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Nonalcoholic fatty liver disease (NAFLD) is the hepatic manifestation of metabolic syndrome as an arising health issue globally. Obesity and diabetes seem coherent with high NAFLD occurrence. Currently, its underlying mechanism of cause and progression remains to be elicited and effective therapy is unavailable. Huge investment on studying its pathogenesis and innovative therapy is underway. As mechanistic studies are more difficult or unethical to be performed in humans, animal model that recapitulates the human disease is urgently needed. Nonhuman primates (NHPs) with NAFLD can highly mimic the human disease. This study aimed to induce NAFLD by various diets (Table 1) in normoglycemic and diabetic NHPs with biomarker and histopathology assays to quantify NAFLD occurrence. Our data show that diabetic monkeys with high-fat-high-fructose (HFHF) diet developed NAFLD which could assemble to the human disease and be highly valuable for NAFLD research. However, low choline (LC), LC+10% fructose, LC+low methionine (LCM)+10% fructose diets for 31 weeks had no significant effects on liver function, except significant decrease in body weight and food intake in LCM+10%F group and some increase in liver histological scores in LC+10%F group (Table 1). In summary, normal diet with LC or LCM ± high fructose is hardly to induce NAFLD in normoglycemic NHPs, at least in 31 weeks. HFHF could induce or enhance NAFLD in diabetic NHPs. Disclosure J. Gao: Employee; Self; Crown Bioscience. D. Wang: Employee; Self; Crown Bioscience. Y. Ji: Employee; Self; Crown Bioscience. X. Zhang: Employee; Self; Crown Bioscience. C. Yang: None. J. Zhang: Employee; Self; Crown Bioscience. W. Ye: Employee; Self; Crown Bioscience. F. Shi: Employee; Self; Crown Bioscience. M. Jiang: Employee; Self; Crown Bioscience. Q. Jin: Employee; Self; Crown Bioscience. D. Zhang: None. X. Wang: None. W. Song: Employee; Self; Crown Bioscience. K. Chng: None. H. Jiang: Employee; Self; SYSE Bio-tec Co., Ltd. Y. Wang: None. Y. Xiao: Employee; Self; Crown Bioscience.

MS-NASH mouse is a leptin pathway intact model of spontaneous metabolic diseases. Our previous work developed a nonalcoholic steatohepatitis (NASH) model in MS-NASH mice fed with western diet and fructose drinking water (WDF). This study showed aggravated liver cholesterol content and fibrosis score by using a modified Amylin diet (GAN). Liver Cholesterol was increased from 10.11±1.58 mg/g to 14.07±0.81 mg/g and fibrosis score increased from 1.5±0.3 to 2.1±0.2 when comparing GAN with WDF and with C57Bl/6 mice. Administration of 30 mg/kg Obeticholic Acid (OCA, QD, PO) or 30 mg/kg Elafibranor (GFT-505, QD, PO) for 8 weeks significantly reduced serum liver enzymes, liver lipids and fibrosis scores. Conclusion: The GAN diet fed MS-NASH mice can be a better NASH model for novel compound efficacy. Disclosure G. Zhang: None. K. Chng: None. Y. Wang: None.

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.

The FATZO mouse is a polygenic model with an intact leptin system and was developed by cross breeding of C57BL6/J (C57) with AKR/J. FATZO mice have been used in many dysmetabolic studies. The epidemic health issue of nonalcoholic fatty liver disease (NAFLD, including NASH) is the organ dysmetabolic manifestation. This study was to investigate a high fat/fructose diet (HFFD) on liver function in FATZO mice. The body weights of FATZO (n=30) at age of 9 to 10 weeks were significantly heavier than those of age-matched C57 (n=19) mice (39±0.4g vs. 23±0.2g, p Disclosure X. Wang: None. W. Ye: None. Y. Chen: None. X. Zhang: None. Y. Wang: None. Y. Xiao: Employee; Self; Crown Bioscience Inc.

