Genetic association of IDE, POU2F1, PON1, IL1a and IL1bwith type 2 diabetes in Pakistani population

Andleeb Batool • Nusrat Jahan • Yisuo Sun •

Atif Hanif • Hong Xue

Received: 8 July 2013 / Accepted: 16 January 2014

� Springer Science+Business Media Dordrecht 2014

Abstract A number of genes are known to be involved in

glucose homeostasis. Mutations and polymorphisms in

candidate genes may effect insulin production, action or

resistance. This study was designed to report the associa-

tion of genetic polymorphism with the type 2 diabetes

(T2D) in Pakistani population. A total of 458 subjects (case

n = 288, control n = 170) participated in the study. Nine

single nucleotide polymorphisms were investigated in

genes IDE (rs6583813 C[T, rs7910977 C[T), POU2F1

(rs3767434 A[T, rs10918682 A[T, rs2146727 A[G),

WFS1 (rs734312 A[G), PON1 (rs854560 T[A), IL1a(rs1800587 C[T) and IL1b (rs1143634 C[T). Genotyping

was performed by DNA sequencing after nested polymer-

ase chain reaction of targeted regions. Results indicated

that rs7910977 in IDE showed significant association with

the development of T2D [P = 0.012, OR 1.677 (95 % CI

1.112–2.438)]. The rs10918682 in POU2F1 was associated

with T2D [P \ 0.001, OR 3.606 (95 % CI 2.165–6.005)].

The rs854560 in PON1was associated with incidences of

T2D and increased the risk of cardiovascular complications

[P = 0.031, OR 0.663 (95 % CI 0.455–0.965)] in diabet-

ics. The rs734312 from WFS1 gene was associated with

diabetes at genotype level (P \ 0.01). Haplotype analysis

of rs1800587–rs1143634 depicted CC haplotype increased

the susceptibility to diabetes (P \ 0.05). Haplotype GAA

from rs2146727–10918682–rs3767434 was protective

against diabetes (P \ 0.01) and GGA exhibited the asso-

ciation with T2D (P \ 0.01). Haplotype CT from

rs6583813–rs7910977 was protective against diabetes

(P = 0.02). Our study provided evidence to IDE, PON1,

WFS1, POU2F1, IL1a and IL1b associated with T2D in

Pakistanis.

Keywords Complications � Disease associated

haplotype � Single nucleotide polymorphisms (SNPs) �Susceptibility

Introduction

There is rapid increase in prevalence of diabetes within a

short time span throughout the world [1]. The risk for onset of

type 2 diabetes (T2D) increased 2–6 times with positive

family history and it’s known to be a crude way of estimating

the individuals at risk of diabetes [2]. T2D is one of the major

public problems in the south Asian countries such as Paki-

stan, India and Bangladesh with the prediction that by 2025

the diabetes will affect 76 million of adults [3]. Pakistan is a

developing country with a population of 154 million and

10 % of its adults are diabetic. The country has ranked sixth

according to diabetic population countries [4].

