• Wed. Feb 12th, 2025

The remaining authors have nothing to disclose

Byacusticavisual

Jan 19, 2025

The remaining authors have nothing to disclose. Footnotes Abbreviations: ANGPTLangiopoietin-like proteinANOVAanalysis of varianceHFHChigh-fat, high-cholesterolIgGimmunoglobulin GhANGPTL8-mFchuman angiopoietin-like protein 8 expressed with C-terminal mouse immunoglobulin G2a Fc tagHBS-ET10 mM HEPES (pH 7.4), 150 mM NaCl, 3 mM EDTA, and 0.05% volume-to-volume ratio surfactant Tween 20HDL-Chigh-density lipoprotein cholesterolHLhepatic lipaseHPLChigh-performance liquid chromatographyLDL-Clow-density lipoprotein cholesterolLPLlipoprotein lipasemfANGPTL8-mFcmonkey angiopoietin-like protein 8 expressed with C-terminal mouse IgG2a Fc tagSCsubcutaneous(ly)SEMstandard error of the meanTCtotal cholesterolTGtriglyceride.. associated with hypertriglyceridemia, whereas genetic inactivation of reduced plasma TGs (1, 2). ANGPTL8 is usually expressed in liver and adipose tissue and its expression is highly upregulated by feeding (1, 3, 4). For this reason, it has been proposed that ANGPTL8 contributes to TG handling during Ziprasidone hydrochloride refeeding and directs fatty acids to adipose tissue for storage (2). Consistent with this, imaging system (Caliper Life Sciences) as described elsewhere (11). Metabolic cage data were generated using the Oxymax Ziprasidone hydrochloride laboratory animal monitoring system (CLAMS; Columbus Devices). Mice were individually monitored in cages with center feeds for 96 hours. Data generated in the first 24 hours were omitted from the analysis. Food intake was measured constantly and divided into calories consumed per light and dark phase of the light cycle. Oxygen consumption and carbon dioxide production were measured in 17-minute intervals during a 4-day span and plotted over time in hours. Energy expenditure was calculated as a function of the respiratory quotient and the oxygen consumption, normalized to body weight. Additionally, as the groups had divergent body weights at the time of analysis, energy expenditure was expressed as kilocalories per hour per mouse using an adjusted mean body weight of the two groups combined. This was achieved using analysis of covariance with body weight as the covariance, as described (12, 13). Briefly, the adjusted energy expenditure for each animal was calculated as = ? ? is usually single animal adjusted energy expenditure (kcal/h), is single animal energy expenditure (kcal/h), is single animal body weight (kg), is adjusted mean body weight of the two treatment groups combined (kg), and is the slope of the line of energy expenditure plotted vs body weight for each animal and each treatment group calculated by linear regression analysis. Study in cynomolgus monkeys The study was performed at Crown Bioscience. Eighteen spontaneous hypertriglyceridemic monkeys were selected based on their nonfasted serum TG levels and divided into three groups. The monkeys were individually housed, had free access to water, and were fed twice daily with a complete nutritionally balanced diet (Shanghai Shilin Biotechnology), enriched with seasonal fruits and vegetables. All animal procedures were approved by the Crown Bioscience Institutional Animal Care and Use Committee and performed according to guidelines approved by the Association for Assessment and Accreditation of Laboratory Animal Care. On day 0 the monkeys were administrated REGN3776 at 3, 7, or 10 mg/kg. Blood (4 mL) was collected into BD sterile venous blood collection tubes (Accu-Chek Active; Roche) PMCH from nonfasted animals at 1- to 5-day intervals up to day 45. After TG levels for all animals returned to baseline, animals were allowed to washout for at least 2 weeks, and then five animals were selected for the treatment with saline. Blood was collected on consecutive days after saline administration on the same schedule as the REGN3776-injected groups. Serum TG, TC, LDL-C, and HDL-C levels were measured by an Advia 2400 system Ziprasidone hydrochloride (Siemens). Data analysis All data are shown as mean SEM. Statistical analyses were performed utilizing GraphPad Prism 6.0 software. LPL and HL activities in REGN3776 and control antibody-treated mice were compared by a Welch test. All other parameters were analyzed by two-way analysis of variance (ANOVA) with repeated steps. If a significant ratio was obtained with two-way ANOVA, post hoc analysis was conducted between groups with a Bonferroni or Sidak posttest. In Ziprasidone hydrochloride the monkey study, the average of each parameter on day ?15, ?7, and 0 was used as the baseline value. Results characterization of ANGPTL8 blocking antibody The relative affinities of ANGPTL8 blocking antibody REGN3776 to human, mouse, and monkey ANGPTL8 were compared using surface plasmon resonance. REGN3776 binds human and monkey ANGPTL8 with comparable affinities (Table 1; Supplemental Fig. 1). REGN3776 does not bind mouse ANGPTL8 or human or mouse ANGPTL3 or ANGPTL4 (Table 1). Table 1. Summary of Binding Kinetic Parameters for the Conversation of REGN3776 With hANGPTL8-mFc or mfANGPTL8-mFc < 0.01, ***< 0.001, ****< 0.0001. It.