A novel translational model of hypertension-accelerated diet-induced diabetic
kidney disease with declining GFR and advanced pathology
Arianne van Koppen1, Matt Breyer2, Romer Gonzalez Villalobos2, Andrea R. Nawrocki2, Savithri Kota2, Simon Hinke2, Tri Q. Nguyen3, Amelie
Dendooven4, Harry van Goor5, Karen Tseng6, Wei-Chan Hsu6, Aswin Menke1, 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.

Introduction

HFD and FFD induce comparable diabetes development

The prevalence of diabetes and diabetic kidney disease israpidly increasing. Development of novel therapeutics ishampered by the lack of translational animal models
resembling the later stages of the DKD. TNO has developed
a diet-induced hypertension-accelerated DKD model. Thiscan help to better understand the different disease stages,
lead to identification of new therapeutic targets andbiomarkers.

ABody weight

Fasted blood glucose

B

50

start diet4030-2 02468 10 12 14 16 18 20(g)weightglucose
Body Blood(units)

Body weight (g)

60

30

**
***
****
*** Body weight*

Quantitative sclerosis scoring and GBM thickeningA* = p <0.05 vs UNX+chow** = p <0.01 vs UNX+chow***= p <0.005 vs UNX+ chow****= p <0.001 vs UNX+chow

60

UNX+chow

UNX+HFD (n=8)

50

UNX+FFD (n=10)

10

UNX+HFD+LNNA (n=7)
UNX+FFD+LNNA (n=7)

40weeks

816

UNX+FFD

UNX+HFD+LNNA UNX+FFD+LNNA

UNX+chow (n=9)**
204UNX+HFD

20B

IFTA

30

*

UNX+HFD+LNNA 20w (n=10)*weeks3000xC*300****

3020nm

10

10000UNX+HFD

Method6000

Male KKAy mice (8 weeks) underwent uninephrectomy(UNX). After 3 weeks recovery, mice received either highfat diet (HFD) or fast food diet (FFD) with or without LNNA(50mg/L) for 16 weeks. Chow-fed mice were used ascontrols. One group fed HFD+LNNA was continued untilweek 20.

•Body weight, food and water intake was monitoredweekly, blood glucose every 4 weeks.

•GFR was measured transdermally using retro-orbital
injections with FITC-sinistrin.

•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.

E Mesangium expansion

Glomerular size
*

***
***
****

***

50*****

40**

nm4000unx

20000****
30

****
**
****

****

UNX+HFD+LNNA****
20100unxD•

UNX+chowUNX

Reduction in GFR most prominent in high fat diet with LNNA

UNX**200weeks% PAS+ surface

The aim of this study was to determine the time-dependent
progression of the diet-induced DKD model and to evaluate
the effect of diet and hypertension functionally andpathophysiologically.

%

Aim

A: Both HFD and FFD induced increased body weight with-2HFD
being
prominent
LNNA did not affect body
0 24 6the8most
10 1214 16 18 inducer.
20
weight gain. B. Blood glucose steeply increased after diet initiation. No effects of LNNA were observed. Data are mean ± SD7000x

GBM thickness

UNX+HFD+LNNA wk20A. Quantitative scoring of glomeruli showed that HFD+LNNA showed most glomerular damage which wasconfirmed by automated scoring (D and E) and most interstitial fibrosis and tubular atrophy (B)). GBM
thickening was confirmed by electron microscopy (C).

Conclusions
GFR showed several phases of DKD including hyperfiltration and declining GFR (A) whereas HFD+LNNA
showed the quickest decline. GFR decline was comparable in a separate time course study (B).
Albuminuria was strongest increased in HFD+LNNA (C).

Contact information: arianne.vankoppen@tno.nl

DKD development was most prominent in male KKAy mice on HFD andLNNA (50 mg/L) as shown by an initial period of hyperfiltration followed bya continuous decline of GFR and advanced renal pathology. Together withincreased albuminuria this indicates that the mice reach the phase of overt
DKD resembling the clinical phenotype.