A number of genetic studies were performed in different

populations and 40 genetic loci were found associated with

diabetes [5–8]. Different single nucleotide polymorphisms

A. Batool (&) � N. Jahan

Department of Zoology, Government College University,

Lahore, Pakistan

e-mail: bandleeb@yahoo.com

A. Batool � Y. Sun � H. Xue

Division of Life Science and Applied Genomic Centre, Hong

Kong University of Science and Technology, Clear Water Bay,

Hong Kong

Y. Sun � H. Xue

Nano Science and Nano Technology Program, The Hong Kong

University of Science and Technology, Clear Water Bay,

Hong Kong

A. Hanif

Institute of Biochemistry and Biotechnology, University

of Veterinary and Animal Sciences, Lahore, Pakistan

123

Mol Biol Rep

DOI 10.1007/s11033-014-3165-y

(SNPs) from susceptible genes HNF4A, CAPN10, TCF1,

KCNJ11, HHEX, PPARG and ADIPOQ loci (involved in

insulin secretion, regulation and action) were studied in

relevance to association with the onset of T2D [9–13]. It

was previously reported that chromosome 1 is linked with

T2D in different populations. Gene POU2F1 is also located

on chromosomal region 1q24 [8]. The data of international

1q consortium demonstrated the association or linkage of

this region (1q21–25) with T2D from eight populations

[14]. The association studies in the Chinese population

suggested that POU2F1 gene effect on the b-cell function

by inflammation, proliferation and regulation of cellular

differentiation [6]. Candidate gene insulin degrading

enzyme (IDE) located on chromosome 10q23.3 and is

playing a role in the glucose homeostasis and reported to be

associated with diabetes [5]. Variants in WFS1 gene are

associated with insulin secretion in Europeans [8] and

PON1 gene is coded on chromosome 7q21.3-22., and it’s a

calcium dependent esterase and Low level of PON1

activity, thought to increase the risk of atherosclerosis and

play a role in the predisposition of vascular disorder in

diabetes [16, 17].

Diabetes was also recognized as a disease mediated by

inflammatory and immune responses which lead to impaired

signaling of insulin and selective destruction of b-cells, so

cytokines have an important role in this mechanism. Inter-

leukin-1(IL1) has a central role in regulation of inflammatory

and immune responses and IL1a and IL1b are proinflam-

matory cytokines [18]. Polymorphism in IL1 gene was

reported to be associated with obesity which is a risk factor

for the onset of diabetes [19] as well as was also playing a role

in glucose homeostasis and diabetes prevalence [20].

The current study was designed to investigate the vari-

ants in six genes (IDE, POU2F1, WFS1, PON1, IL1a and

IL1b) and their association with T2D. Some SNPs were

reported which were studied to find out association with

T2D in Pakistani Population in comparison to UK popu-

lation [3, 21, 22]. First Cousin marriages are very common

in Pakistan contributing towards the specific genetic pools

for families or casts which increase the emergence of

genetic disorders in a specific population. To our knowl-

edge these genetic variants are not studied in Pakistani

population and it’s the first study to report their association

with T2D.

Materials and methods

DNA sampling

Pakistani subjects (n = 458) were included in this study for

genetic analysis. All of the participants were informed for

the current study and samples were drawn with their

consent. It was a case–control study with 288 T2D patients

(diagnosed as diabetics according to WHO criteria) and 170

were control subjects (non-diabetics, with no family history

of diabetes). Blood sampling to study genetic polymorphism

was carried out from Diabetes Centre (Allama Iqbal Medical

College Lahore) and the data was collected regarding age,

age at the time of diagnosis, body mass index (BMI) and

family history. Blood (5 ml) was collected in EDTA coated

tubes and stored at 4 �C for further process of DNA

extraction. The study was approved by Research Ethical

committee of Government College University Lahore.

DNA extraction and genotyping

DNA was extracted from blood cells by Fermentas Genomic

DNA purification kit (#K0512, USA) as described by the

manufacturer’s protocol. Precipitated DNA was dissolved in

TE (Tris–EDTA buffer) buffer and stored at -20 �C for

further genetic analysis. Primers (forward and reverse) for

first and nested polymerase chain reaction (PCR) were

designed (Table 1). Genomic regions of the interest were

amplified for selected SNPs (Table 1) and nested PCR was

used to increase the specificity of amplification [22, 23]. The

SNPs were genotyped by using genetic analyzer (ABI,

3130X/ Genetic Analyzer) and the genotyping rate was

found over 95 % for all the tested SNPs.

Stastical analysis

All SNPs in control samples were passed from Hardy–

Weinberg Equilibrium (HWE) using the program GENE-

POP 4.0 (P [ 0.05). Fischer’s, Chi square (v2) and Pearson

Chi square tests were used to find the association of SNPs

with diabetes. SNPs with P value \ 0.05 were found to be

associated with diabetes. Frequencies of alleles and geno-

types were calculated in control and diabetes patients

through UNPHASE program (3.1.3 version). Haplotype

association, linkage disequilibrium (LD = D0) and correla-

tion (r2) was studied through SHEsis program: http://analy

sis2.bio-x.cn/myAnalysis.php [24, 25]. Frequency less than

0.01 was ignored for the haplotype analysis. D0 = 1 known

as complete LD and D0\ 1 indicates the complete ancestral

LD has been disrupted. Therefore, statistically significant

values of D0 that are near to one (e.g. 0.9) provide a useful

indication of minimal historical recombination.

