No gene copy number changes in Dupuytren’s contracture
by array comparative genomic hybridization
Sippy Kaura,1, Minna Forsmanb,1, Jorma Ryha ¨nenb,
Sakari Knuutilaa,*, Marcelo L. Larramendya
aDepartment of Pathology, Haartman Institute and HUSLAB, University of Helsinki and University
Central Hospital, POB 21 (Haartmaninkatu 3), Helsinki 00014, Finland
bDepartment of Surgery, Oulu University, Aapistie 5A, POB 5000, 90014, Oulu, Finland
Received 8 December 2007; received in revised form 15 January 2008; accepted 28 January 2008
Dupuytren’s contracture (DC), a benign disease of unknown origin, is characterized by abnormal
fibroblast proliferation and matrix deposition within the palmar and plantar faciae, causing contrac-
ture of the digits. Conventional cytogenetic studies of cultured fibroblast cells from DC nodules
have revealed nonrecurrent, but usually normal, clonal (mainly þ7, þ8, and eY, plus structural
changes) and sporadic (nonclonal) numerical/structural rearrangements. No unique cytogenetic
features of DC are known so far. We used 44K oligonucleotide-based array comparative genomic
hybridization to obtain a wide pattern of chromosomal imbalances in 18 patients with DC. The
genome-wide analysis revealed no changes of DNA copy number sequences. Accordingly, gene
amplifications or deletions are apparently not involved in the progression of abnormal fibroblast
proliferation and matrix deposition that lead to DC.
? 2008 Elsevier Inc. All rights reserved.
Dupuytren’s contracture (DC) (OIMM 126900) is a be-
nign disease of unknown origin. The Office of Rare
Diseases of the National Institutes of Health (NIH, Bethes-
da, MD, USA) lists it as a ‘‘rare disease’’ because it affects
less than 200,000 people in the United States (http://
asp?ID58237). DC is prevalent among Caucasians, and
particularly high frequencies (30e40%) have been reported
in those of Northern European ancestry. DC is also com-
mon among the offspring of immigrants from Northern Eu-
rope, but rare in other ethnic groups, such as Chinese and
Africans, where findings have been sporadic [1,2]. DC is
characterized by abnormal fibroblast growth proliferation
and matrix production (e.g., myofibroblasts with increased
amounts of actin , collagen type III over type I ,
proteoglucan in the extracellular matrix , and enhanced
concentration of oxygen free radicals ).
It is still uncertain whether DC is a complex oligogenic
condition or a simple monogenic Mendelian disorder. The
observed male predominance is related to the expression
of androgen receptors in Dupuytren’s fascia rather than be-
ing of genetic origin . Gene mutations may be involved
in the pathogenesis of DC. A hitherto unknown heteroplas-
mic mutation located within the mitochondrial 16s ribo-
somal RNA has been demonstrated in 90% of DC with a
maternally transmitted inheritance pattern  and an auto-
somal dominant gene for DC at 16q between D16S419 and
Cytogenetic investigations of DC have revealed numer-
ical and structural clones, random numerical and structural
aberrations, prophasing, and premature centromere separa-
tion [10e14]. No unique cytogenetic features of DC are
known so far.
We applied 44K oligonucleotide-based array compara-
tive genomic hybridization (aCGH) on 19 samples of DC
to obtain a genome-wide pattern of chromosomal imbal-
ances and determine aberrations that have not been detected
by conventional cytogenetic and CGH analyses.
2. Materials and methods
Nineteen palmar samples from 18 patients diagnosed
with Dupuytren’s contracture were referred to surgery with
* Corresponding author. Tel.: þ358-9-19126527; fax: þ358-9-
E-mail address: firstname.lastname@example.org (S. Knuutila).
1Both of these authors contributed equally to this report.
0165-4608/08/$ e see front matter ? 2008 Elsevier Inc. All rights reserved.
Cancer Genetics and Cytogenetics 183 (2008) 6e8
preselection (Table 1). An aponeurectomy was carried out
to restore an extension limitation of 30 degrees or more
in the metacarpal joint or to heal an annoying midpalmar
cord. All patients were clinically estimated and evaluated
for surgery by two hand surgeons according to normal op-
erating routine. No immunologic studies were performed
on samples. Surgically selected tissues were obtained from
each patient and maintained in liquid N2until DNA extrac-
tion was performed.
