bluuebunny/crossref_metadata_2025 huggingface dataset:
26-LB: Systemic Inhibition of MicroRNA-451 Impairs Renal Autophagy Process in a Mouse Model of Insulin Resistance and Early Diabetic Nephropathy
Authors: FLUITT MAURICE B. , SHIVAPURKAR NARAYAN , LI LIJUN , ECELBARGER CAROLYN M.
Published: June 2019

Diabetic nephropathy (DN) is a severe complication of diabetes mellitus and a common cause of end stage renal disease (ESRD). Novel effective therapies are needed to combat disease progression. Several studies suggest a renal protective role of microRNA-451 (miR-451) in ameliorating progression of DN. However, the role of miR-451 in DN is not understood. Defects in the autophagy process has been associated with DN. The current study aimed to elucidate the role of miR-451 in the development of DN in a mouse model of insulin resistance. Male TALLYHO/Jng mice were placed on a high-fat diet (60% kCal) and divided into two treatment groups. Mice were injected (intraperitoneal) once a week for 8-weeks with locked nucleic acid (LNA) miR-451 inhibitor or LNA-scramble (2mg/kg/bw; n=8/treatment). Mice were humanely euthanized after 12 weeks and kidneys harvested. LNA-miR-451 inhibitor significantly reduced renal expression of miR-451 by 6-fold. Collagen deposition was higher in inhibitor treated mice, as revealed by Masson’s Trichrome. Period Acid Schiff did not reveal significant glomerular damage between the two groups. Kidney weight (normalized to body weight) was significantly higher in the inhibitor treated mice. Western blotting analyses of cortex homogenates revealed increased expression of markers of renal injury, KIM1 and NGAL (p<0.05), in inhibitor treated mice. The expression of fibrotic signaling molecules mTOR, AMPKα, and TGF-β was significantly higher in the inhibitor treated group. The expression of autophagy markers SQSTM1/p62 and Beclin-1 was higher and lower, respectively, in inhibitor treated mice (p<0.05). The findings of this study suggest that reduced expression of miR-451 in DN impairs autophagy response in the kidney possibly via hyperactivation of the mTOR-signaling pathway. miR-451 may confer protection against DN by targeting key components of the autophagy process in the diabetic kidney.

Disclosure

M.B. Fluitt: None. N. Shivapurkar: None. L. Li: None. C.M. Ecelbarger: None.

Funding

American Diabetes Association (1-18-PMF-002 to M.B.F.); National Institutes of Health (TL1TR001431)
264-OR: Treatment with Human Placental Lactogen (hPL-A) Improves Glucose Homeostasis One Year after Pancreatic Islets Transplantation in Mice Anterior Eye Chamber
Authors: DONADEL GIULIA , ARRIGA ROBERTO , MARCHETTI VALENTINA , PASTORE DONATELLA , COPPOLA ANDREA , PACIFICI FRANCESCA , SCIOLI MARIA GIOVANNA , ORLANDI AUGUSTO , DELLA-MORTE DAVID
Published: June 2019

Type 1 diabetes (T1D) is a chronic disease characterized by destruction of insulin producing beta cells in the pancreas. In addition to conventional insulin treatment, islets transplantation represents an alternative and promising therapeutic approach to treat patients with T1D. Increasing evidences suggest that human placental lactogen (hPL), a polypeptide placental hormone, is able to promote islets survival and proliferation in vitro, however it is unknown its implication in vivo models. Therefore, in the present study, we used hPL isoform A (hPL-A) to evaluate the role of this hormone in murine engrafted islets. T1D was induced in C57BL6/J male mice by a single intraperitoneal high dose (180 mg/kg) of streptozotocin (STZ). Collagenase p and a Histopaque 1110 gradient solution were used to isolate pancreatic islets. The anterior chamber of the eye was used to inject isolated islets. The engraftment was evaluated by confocal microscopy and immunohistochemical staining on section of the right and left eye. Glucose tolerance test (GTT) was used to assess glucose tolerance in mice. First, we evaluated the engraftment of freshly isolated pancreatic islets. Iris dissections showed a positive staining for insulin and glucagon. Second, since 2 months after transplantation, STZ-treated mice injected with islets stimulated with hPL-A, showed an improvement in fed glycaemia compared to STZ-treated mice transplanted with islets alone, and non-transplanted STZ-treated mice, for a follow-up of 1 year. Furthermore, mice treated with hPL-A, showed GTT with a similar pattern to untreated control mice; islets vascularization and angiogenesis was showed using CD31. Taken together, present results provide evidence that hPL-A could have a long-term effect on islet graft function and survival through enhancing glucose homeostasis.

Disclosure

G. Donadel: None. R. Arriga: None. V. Marchetti: Employee; Self; STEMCELL Technologies. D. Pastore: None. A. Coppola: None. F. Pacifici: None. M. Scioli: None. A. Orlandi: None. D. Della-Morte: None.

Funding

Italian Ministry of Instruction University and Research; Fondazione Roma
266-OR: Plasma Membrane sn-1,2 Diacylglycerol Mediates Lipid-Induced Hepatic Insulin Resistance
Authors: LYU KUN , ZHANG YE , ZHANG DONGYAN , KAHN MARIO , NOZAKI YUICHI , BHANOT SANJAY , BOGAN JONATHAN , CLINE GARY , SAMUEL VARMAN , SHULMAN GERALD I.
Published: June 2019