Results

Total of 458 subjects were included in the current study.

Demographic characteristics of the patients are summa-

rised in Table 2. The nine SNPs from six genes (IDE, IL1A,

Mol Biol Rep

123

IL1B, PON1, WFS1 and POU2F1) were analysed and their

frequency data was presented in Table 3 for alleles and

Table 4 for genotypes. No deviation from HWE (P [ 0.05)

was detected in eight SNPs out of nine for control group

(Table 3). However rs3767434 in POU2F1 exhibited

minor deviation (P = 0.03). SNPs rs7910977, rs854560,

rs10918682 from three genes IDE, PON1, POU2F1

respectively showed significant association with diabetes at

single SNP level.

Single SNP association analysis

Nine SNPs in genes IDE (rs6583813, rs7910977), IL1a(rs1800587), IL1b (rs1143634), POU2F1 (rs2146727,

rs10918682, rs3767434), PON1 (rs654560) and WFS1

(rs734312) were sequenced. Results of stastical analysis

(Table 3) depicted that no significant association was

found between rs6583813 and risk of diabetes development

(P [ 0.05) and rs7910977 [P \ 0.05, OR. 1,647 (95 % CI

1.112–2.438)] showed an association with the disease

(Table 4). Allele C was the major allele in rs7910977 and

CC was the homozygous major genotype with 0.712 fre-

quency in patients and 0.592 in controls (Table 4)

increasing the risk of diabetes onset.

POU2F1 gene located on first chromosome and three

SNPs (rs2146727, rs10918682, rs3767434) were targeted

on this locus. The rs2146727 and rs3767434 were with

P [ 0.05 and rs10918682 (POU2F1) [P \ 0.001, OR

3.606 (95 % CI 2.165–6.004)] exhibited association with

diabetes (Table 3). Heterozygous AG was a risk genotype

of rs10918682 associated with disease as compare to AA

and GG in Pakistani population and was significantly

associated with diabetes in Pakistanis (Table 4).

Both genes from interleukin-1, IL1a using rs1800587 as

marker and IL1b using rs1143634 as marker were not

associated with the diabetes in Pakistanis (P [ 0.05). IL1b(rs1143634) was only with two genotypes CC and CT

whereas homozygous TT was absent in this SNP. The

rs1800587 and rs1143634 were associated with diabetes at

genotype level (P \ 0.05). A significant association was

seen of PON1 gene (rs854560) with diabetes [P \ 0.05,

OR 0.663 (95 % CI 0.455–0.965)]. Gene WFS1 was not

associated with the onset of diabetes (P [ 0.05) and there

was no significant difference in distribution of A and G

allele of rs734312 in control and patients (Table 3).

However was associated with T2D at genotype level

(P = 0.0001) (Table 4).

Haplotype association analysis

Haplotype analysis (Table 5) of rs1800587 (IL1-alpha) and

rs1143634 (IL1-beta) determined that haplotype of major

alleles CC was associated with the increase of diabetesTa

ble

1P

CR

pri

mer

sfo

rse

lect

edS

NP

s

Gen

eS

NP

s1

stP

CR

Pro

du

ctsi

ze(b

p)

Nes

ted

PC

RP

rod

uct

size

(bp

)A

nn

eali

ng

tem

p(�

C)

IDE

rs6

58

38

13

F:

AT

TT

GG

CA

AA

AC

AC

AC

AC

G9

97

F:

AC

AG

GG

GG

AA

GG

GT

AT

AT

GG

73

76

0

R:

GA

TC

AC

TC

AC

CC

AC

CT

GG

TC

rs7

91

09

77

R:

GC

AG

AA

TC

TC

CT

TC

CC

TT

CC

PO

U2

F1

rs3

76

74

34

F:

GT

GG

GA

GG

TT

GA

TG

GG

AG

CT

GT

C1

20

4F

:G

GT

AG

GT

CA

TA

GA

GC

CA

AG

GA

GG

C7

98

62

R:

TG

GG

AG

GC

CC

GT

TT

TG

GG

TT

R:

TC

AA

GG

TG

GC

GA

CC

AC

AC

CA

A

PO

U2

F1

rs1

09

18

68

2F

:C

CT

TG

GC

CT

CC

CA

AA

GT

G9

68

F:

