonists, dulaglutide (D) and semaglutide (S), on domestically approved doses in Japan, was assessed by the multicenter RCT to compare the efficacy, safety and patient satisfaction of D at a dose of 0.75mg and S at a dose of 0.25-1.0mg in patients with type 2 diabetes. The study protocol was approved by the IRB of Kawasaki Medical School (No.5276-01). A total 120 patients eligible for the criteria with HbA1c of 7% or more were randomly assigned to groups D and S (D:S = 59:61), and 107 subjects completed the trial for 24 weeks (D:S=53:54). Backgrounds of 107 subjects were age 62.7 ± 10.7 years, disease duration 13.9 ± 7.4 years, BMI 29.3 ± 5.9 kg/m2, HbA1c 8.0 ± 0.6%. There were no differences in these parameters between 2 groups. Dose of S was escalated by 4 weeks from 0.25mg to adequate dose for each, and average dose was 0.63±0.24mg. The primary endpoint was the difference of HbA1c level between 2 groups at 24 weeks. The HbA1c level at 24 weeks was significantly lower in S than D (D: 8.1±0.6→7.4±0.8 vs S: 7.9±0.5→ 6.7±0.5%, p<0.0001). Reduction in BMI and VFA levels were also more significant in S (D vs. S), BMI: 29.2→28.8 vs. 29.4→28.1 kg/m2, VFA: 174.5→164.6 vs. 175.8→153.2cm2, respectively (p< 0.05). The achievement rate of HbA1c <7% was higher in S (D vs S; 30.8% vs 70.4%, p<0.0001). The parameters such as LDL-C, ALT, γGTP, and apo B/A1 were decreased, and the L/S ratio was increased only in S. Compared the changes in L/S ratio less than 0.9 at baseline, which is diagnosed as NAFLD by CT, between two groups, the ratio was significantly elevated only in the S, but not in D at 24weeks. Gastrointestinal symptoms were observed in 13.2% of D and 46.3% of S (p<0.01). DTR-QoL scores related to pain and gastrointestinal symptoms were superior in D. This prospective trial demonstrated that S has more pronounced effects on glucose and BMI lowering, as well as a reduction in liver fat percentage and visceral fat area. Disclosure T. Kimura: Speaker's Bureau; Sanwa Kagaku Kenkyusho, Kowa Research Institute, Inc., Taisho Pharmaceutical Holdings Co., Ltd., Boehringer Ingelheim Japan, Inc., Sumitomo Dainippon Pharma Co., Ltd., Sanofi, MSD Life Science Foundation, Lilly, Daiichi Sankyo, Novo Nordisk. Y. Katakura: Speaker's Bureau; Abbott. M. Shimoda: Speaker's Bureau; Sanofi, Lilly, AstraZeneca, Ono Pharmaceutical Co., Ltd., Kowa Company, Ltd., Novo Nordisk, Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Holdings Co., Ltd., MSD Life Science Foundation, Mitsubishi Tanabe Pharma Corporation. F. Kawasaki: None. M. Yamabe: Speaker's Bureau; Novo Nordisk, Eli Lilly Japan K.K., Abbott Japan Co., Ltd., Sanofi, Boehringer Ingelheim Japan, Inc., Kowa Company, Ltd., Ono Pharmaceutical Co., Ltd., Mitsubishi Tanabe Pharma Corporation, Sumitomo Dainippon Pharma Co., Ltd., Terumo Corporation. F. Tatsumi: Speaker's Bureau; Abbott Japan Co., Ltd., Kowa Company, Ltd., Novo Nordisk. M. Matsuki: None. Y. Iwamoto: Speaker's Bureau; Kowa Company, Ltd. T. Anno: None. Y. Fushimi: None. S. Kamei: None. Y. Kimura: None. J. Sanada: Speaker's Bureau; Lilly, Kowa Company, Ltd. Y. Hirata: None. S. Nakanishi: Speaker's Bureau; Boehringer Ingelheim Japan, Inc., Novo Nordisk, Sanofi, Taisho Pharmaceutical Holdings Co., Ltd., Ono Pharmaceutical Co., Ltd., Daiichi Sankyo, Kowa Company, Ltd., Abbott, Mitsubishi Tanabe Pharma Corporation. T. Mune: None. K. Kaku: Speaker's Bureau; Astellas Pharma Inc., Daiichi Sankyo, Eli Lilly Japan K.K., Boehringer Ingelheim Japan, Inc., Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation. Advisory Panel; Novo Nordisk. Consultant; Sanwa Kagaku Kenkyusho. Speaker's Bureau; Taiho Pharmaceutical Co. Ltd., Takeda Pharmaceutical Co., Ltd. H. Kaneto: Research Support; Sumitomo Dainippon Pharma Co., Ltd., Taiho Pharmaceutical Co. Ltd., Boehringer Ingelheim Japan, Inc. Speaker's Bureau; Lilly, Sanofi, Boehringer Ingelheim Japan, Inc., Novo Nordisk, Sumitomo Dainippon Pharma Co., Ltd. Research Support; Abbott.

886-P: Protective Effects of DPP-4 Inhibitor and SGLT2 Inhibitor on Pancreatic ß-Cell Function—Randomized, Parallel-Group, Multicenter, Open-Label Study (SECRETE-I)

Aims: Effects of DPP-4 inhibitor or SGLT2 inhibitor on human β-cell function are not well understood, and there is no report showing the difference between the two agents. The aim of this study is to compare the effects of these drugs on β-cell function in patients with type 2 diabetes. Methods: The study protocol was approved by the IRB of Kawasaki Medical School (No. jRCTs061190008). A total of 103 patients met the inclusion criteria (age; 20 to 79 years, HbA1c; ≥7.0% and <9.0%). Subjects were randomly assigned to luseogliflozin (L) group (n=49) or teneligliptin (T) group (n=54) and received 24 wks of intervention followed by 1-2 wks of drug washout. The primary endpoint was the change in log-transformed (Ln) disposition index (DI) 0-120 from baseline. Subjects underwent 75gOGTT before and after treatment. Results: The main baseline backgrounds of L and T groups were as follows; age: 60.8±11.1 vs. 62.6±11.2 years (p=0.4), diabetes duration: 10.1±7.9 vs. 9.2±7.6 years (p=0.6), BMI: 27.0±4.2 vs. 27.2±5.4 kg/m2 (p=0.8), HbA1c: 7.6±0.4 vs. 7.5±0.5 % (p=0.5), DI 0-120: 0.80±0.60 vs. 0.92±0.59 (p=0.1). HbA1c levels were significantly decreased in both groups, but the amount of change was greater in the T group (-0.2%, p=0.02). Body weight was significantly decreased only in the L group, with a group difference of -2.5 kg (p<0.001). Ln DI 0-120 were improved in both groups: -0.46±0.66 to -0.15±0.59 (p=0.01) in L group and -0.26±0.60 to -0.02±0.60 (p=0.003) in T group. The change in Ln serum proinsulin/CPR ratio, a marker of β-cell dysfunction, was reduced in L group (1.63±0.63 to 1.56±0.68, p=0.16), but rather increased in T group (1.70±0.75 to 1.90±0.51, p=0.01), with significant difference between the 2 groups (-0.27; p=0.004). Conclusion: Improvement effects on DI 0-120 were almost identical between luseogliflozin and teneligliptin. On the other hand, burden on β-cells was mitigated only in luseogliflozin group. Disclosure M.Shimoda: Speaker's Bureau; Sanofi, Lilly, AstraZeneca, Ono Pharmaceutical Co., Ltd., Kowa Company, Ltd., Novo Nordisk, Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Holdings Co., Ltd., MSD Life Science Foundation, Mitsubishi Tanabe Pharma Corporation. A.Obata: None. T.Kimura: Speaker's Bureau; Sanwa Kagaku Kenkyusho, Kowa Research Institute, Inc., Taisho Pharmaceutical Holdings Co., Ltd., Boehringer Ingelheim Japan, Inc., Sumitomo Dainippon Pharma Co., Ltd., Sanofi, MSD Life Science Foundation, Lilly, Daiichi Sankyo, Novo Nordisk. F.Kawasaki: None. F.Tatsumi: Speaker's Bureau; Abbott Japan Co., Ltd., Kowa Company, Ltd., Novo Nordisk. S.Nakanishi: Speaker's Bureau; Boehringer Ingelheim Japan, Inc., Novo Nordisk, Sanofi, Taisho Pharmaceutical Holdings Co., Ltd., Ono Pharmaceutical Co., Ltd., Daiichi Sankyo, Kowa Company, Ltd., Abbott, Mitsubishi Tanabe Pharma Corporation. T.Mune: None. K.Kaku: Advisory Panel; Novo Nordisk, Consultant; Sanwa Kagaku Kenkyusho, Speaker's Bureau; Astellas Pharma Inc., Daiichi Sankyo, Eli Lilly Japan K.K., Boehringer Ingelheim Japan, Inc., Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation, Taiho Pharmaceutical Co. Ltd., Takeda Pharmaceutical Co., Ltd. H.Kaneto: Research Support; Sumitomo Dainippon Pharma Co., Ltd., Taiho Pharmaceutical Co. Ltd., Boehringer Ingelheim Japan, Inc., Abbott, Speaker's Bureau; Lilly, Sanofi, Boehringer Ingelheim Japan, Inc., Novo Nordisk, Sumitomo Dainippon Pharma Co., Ltd. Y.Katakura: Speaker's Bureau; Abbott. M.Iwamoto: Speaker's Bureau; Mitsubishi Tanabe Pharma Corporation, Novo Nordisk, Boehringer Ingelheim Inc., Eli Lilly Japan K.K., Sumitomo Dainippon Pharma Co., Ltd. Y.Kimura: None. T.Anno: None. H.Isobe: None. Y.Iwamoto: Speaker's Bureau; Kowa Company, Ltd. Y.Fushimi: None. J.Sanada: Speaker's Bureau; Lilly, Kowa Company, Ltd. Funding Taisho Pharmaceutical Holdings Co., Ltd.