The identity and subcellular localization of the bioactive lipid species that causes hepatic insulin resistance remain controversial. Though chronic diacylglycerol acyltransferase 2 (DGAT2) knockdown improves hepatic steatosis and insulin action, we hypothesized that acute DGAT2 knockdown would transiently increase hepatic diacylglycerol (DAG) content to specifically assess the role of hepatic DAG on hepatic insulin action. Regular chow-fed rats were treated with a single dose of DGAT2 antisense oligonucleotide and studied 2 days later. The content and subcellular localization of sn-1,2, sn-2,3 and sn-1,3 DAGs were assessed by LC-MS/MS in the endoplasmic reticulum (ER), mitochondrial (MIT), plasma membrane (PM), lipid droplet (LD) and cytosol (CYT) fractions. Acute knockdown of hepatic DGAT2 induced hepatic insulin resistance with impaired suppression of hepatic glucose production during a hyperinsulinemic-euglycemic clamp. Insulin receptor kinase (IRK) activation was impaired, as reflected by reduced insulin-stimulated IRK Y1162 phosphorylation. This was associated with increased PKCε activation, as reflected by increased translocation from CYT to membrane, and increased sn-1,2 DAG content specifically in the PM. In contrast, there were no changes in hepatic ceramide content or sn-2,3 and sn-1,3 DAGs in the five subcellular fractions tested (ER, MIT, PM, LD, CYT). We next infused [13C16] palmitate in 24-hr-fasted rats to track hepatic DAG synthesis in vivo and observed that sn-1,2 DAGs were the major (73%) product of re-esterification from exogenous fatty acids, compared to sn-2,3 (25%) and sn-1,3 (2%) DAGs. We also found that sn-1,2 and sn-2,3 DAGs were primarily synthesized in the ER before translocating to PM, whereas sn-1,3 DAGs were exclusively produced in the LD by lipolysis.

Conclusion: These data demonstrate that sn-1,2 DAGs in the PM, derived from re-esterification of exogenous fatty acids, are sufficient to induce hepatic insulin resistance at the level of IRK.

Disclosure

K. Lyu: None. Y. Zhang: None. D. Zhang: None. M. Kahn: None. Y. Nozaki: None. S. Bhanot: None. J. Bogan: None. G. Cline: None. V. Samuel: None. G.I. Shulman: Advisory Panel; Self; AstraZeneca, Janssen Research & Development, Merck & Co., Inc. Advisory Panel; Spouse/Partner; Merck & Co., Inc. Board Member; Self; Novo Nordisk A/S. Consultant; Self; Aegerion Pharmaceuticals, IMetabolic BioPharma Corporation, Longitude Capital, Nimbus Discovery, Inc., Staten Biotechnology B.V.

Funding

National Institutes of Health (R01DK116774)
268-LB: Investigating the Role of MPO and ANGPTL6 in Obesity and Diabetes and Their Potential Role in Wound Healing
Authors: QADDOUMI MOHAMMAD G. , ABU-FARHA MOHAMED , THOMAS CHERIAN PREETHI , AL KHAIRI IRINA , MOHAMED CHANNANATH ARSHAD , ALANBAEI MUATH , ABUBAKER JEHAD , AL-MULLA FAHD
Published: June 2019

Purpose: Myeloperoxidase (MPO) is a major neutrophil-derived protein known to play an important role in inflammation, oxidative stress, and atherosclerosis. Like other angiopoietin-like protein (ANGPTL), ANGPTL6 is reported to regulate metabolic processes but is thought to counteract obesity through increased energy expenditure. This study evaluated the plasma level of both MPO and ANGPTL6 in obese and diabetic people to investigate their clinical association.

Methods: A total of 204 people were enrolled in this study (109 nondiabetic and 95 T2D). ANGPTL6, MPO, and other biomarkers plasma levels were measured by ELISA.

Results: MPO plasma level was unchanged between diabetics and nondiabetics, whereas ANGPTL 6 plasma level increased in diabetics (33.73±1.31 pg/mL) compared to nondiabetics (28.74±1.01 pg/mL) (P <0.003). However, MPO plasma level increased in obese diabetics (75.28±3.19 ng/mL) compared to non-obese diabetics (65.71±2.54 ng/mL) (P <0.02). Similarly, ANGPTL6 plasma level was increased in obese diabetics (35.74±1.68 pg/mL) compared to non-obese diabetics (29.05 ± 1.61 pg/mL) (P <0.01). Obese nondiabetics had a higher level of MPO compared to non-obese nondiabetics although it did not reach statistical significance (P = 0.05). In contrast, no changes in the plasma levels of ANGPTL6 were seen in nondiabetics irrespective of weight. Both MPO and ANGPTL plasma levels were higher in obese than non-obese people (P < 0.05). MPO was positively associated with ANGPTL6, TG, BMI, TNF alpha, HsCRP, IL6, and PAI1.

Conclusion: Both MPO and ANGPTL6 seem to regulate obesity, although MPO might do it independently of diabetes status and they might be also involved in wound healing as previously reported. The positive association of MPO with several biomarkers of inflammation and atherosclerosis may underlie its pathogenic mechanism in obesity and diabetes related wound healing and other diabetes related pathways.

Disclosure

M.G. Qaddoumi: None. M. Abu-farha: None. P.T. Cherian: None. I. Al khairi: None. A.M. Channanath: None. M. Alanbaei: None. J. Abubaker: None. F. Al-Mulla: None.
270-OR: Adipocyte Integrin-Linked Kinase Is Necessary for the Development of Diet-Induced Adipose Insulin Resistance
Authors: BUGLER-LAMB AIMEE , HENNAYAKE CHANDANI , HASIB ANNIE , ASHFORD MICHAEL L. , KANG LI
Published: June 2019

The remodeling of the extracellular matrix (ECM) in tissues including adipose tissue, and the expression of collagen binding integrin α2β1 have been associated with the development of insulin resistance. Integrin-linked Kinase (ILK), an adaptor protein and a key component of the IPP (ILK-Parvin-PINCH) complex mediates integrins linking the ECM with intracellular signalling. This study determines the role of ILK in adipose function and insulin resistance in chow and high fat (HF)-fed mice. HF feeding increased adipose ILK expression in mice (2.7 ±0.62 vs. 1±0.23; p=0.01). Adipocyte-specific deletion of ILK resulted in a decrease in fat mass (21.6±1.6% vs. 27.3±1.4%; p=0.009) and epididymal tissue weight (eWAT) (1.04±0.07g vs. 1.62±0.11g; p<0.001) in HF-fed mice, but not in chow-fed mice. During the hyperinsulinemic-euglycemic clamp, HF-fed ILK deficient mice showed increased glucose uptake (66.22±p=0.028) in the brown adipose tissue (BAT) compared with the wild type littermate controls. Increased glucose uptake in BAT is associated with increased phosphorylation of AKT (1.96±0.16 vs. 1±0.28 fold change; p=0.01), and decreased phosphorylation of p38 and JNK (0.23 ± 0.17 vs. 1±0.48; p=0.024 and 0.41 ±0.12 vs. 1±0.17; p=0.025 respectively). The inhibition of lipolysis by insulin was also improved in HF-fed ILK deficient mice determined by plasma NEFA level during the clamp. This could be attributed to decreased cell size (0.85±0.042 vs. 1±0.06; p=0.0001), decreased collagen deposition (22.8±0.98 vs. 30.28± 2.05; p=0.01) and a trend increase in vascularization (3.47±1.26 vs. 1±0.23; p=0.07) in the eWAT. These data suggest that adipocyte ILK plays an important role in regulating adipocyte morphology, signaling and glucose and lipid homeostasis, therefore representing a potential target for obesity, insulin resistance and diabetes.