TG

AG

GT

TT

CA

TC

TC

GT

TT

GT

TG

64

26

0

R:

CA

TT

TG

GT

GG

TT

TC

AT

CA

CT

Grs

21

46

72

7R

:G

TT

CT

CA

GC

AC

GC

AC

TT

CC

WF

S1

rs7

34

31

2F

:T

TC

CC

CA

TC

GC

CA

GC

AA

GG

A1

,33

3F

:C

GC

CT

CC

AT

CG

GC

TA

CT

TC

CT

CT

45

56

0

R:

TC

AC

GC

GG

AC

GT

AC

TT

GA

AG

CG

R:

AT

CT

TC

AC

GT

GC

CG

CC

TG

GT

PO

N1

rs8

54

56

0F

:A

CA

GA

GT

TT

TG

GT

GG

GC

CA

GA

GG

1,8

33

F:

AT

CC

CT

AG

GC

CA

GA

TG

GT

GG

88

85

3

R:

GT

CT

TT

GA

GG

AA

AA

AG

CT

CT

AG

TC

CA

R:

AG

CA

GC

TG

GG

AG

GC

AA

CC

AA

C

IL1

-Ars

18

00

58

7F

:G

TC

AC

AG

GA

TG

GC

TC

CC

AA

CG

C1

,70

4F

:G

CT

GC

TT

TC

CT

CC

CA

GA

TC

CA

TG

C5

41

63

.8

R:

AA

GC

TC

CT

GG

TT

AG

GG

CA

GG

GT

R:

AA

CT

CT

CC

AC

CC

TG

GC

CC

TG

TT

IL1

-Brs

11

43

63

4F

:A

AA

GG

TA

GA

AG

GC

CC

AG

CG

G8

52

F:

AC

CT

GA

AG

CT

GG

AA

CC

CA

TG

57

86

4

R:

TT

CT

CA

GG

GT

CA

CA

CT

CC

TG

TR

:T

CC

TG

GG

AG

TG

CT

TG

AG

CC

GT

Ffo

rwar

d,

Rre

ver

se,

bp

bas

ep

airs

Mol Biol Rep

123

susceptibility [P \ 0.05, OR 1.488 (1.099–2.014)]. More-

over the CT haplotype was higher in frequency in controls

as compare to patients [P \ 0.01, OR 0.298 (0.139–0.639)

indicated protection against diabetes. Haplotype analysis of

IDE (rs6583813 and rs7910977) depicted CT haplotype

from these two SNPs was with higher frequency in controls

(0.252) than patients (0.173) conferring as protective hap-

lotype against diabetes [P = 0.02, OR 0.635 (0.419–

0.931)]. Haplotype GGA from POU2F1 (rs2146727–

rs10918682–rs3767434) was associated with the suscepti-

bility of T2D [P \ 0.001, OR 4.451(1.951–10.196)] while

GAA was protective against the risk of diabetes onset

[P \ 0.001, OR 0.332 (0.186–0.592)].

Linkage disequilibrium test

Analysis of genetic variants depicted the recombination (D0)between the SNPs with in a loci. Recombination in two

genetic variants (rs7910977, rs6583813) of IDE gene was

96 % (D0 = 0.968) and correlation coefficient (r2 = 0.160)

indicated the two SNPs are not good predictor of each other.

Strong LD was described in POU2F1 gene (rs3767434,

rs10918682, rs2146727) with 93 % (D0 = 0.937) recombi-

nation. Correlation between rs10918682 and rs2146727

revealed that these two SNPs are good predictor of each

other (r2 = 0.419). No correlation was detected in IL1a(rs1143634) and IL1b (rs1800587).

Discussion

There is a rapid increase in diabetes in Pakistani popula-

tion; higher BMI, sedentary life style and positive family

history are identified as major risk factors for the disease

onset. T2D is a multifactorial disease caused by the mutual

interaction of genetics and environmental factors [3]. In

Punjab, Pakistan first cousin marriages are very common.