850-P: The Efficacy of Imeglimin in Diabetes Subgroups Stratified by Data-Driven Cluster Analysis—A Post Hoc Analysis of Imeglimin Clinical Trial Data

Objectives: We performed a post-hoc analysis of clinical trial data with the goal of investigating whether diabetic subgroups identified by data-driven clustering respond differently to imeglimin. Methods: Data from randomized, double-blind clinical trials of imeglimin as monotherapy or add-on to insulin therapy were included in the analysis. For monotherapy, 1) duration of type 2 diabetes, 2) baseline BMI, 3) baseline HbA1c, and 4) baseline HOMA-β, and for add-on to insulin therapy, the first three and 4) insulin total daily dose were applied as coordinates to form clusters by the non-hierarchical k-means method. The efficacy of imeglimin were examined for each cluster. Results: We identified four clusters of patients with diabetes, which had significantly different patient characteristics. Cluster 1 was a group with lower values of all four indices compared to the total patient population before cluster segregation, cluster 2 was a group with longer duration of diabetes, cluster 3 was a group with higher baseline BMI and higher HOMA-β (for monotherapy) or higher insulin total daily dose (for add-on to insulin therapy), and cluster 4 was a group with higher baseline HbA1c. The improvement in HbA1c with imeglimin differed significantly among the four clusters. The mean change differences versus placebo for imeglimin in monotherapy ranged from -0.64% to -1.27%, and add-on to insulin therapy ranged from -0.31% to -0.82%. In all clusters except for cluster 3 in the add-on to insulin therapy, imeglimin showed statistically significant HbA1c improvement compared to placebo. Conclusion: Data-driven clustering of patients with type 2 diabetes allowed us to identify subgroups that respond differently to imeglimin in improving HbA1c. This new stratification might help to tailor and target the imeglimin treatment to patients who would benefit most. Disclosure K.Hagi: Employee; Sumitomo Pharma. K.Kochi: Employee; Sumitomo Dainippon Pharma Co., Ltd., Sumitomo Pharma Co., Ltd. H.Watada: Research Support; Boehringer Ingelheim Japan, Inc., Mitsubishi Tanabe Pharma Corporation, LifeScan Diabetes Institute, Eli Lilly Japan K.K., Sun Pharmaceutical Industries Ltd., Teijin Pharma Limited, Taisho Pharmaceutical Holdings Co., Ltd., Ono Pharmaceutical Co., Ltd., Kowa Company, Ltd., Merck Sharp & Dohme Corp., Sanwa Kagaku Kenkyusho, Speaker's Bureau; Mitsubishi Tanabe Pharma Corporation, Taiho Pharmaceutical Co. Ltd., Novo Nordisk, Abbott Japan Co., Ltd., Astellas Pharma Inc., Merck Sharp & Dohme Corp., Kissei Pharmaceutical Co., Ltd., AstraZeneca, Ono Pharmaceutical Co., Ltd., Boehringer Ingelheim Japan, Inc., Sanofi, Sun Pharmaceutical Industries Ltd., Eli Lilly Japan K.K. K.Kaku: Advisory Panel; Novo Nordisk, Consultant; Sanwa Kagaku Kenkyusho, Speaker's Bureau; Astellas Pharma Inc., Daiichi Sankyo, Eli Lilly Japan K.K., Boehringer Ingelheim Japan, Inc., Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation, Taiho Pharmaceutical Co. Ltd., Takeda Pharmaceutical Co., Ltd. K.Ueki: Advisory Panel; Abbott Japan Co., Ltd., Research Support; Abbott Japan Co., Ltd., Boehringer Ingelheim Japan, Inc., Eli Lilly Japan K.K., Sumitomo Dainippon Pharma Co., Ltd., Novo Nordisk, Sanofi K.K., MSD Life Science Foundation, Takeda Pharmaceutical Co., Ltd., Speaker's Bureau; Boehringer Ingelheim Japan, Inc., Eli Lilly Japan K.K., Astellas Pharma Inc., AstraZeneca, Sumitomo Dainippon Pharma Co., Ltd., Novo Nordisk, Taiho Pharmaceutical Co. Ltd., Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation, Ono Pharmaceutical Co., Ltd.

1604-P: DPP-4 Inhibitor and SGLT2 Inhibitor Additively Ameliorate Hepatic Steatosis in NAFLD Model Mice through Different Action Mechanisms

Nonalcoholic fatty liver disease (NAFLD) is closely associated with the development and pathogenesis of type 2 diabetes mellitus (T2DM). Luseogliflozin (Luseo) has been reported to improve NAFLD in T2DM model mice by affecting factors related to fatty acid synthesis and beta oxidation. Anagliptin (Ana) improves NAFLD in mice by altering the M1/M2 macrophage ratio. The aim of this study is to assess the molecular mechanisms of DPP4 and SGLT2 inhibitors, which are often used in combination clinically, on NAFLD. Six-week-old male C57BL/6J mice were fed a high fat diet (HFD) to create a diet-induced NAFLD model, and at 14 weeks of age, the mice were divided into 4 groups: control, Luseo (15 mg/kg/day: mixed diet), Ana (300 mg/kg/day: mixed water), and combination group. After the 4 weeks intervention, various metabolic parameters were evaluated. In addition, the expression of mRNAs was analyzed by qPCR. Luseo and combination groups showed a significant reduction in body weight. HE and Oil-Red-O staining demonstrated an improvement of hepatocyte ballooning and decreased fat staining area in the combination compared to the control. Liver qPCR showed that the expression levels of genes related to lipid synthesis and glycogenesis such as SCD1 and PEPCK in Luseo were decreased and inflammatory response such as IL6 were decreased in Ana. The expression levels of chemokine-related genes including CCL3, CCL4, and CCL5 were also decreased in Ana. Those findings observed in Luseo and Ana groups were also confirmed in the combination. In summary, Luseo improved lipid synthesis and glycogenesis in addition to body weight reduction, and reduced inflammation-related factors, while Ana suppressed inflammatory cytokines and chemokine-related pathways. Combination of two drugs markedly ameliorated hepatocyte fat deposition. Thus, it is likely that both DPP-4 inhibitor and SGLT2 inhibitor contributed to the improvement of NAFLD through different action mechanisms. Disclosure Y. Iwamoto: Speaker's Bureau; Kowa Company, Ltd. T. Kimura: Speaker's Bureau; Sanwa Kagaku Kenkyusho, Kowa Research Institute, Inc., Taisho Pharmaceutical Holdings Co., Ltd., Boehringer Ingelheim Japan, Inc., Sumitomo Dainippon Pharma Co., Ltd., Sanofi, MSD Life Science Foundation, Lilly, Daiichi Sankyo, Novo Nordisk. H. Iwamoto: None. Y. Fushimi: None. J. Sanada: Speaker's Bureau; Lilly, Kowa Company, Ltd. Y. Katakura: Speaker's Bureau; Abbott. M. Shimoda: Speaker's Bureau; Sanofi, Lilly, AstraZeneca, Ono Pharmaceutical Co., Ltd., Kowa Company, Ltd., Novo Nordisk, Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Holdings Co., Ltd., MSD Life Science Foundation, Mitsubishi Tanabe Pharma Corporation. T. Mune: None. K. Kaku: Speaker's Bureau; Astellas Pharma Inc., Daiichi Sankyo, Eli Lilly Japan K.K., Boehringer Ingelheim Japan, Inc., Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation. Advisory Panel; Novo Nordisk. Consultant; Sanwa Kagaku Kenkyusho. Speaker's Bureau; Taiho Pharmaceutical Co. Ltd., Takeda Pharmaceutical Co., Ltd. H. Kaneto: Research Support; Sumitomo Dainippon Pharma Co., Ltd., Taiho Pharmaceutical Co. Ltd., Boehringer Ingelheim Japan, Inc. Speaker's Bureau; Lilly, Sanofi, Boehringer Ingelheim Japan, Inc., Novo Nordisk, Sumitomo Dainippon Pharma Co., Ltd. Research Support; Abbott.