Disclosure

A. Bugler-Lamb: None. C. Hennayake: None. A. Hasib: None. M.L. Ashford: None. L. Kang: None.

Funding

Diabetes UK; Diabetes Research & Wellness Foundation
282-LB: Dysregulated FGF21 Links Hepatic Insulin Resistance to Dysfunctional BAT
Authors: STOEHR OLIVER , TAO RONGYA , COPPS KYLE D. , WHITE MORRIS F.
Published: June 2019

LDKO (hepatic Irs1/Irs2 double knockout) mice show a severe diabetic phenotype owing to the impaired communication of the insulin resistant liver throughout the body by the dysregulated production and secretion of hepatokines. Excess Fst (follistatin) secreted by the liver of LDKO-mice inhibits insulin action in WAT (white adipose tissue) and promotes hepatic glucose production; however, Fst does not dysregulate BAT (brown adipose tissue). Regardless, BAT in LDKO-mice lacked typical BAT characteristics on the anatomical and molecular level. Glucose uptake was severely impaired, which might contribute to the diabetic phenotype of LDKO-mice. Succinate stimulates BAT and protects control mice against obesity and hyperglycemia during HFD feeding; however, succinate has no beneficial effect in LDKO-mice revealing non-responsive/dysfunctional BAT in these mice. Transplantation of healthy BAT from wild type mice into LDKO-mice was unbeneficial, whereas transplantation of healthy BAT into control mice improved their whole-body glucose metabolism and glucose tolerance. Next, we investigated whether dysregulated hepatokines might impair BAT function in LDKO-mice. Among the dysregulated hepatokines in LDKO-mice FGF-21 (fibroblast growth factor 21) was reduced significantly. Re-expression of FGF-21 in the liver via adenoviral infection increased FGF-21 serum levels into the normal range and restored normal glucose uptake into BAT and improved glucose tolerance during chow or HFD feeding. These results suggest that reduced FGF-21 promotes BAT dysfunction during hepatic insulin resistance, whereas re-expression of FGF-21 restored glucose metabolism regardless of the persistent hepatic insulin resistance and uncontrolled gluconeogenesis owing to elevated hepatic Fst.

Disclosure

O. Stoehr: None. R. Tao: None. K.D. Copps: None. M.F. White: Advisory Panel; Self; Housey Pharmaceutical Research Laboratories.
282-OR: Impact of a Hybrid Closed-Loop Insulin Delivery System on Hypoglycemia Awareness in Individuals with Type 1 Diabetes
Authors: BURCKHARDT MARIE-ANNE , DART JULIE , SMITH GRANT J. , ABRAHAM MARY B. , PARAMALINGAM NIRUBASINI , O'DEA JOANNE M. , BOCK MARTIN DE , DAVIS ELIZABETH A. , JONES TIMOTHY
Published: June 2019

Background: Individuals with impaired awareness of hypoglycemia (IAH) are at risk of severe hypoglycemia. Although it is known that elimination of hypoglycemia can restore hypoglycemia awareness in this high-risk group, this has been challenging in real life. Technological advancements like closed loop system provide an opportunity to minimize hypoglycemia. Hence, the aim of this study was to evaluate whether a hybrid closed loop insulin delivery system can improve hypoglycemia awareness in individuals with IAH.

Methods: Participants with type 1 diabetes and IAH (Gold Score ≥4) were randomized to either a hybrid closed loop system (Medtronic 670G) (intervention) or standard insulin pump therapy (control) for 8 weeks. Gold scores and hyperinsulinemic hypoglycemic clamp studies were undertaken at baseline and at the end of the study with evaluation of the hormonal and symptomatic responses to hypoglycemia.

Results: A total of 17 participants completed the study (mean (SD) age 35.8 years (11.2), diabetes duration 24.2 years (11.3), HbA1c 7.8% (1.2)). 8 participants were randomized to control and 9 participants to intervention. The baseline (Median, IQR) Gold score was 6.0 (5.0; 6.0) in the overall cohort and did not differ between groups. At the study end, Gold score improved in the intervention group: control vs. intervention 6.0 (4.5; 6.0) vs. 3.5 (3.0; 4.5); p = 0.027. Symptom scores (adrenergic and neuroglycopenic) during the clamp were not different between the control and intervention groups.

Conclusions: The use of a hybrid closed loop system over 8 weeks improved self-reported hypoglycemia awareness, however there was no evidence for improvement in the symptomatic responses to experimentally induced hypoglycemia.

Disclosure

M. Burckhardt: None. J. Dart: None. G.J. Smith: None. M.B. Abraham: None. N. Paramalingam: None. J.M. O'Dea: None. M. de Bock: None. E.A. Davis: None. T. Jones: Research Support; Self; Dexcom, Inc., Medtronic MiniMed, Inc. Speaker's Bureau; Self; Eli Lilly and Company, Roche Diabetes Care.
284-OR: How Do Patients and Caregivers Prioritize Outcomes for Type 1 Diabetes (T1D) Treatment?
Authors: MARINAC MARJANA , HUTTON CAMPBELL T. , MANSFIELD CAROL , SUTPHIN JESSIE , GALLAHER KATHLEEN , SULLIVAN SEAN
Published: June 2019

Background: Currently, HbA1c is the primary measure of treatment effectiveness for type 1 diabetes (T1D). We investigated patient priorities for measures of T1D control in addition to HbA1c.