According to our knowledge, current study is first to report

the association of rs7910977 (IDE), rs10918682 (POU2F1)

and rs854560 (PON1) with diabetes. The IDE gene was

identified for the intracellular proteolysis of insulin. This

gene is located on chromosome 10 that was suggested to be

linked with diabetes. In this research IDE (rs7910977) was

involved in pathogenesis of diabetes (P = 0.012) and

genotypes for rs7910977 were CC, CT and TT where CC

was with higher frequency (0.712) in the patients than

Table 2 Demographic characteristics

Characteristics Type 2 diabetes

patients

Control

n 288 170

Age (years) 50.45 ± 14.863 39.504 ± 18.185

Age at diagnosis

(years)

41.871 ± 11.71 –

BMI 26.723 ± 7.115 26.481 ± 6.183

Positive family history 191 (66.32 %) –

Negative family history 97 (33.68 %) –

Hypertension 144 (50 %) –

Duration

1–5 129 (44.79 %) –

6–10 58 (20.14 %)

[10 101 (35.06 %)

Complications

Nephropathy 32 (11.11 %) –

Retinopathy 27 (9.37 %)

Cancer 14 (4.86 %)

Cardiovascular 42 (14.58 %)

Data is presented as mean ± std

– nil, n number (% age)

Table 3 Association analysis at allele level

Gene SNPs Allele frequency RR (95% CI) OR (95% CI) P values

M (case/

control)

m (case/control) Fisher’s

exact

v2 Pearson

v2

IDE rs6583813 C (0.579/0.639) T (0.421/0.361) 0.902 (0.795, 1.024) 0.767 (0.551, 1.068) 0.132 0.1367 0.116

rs7910977 C (0.823/0.748) T (0.177/0.252) 1.113 (1.019, 1.215) 1.647 (1.112, 2.438) 0.014* 0.016* 0.012*

POU2F1 rs2146727 A (0.640/0.655) T (0.360/0.345) 1.043 (0.841, 1.292) 1.067 (0.767, 1.484) 0.736 0.7644 0.701

rs10918682 A (0.760/0.919) G (0.240/0.081) 2.980 (1.892, 4.691) 3.606 (2.165, 6.004) 0.0001* 0.0001* 0.0001*

rs3767434 A (0.857/0.908) G (0.143/0.092) 1.551 (0.951, 2.582) 1.643 (0.949, 2.846) 0.080 0.099 0.074

WFS1 rs734312 A (0.528/0.560) G (0.461/0.349) 0.911 (0.809, 1.025) 0.804 (0.606, 1.066) 0.133 0.148 0.129

PON1 rs854560 T (0.806/0.734) A (0.126/0.174) 0.708 (0.518, 0.969) 0.663 (0.455, 0.965) 0.0384* 0.0389* 0.0312*

IL1a rs1800587 C (0.615/0.577) T (0.385/0.423) 0.892 (0.743, 1.075) 0.830 (0.611, 1.127) 0.2429 0.263 0.2315

IL1b rs1143634 C (0.865/0.860 T (0.117/ 0.134) 1.009 (0.958, 1.062) 1.072 (0.716, 1.605) 0.7563 0.815 0.735

OR odd ratio, RR relative risk, v2 Chi Square, allele frequency, M major allele, m Minor allele

* Significant SNPs with P \ 0.05

Mol Biol Rep

123

control (0.592). In line to our findings IDE locus was

reported to be involved in decrease of function in pancre-

atic beta-cells in Europeans [26]. It was reported that

genetic variants at IDE confer clinically significant risk of

diabetes in Caucasians [27]. A study in China (Shanghai/

Beijing) described that it’s a risk for diabetes in Shanghai

(P = 0.008) where as it was not a risk genetic variant for

individuals of Beijing (Wu et al. [7]). Data also suggested

that the variants in IDE were associated with onset of

diabetes by its influences on pediatric BMI [28].

In our study out of three SNPs from POU2F1 one SNP

(10918682) reported to be associated with T2D

[P \ 0.001, OR 3.606 (2.165, 6.004)]. The other two SNPs

(rs2146727 and rs3767434) were not associated with T2D.

Still this gene is only studied in Chinese population by Ng

et al. [6] and the association of POU2F1 (rs10918682) with

pathogenesis of diabetes was reported in East Asians that’s

inline to the present study. It was reported in an interna-

tional 1q consortium that the linkage of 1q21–25 region

was linked with T2D in eight populations [14].

