DEPTOR 5'UTR polymorphisms and

insulin resistance in obese children

and adolescents
Jernej Kova1, Tamara utu1, Simon Horvat2, Jasmina Beltram2,
Katarina Trebuak Podkrajek1, Tadej Battelino3,4, Primo Kotnik3
1. Unit for Special Laboratory Diagnostics, University Children's Hospital, UMC Ljubljana,
Ljubljana, Slovenia
2. National Institute of Chemistry, Ljubljana, Slovenia
3. Department of Endocrinology, Diabetes and Metabolic Diseases, University Children's
Hospital, UMC Ljubljana, Ljubljana, Slovenia
4. Department of Paediatrics, Faculty of Medicine, University of Ljubljana, Ljubljana,
Slovenia

Presenter Disclosure Information

Primo Kotnik

Disclosed no conflict of interest.

Background

Obese children and adolescents are prone to become overweight or obese

adults and will present with obesity complications at that time, however
selected will develop them early.

Impaired sensitivity of tissues to insulin (especially liver, skeletal muscle

and adipose tissue) is possibly the main metabolic complication of obesity.

Goal is to early recognize those at risk of obesitys complications and

aplly efficient therapeutic strategies to battle obesity and its complications.

Genetic background has an important role in the development of IR and

T2D genetic markers for the determination of increased risk?

Reilly 2011, Weiss 2013

mTOR (Mammalian Target of

Rapamycin)

The mTOR signaling pathway

senses growth factors and
nutrients to regulate many
biological processes involved in
the promotion of cell growth.

mTORC1:
affects lipogenesis and
adipogenesis.
directly involved in insulin
downstream signalling through the
feedback control of insulin
receptor substrate 1 (IRS-1).

mTOR signalling deregulation

leads to obesity and increased risk
of insulin resistance (IR) and type
2 diabetes.

Laplante 2012, Cornu 2013

http://www.cellsignal.com/

DEPTOR (DEP domain-containing

mTOR-interacting protein)

DEPTOR represses mTOR

signaling.

Transgenic mice overexpressing

DEPTOR accumulate more WAT.
In humans, DEPTOR expression
in WAT correlates with the degree
of obesity.

Overexpression of DEPTOR in
muscles and activation of Akt
signalling cascade is associated
with improved insulin sensitivity
and glucose tolerance in mice
models.

Role in the regulation of

physiological processes in vivo in
humans is unknown.

Laplante 2009, Horvat 2013, Meng 2013

http://www.cellsignal.com/

Aims

No mutations in the coding region of DEPTOR gene have been determined

to be associated with obesity or its complications.

DEPTORs actions are mainly regulated by its expression levels + has

similar expression profile to genes known to be regulated by epigenetic
mechanisms.

Hypothesis - gene variants in the promoter region of DEPTOR

gene could be associated with the development of IR in obese
children and adolescents.

Subjects (meanSD)
N

Sex
(F/M)

Age
(years)

BMI-SDS

HOMA-IR*

WBISI*

IR

79

44/35

14.2 2.6

2.9 0.7

5.1 2.4

1.89 0.65

Non-IR

117

60/57

12.4 3.7

2.7 0.7

1.7 0.9

5.97 2.96

*p < 0.0001

12

450
400

10

300

IR
Non-IR

Insulin (mU/l)

Glucose (mmol/l)

350

250

IR
Non-IR

200
150

100

2
50
0

0
0

20

40

60

80

Time (min)

100

120

140

20

40

60

80

Time (min)

100

120

140

Methods
Genotyping of DEPTOR 5'UTR region was performed by High
Resolution Melting analysis and Sanger sequencing.

Chi-square test with Yates correction was used to detect statistical

significance in the frequency of analysed genotypes.

Results
Four polymorphisms (SNP) were identified:

rs7840156 (MAF:32.7%)
rs75781905 (MAF:7.33%)
rs117543860 (MAF:2.62%)
rs140142743 (MAF:0.26%)

CORE PROMOTER

5UTR

rs117543860 (c.-12G>A)
rs7840156 (c.-143T>C)
rs75781905 (c.-33C>A)

rs140142743 (c.-36C>T)

EXON_1

Results
SNP rs7840156 (c.-143T>C) (n=196).
HOMA-IR

WBISI

IR

27.7% 72.3%

IR

32.4% 67.6%

NonIR

38.3% 61.7%

NonIR

45.0% 55.0%

OR 0.63 (95%CI = 0.41-0.87; p=0.038)

OR 0.58 (95%CI = 0.38-0.90; p=0.018)

Results
(pubertal and normoglycemic only)
SNP rs7840156 (c.-143T>C) (n=138).
HOMA-IR

WBISI

IR

28.7% 71.3%

IR

25.9% 74.1%

NonIR

41.4% 58.6%

NonIR

40.3% 59.7%

OR 0.57 (95%CI = 0.35-0.94; p=0.039)

OR 0.52 (95%CI = 0.30-0.88; p=0.020)

Bioinformatic Analysis
REGULATORY FEATURE:

rs7840156

RESOURCE:

Transcription Factor
Binding Sites

Project ENCODE

Predicted
Transcription Factors

Algorithm ALIBABA
Core promoter

Predicted Promoter
Evolutionary Constrained
Unmethylated DNA

5UTR

UCSC
Eukaryotic Promoter
Database

ENSEMBL
ENSEMBL

CpG island

MethPrimer, EMBOSS CpGplot

USCS, CpG Island Searcher

Open chromatin

ENSEMBL
H3K4me2

Histone
modifications

H3K4me3
H2AZ
H3K9ac
H3279ac
H3K79me3

Project ENCODE

Conclusions
SNP rs7840156 (c.-143T>C) possibly a useful genetic biomarker in
the determination of obese youth at risk of IR and diabetes.
Further suggest a role of mTOR signaling pathway in the
development of diabetes.

http://www.cellsignal.com/

Acknowledgments

University Children's Hospital

Ljubljana, Slovenia:

Unit for Special Laboratory

Diagnostics:
Jernej Kova
Tamara utu
Katarina Trebuak Podkrajek

Department of Endocrinology,
Diabetes and Metabolic Diseases:
Tadej Battelino

National Institute of Chemistry,

Ljubljana, Slovenia:
Jasmina Beltram
Simon Horvat