CYTOGENETIC EVIDENCE OF INVOLVEMENT OF CHROMOSOME REGION-15Q12 AND REGION-12Q15 IN CONDITIONS WITH ASSOCIATED OVERGROWTH

MaryAnnLiebert, Inc., Publishers

Pp. 227-231

Cytogenetic

Evidence of Involvement of Chromosome

Regions

15ql2

and

_12ql5

in

Conditions

with

Associated

_Overgrowth

ANITA

WAJNTAL,1

DANILO

MORETTI-FERREIRA,2

DEISE HELENA DE

SOUZA,1

and

CELIA

PRISZKULNIK KOIFFMANN1

ABSTRACT

Syndromes

with associated

_overgrowth

are

poorly

understood. Besides their modeof

inheritance,

nothing

is

known

_regarding

the basic

_genetic

alterations that lead to their abnormal

phenotypic

manifestations. The

chromosome localization of the genes involved remainsunknown forthisgroup of

syndromes,

withthe

_only

exception

being

the Wiedemann-Beckwith

syndrome.

INTRODUCTION

An

increased propensity for tumor

_development

has

been described in mostof the

overgrowth

syndromes.

This

_propensity

has been well established in the Wiede-mann-Beckwith

_{syndrome (macrosomia, macroglossia,}

omphalocele,

ear

creases). Bannayan-Zonana

_syndrome

(neonatal

macrosomiawith

_{postnatal growth}

deceleration,

macrocephaly, hamartomas).

Ruvalcaba-Myhre-Smith

syndrome (macrosomia, macrocephaly, polyposis

of

colon,

pigmentary changes

of

_penis),

and _Proteus syn-drome

_{(macrosomia, hemihypertrophy, macrodactyly,}

hamartomas),

andalso has been

_suggested

by

Maldonado

etal

(1984)

and Cole and

_Hughes

(1990)

for the Sotos syn-drome

_{(macrosomia, macrocephaly,}

_poor coordination

during infancy).

This

_{tendency points}

toward the

possibil-ity

thatdifferent cellular

_growth

factors

_{might play}

an im-portant role in this_group of

_syndromes.

The localization of thegenesthatare

_responsible

for the

overgrowth syndromes

is a _very

_important

clue toward

understanding

the basic mechanisms that leadtothe clini-cal

manifestations;

consequently,

this_{group of}

_syndromes

might

throw some

_light

on a few of the various

mecha-nisms of

_{tumorigenesis.}

Chromosome aberrations have been avaluable tool for

disease_gene

_assignment.

Among

the

_syndromes

_leading

to

overgrowth, only

the Wiedemann-Beckwith

_syndrome

was

assigned

toa

_specific

chromosomal

_region;

rare

karyotypic

abnormalities,

suchas

deletions,

_{duplications,}

or

transloca-tions at

_llpll-pl5,

were the initial hints

leading

to the final localization at

llpl5.5.

Previously

we have

suggested

the association of two chromosome

_{regions (15ql2}

and

_12ql5)

with

overgrowth-associated

_{syndromes (Wajntal}

and

Koiffmann, 1991)

and

tumorigenesis (Wajntal

et

_{al., 1991); however,}

clinical data and

_cytogenetic

evidence remain

_unpublished.

We _present

the

_cytogenetic

dataonstructural chromosome aberrations

detected in three

_proposita

studied

by

use, twowith

over-growth

and malformations that are not

_typical

_{of any of} the

_already

well-defined

_syndromes,

and one with Sotos

syndrome

whose father is also affected and has the same

chromosome

_abnormality.

The clinical features ofthe pa-tients who remained

_undiagnosed

will also be described

and discussed in view of their relevance to

_cytogenetic

data.