A significant association was seen between PON1 gene

(rs854560) with diabetes [P \ 0.05, OR 0.663 (0.455,

0.965)] and TT homozygous was with 0.690 frequency in

cases that is higher in comparison control (0.571). In

contrast to current study results there was no evidence of

association of rs854560 polymorphism with disease

(P = 0.370) OR 0.97 [95 % CI 0.91–1.04] [15]. L55M

(rs854560) influence activity of PON1 gene, antioxidant

activity of this gene play a role in prevention of oxidative

stress, its role in pathophysiological process associated

with atherosclerosis and diabetes mellitus. This SNP

associated with changes in concentration of the enzyme

[16]. Polymorphism in PON1 gene was also reported to be

associated with diabetic retinopathy [29].

We did not find any association between rs734312

(WFS1) with T2D at allele level (P = 0.1337) but was

associated at genotype level in Pakistanis, a south Asian

population. Minton et al. [30] provide the evidence that

variants in WFS1 gene associated with the susceptibility of

T2D. This SNP was identified as associated with diabetes

risk in the population of UK, French and Ashkenazi [15,

31].

IL1a (rs1800587) and IL1b (rs1143634) at allele and

genotype levels did not show any relationship with the

development of T2D (P [ 0.05). However, rs1143634 in

IL1b was with two genotypes (CC, CT); no association was

observed with diabetes at genotype level (P [ 0.05). The

rs1800587 and rs1143634 showed association with T2D at

haplotype level (Table 5). It was reported that insulin

producing b-cells in pancreatic islets are specifically prone

Table 4 Association analysis at genotype level

SNP_ID M [ m HWE P value Genotype frequency Overall genotype P value

MM (case/control) Mm (case/control) mm (case/control)

rs6583813 C[T 0.85 CC (0.369/0.418) CT (0.421/0.443) TT (0.210/0.139) 0.27

rs7910977 C[T 0.09 CC (0.712/0.592) CT (0.223/0.312) TT (0.065/0.096) 0.08

rs2146727 A[G 0.12 AA (0.430/0.460) AG (0.420/0.389) GG (0.150/0.151) 0.84

rs10918682 A[G 0.15 AA (0.597/0.855) AG (0.325/0.129) GG (0.078/0.016) 0.0001*

rs3767434 A[G 0.03 AA (0.734/0.849) AG (0.245/0.118) GG (0.021/0.034) 0.07

rs734312 A[G 0.06 AA (0.253/0.383) AG (0.552/0.354) GG (0.185/0.171) 0.0001*

rs854560 T[A 0.07 TT (0.690/0.571) AT (0.231/0.326) AA (0.011/0.011) 0.05

rs1800587 C[T 0.87 CC (0.314/0.320) CT (0.603/0.514) TT (0.083/0.166) 0.06

rs1143634 C[T 0.08 CC (0.747/0.726) CT (0.235/0.269) TT (0.00/0.00) 0.41

HWE Hardy–Weinberg equilibrium test (performed only for controls), M major allele, m minor allele

* Association with T2D (P \ 0.05)

Table 5 Aaaociation analysis at haplotype level

Haplotype Case

(freq)

Control

(freq)

P value OR (95% CI)

Loci: 6583813–rs7910977

CC 0.406 0.391 0.676 1.076 (0.769–1.499)

CT 0.173 0.252 0.020 0.635 (0.419–0.931)

TC 0.416 0.357 0.133 1.294 (0.924–1.813)

Loci: rs1800587–rs1143634

CC 0.595 0.497 0.010 1.488 (1.099–2.014)

CT 0.026 0.084 0.001 0.298 (0.139–0.639)

TC 0.307 0.372 0.071 0.746 (0.543–1.026)

TT 0.072 0.047 0.178 1.556 (0.184–2.975)

Loci: rs2146727–rs10918682–rs3767434

AAA 0.567 0.561 0.916 1.024 (0.662–1.583)

AAG 0.077 0.067 0.732 1.158 (0.499–2.685)

GAA 0.109 0.269 0.001 0.332 (0.186–0.592)

GGA 0.199 0.053 0.001 4.451 (1.951–10.156)

Bold values depicted the significantly associated haplotypes with

diabetes P \ 0.05

Mol Biol Rep

123

to IL1b induced destruction and loss of function as well as

insulin resistance in obesity [32]. Loutola et al. [33]

reported in genetic analysis that rs1800587 (IL1a) and

rs1143634 (IL1b) with P values 0.002 and 0.023 respec-

tively were associated with risk of developing diabetes

(gender specific). IL1-alpha and IL1-beta were associated

with central obesity (P \ 0.05) in western Australian

population [20]. In a European study it was reported that

IL1b was associated with diabetes and its minor allele was

found to be associated with higher level of blood glucose in

comparison to major allele [21].