Introduction: Cardiomyopathy is the leading cause of morbidity and mortality among all complications of type 2 diabetic (T2D) and obese patients. Diabetic cardiomyopathy (DC) is characterized by an initial cardiac hypertrophy followed by thinning of the cardiac walls with declines in both systolic and diastolic functions, which ultimately leads to heart failure. No rodent models fully captured phenotypes of DC. The ZDSD rat, a new generation of T2D rat model with intact leptin signaling features with slow onset of diabetes and obesity, which closely mimics the development of the disease in patients. Here we sought to evaluate the cardiac function during the development of metabolic syndromes in ZDSD rats. Methods: 12 male ZDSD rats and age-matched SD controls were monitored for blood pressure, glucose, and cardiac function using echocardiography. Animals were also challenged with 1 mg/kg dobutamine for the assessment of cardiac reserve. Results: ZDSD rats developed hypertension from age of 18 weeks. Their left ventricular (LV) functions were compromised along with changes in cardiac morphology. At resting state, ZDSD rats showed LV hypertrophy from age of 18 to 22 weeks after which cardiac walls became thinner with larger LV volume. At 34 weeks old, both ejection fraction (EF) and transmitral E/A ratio declined. Upon treatment with dobutamine for 5 minutes, SD rats reached almost 98% EF, while the values of ZDSD were 91% at 30 and 34 weeks old, suggesting the loss of LV contractility the animals. Conclusion: ZDSD rats which carry multiple dysmetabolic phenotypes are spontaneously hypertensive with reduction in LV function and cardiac reserve which resembles ultrasonic symptoms of diabetic cardiomyopathy patients. Therefore, ZDSD rats may serve as a suitable preclinical model to study potential therapeutic approaches to treat cardiomyopathy with presence of metabolic syndromes. Disclosure G. Sun: None. G. Zhang: None. Y. Wang: None.

Introduction: Type 2 diabetes (T2D) and obesity are strongly associated with peripheral immune cell responses. Subsets of T cells in the circulation are correlated with the severity of the diseases, which may serve as novel targets for antidiabetic or obese treatment. Higher numbers of blood Th1, Th17 and Th22 cells of T helper cells were found in disease patients, while Th2 and Treg cells were in reverse correlation of the severity of the disease. FATZO mouse is a new generation of T2D mouse model with intact leptin signaling. The model is featured with slow onset of diabetes and obesity. Here, we aim to characterize the peripheral immune cell responses in FATZO mice for the use of preclinical drug development targeting diabetes or obesity related inflammation. Methods: Freshly collected blood from 12 FATZO mice at age 6, 12, 22 and 34 weeks were analyzed for the presence of monocytes, B, T, NK, NK-T, CD4+, CD8+ T cells and subsets of T helper cells (Th1, Th2, Th17, Th22, Treg) using FACS. Serum inflammatory cytokines such as IL1, IL2, IL6, IL17, TNFα and IFNγ were also measured. Results: FATZO mice developed obesity and hyperglycemia from age of 12 weeks. Concomitantly, higher numbers of leukocytes were noticed with an increase and decrease in frequency of B and NK cells respectively. Within T cells, numbers of CD4+ T cells decreased by 20%, while CD8+ T cells increased by 25%. When subsets of T helper cells were assessed, percentages of Th1 and Th17 cells significantly elevated during the development of diabetes and obesity, while frequencies of Th2 and Treg cells declined. Serum inflammatory cytokine measurements showed a tendency in higher levels of serum IL1a when the animals became diabetic. Conclusion: FATZO mice develop diabetes and obesity with changes in blood immune cell responses that closely mimic disease patients. The results suggest that FATZO might serve as an ideal model for preclinical research for the diabetes and obesity related inflammation. Disclosure G. Sun: None. W. Ye: Employee; Spouse/Partner; Crown Bioscience Inc.. X. An: None. Y. Xiao: None. Y. Wang: None.