Method: We developed discrete-choice experiment surveys for adult patients and caregivers of children with T1D that presented choices between hypothetical treatments described by 6 attributes with varying levels: HbA1c, time in range, weekly number and severity of hypoglycemia and hyperglycemia events, additional disease management time, and additional treatment cost. The clinical outcomes were defined based on a 2017 consensus report standardizing the definition of outcomes beyond HbA1c. Respondents answered nine treatment choice questions, and the data were analyzed using random-parameters logit.

Results: Recruited through an online market research panel, 300 adult patients and 400 caregivers completed the survey. Patients and caregivers placed the most importance on reducing low and high events. Additional time in range was as important as a reduction of more than ½ point in HbA1c, but both were less important than avoiding weekly lows and highs. Patients who did not use an insulin pump or CGM placed relatively more weight on reducing lows and highs and on improvements in HbA1c and time in range compared to patients who use technology. For patients, avoiding 1 to 5 lows (54-69 mg/dL) per week was 5 times more important than achieving target HbA1c from ½ point above. Avoiding 1 to 5 highs (181-250 mg/dL) per week was 7 times more important than achieving target HbA1c from ½ point above. Caregivers of children < 12 years placed relatively more weight on avoiding lows and highs than those with older children.

Conclusion: Overall, patients and caregivers prioritized control of hypoglycemia and hyperglycemia events, including mild to moderate events. The research has implications for outcomes used to measure the benefits of treatment in research, development, and reimbursement of therapies for T1D.

Disclosure

M. Marinac: None. C.T. Hutton: None. C. Mansfield: Other Relationship; Self; Multiple companies. J. Sutphin: Other Relationship; Self; Multiple Companies. K. Gallaher: Other Relationship; Self; Multiple companies. S. Sullivan: None.

Funding

The Leona M. and Harry B. Helmsley Charitable Trust; JDRF International
288-LB: Effects of the Hepatic Mitochondrial Calcium Uniporter on Hepatic Gluconeogenesis and Mitochondrial Metabolism in Awake Mice
Authors: LEE JI EUN , GOEDEKE LEIGH , ZHANG YE , PERRY RACHEL J. , GOODMAN RUSSELL , MOOTHA VAMSI , SHULMAN GERALD I.
Published: June 2019

Increased hepatic glucose production, due to increased rates of gluconeogenesis, is a contributor to both fasting and postprandial hyperglycemia in patients with T2D. Recent evidence implicates a role for hepatic calcium signaling in mediating glucagon's effect to stimulate glucose production and mitochondrial oxidation; however, the cellular and molecular mechanisms by which glucagon promotes these alterations remain poorly understood. We hypothesize that an acute physiological increase in plasma glucagon concentrations will increase cytosolic and mitochondrial calcium concentrations, promoting increases in rates of hepatic mitochondrial oxidation and gluconeogenesis in a transcription-independent manner. To test this hypothesis, we used mice with a liver-specific loss of the mitochondrial calcium uniporter (MCU), which is the main transporter that allows rapid transport of calcium into the mitochondrial matrix. We found that glucagon increased oxygen consumption rate by 80% (p<0.0001) in WT hepatocytes, while this effect was abrogated in MCU KO hepatocytes. We also measured hepatic mitochondrial oxidation (VCS) in vivo using Positional Isotopomer NMR Tracer Analysis (PINTA) and found a 74% decrease in VCS (p=0.05) in MCU KO mice compared to WT mice. This suggests that tightly regulated mitochondrial calcium signaling is necessary for regulating hepatic mitochondrial oxidation. Next, we measured intrahepatic lipolysis and hepatic gluconeogenesis in vitro and in vivo. We found that glucagon increased lipolysis (p=0.05) and glucose production (p=0.024) by ∼1.5 fold in MCU KO compared to WT hepatocytes. Furthermore, in vivo PINTA results showed that glucagon increased pyruvate carboxylase flux (VPC) by 68% (p=0.08) in MCU KO mice. Taken together, these results suggest that mitochondrial calcium signaling through MCU is necessary to regulate the effect of glucagon to modulate hepatic gluconeogenesis and mitochondrial oxidation.

Disclosure

J. Lee: None. L. Goedeke: None. Y. Zhang: None. R.J. Perry: Research Support; Self; AstraZeneca. R. Goodman: None. V. Mootha: Advisory Panel; Self; Janssen Pharmaceuticals, Inc. Consultant; Self; 5AM Ventures. Other Relationship; Self; Raze Therapeutics. G.I. Shulman: Advisory Panel; Self; AstraZeneca, Janssen Research & Development, Merck & Co., Inc. Advisory Panel; Spouse/Partner; Merck & Co., Inc. Board Member; Self; Novo Nordisk A/S. Consultant; Self; Aegerion Pharmaceuticals, IMetabolic BioPharma Corporation, Longitude Capital, Nimbus Discovery, Inc., Staten Biotechnology B.V.

Funding

National Institutes of Health
289-LB: Hepatic Gamma-Amino Butyric Acid Release Drives Hyperinsulinemia and Insulin Resistance
Authors: GEISLER CAROLINE , GHIMIRE SUSMA , RENQUIST BENJAMIN J.
Published: June 2019