Our study demonstrated that the genetic variants in IDE,

PON1, WFS1, IL1a, IL1b and POU2F1 play a role in the

development of T2D in Pakistani population. There is a

need for replicative studies at large scale to determine the

role of genetic variants in pathology of diabetes which can

be helpful in early prediction as well as control and pre-

vention of this disease.

Acknowledgments We are grateful to our colleagues at Applied

Genomic Centre of Hong Kong University of Science and Technol-

ogy for their assistance during this work.

References

1. Yang W, Lu J, Weng J, China National Diabetes and Metabolic

Disorders Study Group (2010) Prevalence of diabetes among men

and women in China. N Engl J Med 362:1090–1101

2. Steyn NP, Bennett J, Temple N et al (2004) Diet, nutrition and

the prevention of type 2 diabetes. Public Health Nutr 7(1A):

147–165

3. Rees SD, Hydrie MZ, Shera AS et al (2011) Replication of 13

genome-wide association (GWA)-validated risk variants for type 2

diabetes in Pakistani populations. Diabetologia 54(6):1368–1374

4. Shera AS, Rafique G, Khwaja IA et al (1995) Pakistan national

diabetes survey: prevalence of glucose intolerance and associated

factors in Shikarpur, Sindh Province. Diabet Med 12:1116–1121

5. Groves JC, Wiltshire S, Smedley D et al (2003) Association and

haplotype analysis of the insulin-degrading enzyme (IDE) gene, a

strong positional and biological candidate for type 2 diabetes

susceptibility. Diabetes 52:1300–1305

6. Ng MC, Park KS, Oh B et al (2008) Implication of genetic

variants near TCF7L2, SLC30A8, HHEX, CDKAL1, CDKN2A/

B, IGF2BP2, and FTO in type 2 diabetes and obesity in 6,719

Asians. Diabetes 57:2226–2233

7. Wu Y, Li H, Loos RJ et al (2008) Common variants in CDKAL1,

CDKN2A/B, IGF2BP2, SLC30A8, and HHEX/IDE genes are

associated with type 2 diabetes and impaired fasting glucose in a

Chinese Han population. Diabetes 57:2834–2842

8. McCarthy MI (2003) Growing evidence for diabetes suscepti-

bility genes from genome scan data. Curr Diab Rep 3:159–167

9. Groves CJ, Zeggini E, Minton J et al (2006) Association analysis

of 6,736 U.K. subjects provides replication and confirms TCF7L2

as a type 2 diabetes susceptibility gene with a substantial effect

on individual risk. Diabetes 55(9):2640–2644

10. Barroso I, Luan J, Middelberg PSR et al (2003) Candidate gene

association study in type 2 diabetes indicates a role for genes

involved in b-cell function as well as insulin action. PLoS Biol

1(1):20

11. Hoek M, Dehghan A, Witteman CMJ et al (2008) Predicting type

2 diabetes based on polymorphisms from genome-wide associa-

tion studies a population-based study. Diabetes 57:3122–3128

12. Moore AF, Florez JC (2008) Genetic susceptibility to type 2

diabetes and implications for antidiabetic therapy. Annu Rev Med

59:95–111

13. Sangheraa DK, Demircib FY, Beena L et al (2010) PPARG and

ADIPOQ gene polymorphisms increase type 2 diabetes mellitus

risk in Asian Indian Sikhs: Pro12Ala still remains as the strongest

predictor. Metab Clin Experimental 59:492–501

14. Prokopenko I, Zeggini E, Hanson RL et al (2009) Linkage dis-

equilibrium mapping of the replicated type 2 diabetes linkage

signal on chromosome 1q. Diabetes 58:1704–1709

15. Cheurfa N, Brenner GM, Reis AF et al (2011) Decreased insulin

secretion and increased risk of type 2 diabetes associated with

allelic variations of the WFS1 gene: the data from epidemiolog-