Introduction: Progressive albuminuria and bi-phasic changes in glomular filtration rate (GFR) are hall marks of functional deterioration of the kidney in diabetic nephropathy. However, not all diabetic rodent models can capture the development of such changes in the kidney. FATZO, a new generation of diabetic mouse model with intact leptin signaling pathway shows all the metabolic dysfunctions that mimic human patients such as obesity, dyslipidemia, diabetes and NASH etc. Here, we sought to further characterize the model for the evaluation of its development in diabetic nephropathy. Method: Male FATZO mice on 50diet or on western diet with addition of 10% fructose (WD+fru) from age of 11 weeks were subject to monthly urine collection for albumin and creatinine measurement for 28 weeks, after which animals were treated with Lisinopril (15 mg/kg) and pioglitazone (20 mg/kg) daily for 6 weeks (n=8 for each group). Animals were also monitored for GFR using a subcutaneous monitoring device (Medibeacon, US). Results: Animals on WD+fru were heavier than those on 50diet. Notably, WD+fru markedly elevated urine albumin levels compared to 50diet in FATZO mice which resulted in higher albumin/creatinine ratio and urine albumin excretion rate (UAE) from 2 weeks on diet, while lisinopril treatment for 7 weeks could reduce the levels respectively. In addition, GFR was slightly elevated after 6 weeks of diet induction, which could be normalized by either pioglitazone or lisinopril treatment. Conclusion: We have created a mouse model with progressive albuminuria and elevated GFR that can be reversed by angiotensin-converting-enzyme (ACE) inhibitor. The model with associated dysmetabolic phenotypes including obesity and dyslipidemia can be used as an ideal tool for studying antidiabetic nephropathy treatment. Disclosure G. Sun: None. G. Zhang: None. Y. Wang: None.

Pancreatic β-cell regeneration is considered as a possible therapy for cure of type 2 diabetes (T2D). It has been reported that administration of FTY720, a sphingosine 1-phosphate (S1P) receptor modulator, to db/db mice led to sustained normalization of hyperglycemia by stimulating β-cell in vivo regeneration through PI3K-dependent regulation of cyclin D3 and p57KIP2. To further evaluate the therapeutic potential of FTY720 in the treatment of T2D, we examined the effects of FTY720 on glucose homeostasis in spontaneous diabetic NHP model. NHPs with mean fasting glucose (FG) level of 249 mg/dL were randomly divided into the control (vehicle) and the FTY720-treatment group. FTY720 was orally administered at 5 mg/kg to diabetic NHPs once per day and the FG and HbA1c levels were measured biweekly. Here we show that FTY720 treatment of diabetic NHPs significantly lowered their FG and HbA1c in two weeks. After 10 weeks of treatment, their FG and HbA1c levels remained low and their glucose tolerance was significantly improved. To determine the effect of FTY720 on β-cell function, IVGTT was performed. Following glucose injection, the insulin levels in the control NHPs barely changed. In contrast, the FTY720-treated NHPs secreted large amount of insulin in response to glucose stimulation, consistent with β-cell in vivo regeneration observed in mice. We also evaluated the cardiac function in these NHPs by echocardiography, from which ejection fraction (EF) and fractional shortening (FS) are found abnormally lower in diabetic NHPs (EF=60% and FS=30%) than that in normal NHPs (EF=68% and FS=43%). After 10 weeks of treatment, however, EF in the treatment group was significantly increased from 60% to 70% and FS from 30% to 38% while these values remained abnormally low in the control group. Our data strongly suggest that FTY720 has a great therapeutic promise for the treatment of T2D with benefit to improve the cardiac function in the patients. Disclosure Y. Wang: None. Y. Liu: None. Q. Wei: Employee; Self; Crownbio.Inc.. X. Wang: None. G. Sun: None. Y. Xiao: None. G. Yu: None. Z.A. Ma: None.