The degree of hepatic lipid accumulation in obesity directly correlates with the severity of hyperinsulinemia and systemic insulin resistance. Here, we propose a mechanism that explains this associative link, whereby, hepatic steatosis dysregulates glucose and insulin homeostasis. Obesity-induced lipid accumulation results in hepatocyte depolarization. We have established that hepatocyte depolarization depresses hepatic vagal afferent nerve (HVAN) firing (22.9 ± 7.6%; P < 0.05). The HVAN plays a key role in regulating whole-body glucose homeostasis. Accordingly, we propose that dysregulated hepatocyte-vagal communication underlies the metabolic dysfunction in obesity. Using an ex vivo explant liver slice model, we showed that diet induced obesity nearly triples hepatic GABA release/mg DNA in mice (P < 0.05). We hypothesize that hepatic GABA release decreases HVAN activity to stimulate insulin release and decrease peripheral insulin sensitivity. To establish that hepatocyte depolarization encourages GABA export in obesity, we virally induced expression of Kir2.1, a hyperpolarizing channel, in hepatocytes. Kir2.1 expression decreased hepatic slice GABA release to half of that observed in control obese mice, while protecting against hyperinsulinemia and insulin resistance on a high fat diet. Hepatic GABA is synthesized by GABA-Transaminase (GABA-T). To determine the role of hepatic GABA production in diet induced metabolic dysfunction, we treated obese mice with the irreversible GABA-T inhibitors, ethanolamine-O-sulphate (EOS) or vigabatrin (8mg/day; IP). Within 5 days, GABA-T inhibition ameliorated the hyperinsulinemia and insulin resistance of obesity (P < 0.05). These improvements in glucose homeostasis are dependent on an intact HVAN, as EOS did not affect serum insulin or insulin sensitivity in obese hepatic vagotomized mice. This work establishes that limiting hepatocyte GABA production or release can mute the glucoregulatory dysfunction common to obesity.

Disclosure

C. Geisler: None. S. Ghimire: None. B.J. Renquist: None.

Funding

Arizona Biomedical Research Commission (ADHS14-082986, ADHS18-201472 to B.J.R.); American Heart Association (15BGIA25090300 to B.J.R.)
29-LB: A Multicenter, Randomized Controlled Study of the Efficacy of Once-Weekly DPP-4 Inhibitor Omarigliptin in Patients with Type 2 Diabetes Undergoing Maintenance Hemodialysis
Authors: YOSHIZAWA YUTA , HOSOJIMA MICHIHIRO , KABASAWA HIDEYUKI , TANABE NAOHITO , KITAMURA TADAHIRO , NARITA ICHIEI , SAITO AKIHIKO
Published: June 2019

Background: DPP-4 inhibitors are used widely in patients with type 2 diabetes (T2D) undergoing maintenance hemodialysis (HD), but the efficacy of the once-weekly DPP-4 inhibitor omarigliptin (O) is unknown.

Methods: This open, multicenter, randomized, non-inferiority trial compared glycemic control and safety of O and the daily DPP-4 inhibitor linagliptin (L) (UMIN000024284). We enrolled 33 T2D patients undergoing maintenance HD, who had been treated with L for at least 3 months at Niigata University Medical and Dental Hospital and three affiliated dialysis facilities between April 2017 and March 2018. The patients were randomized to receive O (n = 16) or L (n = 17). Primary endpoints were changes in HbA1c and glycoalbumin (GA) over 24 weeks from baseline (with non-inferiority margins of 0.25% and 0.75%, respectively). Secondary endpoints were changes in blood glucose and plasma glucagon measured by sandwich ELISA after single HD and the incidence of adverse events including hypoglycemia.

Results: In the full analysis set (n = 30), the difference in the mean change in primary endpoint values between the O and L groups was −0.6% for HbA1c and −1.7% for GA, with one-tailed upper 95% limits of −0.17% and 0.4%, respectively, below the respective non-inferiority limits. HbA1c reduction was significantly greater in the O group (−0.2% ± 0.6%) than in the L group (0.4% ± 0.8%, two-tailed p = 0.027). Blood glucose reduction after single HD was significantly greater in the O group (−18.4 ± 31.4 mg/dL) than in the L group (25.2 ± 59.5 mg/dL, two-tailed p = 0.025), but plasma glucagon reduction after single HD was not greater in the O group (−6.8 ± 14.1 pg/dL) than in the L group (2.6 ± 16.6 pg/dL, two-tailed p = 0.11). No subjects in the O group experienced hypoglycemia.

Conclusion: Omarigliptin is feasible for glycemic control in T2D patients undergoing maintenance HD. Further studies are needed to determine the mechanisms of its efficacy.

Disclosure

Y. Yoshizawa: None. M. Hosojima: Research Support; Self; Astellas Pharma Inc., Biotech Japan Corporation, Daiichi Sankyo Co.,Ltd., Eli Lilly Japan K.K., Forica Foods Co.,Ltd., Kameda Seika Co.,Ltd., Kyowa Hakko Kirin Co., Ltd., Mitsubishi Tanabe Pharma Corporation, MSD K.K., Novo Nordisc Pharma Ltd., Sato Foods Industries Co.,Ltd., Taisho Toyama Pharmaceutical.Co.,Ltd., Takeda Pharmaceutical Co.,Ltd. H. Kabasawa: Research Support; Self; Astellas Pharma Inc., Biotec Japan Corporation, Daiichi Sankyo Company, Limited, Forica Foods CO.,LTD., KAMEDA SEIKA CO., LTD., Kyowa Hakko Kirin Co., Ltd., Mitsubishi Tanabe Pharma Corporation, MSD K.K, Novo Nordisc Pharma Ltd, Sato Foods Industries CO.,LTD., Taisho Toyama Pharmaceutical.CO.,LTD., Takeda Pharmaceutical Company Limited. N. Tanabe: None. T. Kitamura: None. I. Narita: None. A. Saito: None.
29-OR: High Prevalence of Pathogenic Mutations in Genes Causing Monogenic Diabetes among Patients with Common Type 2 Diabetes
Authors: BONNEFOND AMELIE , FROGUEL PHILIPPE ,   LILLE-D.E.S.I.R. GENETICS STUDY
Published: June 2019

