The effects of liraglutide and dapagliflozin on cardiac function and structure in a multi-hit mouse model of heart failure with preserved ejection fraction

Abstract

Aims: Heart failure with preserved ejection fraction (HFpEF) is a multifactorial disease that constitutes several distinct phenotypes, including a common cardiometabolic phenotype with obesity and type 2 diabetes mellitus. Treatment options for HFpEF are limited, and development of novel therapeutics is hindered by the paucity of suitable preclinical HFpEF models that recapitulate the complexity of human HFpEF. Metabolic drugs, like glucagon-like peptide receptor agonist (GLP-1 RA) and sodium-glucose co-transporter 2 inhibitors (SGLT2i), have emerged as promising drugs to restore metabolic perturbations and may have value in the treatment of the cardiometabolic HFpEF phenotype. We aimed to develop a multifactorial HFpEF mouse model that closely resembles the cardiometabolic HFpEF phenotype, and evaluated the GLP-1 RA liraglutide (Lira) and the SGLT2i dapagliflozin (Dapa).

Methods and results: Aged (18-22 months old) female C57BL/6J mice were fed a standardized chow (CTRL) or high-fat diet (HFD) for 12 weeks. After 8 weeks HFD, angiotensin II (ANGII), was administered for 4 weeks via osmotic mini pumps. HFD + ANGII resulted in a cardiometabolic HFpEF phenotype, including obesity, impaired glucose handling, and metabolic dysregulation with inflammation. The multiple hit resulted in typical clinical HFpEF features, including cardiac hypertrophy and fibrosis with preserved fractional shortening but with impaired myocardial deformation, atrial enlargement, lung congestion, and elevated blood pressures. Treatment with Lira attenuated the cardiometabolic dysregulation and improved cardiac function, with reduced cardiac hypertrophy, less myocardial fibrosis, and attenuation of atrial weight, natriuretic peptide levels, and lung congestion. Dapa treatment improved glucose handling, but had mild effects on the HFpEF phenotype.

Conclusions: We developed a mouse model that recapitulates the human HFpEF disease, providing a novel opportunity to study disease pathogenesis and the development of enhanced therapeutic approaches. We furthermore show that attenuation of cardiometabolic dysregulation may represent a novel therapeutic target for the treatment of HFpEF.

Keywords: Cardiometabolic; Dapagliflozin; HFpEF; Liraglutide; Mouse model.

Graphical abstract

Figure 1

HFD induces obesity and impaired glucose; this phenotype is not affected by addition of ANGII. (A) Overview of experimental design and an example of a CTRL and HFD + ANGII mouse. (B) Body mass composition (fluid, fat, and lean mass) measured by minispec LF90_II body composition analyser (n = 7–11 mice). (C) Weight development over time (weeks) (n = 7–11 mice per group). (D) Plasma glucose levels over time (min) after glucose loading (2 g/kg body weight) in an oral glucose tolerance test (n = 7–9 mice per group). To determine glucose tolerance, we calculated AUC per treatment group. (E) Fasting glucose levels per group (n = 6–12 mice per group). AUC, area under the curve; LBM, Lean, fat and fluid Body mass Measurements; CTRL, control chow; CTRL + ANGII, angiotensin II treated group on control chow; HFD, high-fat diet; HFD + ANGII, angiotensin II treated group on high-fat diet. Data are presented as mean + standard errors of the mean. *Kruskal–Wallis test followed by Mann–Whitney U test P <0.05 is considered significant.

Figure 2

Combination of HFD + ANGII results in a phenotype of concentric LV hypertrophy with preserved FS and diastolic dysfunction. (A) FS (systolic function) (n = 7–11 mice per group). (B) LVAWd (n = 7–11 mice per group). (C) Representative M-mode echocardiographic images of LV. (D) GLS as marker of myocardial deformation per treatment group (n = 6–9 mice per group). (E) Quantification of RPLSR as marker of diastolic function (n = 6–10 mice per group). (F) Pmax aorta measured by intracardiac pressure measurements (n = 5–9 mice per group). FS, fractional shortening; LVAWd, left ventricular anterior wall thickness in diastole; RPLSR, reverse peak longitudinal strain rate; GLS, global longitudinal strain; CTRL, control chow; CTRL + ANGII, angiotensin II treated group on control chow; HFD, high-fat diet; HFD + ANGII, angiotensin II treated group on high-fat diet; Data are presented as mean + standard errors of the mean. *Kruskal–Wallis test followed by Mann–Whitney U test P <0.05 is considered significant.