1736-P: Effect of Glucokinase Haploinsufficiency on Pancreatic Beta-Cell Function and Mass in High-Fat, High-Sucrose Diet-Fed Mice

Background and Aim: We previously found in db/db mice that glucokinase haploinsufficiency reduces excessive glucose signaling in beta-cells and optimizes intracellular metabolic patterns, thereby preserving pancreatic beta-cell function and mass, ultimately improving glucose tolerance. In the present study, we investigated the effect of glucokinase haploinsufficiency on beta-cell function and mass in high-fat, high-sucrose diet (HFHS)-fed mice. Method: Four-week-old male wild-type (Gck+/+) and glucokinase haploinsufficient (Gck+/−) mice were fed normal chow (NC) or HFHS. Blood glucose levels were measured, and glucose tolerance, glucose-stimulated insulin secretion (GSIS), and pancreatic beta-cell mass were evaluated over time (16, 40, and 60 weeks of age). Results: At 16 weeks of age, no difference was found in glucose tolerance between NC and HFHS feeding in both Gck+/+ and Gck+/− mice. At 40 weeks of age, Gck+/+ HFHS-fed mice exhibited increases in GSIS and beta-cell mass versus Gck+/+ NC-fed mice (7.0 ± 1.2 mg vs. 3.3 ± 1.6 mg; P < 0.01), and glucose tolerance was preserved. Conversely, Gck+/− HFHS-fed mice displayed no increase in beta-cell mass (4.1 ± 0.5 mg vs. 3.9 ± 1.5 mg), a slight increase in GSIS, and worsening of glucose tolerance compared with Gck+/− NC-fed mice. At 60 weeks of age, Gck+/+ HFHS-fed mice displayed further increases in beta-cell mass (14.7 ± 2.0 mg) and GSIS with no worsening of glucose tolerance. In Gck+/− HFHS-fed mice, beta-cell mass (5.5 ± 1.4 mg) and GSIS were partially increased, and no further worsening of glucose tolerance was observed after 40 weeks of age. Conclusion: Gck+/− mice fed HFHS displayed insufficient increases in beta-cell mass and GSIS, resulting in worsening of glucose tolerance. In the longer term, however, these mice did not exhibit subsequent declines in beta-cell mass and GSIS nor progressive worsening of glucose tolerance unlike the non-compensatory phase in people with type 2 diabetes. Disclosure I.Shigesawa: None. T.Atsumi: Speaker's Bureau; Eli Lilly Japan K.K., Kowa Company, Ltd. A.Nakamura: Research Support; Abbott Japan Co., Ltd., Boehringer Ingelheim Japan, Inc., Daiichi Sankyo, Taisho Pharmaceutical Holdings Co., Ltd., Teijin Pharma Limited, Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation. Y.Yamauchi: None. S.Kawata: None. A.Miyazaki: None. H.Nomoto: Speaker's Bureau; Novo Nordisk, Sumitomo Pharma, Co., Ltd. H.Kameda: None. H.Miyoshi: Research Support; Abbott, LifeScan Diabetes Institute, Taisho Pharmaceutical Holdings Co., Ltd., Speaker's Bureau; AstraZeneca, Boehringer Ingelheim Japan, Inc., Eli Lilly Japan K.K., Kowa Company, Ltd., MSD Life Science Foundation, Novo Nordisk, Ono Pharmaceutical Co., Ltd., Sanofi K.K., Mitsubishi Tanabe Pharma Corporation. Y.Terauchi: Advisory Panel; Novo Nordisk, Eli Lilly Japan K.K., Sanofi, AstraZeneca, MSD Life Science Foundation, Consultant; Astellas Pharma Inc., Research Support; Abbott Japan Co., Ltd., Boehringer Ingelheim Japan, Inc., Mitsubishi Tanabe Pharma Corporation, Daiichi Sankyo, Novo Nordisk, Eli Lilly Japan K.K., Sanofi, Sumitomo Dainippon Pharma Co., Ltd., Kyowa Kirin Co., Ltd., Speaker's Bureau; Kowa Company, Ltd., Merck Sharp & Dohme Corp., Ono Pharmaceutical Co., Ltd., Abbott Japan Co., Ltd., Boehringer Ingelheim Japan, Inc., Novartis Pharmaceuticals Corporation, Mitsubishi Tanabe Pharma Corporation, Daiichi Sankyo, Sanwa Kagaku Kenkyusho, Novo Nordisk, Eli Lilly Japan K.K., Sanofi, Sumitomo Dainippon Pharma Co., Ltd., Astellas Pharma Inc., AstraZeneca, Taisho Pharmaceutical Holdings Co., Ltd., Medtronic, Teijin Pharma Limited, Roche Diagnostics, LifeScan Diabetes Institute, Terumo Corporation, Bayer Inc., Eisai Co., Ltd., Takeda Pharmaceutical Co., Ltd., Kyowa Kirin Co., Ltd.

1216-P: Effects of Aging after Age 65 on Glucose Tolerance, Insulin Sensitivity, and ß-Cell Function in Japanese—The Bunkyo Health Study

The number of new-onset diabetes cases in elderly adults is on the rise, and it is particularly prominent in Japan in which the ageing rate is the highest in the world. With ageing, both insulin sensitivity and insulin secretion deteriorate, however, it remains unclear whether these factors are further exacerbated with ageing in persons older than 65 years. The purpose of this study was to determine age-related changes in glucose tolerance and its related factors in Japanese elderly adults after age 65. Here, we studied 1438 old adults who were not taking oral hypoglycemic agents and were not diagnosed with diabetes. All subjects were assessed for glucose tolerance, insulin sensitivity (Matsuda index and Adipo-IR (fasting insulin x fasting free fatty acid (FFA)) and β-cell function (insulinogenic index) by 75g oral glucose tolerance test (OGTT). We divided participants according to age (65-69 years (n=465), 70-74 years (n=422), 75-79 years (n=326), and 80-84 years (n=225)) and compared clinical characteristics by ANOVA and post-hoc analysis. The mean BMI of all subjects was normal range (22.7±3.0kg/m2). The prevalence of newly diagnosed diabetes increased with age (7.5%, 11.1%, 12.0%, 18.7%, respectively). Fasting plasma glucose, serum insulin levels and insulinogenic index were comparable among the groups; however, area under the curve (AUC)-glucose and AUC-insulin during the 75g OGTT were significantly higher and Matsuda index and disposition index were significantly lower in the 70-74, 75-79 and 80-84 years groups than in the 65-69 years group. Similarly, percent body fat, fasting FFA and Adipo-IR were significantly higher in the 75-79 and 80-84 years groups than in the 65-69 years group. In conclusion, ageing in Japanese after age 65 is associated with exacerbation of body fat accumulation, adipose tissue and systemic IR, which, combined with decreased β-cell compensatory function, leads to worsening glucose tolerance. Disclosure H.Naito: None. M.Sato: None. S.Kadowaki: None. T.Funayama: Speaker's Bureau; Daiichi Sankyo, Boehringer Ingelheim Japan, Inc., Novo Nordisk. H.Watada: Research Support; Boehringer Ingelheim Japan, Inc., Mitsubishi Tanabe Pharma Corporation, LifeScan Diabetes Institute, Eli Lilly Japan K.K., Sun Pharmaceutical Industries Ltd., Teijin Pharma Limited, Taisho Pharmaceutical Holdings Co., Ltd., Ono Pharmaceutical Co., Ltd., Kowa Company, Ltd., Merck Sharp & Dohme Corp., Sanwa Kagaku Kenkyusho, Speaker's Bureau; Mitsubishi Tanabe Pharma Corporation, Taiho Pharmaceutical Co. Ltd., Novo Nordisk, Abbott Japan Co., Ltd., Astellas Pharma Inc., Merck Sharp & Dohme Corp., Kissei Pharmaceutical Co., Ltd., AstraZeneca, Ono Pharmaceutical Co., Ltd., Boehringer Ingelheim Japan, Inc., Sanofi, Sun Pharmaceutical Industries Ltd., Eli Lilly Japan K.K. Y.Tamura: None. H.Kaga: None. Y.Someya: None. H.Tabata: None. S.Kakehi: None. Y.Yoshizawa: None. M.Kiya: None. T.Tajima: None. N.Ito: None. Funding Strategic Research Foundation at Private Universities (S1411006); Ministry of Education, Culture, Sports, Science and Technology of Japan (18H03184); Mizuno Sports Promotion Foundation; Mitsui Life Social Welfare Foundation