In contrast to monogenic diabetes, the genetic investigations of common forms of type 2 diabetes (T2D) have led to modest advances in precision medicine. We aimed to assess the prevalence in common T2D of pathogenic mutations across 33 genes causing monogenic diabetes, and to compare it to the prevalence found in normoglycemic individuals. Through next-generation sequencing, genes were sequenced in a case-control study including more than 6,000 individuals. We then applied stringent quality control steps before the analyses. The pathogenicity of variants was assessed using the American College of Medical Genetics and Genomics criteria. Primary outcome was the prevalence of pathogenic or likely pathogenic mutations in the case-control study. Secondary outcomes included age of T2D diagnosis, body mass index, T2D treatment and T2D family history. We found a significant association between pathogenic or likely pathogenic variants and T2D risk (p<0.001; 6.5% of carriers among cases). This association was explained in part by pathogenic mutations of two actionable genes, GCK and HNF1A. Overall, among cases, carriers were leaner (β=-1.5±0.5 kg/m²; p<0.01) and tended to develop T2D earlier (β=-1.2±0.7 years; p=0.05) compared to non-carriers, but use of antidiabetic drugs and T2D family history were not different. We reported a high prevalence of pathogenic mutations in monogenic diabetes genes among common T2D patients who did not present with clinical characteristics of monogenic diabetes (i.e., MODY). These results could lead to precision diabetic medicine, and to significant savings in health expenditure via systematic molecular diagnosis of newly diagnosed patients.

Disclosure

A. Bonnefond: None. P. Froguel: None.

Funding

European Research Council; French National Research Agency
298-LB: GLP-1 Secretion Enhancement after H. Pylori Eradication Treatment Is Related to Gut Microbiota Modification
Authors: CORNEJO-PAREJA ISABEL , MARTIN-NUÑEZ GRACIA MARÍA , MORENO-INDIAS ISABEL , CARDONA-DÍAZ FERNANDO F. , ROCA-RODRIGUEZ M. MAR , COIN-ARANGüEZ LETICIA , MANCHA-DOBLAS ISABEL , TINAHONES FRANCISCO
Published: June 2019

Introduction: Antibiotic treatments modify gut microbiota. Microbiota alterations (in composition and biodiversity) are fundamental pieces in the development of metabolic disorders (T2DM and obesity).

Objective: To evaluate the relationship between microbiota changes due to antibiotic treatment in patients colonized by H. pylori and GLP-1 secretion.

Methods: A prospective case control study was assessed. We evaluated a H. pylori positive patient model before and two months after a short-term eradication therapy and a control group. Anthropometric measurements, carbohydrate and lipid metabolisms, GLP-1 levels and fecal microbiota composition (determined by 16S rRNA gene (V3-V4) sequencing with an Illumina Miseq) were analyzed.

Results: We studied 40 cases and 20 controls (60% women, respectively). Average age was 47.0±2 vs. 43.6±2.7 years. 42.5% vs. 40% had clinical history of gastrointestinal disease. After eradication treatment, we found significant changes in microbiota profile at phylum, family, genus and species levels. The Chao1 and Shannon indices showed a decreased in bacterial richness and diversity in patients (pre and post H. pylori eradication) compared to controls. GLP-1 secretion and carbohydrate metabolism were improved after antibiotic treatment. GLP-1 changes were related to microbial community disturbances through correlation analysis, specifically with Bifidobacterium adolescentis (r=0.354, p=0.034), Lachnobacterium (r=-0.332, p=0.048) and Coriobacteriaceae (r=0.372, p=0.026).

Conclusions: Antibiotic treatment to eradicate H. pylori increased the secretion of GLP-1 and improved the carbohydrate metabolism. This GLP-1 secretion improvement was related to changes in microbial community because of the antibiotic treatment, particularly in Bifidobacterium adolescentis, Lachnobacterium and Coriobacteriaceae. These taxa could play their roles through the production of short chain fatty acids (SCFA) and/or bile acid metabolism.

Disclosure

I. Cornejo-Pareja: None. G. Martin-Nuñez: None. I. Moreno-Indias: None. F.F. Cardona-Díaz: None. M. Roca-Rodriguez: None. L. Coin-Arangüez: None. I. Mancha-Doblas: None. F. Tinahones: Advisory Panel; Self; Lilly Diabetes, Novo Nordisk A/S, Regeneron Pharmaceuticals, Sanofi-Aventis. Consultant; Self; AstraZeneca. Research Support; Self; AstraZeneca. Speaker’s Bureau; Self; Danone Nutricia Research, Lilly Diabetes, Novo Nordisk A/S, Regenerative Medical Solutions, Sanofi-Aventis.
30-LB: Diabetes Complications Including Nephropathy, Neuropathy, and Retinopathy Can Now Be Studied in a New Spontaneous Autoimmune Diabetes Transgenic Model
Authors: IMAM SHAHNAWAZ , JAUME JUAN C. , ALFONSO-JAUME MARIA A.
Published: June 2019

We have developed a spontaneous humanized transgenic mouse model of autoimmune diabetes chronically develops diabetic complications.

Our humanized mouse develop diabetic nephropathy. The basal glomerular membrane thickness was doubled in size at 4 months of age as shown by electron microscopy. The manifestation of glomerulopathy subsequently correlates with microalbuminuria. At 6 month, mice show advanced features of diabetic nephropathy like focal segmental/global glomerular sclerosis and focal tubular atrophy. The typical features of Kimmelstiel-Wilson are seen when severe chronic kidney disease develops.

Our humanized mice develop diabetic retinopathy. We have noted a significant increase in acellular capillaries of the retina at the age of 6 months. Ocular tomography revealed a significant reduction in total retinal thickening. Fundoscopic retinal images revealed a classic vascular leakage as observed in humans.

Our humanized mice also show diabetic neuropathy. We have observed an impaired sciatic and optic nerve conduction. Specifically, optic nerves at 6-months old diabetic mice showed an increased refractory period of the second compound action potential (CAP). This increase in the refractory period of CAP in the diabetic optic nerves is consistent with the reduced conduction velocity. However, a limb vasculopathy has not been observed so far.

In our mouse model, the mouse MHC-II was replaced with human DQ8 in all antigen-presenting cells. The mouse also expresses human GAD65 in pancreatic beta cells. The diabetic phenotype of our mouse mimics the human disease progression. Importantly, our mouse model has a genetic background that compromises beta-cell neogenesis and/or regeneration (1).

Our humanized mice model has the closest known phenotype to human diabetes and is most suitable for addressing physiopathology and treatment of diabetes complications. (U.S. Provisional Patent #62/713,827).

