2008 Wiley-Liss, Inc. American Journal of Medical Genetics Part
A 146A:18651870 (2008)
A Multiplex Family With Possible MetaphysealSpahr-Type Dysplasia
and Exclusion of RMRP and
COL10A1 as Candidate Genes
Andre Megarbane,1* Eliane Chouery,1 and Ismat Ghanem21Unite de
Genetique Medicale, Faculte de Medecine, Universite Saint Joseph,
2Service dOrthopedie, Hotel-Dieu de France, Beirut, Lebanon
Received 29 January 2008; Accepted 9 April 2008
We report a consanguineous Lebanese family where sixindividuals
had disproportionate short stature, short limbs,and bilateral genu
varum that were apparent after 2 years ofage. Major radiographic
features in infancy included flared,cupped, and ragged metaphyses
of all long bones, delayedmaturation of the scaphoid, trapezoid,
and trapezium bones,wide and slightly irregular ribs, short femoral
necks, flatfemoral heads, and irregularity of the iliac crest.
Radiographsof an affected adult showed diminished
metaphysealchanges and slightly short femoral necks, when
to the younger patients. We suggest that this family isaffected
with the Spahr type of metaphyseal chondrodys-plasia. Sequencing of
RMRP, and a haplotype analysis usinghighly informative markers
around the COL10A1 excludedboth genes from being pathogenic in this
family. We areaware of only two previous reports of families with
clinicalfeatures similar to the Spahr type of metaphyseal
chondro-dysplasia. 2008 Wiley-Liss, Inc.
Key words: dysplasia; metaphysis; genu varum; short stature
How to cite this article: Megarbane A, Chouery E, Ghanem I.
2008. A multiplex family with possiblemetaphyseal Spahr-type
dysplasia and exclusion of RMRP and COL10A1 as candidate genes.
Am J Med Genet Part A 146A:18651870.
The metaphyseal dysplasias are a clinically hetero-geneous group
of disorders characterized by meta-physeal irregularities and,
usually, with short stature.They can be distinguished by clinical
and radio-graphic findings and genetic testing. We
recentlyevaluated a consanguineous Lebanese family wheresix
individuals of different ages presented withpostnatal
disproportionate short stature, short limbs,genu varum, and
metaphyseal abnormalities. Clin-ical analysis and candidate gene
testing were used toevaluate the family.
MATERIALS AND METHODS
Members of this kindred (Fig. 1) originate fromBeirut, capital
of Lebanon, and belong to the SunniMuslim community. Three patients
have beenexamined. The patients underwent an extensivework-up
including a thorough clinical evaluation,total body X-rays, and
routine blood tests (completeblood count, serum electrolytes, blood
glucoselevels, white blood cells enzyme assays
screening,triglycerides, cholesterol, thyroid, liver and
renalfunction tests, PTH and phosphatase alkaline levels).
Peripheral blood was obtained after informedconsent was signed
from individuals IV-4 and IV-5and their three sibs (Fig. 1).
Genomic DNA wasextracted from leucocytes by standard
Genomic and cDNA sequences of RMRP(AF334829, M29916.1) were
obtained from NCBIdatabase. The entire transcribed region and500
bppromoter region were amplified by polymerasechain reaction from
genomic DNA, and werescreened for mutations through direct
sequencing.Primers sequences were chosen according to Naka-shima et
al. . Mutations were numbered fromthe putative translational
start site of M29916.1.
PCR was performed using Taq DNA polymerase(Invitrogen Life
Technologies, Carlsbad, CA). Theamplification conditions for each
PCR were 958C for5 min, followed by 35 cycles of 958C for 60 sec,
*Correspondence to: Andre Megarbane, M.D., Ph.D., Unite
deGenetique Medicale, Faculte de Medecine, Universite Saint Joseph
deBeyrouth, 42, rue de Grenelle, 75007 Paris, France.E-mail:
for 60 sec and 728C for 60 sec, with a final extensionof 10 min
at 728C. The amplified PCR fragments wereanalyzed on 0.8% agarose
PCR products from genomic DNA were purifiedusing the Ultra PCR
Clean-Up Kit (Abgene, Surrey,UK), and the products were sequenced
for bothstrands using the BigDye1 Terminator v1.1 CycleSequencing
Kit (Applied Biosystems, Foster City,CA). A total volume of 7.5 ml
sequencing reactionsincluding 2 ml Big Dye, 1023 ng of purified
DNAtemplate, and 1.6 pmol of either forward or reverseprimerswas
incubated for 25 cycles at 968C for 10 sec,508C for 5 sec, and 608C
for 4 min. Sequencereactions were purified on sephadex G50
(Amer-sham Pharmacia Biotech, Foster City, CA). Thelabeled products
were dissolved in HiDi formamideprior to loading into an ABI 3100
system. The sameprimers as for the DNA amplification were used
forsequencing. Electrophoregrams were analyzedusing Sequence
Analysis Software version 5.2(Applied Biosystems) and compared to
referencesequences using ChromasPro version 1.22 (Techne-lysium,
Haplotype analysis was performed using eighthighly informative
markers (D6S302, D6S416,D6S418, D6S432, D6S304, D6S262, D6S292,
andD6S308) flanking the COL10A1 gene. The positionof the gene and
markers were determined by UCSCHuman Genome Browser
(http://www.genome.ucsc.edu). Genotyping was performed as
describedelsewhere [Delague et al., 2000].
