Cardiac damage in DKD/CKD mouse model resembles HFpEF and can be reducedby Standard-of-care treatment
Arianne 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, Elsbet Pieterman1, Anke M. Smits7, 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, 6BPM, Taiwan, 7 LUMC, the Netherlands.

•Pathology assessment includes quantitatively scoring offibrosis using image analysis.

1.0

2

heart weightlung wet weight

0.35
0.30
0.25

*
0.20

**

*
**

1000

0.20
0.15

0.15

Combination therapy with Lisinopril andDapagliflozin reduces heart weight andcardiac fibrosis.
Histopathological evaluation showed
macroscopical damage and microscopic
myocardial fibrosis and calcification.

0UNX+HFD+LNNAACEi +dapa 5
ACEi +dapa 20* p<0.05** P<0.01

Contact information: arianne.vankoppen@tno.nl48B Stroke volume
50

GFR (ul/min)

200100
0Ejection fraction****

End systolic volume

**

40
30

60

600

4030

p=0.056

***

400
200
01280

10chow
UNX+HFD
UNX+HFD+LNNA**

UNX+HFD+PCSK9UNX+HFD+LNNA+PCSK920

A: PCSK9 AAV
increases plasma
0cholesterol, induces
LV posterior wall LV inner diameter LV inner diameter atherosclerosis with noLV posterior wall
systolic
diastolicthicknessdiastolicthickness systolic
additional effect on
*
3.05
2.0
GFR B: Cardiac
2.5*
imaging showed
*
2.0
2.5
1.5
reduced ejection
41.5
fraction and increased
2.0
1.0
end systolic volume
1.0
3
after PCSK9 AAV
1.5
0.5
0.5
treatment. Data shownas mean ±.SD.
2
1.0
0.0
0.0
40

20

cardiac fibrosis

2000

-1300

weeks of HFD

3000

0.25

Total lesion area (*103 um2)

0Combination therapy reduces cardiac damage
0.40

***

20400

****
**
****

10mm0.0340800

*****

500

Volume (ul)1.5***mm

0.5***

Fraction (%)

0.0mm

Volume (ul)mmFraction (%)

mm

mm

0.5

8060

GFR

Atherosclerotic lesion
size area

Plasma cholesterol

Amm

Cardiac imaging
showed3040
UNX+HFD
preserved
UNX+HFD+LNNA ejection fraction,2060
low end systolic30* p<0.05
volume and left
10ventricle inner40
20
diameter and0posterior wall
Left ventricle inner
Left ventricle posterior Left vetricle posterior
Left ventricle inner
thickening,wall thickness systolic wall thickness diastolic diameter systolic
diameter diastolicsuggesting**3.02.052.5**
concentric*
hypertrophy and
2.0
2.5
1.5
HFpEF. Data4
1.5
shown as mean2.0
1.0± SD.
1.0UNX+chowmmMale KKAy mice underwent uninephrectomy (UNX). Afterrecovery, mice received high fat diet (HFD) with or without
the vasoconstrictor LNNA (50mg/L) for 16 weeks. In thestandard-of-care intervention group, at wk 4 Lisinopril (2.5mg/kg/day) was started. At week 8 Dapagliflozin (5 and 20mg/kg/day). 1 group of mice received AAV-mPCSK9 (gainof function mutation) injection which reduces liver LDL
receptor levels leading to high circulating cholesterol andincreased susceptibility to atherosclerosis. At week 12, a
subset of mice underwent cardiac imaging. GFR wasmeasured using a transdermal measurement system.

p=0.0506*% area *1000

•

4080

weight (g)Methods*p=0.0502

Cholesterol (mM)

50

End systolic volume

Ejection fraction

Volume (ul)Stroke volume

Aim

The aim of this study was 1) to functionally characterizecardiac damage in TNO’s DKD/CKD model 2) to determine
efficacy of standard-of-care combination therapy of a low
dose of Lisinopril followed by on-top-off Dapagliflozin
treatment on cardiac histopathology and 3) to induce
dyslipidemia to resemble the cardiovascular-kidney-metabolic
syndrome using an AAV-PCSK9 gain of function mutation
injection.

Combining dyslipidemia with obesity, diabetes andhypertension leads to CKD and suggests HFrEF

Combining obesity, diabetes and hypertension induces HFpEF

weight (g)Cardiovascular damage is the major cause of death in CKD.
TNO developed a diet-induced hypertension-accelerated
DKD/CKD model resembling cardiovascular-kidney-metabolic
syndrome including obesity, diabetes and hypertension andallows the study how the different components of thissyndrome contribute to the development of cardiac damage.

Volume (ul)Introduction

Conclusions
TNO’s translational DKD/CKD mouse model with UNX+HFD and LNNA
exhibits features of HFpEF. Cardiac damage is reduced by combination
therapy with a low dose of Lisinopril and Dapagliflozin. When inducing
dyslipidemia the heart develops HFrEF. This shows the usability of theKKAy DKD/CKD mouse model for cardiac efficacy studies.