Disclosure

S. Imam: None. J.C. Jaume: None.

Funding

University of Toledo
302-OR: Causes of Blinded CGM Failure during Clinical Trials in Patients with T2D
Authors: SALDIVAR-SALAZAR SERGIO , TESTA MARCIA A. , HAYES JOHANNA F. , SIMONSON DONALD C.
Published: June 2019

Blinded CGM is useful in clinical trials to preserve the validity of outcomes. However, lack of real-time feedback about performance of the system generates a significant number of failed sessions. We analyzed causes of such failed CGM sessions in 453 T2D subjects (mean age 55.2±9.6 years, 51.9% males) using the Medtronic iPro2 to obtain a 6-day baseline CGM during two randomized clinical trials of oral agents and insulin. At the end of the CGM session, non-calibrated data (ISIG - electric signals) were immediately reviewed by a third party to determine the completeness and quality of the dataset (at least 72 continuous hours of ISIG between 10-200 nA) and the need for a new CGM session. If data were not acceptable, clinical sites were promptly contacted to ascertain the conditions of the study subject, insertion technique and technical problems that could cause the failed session, and to recommend other approaches to CGM reinsertion. Subjects who failed the first CGM sessions were reinserted a second or third time when possible. Eighty-two of 453 subjects failed their first CGM insertion, 9 of 56 failed the second insertion, and one of four subjects failed all three insertions (cumulative success rate = 99.8%). The causes of failure were discernible in 38 (41.3%) of the 92 failed cases, including sensor detachment (19.6%), insufficient BG for calibration (17.4%), user error (3.3%), and intentional removal (1.1%). For the remaining 58.7% of cases, the non-calibrated ISIG patterns were analyzed and most likely causes found to be undetermined (25.0%), disconnection (20.7%), sensor failure (8.7%) and device failure (4.3%). Upon clinical review, most failed sessions of undetermined cause or probable disconnection were attributable to patient characteristics (morbid obesity, profuse sweating) or improper insertion technique.

Conclusions: 1) Most identifiable causes of failed CGM can be potentially corrected by proper user/subject training, and 2) only a minimal portion of failed sessions are secondary to faulty devices.

Disclosure

S. Saldivar-Salazar: Speaker's Bureau; Self; Medtronic MiniMed, Inc. M.A. Testa: Stock/Shareholder; Spouse/Partner; GI Windows, Inc. Stock/Shareholder; Self; Phase V Technologies, Inc. J.F. Hayes: None. D.C. Simonson: Stock/Shareholder; Self; GI Windows, Inc. Stock/Shareholder; Spouse/Partner; Phase V Technologies, Inc.
303-LB: Accelerated Hepatic Fibrosis in Western Diet-Fed Nonalcoholic Steatohepatitis Mice Model by Small Dosage of Carbon Tetrachloride
Authors: ZHANG GUODONG , CHNG KEEFE , WANG YIXIN
Published: June 2019

Carbon Tetrachloride (CCl4) is a common hepatotoxin used in liver injury and fibrosis animal models, but it is rare to observe obesity and liver fat accumulation in CCl4-induced liver fibrosis models since weight loss is also provoked. Nonalcoholic Steatohepatitis (NASH) is an epidemic disease coincides with obesity and type 2 diabetes (T2D) causing liver steatosis and inflammation. There are no perfect animal models that capture all steatosis, inflammation and following fibrosis stages of NASH disease, our recent study developed an accelerated fibrotic model in previously established NASH model of western diet fed FATZO/pco mice. Using a dose of 0.08 mL/kg (IP, 2/week) CCl4 on 8-week western diet fed FATZO/pco mice, the average histology fibrosis score increased from 1.2±0.2 to 3.0±0.0, average lobular inflammation score increased from 1.4±0.7 to 1.9±0.1, while average steatosis score remained stable. Hepatotoxicity of CCl4 affected the hepatocyte ballooning score due to increased necrosis, reducing ballooning score from 1.6±0.4 to 1.0±0.0. It is also interesting to find out the FATZO/pco strain tolerated CCl4 challenges better than the control C57Bl/6 strain.

Conclusion: The study developed an accelerated NASH mice model with balanced performance in all stages of histopathology and provides better translational values in comparison with human patients.

Disclosure

G. Zhang: None. K. Chng: None. Y. Wang: None.
304-OR: Polygenic Risk Score Predicts Type 2 Diabetes Susceptibility in a Diverse Consumer Genetic Database
Authors: MULTHAUP MICHAEL L. , KITA RYOSUKE , ERIKSSON NICHOLAS , ASLIBEKYAN STELLA , SHELTON JANIE , TENNEN RUTH I. , KIM ESTHER , KOELSCH BERTRAM
Published: June 2019

With the rise in the prevalence of type 2 diabetes (T2D), as well as undiagnosed cases of T2D and prediabetes (25% and 90%, respectively), early detection is imperative to minimize individual and societal burden. T2D is highly heritable, and personal genetic information is increasingly available to the general public. Studies have suggested that T2D risk reduction strategies may be more effective for individuals with high T2D genetic risk, supporting the use of genetics as a screening tool to inform cost-effective interventions.

We trained a polygenic risk score (PRS) for T2D based on >1,200 genotyped variants in >600,000 European consented research participants from a consumer genetic database who self-reported if they had been diagnosed with T2D. We tested the PRS' performance in separate sets of participants covering five different ancestries (African-American, East-Asian, European, Latino, and South-Asian; ~ 600,000 participants).

The area under the receiver-operator curve of this PRS varied from 0.65 to 0.57, performing best in European and worst in African ancestries. The PRS was calibrated separately in each ancestry to account for differences in T2D prevalence. European participants with a PRS in the top 5% of the distribution have a T2D odds ratio of more than 3, and lifetime risk for this group exceeds 65%. Our PRS is strongly correlated with an independently derived T2D PRS from (Scott et al. 2017 GWAS, Spearman rho=0.44, p < 1x10^-200) but is more predictive in our dataset (AUC 0.65 vs. 0.59). Lastly, we defined an "increased likelihood" result based on the PRS threshold at which risk of T2D from genetics alone exceeds the risk of T2D due to being overweight. In our database, 19% of individuals met this criterion.