The proband (Fig. 1: V-5) was born at term to a 27-year-old
mother and the 30-year-old father. Her birthweight was 3,500 g
(50th centile), and length 48 cm(25th centile). Head circumference
(OFC) was notrecorded but considered normal by the parents. At
the age of 4 months, the parents noticed the presenceof short
limbs and genu varum. Developmentalmilestones were within the
This girl was seen at the age of 78/12 years, whenshe was
referred for familial short stature. She hadnormal school
performance. Her height was 113 cm(3rd centile), weight 23 kg (50th
FIG. 1. Pedigree of the family. Affected patients are indicated
by darkened symbols.
FIG. 2. Photograph of Patient V-5. Note the short stature and
the genu varum.[Color figure can be viewed in the online issue,
which is available atwww.interscience.wiley.com.]
1866 MEGARBANE ET AL.
American Journal of Medical Genetics Part A
- circumference 52 cm (>50th to
- At examination, her age was 35 years. Her height was140 cm
malformations that consist mainly of early metaphy-seal changes
with apparent regression. Because oneof the most striking features
was the metaphysealdysplasia, differential diagnoses were thus
consid-ered with skeletal dysplasias featuring
The overall pattern of abnormalities seen in thepatients
described herein is distinct from themetaphyseal dysplasia Jansen
type (OMIM 156400),the VaandragerPina syndrome [Kozlowski
andSikorska, 1970] (OMIM 250300), the metaphysealdysplasia
Shwachman type [Makitie et al., 2004](OMIM 260400), the entity
described by Castriota-Scanderbeg et al. , and the child
described byHoeffel et al. . The anadysplasia type I and
II[Maroteaux et al., 1991; Le Merrer and Maroteaux,1998], and the
entity reported by Wiedemann andSpranger  were ruled out as
possible diagnosesby the fact that there is only a mild decrease in
finallength or a normal stature in those syndromes, and adifferent
mode of inheritance.
The cartilage-hair-hypoplasia dysplasia (CHH)(OMIM 250250) is an
autosomal recessive chondro-dysplasia characterized by metaphyseal
dysplasia,short stature, and often, fine and sparse hair,transient
macrocytic anemia, and immunodefi-ciency. It is differentiated from
the patients reportedhere by the fact that in the latter the length
is normalat birth, the hands are not pudgy, and there are noround
epiphyses. Nevertheless, as carpal abnormal-ities had been reported
in CHH and found in thepresent family, and because of the marked
clinicalheterogeneity found in those with RMRP mutations[Ridanpaa
et al., 2001], we sequenced this gene butno mutations were
The Schmid metaphyseal dysplasia (OMIM156500) is a well known
type X collagen chondro-dysplasia. Its manifestations are similar
to the presentfamily including anormal appearance at birth, a
shortadult stature, bowed legs, and a coxa vara. Never-theless, it
is an unlikely diagnosis for the presentfamily because it has an
autosomal dominant modeof inheritance. In addition, in the Schmid
type,metaphyseal irregularity is most pronounced at theknees,
epiphyses are wide, and mild platyspondylyin infancy and early
childhood, and mild handinvolvement such as shortening of tubular
bonesand metaphyseal cupping of metacarpals andproximal phalanges
can be seen [Lachman et al.,1988; Nishimura et al., 2001; Elliot et
al., 2005].Haplotype analysis excluded linkage of COL10A1 tothis
Spahr and Spahr-Hartmann  described aconsanguineous family
with an autosomal recessivemetaphyseal dysplasia very similar to
Schmid meta-physeal dysplasia. Farag and Teebi  reportedfour
affected sibs who probably had the samemetaphyseal dysplasia.