1758-P: Intracellular Metabolic Change in Rodent Pancreatic ß-Cells under Diabetic Condition Is Flexible and May Compensate for Glucose Intolerance

Background: In pancreatic β-cells under diabetic conditions, metabolic stressors cause mitochondrial dysfunction and reduce oxidative phosphorylation, resulting in impaired intracellular metabolism. It has been suggested that aberrant activation of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) plays a role in this impairment. However, the pathological significance of PFKFB3 activation and its flexibility are not clear. Methods: Eight-week-old ob/ob mice were fed a standard chow ad lib or the same diet restricted to 2 g/day, or tofogliflozin added chow ad lib for 4 weeks. The body mass and glucose tolerance were compared among the groups. The expression of glycolytic enzymes in the pancreatic islets was evaluated using western blotting, immunohistochemistry and DNA microarray analysis. Next, INS-1 cells were cultured in medium containing a low (5.5 mM; low-G) or a high (22 mM: high-G) glucose concentration, or high-G with PFKFB3 inhibiter (3PO) for 48 hours. The expression of mafa and pdx-1 were evaluated using real time PCR. Results: Ad lib-fed mice showed hyperglycemia and greater weight gain than food restricted mice and tofogliflozin treated mice. The expression of PFKFB3 and other glycolytic enzymes were higher in ad lib-fed mice, whereas such aberrant expression were suppressed by restricted feeding and tofogliflozin. DNA microarray analysis revealed increased expression of genes related to glycolytic pathway in ad lib-fed mice. However, these changes were corrected in a restricted diet and tofogliflozin groups. In INS-1 cells, high-G caused elevated pfkfb3 and decreased mafa and pdx-1 expression compared with low-G exposure. Inhibition of glycolytic pathway by 3PO in high-G environment did not normalize the defect of mafa and pdx-1. Conclusion: The intracellular metabolic changes in pancreatic β cells associated with obesity and diabetes are flexible and may compensate for glucose intolerance. Disclosure K.Chiba: None. H.Nomoto: Speaker's Bureau; Novo Nordisk, Sumitomo Pharma, Co., Ltd. R.Izumihara: None. H.Kameda: None. H.Miyoshi: Research Support; Abbott, LifeScan Diabetes Institute, Taisho Pharmaceutical Holdings Co., Ltd., Speaker's Bureau; AstraZeneca, Boehringer Ingelheim Japan, Inc., Eli Lilly Japan K.K., Kowa Company, Ltd., MSD Life Science Foundation, Novo Nordisk, Ono Pharmaceutical Co., Ltd., Sanofi K.K., Mitsubishi Tanabe Pharma Corporation. A.Nakamura: Research Support; Abbott Japan Co., Ltd., Boehringer Ingelheim Japan, Inc., Daiichi Sankyo, Taisho Pharmaceutical Holdings Co., Ltd., Teijin Pharma Limited, Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation. T.Atsumi: Speaker's Bureau; Eli Lilly Japan K.K., Kowa Company, Ltd.

1381-P: Effect of Dietary Potassium Intake and Its Interaction with Sodium Intake on Risk of Developing Cardiovascular Disease (CVD) in Persons with Type 2 Diabetes (T2D)—Japan Diabetes Complication and Its Prevention Prospective Study (JDCP Study)

Many guidelines recommend increased potassium intake. However, the relationship between dietary potassium intake and incidence of CVD in persons with T2D has not been explored, nor is it known if this relationship varies with sodium intake in such persons. Therefore, we investigated these relationships as part of the JDCP study, a nationwide study launched in 2007. A total of 1477 persons with T2D aged 40-75 years completed a brief self-administered Diet History Questionnaire at baseline, and resultant data were analyzed. Primary outcome was the 7-year risk of a CVD event. Hazard ratios (HRs) for CVD according to tertiles of dietary potassium and sodium intake were estimated by Cox regression adjusted for confounders in addition to a stratified analysis of tertiles of sodium intake. Mean daily potassium and sodium intakes in tertiles 1 to 3 were 1.9, 2.6, and 3.5 g and 3.3, 4.2, and 5.3 g, respectively. HbA1c, BMI, triglycerides, and blood pressure were well controlled. No significant associations between potassium and sodium intakes and incidence of CVD were shown for the second and third tertiles relative to the first tertile (potassium: 0.64 [95% CI, 0.34-1.23] and 0.59 [0.28-1.25]; sodium: 1.02 [0.51-2.04] and 1.15 [0.58-2.30]). In the stratified analysis of tertiles of sodium intake, in the high sodium intake groups (T2 and T3) HRs for CVD in the high potassium intake group were significantly lower compared to the low potassium intake group (T2: 0.40 [0.12-1.35] and 0.06 [0.01-0.69]; T3: 0.23 [0.07-0.82] and 0.26 [0.07-0.88]). Associations of potassium intake with CVD in those with low sodium intake (T1) were not significant. Findings suggested that high potassium intake under high sodium intake is associated with a deceased incidence of CVD in persons with T2D although potassium intake under low sodium intake was not associated with incident CVD. Disclosure C.Horikawa: None. H.Shimano: None. J.Satoh: None. Y.Hayashino: Advisory Panel; Novo Nordisk A/S, Ono Pharmaceutical Co., Ltd., Speaker's Bureau; Boehringer Ingelheim Japan, Inc., Sumitomo Dainippon Pharma Co., Ltd., Santen, Daiichi Sankyo, Kowa Company, Ltd. N.Tajima: None. R.Nishimura: Speaker's Bureau; Sanofi K.K., Medicure Inc., Boehringer Ingelheim Inc., Takeda Pharmaceutical Company Limited, Kissei Pharmaceutical Co., Ltd., Novartis Pharma K.K., Eli Lilly Japan K.K., Novo Nordisk, Merck & Co., Inc., Abbott Japan Co., Ltd., Astellas Pharma Inc., Teijin Pharma Limited, Taisho Pharmaceutical Holdings Co., Ltd., Ono Pharmaceutical Co., Ltd., AstraZeneca. Y.Yamasaki: None. H.Sone: Research Support; Novo Nordisk, Astellas Pharma Inc., Kyowa Kirin Co., Ltd., Taisho Pharmaceutical Holdings Co., Ltd., Ono Pharmaceutical Co., Ltd., Eisai Co., Ltd., Takeda Pharmaceutical Co., Ltd. Jdcp study group: n/a. M.Takahara: None. N.Katakami: Speaker's Bureau; Abbott Japan Co., Ltd., Daiichi Sankyo, MSD, Mitsubishi Tanabe Pharma Corporation, Kowa Company, Ltd., Ono Pharmaceutical Co., Ltd., Novo Nordisk, Sumitomo Dainippon Pharma Co., Ltd., Bayer Inc., Lilly, Teijin Pharma Limited, Kyowa Kirin Co., Ltd., Boehringer Ingelheim Japan, Inc., Sanofi. Y.Takeda: None. M.Takeuchi: None. K.Fujihara: None. H.Suzuki: None. K.Tsuda: None. N.Yoshioka: None.