We find that personalized PRS' have the potential to identify large numbers of individuals with increased T2D susceptibility equal to or greater than known risk factors and could prove useful in evaluating T2D risk at individual as well as population levels.

Disclosure

M.L. Multhaup: Employee; Self; 23andMe. R. Kita: Employee; Self; 23andMe. N. Eriksson: None. S. Aslibekyan: Employee; Self; 23andMe. J. Shelton: None. R.I. Tennen: Employee; Self; 23andMe. E. Kim: Employee; Self; 23andMe, inc. B. Koelsch: Employee; Self; 23andMe.
305-OR: Genomic Basis to Shared Influences on Pubertal Timing and Type 2 Diabetes
Authors: COUSMINER DIANA L. , MISHRA RAJASHREE , ARGENZIANO MARIANA A. , MANDUCHI ELISABETTA , HODGE KENYAITA M. , SU CHUN , LEONARD MICHELLE , LU SUMEI , PIPPIN JAMES A. , JOHNSON MATTHEW , WELLS ANDREW D. , CHESI ALESSANDRA , VOIGHT BENJAMIN F. , GRANT STRUAN F.
Published: June 2019

Earlier pubertal onset in girls is associated with later-life type 2 diabetes (T2D) risk. Studies have shown that age at menarche (AAM), a marker of girls’ pubertal timing, and T2D are genetically correlated genome-wide. Since both AAM and T2D also correlate with BMI, the biological link between the two traits could be mediated via BMI. As the most recent genome-wide association study (GWAS) for T2D also adjusted for BMI, we queried if AAM and T2D remain correlated after removing the effect of BMI on T2D. Leveraging such large-scale GWAS data, LD Score Regression revealed that the genetic correlation between AAM and T2D (rg (SE)=-0.24 (0.02) P=2.2x10-22) was only partly attenuated after BMI adjustment (rg (SE)=-0.1 (0.03) P=0.0002), suggesting that BMI does not entirely explain the puberty-T2D link. Five of the 13 significant GWAS-implicated loci associated with both AAM and T2D do not associate with BMI, further supporting that BMI is not the only mediator between the two traits. Given that causal effector genes are unknown at most GWAS loci, identification of target genes at these shared loci should provide key biological insights. To implicate effector genes, we first identified all SNPs in LD with the sentinel SNPs. We then extracted the subsets of proxy SNPs in open chromatin, determined by ATAC-seq, in primary cells or cell lines relevant to this trait area [beta cells (EndoC-βH1), adipose (SGBS and MSC-derived adipocytes), and—given recent findings in both AAM and BMI studies—brain (neural precursors, microglia, astrocytes and hypothalamic neurons)]. Using high resolution Capture-C, we detected consistent physical contacts between open-proxy SNPs and candidate effector genes. Our results support direct SNP-to-gene promoter contacts for at least four of these shared loci (one not associated with BMI) including TSPAN3 at ‘AC046168.1’ and TFAP2D at ‘TFAP2B’, along with leads at the ‘MTCH2/CELF1’ and ‘MAP2K5’ loci. Follow-up experiments are needed to determine the physiological roles of implicated target genes.

Disclosure

D.L. Cousminer: None. R. Mishra: None. M.A. Argenziano: None. E. Manduchi: None. K.M. Hodge: None. C. Su: None. M. Leonard: None. S. Lu: None. J.A. Pippin: None. M. Johnson: None. A.D. Wells: Research Support; Self; GlaxoSmithKline plc. A. Chesi: None. B.F. Voight: None. S.F. Grant: None.

Funding

American Diabetes Association (1-17-PDF-077 to D.L.C.); National Institutes of Health
309-LB: Cell-Permeable Truncated SOCS3 Competitively Inhibits the Interaction Between Endogenous SOCS3 and Leptin Receptor (ObR) to Overcome Leptin Resistance on JAK/STAT Signaling
Authors: LEE KUYSOOK
Published: June 2019

Aim: The abnormal control of leptin, an adipocyte-derived hormone controlling appetite, chronically enhances obesity. On binding to leptin receptor (ObR) expressed in hypothalamic neurons, leptin evokes JAK/STAT signaling, and induces the expression of suppressor of cytokine signaling 3 (SOCS3) as a negative feedback regulator to maintain the homeostatic balance between food intake and energy expenditure. However, due to excess leptin in obesity, increased levels of endogenous SOCS3 promotes leptin resistance and disables the appetite-control by leptin. We aim to develop cell-/tissue-permeable form of a truncated, trans-dominant SOCS3 protein (CP-?SOCS3) to antagonize leptin resistance caused by binding of endogenous SOCS3 to ObR and test its use as an anti-obesity and antidiabetic agent.

Method: We have developed CP-?SOCS3 by fusing truncated SOCS3 with a hydrophobic cell-penetrating peptide, namely advanced macromolecule transduction domain. CP-?SOCS3 was tested using in vitro and in vivo leptin resistance models (in which endogenous SOCS3 is highly expressed) for the ability to intervene in JAK/STAT signaling and promote leptin sensitivity.

Results: CP-?SOCS3 competitively bound at the p-Y985 of ObR against endogenous SOCS3 and significantly promoted leptin-induced JAK/STAT signaling in vitro. Diet-induced obese (DIO) mice treated with CP-?SOCS3 showed 12.1% body weight loss in 14 days (15 mg/kg, IV, daily injection). In addition, CP-?SOCS3 reduced blood cholesterol by 31% and improved glycemic control by 67% in the DIO mice. CP-?SOCS3 was delivered into brain by penetrating brain blood barrier.

Conclusion: CP-?SOCS3 competitively suppressed the inhibitory action of endogenous SOCS3 on JAK/STAT signaling to enhance leptin sensitivity. Therefore, these results suggest that CP-?SOCS3 has a therapeutic potential as a protein-based anti-obesity and antidiabetic agent.

Disclosure

K. Lee: None.