ical study on the insulin resistance syndrome (DESIR) prospec-

tive study. Diabetologia 54(3):554–562

16. Luı0sa Veiga, Silva-Nunes J, Melao A et al (2011) Q192R

polymorphism of the paraoxonase-1 gene as a risk factor for

obesity in Portuguese women. Eur J Endocrinol 164:213–218

17. Zhang G, Li W, Li Z, Lv H et al (2013) Association between

paraoxonase gene and stroke in the Han Chinese population.

BMC Med Genet 14(16):1–10

18. Banerjee M, Saxena M (2012) Interleukin-1 (IL-1) family of

cytokines: role in type 2 diabetes. Clin Chim Acta 413(15–16):

1163–1170

19. Carter WK, Hung J, Powell LB et al (2008) Association of

Interleukin-1 gene polymorphisms with central obesity and

metabolic syndrome in a coronary heart disease population. Hum

Genet 124(3):199–206

20. Luotola K, Paakkonen R, Alanne M et al (2009) Association of

variation in the interleukin-1 gene family with diabetes and

glucose homeostasis. J Clin Endocrinol Metab 94(11):4575–4583

21. Syed AA, Julie EI, Redfern CPF et al (2004) Low prevalence of

the N363S polymorphism of the glucocorticoid receptor in South

Asians Living in the United Kingdom. J Clin Endocrinol Metab

89:232–235

22. Rees SD, Islam M, Hydrie MZI et al (2011) An FTO variant is

associated with type 2 diabetes in South Asian populations after

accounting for body mass index and waist circumference. Diabet

Med 28:673–680

23. W-S Lo, Harano M, Gawlik M et al (2007) GABRB2 association

with schizophrenia: commonalities and differences between eth-

nic groups and clinical subtypes. Biol Psychiatry 61:653–660

24. Shi YY, He L (2005) SHEsis, a powerful software platform for

analyses of linkage disequilibrium, haplotype construction,

and genetic association at polymorphism loci. Cell Res 15(2):

97–98

25. Li Z, Zhang Z, He Z et al (2009) A partition-ligation-combina-

tion-subdivision EM algorithm for haplotype inference with

multiallelic markers: update of the SHEsis (http://analysis.bio-x.

cn). Cell Res 19(4):519–523

26. Pascoe L, Tura A, Patel KS et al (2007) Common variants of the

novel type 2 diabetes genes CDKAL1 and HHEX/IDE are asso-

ciated with decreased pancreatic-cell function. Diabetes

56:3101–3104

27. Florez JC, Jablonski AK, Bayley N et al (2006) TCF7L2 poly-

morphisms and progression to diabetes in the diabetes prevention

program. N Engl J Med 355(3):241–250

28. Zhao J, Bradfield PJ, Zhang H et al (2010) Examination of all

type 2 diabetes GWAS loci reveals HHEX-IDE as a locus influ-

encing pediatric BMI. Diabetes 59:751–755

29. Yan-Lin Kao, Donaghue K, Chan A et al (1998) A variant of

paraoxonase (PON1) gene is associated with diabetic retinopathy

in IDDM. J Clin Endocrinol Metab 83(7):2589–2592

Mol Biol Rep

123

30. Minton ALJ, Hattersley TA, Owen K et al (2002) Association

studies of genetic variation in the WFS1 gene and type 2 diabetes

in U.K. populations. Diabetes 51:1287–1290

31. Sandhu MS, Weedon MN, Fawcett KA et al (2007) Common

variants in WFS1 confer risk of type 2 diabetes. Nat Genet

39(8):951–953

32. Maedler K, Dharmadhikari G, Schumann DM, Størling J (2009)

Interleukin-1 beta targeted therapy for type 2 diabetes. Expert

Opin Biol Ther 9(9):1177–1188

33. Luotola K, Pietila A, Zeller T et al (2011) Associations between

interleukin-1 (IL-1) gene variations or IL-1 receptor antagonist

levels and the development of type 2 diabetes. J Intern Med

269(3):322–332

Mol Biol Rep

123