843-P: Effects of Pemafibrate on Glucose Metabolism in Adults with Type 2 Diabetes and Hypertriglyceridemia—Subanalysis of the PARM-T2D Study

Background and Aims: Pemafibrate, a novel selective peroxisome proliferator-activated receptor modulator, has been demonstrated to have potent effects on abnormalities of lipid metabolism in adults with type 2 diabetes (T2DM) and hypertriglyceridemia in real-world clinical practice (PARM-T2D study). However, the effects of pemafibrate on glucose metabolism have not been fully clarified. Materials and Methods: In this multicenter, open-labeled, prospective, observational study, adults with T2DM were treated with pemafibrate (PEMA group) or continued on their current therapy (CTRL group) for dyslipidemia for 52 weeks. This subanalysis consisted of participants who did not change their treatment for diabetes and who did not receive insulin or insulin secretagogues during the study period. Glycemic control, insulin secretion, and insulin resistance were assessed by HbA1c levels, HOMA2-β, and HOMA2-R, respectively. The changes of these parameters between baseline and the end of the study were compared between the groups. Results: In total, 279 of 650 participants met the criteria for this analysis. There were no statistical differences in baseline characteristics between PEMA (n = 141) and CTRL groups (n = 138). There were no significant changes in HbA1c levels and beta-cell function over 52 weeks between the groups, whereas pemafibrate significantly improved insulin resistance assessed by HOMA2-R versus the CTRL group (−0.20 vs. 0.01; estimated treatment difference, −0.23% [95% confidence interval = −0.44, −0.02]; p<0.05). Pemafibrate also ameliorated lipid profiles and liver dysfunction. Changes in serum triglyceride, HDL-cholesterol, γ-GTP, and ALP levels were significantly correlated with those in HOMA2-R (p = 0.024, p = 0.002, p < 0.001, and p = 0.032, respectively). Conclusions: Pemafibrate reduced insulin resistance and improved lipid profiles and liver dysfunction in individuals with T2DM and hypertriglyceridemia. Disclosure H.Nomoto: Speaker's Bureau; Novo Nordisk, Sumitomo Pharma, Co., Ltd. T.Atsumi: Speaker's Bureau; Eli Lilly Japan K.K., Kowa Company, Ltd. H.Miyoshi: Research Support; Abbott, LifeScan Diabetes Institute, Taisho Pharmaceutical Holdings Co., Ltd., Speaker's Bureau; AstraZeneca, Boehringer Ingelheim Japan, Inc., Eli Lilly Japan K.K., Kowa Company, Ltd., MSD Life Science Foundation, Novo Nordisk, Ono Pharmaceutical Co., Ltd., Sanofi K.K., Mitsubishi Tanabe Pharma Corporation. K.Kito: None. A.Nakamura: Research Support; Abbott Japan Co., Ltd., Boehringer Ingelheim Japan, Inc., Daiichi Sankyo, Taisho Pharmaceutical Holdings Co., Ltd., Teijin Pharma Limited, Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation. H.Kameda: None. K.Omori: None. S.Yanagiya: None. H.Iesaka: None. Y.Oe: None. S.Kawata: None.

1715-P: Osteopenic Obesity Is Associated with Small Vessel Disease in Old Adults—The Bunkyo Health Study

Obesity is one of the most important causes of metabolic disorders and cardiovascular events. Indeed, while the presence of cerebral small vessel disease (SVD) that includes silent brain infarcts, cerebral white matter lesions, and microhemorrhage increased the risk of stroke > 3-fold, and dementia > 2-fold, respectively, obesity has been identified as a risk factor for SVD. On the other hand, low bone mineral density (BMD), is also recognized as a risk factor for SVD. Whereas obesity is generally associated with high BMD, it has recently been reported that there is a special subtype of osteopenic obesity in which obesity and low BMD coexist. Thus, we hypothesized that this osteopenic obesity is at high risk for SVD. To test this hypothesis, we studied 1531 (641 male and 890 female) old adults aged 65 to 85 years without a history of stroke (Bunkyo Health Study). All subjects underwent dual-energy X-ray absorptiometry, brain MRI, and blood sampling, and they were classified into four groups according to the presence or absence of obesity and osteopenia (men) or osteoporosis (women) diagnosed by the BMD at the hip joint. The mean age of the subjects was 73.0 years. The prevalence of osteopenic obesity was 7.8% and 8.0% for men and women, respectively. While overall prevalence of SVD was 25% in men and 22% in women, the prevalence of SVD in osteopenic obesity was 46% and 42%, respectively. Logistic regression analysis revealed that odds ratio [95% CI] for SVD in control, obesity, osteopenia/osteoporosis and osteopenic obesity were 1.00, 1.203 [0.730-1.982], 0.970 [0.670-1.549] and 3.024 [1.560-5.861] in men and 1.00, 1.452 [0.818-2.575], 1.548 [1.012-2.368], 2.747 [1.489-5.069] in women, respectively. In conclusion, osteopenic obesity was independently associated with the presence of SVD in old adults without a history of stroke. Disclosure N.Ito: None. Y.Yoshizawa: None. H.Watada: Research Support; Boehringer Ingelheim Japan, Inc., Mitsubishi Tanabe Pharma Corporation, LifeScan Diabetes Institute, Eli Lilly Japan K.K., Sun Pharmaceutical Industries Ltd., Teijin Pharma Limited, Taisho Pharmaceutical Holdings Co., Ltd., Ono Pharmaceutical Co., Ltd., Kowa Company, Ltd., Merck Sharp & Dohme Corp., Sanwa Kagaku Kenkyusho, Speaker's Bureau; Mitsubishi Tanabe Pharma Corporation, Taiho Pharmaceutical Co. Ltd., Novo Nordisk, Abbott Japan Co., Ltd., Astellas Pharma Inc., Merck Sharp & Dohme Corp., Kissei Pharmaceutical Co., Ltd., AstraZeneca, Ono Pharmaceutical Co., Ltd., Boehringer Ingelheim Japan, Inc., Sanofi, Sun Pharmaceutical Industries Ltd., Eli Lilly Japan K.K. Y.Tamura: None. H.Kaga: None. Y.Someya: None. H.Tabata: None. S.Kakehi: None. M.Kiya: None. T.Tajima: None. H.Naito: None. Funding Strategic Research Foundation at Private Universities (S1411006); Ministry of Education, Culture, Sports, Science and Technology of Japan (18H03184); Mizuno Sports Promotion Foundation; Mitsui Life Social Welfare Foundation

1759-P: Relation between Metabolic Remodeling in Pancreatic ß-Cells and Pancreas Histology in Autopsy Cases with Type 2 Diabetes

Background and Aims: Metabolic remodeling from oxidative phosphorylation to glycolysis via activation of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) occurs in pancreatic β-cells of non-Asian individuals with type 2 diabetes (T2D), resulting in impaired β-cell function. To elucidate the underlying mechanism of metabolic remodeling, we investigated the relationship between PFKFB3 expression in β-cells and histological features of the pancreas in Japanese adults with T2D. Methods: Autopsy pancreatic sections from cadaveric donors with T2D and those without diabetes (ND) were stained with antibodies against PFKFB3, insulin, and glucagon. Ratios of PFKFB3-positive pancreatic β-cells and α-cells were compared between T2D and ND groups. Next, the relationship between PFKFB3 positivity in β-cells and histological features of the islets and pancreas were explored. We conducted thioflavin-T staining to identify amyloid deposition in islets of the T2D group and Azan staining was applied to evaluate the extent of fibrosis in ​​the pancreas. Results: In total, 36 cases (18 cases each from T2D and ND) were analyzed and 100 islets/case were evaluated. Mean ages of T2D and ND individuals were 70.3 ± 7.7 and 64.3 ± 12.8 years, and body mass indexes were 22.7 ± 4.4 and 23.3 ± 4.5 kg/m2, respectively. The PFKFB3-positive β-cell ratio was significantly higher in the T2D group (23.9 ± 16.1% vs 8.7 ± 6.3%, p < 0.05), whereas the PFKFB3-positive α-cell ratio did not differ between groups. A negative correlation between PFKFB3-positive β-cell ratio and pancreatic β-cell mass was found (ρ = −0.34, p < 0.05). In the T2D group, amyloid deposition inside the islets and pancreatic parenchymal fibrosis correlated with β-cell PFKFB3 positivity (ρ = 0.50 and ρ = 0.84; p < 0.05, respectively). Conclusion: Intracellular metabolic remodeling mediated by PFKFB3 occurred in Japanese adults with T2D and histological features of the pancreas were correlated with such changes. Disclosure R.Izumihara: None. H.Nomoto: Speaker's Bureau; Novo Nordisk, Sumitomo Pharma, Co., Ltd. K.Chiba: None. H.Kameda: None. A.Nakamura: Research Support; Abbott Japan Co., Ltd., Boehringer Ingelheim Japan, Inc., Daiichi Sankyo, Taisho Pharmaceutical Holdings Co., Ltd., Teijin Pharma Limited, Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation. H.Miyoshi: Research Support; Abbott, LifeScan Diabetes Institute, Taisho Pharmaceutical Holdings Co., Ltd., Speaker's Bureau; AstraZeneca, Boehringer Ingelheim Japan, Inc., Eli Lilly Japan K.K., Kowa Company, Ltd., MSD Life Science Foundation, Novo Nordisk, Ono Pharmaceutical Co., Ltd., Sanofi K.K., Mitsubishi Tanabe Pharma Corporation. H.Mizukami: None. T.Atsumi: Speaker's Bureau; Eli Lilly Japan K.K., Kowa Company, Ltd. Funding Japan Society for the Promotion of Science (JP20281167); Japan Diabetes Society; MSD Life Science Foundation; Public Interest Incorporated Foundation

786-P: Effects of Switching from Liraglutide or Dulaglutide to Subcutaneous Semaglutide on Metabolic Parameters in Older People with Type 2 Diabetes Mellitus—SWITCH-SEMA1 Subgroup Analysis