2.2. Oligonucleotide aCGH and data analyses
aCGH was performed using Agilent’s 60-mer oligonu-
cleotide-based microarray according to the manufacturer’s
instructions (Human Genome CGH Microarray Kit 44B;
Agilent Technologies, Palo Alto, CA, USA). DNA extrac-
tion from all samples was performed following standard
methods. Reference male and female DNA were extracted
from pooled peripheral blood cells of healthy donors. After
hybridization and post-hybridization washes, arrays were
scanned using the Agilent laser confocal scanner and the
results were analyzed using the CGH Analytics software
(version 3.1; Agilent Technologies) .
3. Results and discussion
Original aCGH data of all DC samples are accessible at
http://www.cangem.org/. We found no chromosomal imbal-
ances in any of the 19 samples, which represent the largest
cohort of DC patients analyzed by aCGH so far.
The gene expression profile of DC has been analyzed
recently [16e19]. Although no common pattern of gene
expression was detected, 46 targets with an altered expres-
sion pattern were found. Most of them were involved in tis-
sue development and cellular differentiation (e.g., collagen
degradation, ossification, and myofibroblast differentiation)
[16e19]. Twenty-four of the altered genes were found to be
overexpressed (i.e., CSF1, TMSB10, COL5A2, WNT5A,
RHOA, P4HA2, CTNNA1, RUNX2, HSPB1, LRRC17, PTN,
SHANK2, ARCN1, CTN1, TUBA1A, DAD1, MMP2, TCF4,
MAFB, GNAS, APP, TMSB4X, PRKX, and GDI1) [16e19].
On the other hand, the remaining 22 targets (i.e., ECHS1,
CLEC3B, CD14, TNF, TFPI2, EPB49, AKR1C1, CD81,
LSP1, WNT11, GDP1, DCN, ALDH2, ATP7B, TNFSF12,
CCL5, LAMA3, LIPE, CYB5A, ACP5, SYMPK, and ICAM1)
were underexpressed [16,18,19]. However, none of these
genes was affected either by DNA copy number gains or
losses within our samples (http://www.cangem.org/).
Conventional cytogenetic and interphase in situ hybrid-
ization studies of fibroblast cell cultures from DC nodules
revealed the presence of nonrecurrent numerical/structural
clonal and sporadic (nonclonal) numerical/structural changes
in up to 50% of the metaphases analyzed. Among them, þ7
and þ8 are the most frequently reported alterations, although
their presence has also been observed in normal palmar fas-
cia cultures of the patients [10e14,20e23]. Thus, it has been
suggested that these alterations are most probably a result of
abnormal growth of trisomic fibroblasts with a selective
growth advantage over cytogenetically normal fibroblasts
in the in vitro condition, without commitment to the origin
of the contracture [10,13]. Taking into account these obser-
vations and the lack of DNA copy number changes we
found, it could be suggested that the presence of trisomies
in DC is most probably related to a Darwinian in vitro
Clinical characteristics of 18 patients with palmar DC studied by aCGH
Coronary heart disease, hypertension
Hypertension, bronchial asthma, hypercholesterolinemia,
diabetes mellitus (oral glucose lowering drugs)
Bronchial asthma, hypercholesterolinemia, diabetes
mellitus (on diet), prostatic hyperplasia
Charcot-Marie-tooth disease, Mb Menier
Coronary heart disease, hypercholesterolinemia
Brother, sister, father
M, male; F, female; *, years at sampling; e, negative; þ, positive
S. Kaur et al. / Cancer Genetics and Cytogenetics 183 (2008) 6e8
selection of trisomy-carrying cells within the samples under Download full-text
culturing with a higher proliferative capacity over that of
cells with a normal karyotype. Although aCGH can reveal
numerical chromosomal changes (gains and losses) of
approximately 35 kilobases in the whole genome , the
alteration detected by this methodology should be present
in at least 50% of the cells [25,26]. Accordingly, we cannot
rule out the possibility that such alterations could exist out-
side the resolution threshold used in the present study or
that the frequency of aberrant cells was lower than 50%
within the samples. Further molecular studies on more sen-
sitive platforms of analysis would be required to clarify
these putative possibilities.
This work was supported by grants from the Sigrid
Juse ´lius Foundation and the Helsinki University Hospital
Research Funds in Finland, and by National University of
La Plata (grant no. 11/N493), National Agency of Scientific
and Technological Promotion (contract grant no. PICT
2004 no. 26116), and the National Council of Scientific
and Technological Research (CONICET, PIP 6386), all in
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