Unfortunately, in the Faragand Teebi report, the clinical history
was not well
documented and the radiological photos were notinsufficient for
further comparison. Similaritiesbetween the Spahr and SpahrHartmann
reportand the patients reported here include a normalfacial
appearance, a short stature that appearedprogressively after the
age of 2 years with a finalheight at around 140 cm, a mild form of
metaphysealdysplasia affecting the bones and that
apparentlyimproved with age, no metaphyseal irregularity ofthe
fingers, and normal joint mobility. The pheno-type is variable as
the patients reported here hadadditional features such as carpal
bone hypoplasia,abnormal ribs, slightly short femoral necks, and
iliaccrest irregularity (patient V-6), but not the abnormalgait,
lordosis, osteoporosis and large articulationsseen in the patients
reported by Spahr and Spahr-Hartmann . Other similar reports
and identi-fication of the gene will further characterize
theclinical spectrum of this disorder and clarify itsetiology.
Castriota-Scanderbeg A, Dallapiccola B, Mingarelli R,
KozlowskiK. 2001.Distinctive metaphyseal chondrodysplasia
simulatingcartilage hair hypoplasia. Am J Med Genet 99:289293.
Delague V, Bareil C, Tuffery S, Bouvagnet P, Chouery E, Koussa
S,Maisonobe T, Loiselet J, Megarbane A, Claustres M. 2000.Mapping
of a new locus for autosomal recessive demyelinat-ing
Charcot-Marie-Tooth disease to 19q13. 1-13.3 in a
largeconsanguineous Lebanese family: Exclusion of MAG as acandidate
gene. Am J Hum Genet 67:236243.
Elliot AM, Field FM, Rimoin DL, Lachman RS. 2005.
Handinvolvement in Schmid metaphyseal chondrodysplasia. Am JMed
Genet Part A 132A:191193.
Farag TI, Teebi AS. 1990. The second family of
Spahr-typemetaphyseal chondrodysplasia: Autosomal recessive
inher-itance confirmed. Clin Genet 38:237239.
Hoeffel JC, Didier F, Chantereau D, Medoc JM, Muller JP.
1987.Metaphyseal dyschondroplasia with cone-shaped epiphyses.Br J
Kozlowski K, Sikorska B. 1970. Dysplasia metaphysaria,
typeVaandrager-Pena. Z Kinderheilkd 108:165170.
Lachman RS, Rimoin DL, Spranger J. 1988.
Metaphysealchondrodysplasia, Schmid type. Clinical and
radiographicdelineation with a review of the literature. Pediatr
Le Merrer M, Maroteaux P. 1998. Metaphyseal anadysplasia typeII:
A new regressive metaphyseal dysplasia. Pediatr
Makitie O, Ellis L, Durie PR, Morrison JA, Sochett EB,
RommensJM, Cole WG. 2004. Skeletal phenotype in patients
withShwachman-Diamond syndrome and mutations in SBDS. ClinGenet
Maroteaux P, Verloes A, Stanescu V, Stanescu R. 1991.
Meta-physeal anadysplasia: A metaphyseal dysplasia of early
onsetwith radiological regression and benign course. Am J MedGenet
Nakashima E, Mabuchi A, Kashimada K, Onishi T, Zhang J,Ohashi H,
Nishimura G, Ikegawa S. 2003. RMRP mutations inJapanese patients
with cartilage-hair hypoplasia. Am J MedGenet Part A
Nishimura G, Manabe N, Kosaki K, Haga N, Ohashi H, NakamuraK,
Ikegawa S. 2001. Spondylar dysplasia in type X collagen-opathy.
Pediatr Radiol 31:7680.
Ridanpaa M, van Enennaam H, Pelin K, Chadwick R, JohnsonC, Yuan
B, vanVenrooij W, Pruijn G, Salmela R, Rockas
METAPHYSEAL SPAHR-TYPE DYSPLASIA 1869
American Journal of Medical Genetics Part A
S, Makitie O, Kaitila I, de la Chapelle A. 2001. Mutations inthe
RNA component of RNase MRP cause a pleiotropichuman disease,
cartilage-hair hypoplasia. Cell 104:195203.
Spahr A, Spahr-Hartmann I. 1961. Dysostose
metaphysairefamiliale: etude de 4 cas dans une fratrie. Helv
Thiel CT, Mortier G, Kaitila I, Reis A, Rauch A. 2007. Type
andlevel of RMRP functional impairment predicts phenotype inthe
cartilage hair hypoplasia-anauxetic dysplasia spectrum.Am J Hum
Wiedemann HR, Spranger J. 1970. Metaphyseal chondrodyspla-sia
(metaphyseal dysostosis)A new type? Z Kinderheilkd108:171186.
1870 MEGARBANE ET AL.
American Journal of Medical Genetics Part A