Combination therapy with Lisinopril and Dapagliflozin rescues GFR decline andglomerular damage in the advanced DKD/CKD KKAY mouse modelArianne van Koppen1, Romer A. Gonzalez-Villalobos2, Andrea R. Nawrocki2, Simon Hinke2, Tri Q. Nguyen3, Amelie Dendooven4, Ingeborg Bajema5,Harry van Goor5, Karen Tseng6, Wei-Chan Hsu6, Aswin Menke1, Matt Breyer2, Reinout Stoop1
1 Department of Metabolic Health Research, TNO Leiden, The Netherlands. 2 Janssen Pharmaceuticals, Boston, United States 3 UMCU, Utrecht, The Netherlands, 4 UZA, Antwerp and UZ Ghent, Belgium, 5 UMCG Groningen, The Netherlands, 6 BPM, Taiwan.

Dapagliflozin reduces blood glucose and increases water intake

Combination therapy reduces renal damage

AA

Unfasted blood glucose

20

103dapa

681012** to ****

2530

lisinopril

Water intakeB* to ****

Water intake (ml/day)

The prevalence of diabetic kidney disease is rapidly
increasing. Development of novel therapeutics is hamperedby the lack of translational animal models resembling all
stages of DKD. TNO developed a diet-induced hypertensionaccelerated DKD model which can be used understand thedifferent disease stages, lead to identification of new
therapeutic targets and biomarkers. Response to standardof-care (SOC) therapy will indicate usability of the model forefficacy studies.

Blood glucose (mmol/L)

Introduction

1420start
diet

dapa

lisinopril

UNX+HFD+LNNA
ACEi +dapa 5
ACEi +dapa 20

15* to **** p<0.05 vs
UNX+HFD+LNNA

10160weeks of diet

2468101214

weeks of diet

A: Dapagliflozin immediately reduced unfasted blood glucose levels dose-independently. B: Dapagliflozin••

GFR was measured using a transdermal GFR
Measurement system.
Pathology assessment includes quantitatively scoring ofglomerular and tubular damage by a team of renal
pathologists, GBM thickening by EM microscopy andautomated mesangium expansion using image analysis.

*100

*

*

UNX+HFD+LNNA

Albumin excretion

660
468

1216

weeks of diet

ACEi+ dapa 20* p<0.05 vs
UNX+HFD+LNNA*2

A: Combination therapy rescued GFR decline and was not different 0
compared to non-induced controls. B:Albuminuria was reduced bycombination therapy. Data shown as mean ± SEM.

Contact information: arianne.vankoppen@tno.nl

UNX+HFD+LNNA
ACEi +dapa 54lisinopril

4dapa

681012weeks of diet141610*50**

number of affected glomeruli

20
100

Segmental sclerosis

ACEi +dapa 5*p<0.05 dapa 5 vsB

50UNX+HFD+LNNA

80

number of affected glomeruli

UNX+chowdapa

1030

Global sclerosis
1086420p=0.0548

2.52.0ACEi +dapa 5
ACEi +dapa 20

1.0* to **** p<0.05 vs
UNX+HFD+LNNA

0.50.0CIFTA

6420

normal glomeruli
diffuse mesangial expansion
nodular mesangial expansion
segmental sclerosis
global glomerulosclerosis

UNX+HFD+LNNA

1.5% affected cortical area

Body weight, food and water intake was monitored
weekly, blood glucose every 4 weeks.

lisinopril

p=0.065

Nodular sclerosis

Mesangial expansion

number of affected glomeruli

120

GFR (ul/min)•Male KKAy mice underwent uninephrectomy (UNX). After
recovery, mice received high fat diet (HFD) and thevasoconstrictor LNNA (50mg/L) for 16 weeks. At wk 4
Lisinopril (2.5 mg/kg/day) was started. At week 8
Dapagliflozin (5 and 20 mg/kg/day).

% GFR reduction

A15

number of glomeruli

Combination therapy rescues GFR decline and reduces
albuminuria

Method•B Normal glomeruli

number of affected glomeruli

The aim of this study was to determine efficacy of standardof-care combination therapy of initial low dose Lisinopril
treatment followed by on-top-off Dapagliflozin treatment onrenal function and histopathology.

increased water intake dose-independently. Data shown as mean ± SEM.

Albumin (mg/3 hours)

Aim**A + B: Quantitative scoring ofglomeruli showed that combination
therapy increased the number ofhealthy glomeruli and decreased
the number of glomeruli withsegmental and global sclerosis. C:
Interstitial fibrosis and tubular
atrophy were significantly reducedby combination therapy. Datashown as mean ± SEM.

Conclusions
Combination therapy with a low dose of Lisinopril and Dapagliflozin (5mg/kg/day) rescues GFR decline and prevents increase of albuminuria inthe KKAy DKD/CKD mouse model. Glomerular and tubular damage werereduced by standard-of-care combination therapy. This further supports
the usability of the KKAy DKD/CKD mouse model for efficacy studies.