Figure 3

Combination of HFD + ANGII increases left ventricular, atrial and pulmonary weights and increased myocardial cross-sectional area, and cardiac stress marker ANP. (A) LV/tibia, left ventricle weight corrected for tibia length (n = 7–10 mice per group). (B) Atria weight/tibia length (n = 6–11 mice per group). (C) Lung weight/tibia length (n = 7–10 mice per group). (D) Cross-sectional area of cardiomyocytes quantified by WGA-FITC staining (30–50 cells per animal, n = 6–9 mice per group). (E) Representative images of histological staining depicting the cardiomyocyte size (WGA), capillary density (CD31+), and fibrosis (Masson). (F) LV mRNA expression levels of ANP (n = 7–9 mice per group). ANP, atrial natriuretic peptide; CTRL, control chow; CTRL + ANGII, angiotensin II treated group on control chow; HFD, high-fat diet; HFD + ANGII, angiotensin II treated group on high-fat diet. Data are presented as mean + standard errors of the mean. *Kruskal–Wallis test followed by Mann–Whitney U test P <0.05 is considered significant.

Figure 4

Effects of Lira and Dapa on body weight, glucose tolerance test, and food intake. (A) Body weight from start of treatment over time (days) (n = 13–14 mice per group). (B) Body mass composition (fluid, fat, lean mass) measured by minispec LF90_II body composition analyser (n = 13–14 mice per group). (C) Total food intake per 3 days (n = 13–14 mice per group). (D) Fasting glucose levels per group measured 2 days before sacrifice (n = 13–14 mice per group). Dotted line represents control reference value. (E) Plasma glucose levels over time (min) after glucose loading (2 g/kg body weight) in an oral glucose tolerance test (n = 9–10 mice per group). To determine glucose tolerance, we calculated area under the curve (AUC) per treatment group. LBM, Lean, fat and fluid Body Mass Measurements; dotted lines represents control reference values; HFD + ANGII + Dapa, high-fat diet + angiotensin II with dapagliflozin treatment; HFD + ANGII + Lira, high-fat diet + angiotensin II with daily liraglutide injection; HFD + ANGII + Saline, high-fat diet + angiotensin II with saline injections. Data are presented as mean + standard errors of the mean. *Kruskal–Wallis test followed by Mann–Whitney U test P <0.05 is considered significant.

Figure 5

Effects of treatment with Lira or Dapa on cardiac function, cardiac structure, and cardiac remodelling. (A) FS (systolic function) (n = 8–13 mice per group). (B) LVAWd (n = 12–14 mice per group). (C) GLS as marker of myocardial deformation (n = 9–13 mice per group). (D) LV/tibia, left ventricle weight corrected for tibia length (n = 13 mice per group). (E) Atria/tibia = atria weight corrected for tibia length (n = 13–15 mice per group). (F) Lung weight/tibia length (n = 13–14 mice per group). FS, fractional shortening; GLS, global longitudinal strain; LVAWd, left ventricle anterior wall thickness in diastole; dotted lines represents control reference values; HFD + ANGII + Dapa, high-fat diet + angiotensin II with dapagliflozin treatment; HFD + ANGII + Lira, high-fat diet + angiotensin II with daily liraglutide injection; HFD + ANGII + Saline, high-fat diet + angiotensin II with saline injections . Data are presented as mean + standard errors of the mean. *Kruskal–Wallis test followed by Mann–Whitney U test P <0.05 is considered significant.

Figure 6

Treatment effects of Lira or Dapa on cardiac fibrosis and cardiac hypertrophy. (A) Histological analysis of fibrosis, expressed in fold-change compared to reference control values (n = 10–11 mice per group). (B) Capillary density determined by number of capillaries per mm² positive stained by CD31+ (n = 10 mice per group). (C) Cross-sectional area of cardiomyocytes quantified by WGA-FITC staining (30–50 cells per animal, n = 9–13 mice per group. (D) Representative images of histological staining depicting the cardiomyocyte size (WGA), capillary density (CD31+), and fibrosis (Masson). Dotted lines represents control reference values; HFD + ANGII + Dapa, high-fat diet + angiotensin II with dapagliflozin treatment; HFD + ANGII + Lira, high-fat diet + angiotensin II with daily liraglutide injection; HFD + ANGII + Saline, high-fat diet + angiotensin II with saline injections. Data are presented as mean + standard errors of the mean. *Kruskal–Wallis test followed by Mann–Whitney U test P <0.05 is considered significant.