Background: Once-weekly treatment with the subcutaneous glucagon-like receptor-1 receptor agonist (GLP-1RA) semaglutide has been reported to have potent beneficial effects on metabolic abnormalities. However, little is known about the usefulness of switching from other GLP-1RAs to semaglutide, especially in older people with type 2 diabetes. Methods: This study is a subgroup analysis of the SWITCH-SEMA1 study, a multicenter, prospective, open-label, parallel-group, randomized controlled study in which people with type 2 diabetes continued liraglutide or dulaglutide therapy or switched to semaglutide for 24 weeks (jRCTs1011200008). One hundred people enrolled in this study were divided into two groups: an aged group consisting of people 65 years or older (n = 49; 72.8 ± 4.4 years old) and a non-aged group (n = 51; 51.2 ± 6.8 years old). The changes of metabolic parameters between the two treatment arms (continuation vs switching to semaglutide) were compared between an aged and a non-aged group. Results: At baseline, BMI, fatty liver index (FLI), C-peptide index (CPI), HOMA2-β, HOMA2-R, eGFR, and triglyceride levels were significantly lower in the aged group (all P < 0.05). After 24 weeks, switching to semaglutide significantly ameliorated HbA1c in both groups (aged: −0.7%, P<0.001; non-aged: −0.7%, P < 0.01). In the aged group, FLI; CPI; HOMA2-β; AST, γ-GTP, and uric acid (UA) levels; and the Diabetes Treatment Satisfaction Questionnaire (DTSQ) score significantly improved after switching to semaglutide (all P < 0.05). The magnitude of changes in BMI and UA levels after switching to semaglutide was greater in the aged group than in the non-aged group (BMI: −1.1 vs. −0.5 kg/m2, P < 0.01; UA: −0.4 vs. −0.1 mg/dL, P <0.05). Conclusions: The effects on metabolic parameters of switching from other GLP-1RAs to once-weekly semaglutide might be greater in aged people than in non-aged people. Disclosure Y.Takahashi: None. H.Nomoto: Speaker's Bureau; Novo Nordisk, Sumitomo Pharma, Co., Ltd. H.Yokoyama: None. A.Miya: None. A.Miyazaki: None. H.Kameda: None. T.Atsumi: Speaker's Bureau; Eli Lilly Japan K.K., Kowa Company, Ltd. A.Nakamura: Research Support; Abbott Japan Co., Ltd., Boehringer Ingelheim Japan, Inc., Daiichi Sankyo, Taisho Pharmaceutical Holdings Co., Ltd., Teijin Pharma Limited, Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation. H.Miyoshi: Research Support; Abbott, LifeScan Diabetes Institute, Taisho Pharmaceutical Holdings Co., Ltd., Speaker's Bureau; AstraZeneca, Boehringer Ingelheim Japan, Inc., Eli Lilly Japan K.K., Kowa Company, Ltd., MSD Life Science Foundation, Novo Nordisk, Ono Pharmaceutical Co., Ltd., Sanofi K.K., Mitsubishi Tanabe Pharma Corporation.

298-OR: Safety and Efficacy of Immunomodulatory Therapy Using Antithymocyte Globulin and Pegfilgrastim in Japanese Patients with Type 1 Diabetes—A Pilot Randomized Clinical Trial

While immunomodulatory therapy using low-dose anti-thymocyte globulin (ATG) has been shown to be efficacious in preserving pancreatic beta-cell function in patients with type 1 diabetes (T1D) in the United States, no clinical trials of immunomodulatory therapy for T1D have been reported in Asia. In this study, we first evaluated the safety and efficacy of ATG/pegfilgrastim (PEG) therapy in Japanese patients with T1D. This was a pilot, open-label, randomized clinical trial. Twelve patients, with the age ≥ 20 years, T1D duration < 12 months, and serum CPR ≥ 0.3 ng/ml, were randomized into the ATG/PEG group (A/P; n = 6) and control group (Ctrl; n = 6). The A/P was administered 2.0 mg/kg ATG and six times of 3.5 mg PEG. The primary endpoint was the frequency of adverse events (AEs), which were evaluated per the CTCAE v5.0, and secondary endpoints included the percentages of decline (%Dec) in CPR-area under the curve (AUC) after mixed-meal loading, the SUIT index, and changes in glycemic control during an observation period of 104 weeks. Regarding safety, no grade 4 and 5 AEs were reported in either group. Grade 3 lymphocytopenia, which is considered to be the clinical effect of ATG, was noted in all 6 patients in the A/P. One severe AE, cellulitis, was reported in the Ctrl. The frequencies of infection were not significantly different between the two groups. Regarding efficacy, %Dec in CPR-AUC in 52 weeks was less than the median value in 5 out of 6 patients in the A/P, whereas 1 out of 5 patients in Ctrl (P = 0.03). Additionally, %Dec in the SUIT index in 104 weeks tended to be lower in the A/P than in the Ctrl (P = 0.06). Changes in HbA1c and glycated albumin levels and the amount of insulin administered were not significantly different between the two groups. In conclusion, ATG/PEG therapy was safely administered to Japanese patients with T1D. Larger scale trials and mechanistic analyses are required to confirm the efficacy of this treatment. Disclosure D.Chujo: None. N.Ohmagari: None. H.Ohyanagi: None. M.Shimoda: None. K.Ueki: Advisory Panel; Abbott Japan Co., Ltd., Research Support; Abbott Japan Co., Ltd., Boehringer Ingelheim Japan, Inc., Eli Lilly Japan K.K., Sumitomo Dainippon Pharma Co., Ltd., Novo Nordisk, Sanofi K.K., MSD Life Science Foundation, Takeda Pharmaceutical Co., Ltd., Speaker's Bureau; Boehringer Ingelheim Japan, Inc., Eli Lilly Japan K.K., Astellas Pharma Inc., AstraZeneca, Sumitomo Dainippon Pharma Co., Ltd., Novo Nordisk, Taiho Pharmaceutical Co. Ltd., Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation, Ono Pharmaceutical Co., Ltd. H.Kajio: None. K.Tobe: Other Relationship; MSD K.K., Novo Nordisk Pharma Ltd., Takeda Pharmaceutical Company Limited, Daiichi Sankyo, Mitsubishi Tanabe Pharma Corporation, Suntory Global Corporation, Ltd.,, Taiho Pharmaceutical Co. Ltd., Japan Diabetes Foundation, Japan Association for Diabetes Education and Care. A.Enkaku: None. M.Kamigishi: None. A.Takikawa: None. M.Matsushita: None. M.Kobayashi: None. K.Yamamoto: None. T.Sato: None. Y.Yamamoto: None. Funding National Center for Global Health and Medicine (28A1204); Japan IDDM Network

1730-P: Stochastic Differentiation of α-Cell Lineage Revealed by Single-Cell RNA Sequencing in Time-Resolved Reporter System

Pancreatic α cells have attracted attention for their plasticity to be transdifferentiated into β cells that could potentially cure diabetes. Although such findings highlight the need for better understanding of α-cell biology, it remains unclear how each α cell originates from endocrine progenitors. We have recently generated a new α-cell reporter mouse, “Gcg-Timer” in which newly generated α (early α) cells can be sorted by FACS separately from more differentiated α (late α) cells. We investigated transcriptional heterogeneity between early and late α cells by bulk RNA sequencing (RNA-seq). To further elucidate α-cell heterogeneity at single-cell resolution, we sorted early and late α cells by FACS from E17.5 Gcg-Timer embryos, and performed single-cell RNA sequencing (scRNA-seq). Consistent with the results of bulk RNA-seq, violin plot analysis demonstrated that the mRNA expression levels of Ins1, Ins2, and Sst were significantly higher in early α cells than in late α cells. Surprisingly, pseudotime analysis demonstrated that the clusters of early α cells overlapped with those of late α cells, which means that the fluorescent reporter-based α-cell maturation does not reflect transcriptional maturation, which is in contrast with our recent finding that the fluorescent reporter-based β-cell maturation reflects transcriptional maturation based on scRNA-seq in “Ins1-eGFP;Timer” embryo. This unexpected finding suggests two possibilities: [1] each α cell stochastically originates from the endocrine progenitor pool, or [2] early α cells get differentiated in a stochastic manner. Thus, differentiation in α-cell lineage appears to be different from sequential differentiation in β-cell lineage, as we and others have demonstrated. Disclosure N.Shimizu: None. S.Sasaki: None. T.Taguchi: None. M.Himuro: Research Support; Japan Diabetes Society Junior Scientist Development Grant, Japan Society for the Promotion of Science. H.Watada: Research Support; Boehringer Ingelheim Japan, Inc., Mitsubishi Tanabe Pharma Corporation, LifeScan Diabetes Institute, Eli Lilly Japan K.K., Sun Pharmaceutical Industries Ltd., Teijin Pharma Limited, Taisho Pharmaceutical Holdings Co., Ltd., Ono Pharmaceutical Co., Ltd., Kowa Company, Ltd., Merck Sharp & Dohme Corp., Sanwa Kagaku Kenkyusho, Speaker's Bureau; Mitsubishi Tanabe Pharma Corporation, Taiho Pharmaceutical Co. Ltd., Novo Nordisk, Abbott Japan Co., Ltd., Astellas Pharma Inc., Merck Sharp & Dohme Corp., Kissei Pharmaceutical Co., Ltd., AstraZeneca, Ono Pharmaceutical Co., Ltd., Boehringer Ingelheim Japan, Inc., Sanofi, Sun Pharmaceutical Industries Ltd., Eli Lilly Japan K.K. F.C.Lynn: None. T.Miyatsuka: Speaker's Bureau; Eli Lilly Japan K.K., Novo Nordisk, Sanofi K.K., Mitsubishi Tanabe Pharma Corporation, Sumitomo Dainippon Pharma Co., Ltd., Teijin Pharma Limited, Sanwa Kagaku Kenkyusho, Daiichi Sankyo. Funding Japan Society for the Promotion of Science (21K16345, 20K08895, 19H04060); Japan Insulin-Dependent Diabetes Mellitus Network

896-P: Effectiveness of Newly Prescribed SGLT2 Inhibitors as Second-Line Therapy following DPP-4 Inhibitors or Metformin as First-Line Therapy in Type 2 Diabetes Patients

Prescriptions of SGLT2 inhibitors (SGLT2i) for patients with type 2 diabetes (T2DM) are increasing. In this study real-world data were used to evaluate the long-term benefits of SGLT2i as a second-line drug for glycemic control in people with T2DM being treated with DPP-4 inhibitor (DPP4i) or metformin (MET) monotherapy. People with T2DM patients on DPP4i or MET monotherapy were followed for at least 12 months after receiving a new SGLT2i prescription. Patients were divided into DPP4i and MET groups, and patient background factors were controlled for using propensity score matching (PM). Changes in HbA1c and other indexes during the 12 months after the new prescription were compared using t-tests, and the factors affecting changes in HbA1c were examined using multiple regression analysis. After PM, there were 192 patients in the DPP4i (n=96) and MET (n=96) groups, with no significant differences in patient backgrounds. HbA1c levels 12 months after a new SGLT2i prescription were significantly lower in both groups compared to base line (8.1% to 7.0%, p<0.001; 8.1% to 7.1%, p<0.001), but there were no significant differences between the two groups in the change in HbA1c level (p=0.271), body weight (p=0.371), systolic blood pressure (p=0.828), eGFR (p=0.642), and Fib-4 index (p=0.674). The change in HbA1c was greater for those with initially higher HbA1c levels (-0.629 [-0.722-0.536], p<0.001) and those with shorter diabetes histories (0.024 [0.001-0.048], p=0.044), but it was not significantly different between the two groups (p=0.236). After controlling for patient background, there was no significant difference in the long-term effects of a new SGLT2i prescription between those previously receiving either DPP4i or MET monotherapy, but higher previous HbA1c levels and a shorter diabetes history may be prediction of better HbA1c 12 months after a new SGLT2i prescription, regardless of prior medication. Disclosure H.Takahashi: Speaker's Bureau; Eli Lilly Japan K.K., Boehringer Ingelheim Japan, Inc., Medtronic, Novo Nordisk. Y.Suganuma: None. T.Ohno: None. R.Nishimura: Speaker's Bureau; Sanofi K.K., Medicure Inc., Boehringer Ingelheim Inc., Takeda Pharmaceutical Company Limited, Kissei Pharmaceutical Co., Ltd., Novartis Pharma K.K., Eli Lilly Japan K.K., Novo Nordisk, Merck & Co., Inc., Abbott Japan Co., Ltd., Astellas Pharma Inc., Teijin Pharma Limited, Taisho Pharmaceutical Holdings Co., Ltd., Ono Pharmaceutical Co., Ltd., AstraZeneca.

1501-P: Deletion of miR-494 Induces Adipocyte Browning and Slows Aging in Mice

Adipocyte browning is a potent therapeutic strategy for obesity and metabolic disorders. We previously reported that miR-494 served as a negative regulator of mitochondrial biogenesis during browning in cultured adipocytes. However, the in vivo evidence has not been provided. In this study, we examined the role of miR-494 in adipocyte browning and aging using miR-494 knockout (KO) mice. KO mice were generated using CRISPR/Cas9 system. KO mice had equivalent energy expenditure, physical activity, food intake and body weight compared to wild-type (WT) mice; had lower resting respiratory quotient (0.95±0.006 vs 0.91±0.004), less visceral fat (16.3±1.08 vs 13.4±0.96 g/kg body weight) and 3.7-fold increased expression of Ucp1 in subcutaneous fat. In addition, KO mice showed cold tolerance during 3 hours exposure to 4ºC and increased oxygen consumption than WT mice after administration of a β3 agonist in thermoneutral condition, indicating an increase in non-shivering thermogenesis. In adipose tissues of KO mice, increased expression of Pgc-1α, a target gene of miR-494, and tendency to increase in mitochondrial DNA were observed. Primary adipocytes derived from KO mice showed enhanced mitochondrial respiration driven by fatty acids as determined by extracellular flux analysis. Finally, KO mice showed an extended median lifespan 29% relative to WT mice (from 113.7 to 146.7 weeks). Aged KO mice (≥ 2 years old) showed reduced age-related changes, such as hair graying and low physical activity. In conclusion, deletion of miR-494 demonstrated increased adipocyte browning and survival advantage in mice. Although there are numerous studies that focused on promoting adipocyte browning, its long-term effect has not been well evaluated. Thus, our data showing the association among miR-494, adipocyte browning and longevity provide a novel insight into adipocyte biology and aging study. The gene sequence of miR-494 and its binding site to Pgc-1α are conserved from mice to humans; it is expected to be applied to human research. Disclosure L.Sugawara: None. K.Morino: Research Support; AstraZeneca, Astellas Pharma Inc., Sanwa Kagaku Kenkyusho, Ono Pharmaceutical Co., Ltd. H.Iwasaki: None. S.Ida: None. T.Yanagimachi: None. M.Lemecha: None. H.Maegawa: None. Y.Fujita: Research Support; Terumo Corporation, Boehringer Ingelheim Japan, Inc., Japan Society for the Promotion of Science, Mitsubishi Tanabe Pharma Corporation, Sumitomo Dainippon Pharma Co., Ltd. S.Kume: Research Support; Boehringer Ingelheim International GmbH, Japan Society for the Promotion of Science. Funding Grants-in-Aid for Scientific Research (22K08650)

1594-P: Interorgan Insulin-Leptin Signal Cross Talk Originating in the Liver Enhances Survival under Food-Shortage Conditions

The liver can store energy and distribute it to other tissues/organs, according to the energy states involving hepatic insulin action. In addition, we and others have shown that the liver transmits metabolic information to other organs, via neuronal and humoral mechanisms. Herein, we demonstrate a novel form of inter-organ crosstalk originating in the liver, which links hepatic insulin action with hypothalamic leptin signaling and promotes survival during food shortage. We first observed that tamoxifen-inducible liver insulin receptor (IR) knockout (iLIRKO) mice ate more and consumed less oxygen with decreased hypothalamic leptin action, thereby gaining body weight. Circulating soluble leptin receptor (sLepR) levels were elevated in iLIRKO mice. We then produced tamoxifen-inducible liver leptin receptor (LepR) and IR double-knockout (iLLepRIRKO) mice. In iLLepRIRKO mice, sLepR elevations were not seen. Moreover, increased body weights, hyperphagia and decreased whole-body oxygen consumption seen in iLIRKO mice were reversed, indicating the importance of hepatic sLepR production in the liver-hypothalamus crosstalk. Human hepatic lepr and circulating sLepR levels also correlated negatively with hepatic insulin action indices. Additionally, we restricted food intake in wild type mice. Food restriction decreased hepatic insulin action and whole-body oxygen consumption with increased circulating sLepR. We furthermore performed food restriction in tamoxifen-inducible liver LepR knockout (iLLepRKO) mice. Notably, in iLLepRKO mice, the decreases in basal metabolic rates were blunted and mortality was markedly increased during food restriction. Collectively, the liver translates metabolic cues regarding energy-deficient status, which is reflected by decreased hepatic insulin action, into sLepR, thereby suppressing energy dissipation. This novel inter-organ crosstalk may assure survival of individuals facing food shortages in nature. Disclosure K.Takahashi: Speaker's Bureau; Taisho Pharmaceutical Holdings Co., Ltd. T.Yamada: Speaker's Bureau; MSD, Mitsubishi Tanabe Pharma Corporation, Eli Lilly Japan K.K., Boehringer Ingelheim Japan, Inc., Novo Nordisk, Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Holdings Co., Ltd., Abbott Japan Co., Ltd. H.Katagiri: None. Funding Japan Society for the Promotion of Science (21K08522, 22H03126, 20H05694); Japan Science and Technology Agency (JPMJPS2023)

866-P: Quantitative Analysis of Metformin-Induced Glucose Excretion in the Intestine

The enhancement of the uptake of 18F-labeled fluorodeoxyglucose (FDG) during [18F]FDG PET is a recently found metformin action in the gut. Taking advantage of PET-MRI, which provides better tissue registration and soft-tissue contrast than conventional PET-CT, we revealed that metformin enhanced FDG accumulation in the intestinal lumen. We now quantitatively analyzed the phenomenon with a newly developed protocol of PET-MRI. We also analyzed the intestinal glucose dynamics using 13C-labeled glucose and mass spectrometry in mice. We adopted imaging sequences of MR enterography to create 3D images of the intestinal tract, then superimposed the [18F]FDG PET images to obtain the 3D distribution of radioactivity. We conducted this test on type 2 diabetes subjects treated (N=5) or not (control, N=5) with metformin. Radioactivity was detected in the wall and lumen at similar extents in the control. However, in the metformin group, the radioactivity in the lumen, but not the wall, was three to four-fold greater than that in the control. We mathematically analyzed the data using a compartment model and calculated the glucose excretion rate (GER). GER of the control and metformin groups were 6.8 ± 1.2 mg/min and 26.1 ± 2.9 mg/min, respectively, the latter being comparable to glucose excretion into the urine induced by SGLT2 inhibitors. For mouse experiments, we intravenously injected 13C-glucose into C57/B6 mice and analyzed the feces 1 hour after injection with a gas chromatography-quadrupole mass spectrometry. We could not detect 13C-glucose in the feces, possibly because gut microbiota metabolizes glucose in the intestine. However, the content of 13C in short-chain fatty acids (SCFAs) was increased by the injection of 13C-glucose, and metformin augmented the incorporation of 13C into SCFAs. These results suggest that glucose in the circulation transfers to the intestinal lumen at a considerable rate and is metabolized by microbiota, at least partly, into SCFAs and that metformin augments the processes. Disclosure Y.Hosokawa: None. K.Sakaguchi: Research Support; Sumitomo Dainippon Pharma Co., Ltd. J.Ito: None. K.Sugawara: None. Y.Morita: None. W.Ogawa: Advisory Panel; Abbott Japan Co., Ltd., Research Support; Boehringer Ingelheim Japan, Inc., Eli Lilly Japan K.K., Novo Nordisk, Teijin Pharma Limited, Kowa Company, Ltd., Sumitomo Dainippon Pharma Co., Ltd., Abbott Japan Co., Ltd., Speaker's Bureau; Sumitomo Dainippon Pharma Co., Ltd. Funding Japan Society for the Promotion of Science (21K08578, 22K18393)

462-P: DPP-4 Deficiency Suppresses the Progression of Atherosclerosis via Inducing M2 Macrophage Polarization

Background: Dipeptidyl peptidase-4 (DPP4) inhibitors suppressed progression of atherosclerosis in atherosclerotic mouse models. However, the role of DPP4 on the progression of atherosclerosis is not clear. We investigated the effect of DPP4 deficiency on the progression of atherosclerosis in DPP4 and apolipoprotein-E double knockout (DPP4/ApoE-DKO) mice. Methods: Both ApoE-KO mice and DPP4/ApoE-DKO mice were fed with normal chow until 20 weeks of age. Whole aorta and aortic sinus were stained with Oil red O. Bone marrow-derived macrophages (mφ) from wildtype or DPP4-KO mice were used in in vitro study. Expression of iNOS, MCP-1, TNF-α and IL-1β (M1 markers), and arginase-1, Mgl2 and Ym1 (M2 markers) mRNA were evaluated by real-time RT-PCR, and mφ polarization was evaluated by flowcytometry. Results: There were no differences on food intake, body weight, lipid profiles and the blood glucose levels by insulin tolerance test between each group, but the blood glucose levels by glucose tolerance test were significantly decreased in DPP4/ApoE-DKO mice. Oil-Red-O staining demonstrated that the size of atherosclerotic lesions was significantly smaller in DPP4/ApoE-DKO mice. In in vitro assay, LPS+IFNγ induced the mRNA expression of M1 markers and there was no difference between wildtype mφ and DPP4-KO mφ. On the other hand, the mRNA expression of M2 markers were significantly increased in DPP4-KO mφ. Flowcytometry revealed that the number of iNOS (M1 marker)-positive cells were increased by LPS+IFNγ, which was comparable between wildtype mφ and DPP4-KO mφ. On the other hand, the number of CD206 (M2 marker)-positive cells was higher in DPP4-KO mφ than that in wildtype mφ. Conclusion: This study suggests that DPP4 deficiency suppresses the progression of atherosclerosis in apoE-KO mice. Moreover, DPP4 deficiency increases the polarization of M2 mφ. These findings may indicate the involvement of DPP4 at least in part on the development of diabetic macrovascular complications. Disclosure Y.Zhou: None. T.Matsumura: Research Support; Shimadzu Corporation, Speaker's Bureau; Eli Lilly Japan K.K., Boehringer Ingelheim Japan, Inc. Y.Yagi: None. T.Yoshinaga: None. T.Senokuchi: None. N.Ishii: None. T.Kondo: None. E.Araki: Research Support; Eli Lilly Japan K.K., Nippon Boehringer Ingelheim, Mitsubishi Tanabe Pharma Corporation, Speaker's Bureau; AstraZeneca, Merck & Co., Inc., Ono Pharmaceutical Co., Ltd., Kowa Company, Ltd., Sumitomo Pharma Co., Ltd., Daiichi Sankyo, Novo Nordisk Pharma Ltd.

1345-P: Proinsulin–to–C-Peptide Ratio Is Associated with the Time above the Target Glucose Range and the Duration of Type 2 Diabetes

Background: The proinsulin-to-C-peptide (PI:C) ratio is thought to be an index of pancreatic β-cell dysfunction, and in subjects with prediabetes and type 2 diabetes (T2DM), abnormally high PI:C ratios have been identified. In the present study, we investigated the relationships of the duration of diabetes and glucose variability in subjects with T2DM with the PI:C ratio. Methods: We performed a prospective observational study of 272 Japanese outpatients with T2DM who underwent blinded professional continuous glucose monitoring (CGM) and fasting blood sample collection. Their fasting PI:C ratio and CGM-related indices were calculated. After placing all the participants into four groups on the basis of quartile (Q) of diabetes duration (Q1 <8 years, Q2 8-13 years, Q3 14-21 years, Q4 ≥22 years), the characteristics associated with PI:C ratio were identified. All of the statistical tests were two-sided and P<0.05 was considered to indicate statistical significance. Results: The mean age of the cohort was 68.0 years, their mean HbA1c 7.1%, and their mean body mass index (BMI) 24.9 kg/m2. High PI:C ratio was associated with a short duration of diabetes (P<0.001). In each group, there was a positive correlation between PI:C ratio and the time above the target glucose range (TAR; >180 mg/dL) in Q1, Q2, and Q3. The closest correlation between PI:C ratio and TAR was found for Q1 but there was no significant correlation for Q4 (Q1: ρ=0.38, P=0.003; Q2: ρ=0.28, P=0.019; Q3: ρ=0.24, P=0.045; Q4: ρ=0.03, P=0.811). Conclusions: We have shown that the PI:C ratio was positively associated with TAR in subjects with a short duration of T2DM. Given that insufficient pancreatic β-cell function and hyperglycemia is closely related, the PI:C ratio may play a role as the index of pancreatic β-cell dysfunction in subjects with a short duration of T2DM, but not in those with long-standing T2DM. Disclosure A.Miya: None. A.Nakamura: Research Support; Abbott Japan Co., Ltd., Boehringer Ingelheim Japan, Inc., Daiichi Sankyo, Taisho Pharmaceutical Holdings Co., Ltd., Teijin Pharma Limited, Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation. H.Nomoto: Speaker's Bureau; Novo Nordisk, Sumitomo Pharma, Co., Ltd. H.Kameda: None. T.Atsumi: Speaker's Bureau; Eli Lilly Japan K.K., Kowa Company, Ltd. Funding Japan Society for the Promotion of Science (22K17766)