MATERIAL AND METHODS

We studied 35

_patients

with

_overgrowth

and

_multiple

malformations: Sotos

_syndrome

was

_diagnosed

for 17

pa-tients;

Weaver

syndrome

for

2; Bannayan-Zonana

syn-drome,

Klippel-Trenaunay-Weber

syndrome,

and Proteus

syndrome,

one caseof

each;

13

_patients

remained without

a

syndromic

diagnosis

because,

as far as we

know,

their

phenotypic

characteristics didnotmatch_anyof thecriteria

previously

described. Patients with Wiedemann-Beckwith

syndrome,

endocrinologie

abnormalities,

orother

well-es-•Department ofBiology, Universityof Säo Paulo, USP Caixa Postal 11461, Sào Paulo, Brazil CEP: 05499.

'DepartmentofGenetics, Universidade Estadual Paulista,UNESP Caixa Postal 0529,Botucatu SP, Brazil CEP: 8610.

tablished conditions such as Marfan

_syndrome,

XYY,

XXY,

etc., wereexcluded fromthis

_study.

All

_patients

were

subjected

to

clinical,

_{endocrinologie,}

radiologie,

and

_cytogenetic

studies. Parentsand sibs were

always

examined and the

_{family history}

was recorded.

Cytogenetic

studies were

_performed

on

_lymphocyte

cul-tureswithGTG

_banding

_techniques.

Whenconsidered

nec-essary, other

banding techniques

werealso used.

RESULTS AND DISCUSSION

Among

the 35

_{patients studied,}

2 were _found to have

gross abnormal chromosome constitution. Both

patients

had somatic characteristics different from _{any of} the known

_{overgrowth syndromes.}

Patient 1

_{(Fig. 1)}

ARA,

a

_boy,

wasbornon December

_{3, 1984,}

to

_healthy

nonconsanguineous

parentsof33

_(father)

and30

_(mother)

yearsof age, after normalpregnancyand electivecaesarian

delivery.

Hewashismother'ssecond

delivery;

hisolder sis-teris

_healthy.

Birth

_weight

was

_3,780

_g

_(>97

_centile),

birth

length

50_cm, OFC 37 cm

_{(97 centile).}

APGARwas 5

₍₁

min)

and7

_{(2 min).}

He started

_{sitting by}

_{1 year,}

_{walking by}

2 years, and

_speaking

when 3 years of age; at _{age AVi} he

wasreferredtoa

genetic

_study

because of

_overgrowth

and

psychomotor

retardation. His

_physical

examination showed:

_height

112 cm

_{(>90%), weight}

26

_kg

_(>97%),

a

OFC 54 cm

_{(>98%), peculiar}

faciès with

high

forehead

and

_{temporal receding}

hair,

large posteriorized

ears with

antihelix

_{hypoplasia, hypertelorism (inner}

canthal distance 3.5_{cm; outer}canthal distance 8

cm),

bulbous_nose,

macro-stomia,

thin _upper

_lip,

micro- and

_{retrognathia,}

short broad

neck,

large

hands: middle

_fingers

5.5 cm

(>75

cen-tile), palms

8.5 cm

_{(>97 centile) clinodactyly}

ofthe 5th

fingers, Sydney-type palmar

crease, and bilateral

disloca-tion of triradius t to a t"

position.

He had fair skin and

blond hair

_differing

from_parents and sister.

Chromosome

_analysis

of the

_patient

disclosed the

fol-lowing

constitution:

45,XY,t(15;15) (qter—pll::ql2

or

ql3—qter)

with a _consequent deletion of

_15ql2

or

ql3

(Fig. 2).

The

father, mother,

and sisterare

karyotypically

normal.

Patient 2

(Fig. 3)

JAA,

a

_girl,

wasbornonNovember

26, 1988,

from

heal-thy

nonconsanguineous

parents. The fatherwas 22_years

and themother 18_yearsold.The

mother,

a

_primigesta

and

primípara

young woman, had a normal_{pregnancy except}

for one

_episode

of

_{bleeding during}

the fourth month of

gestation. Delivery

was

_by

caesarian section because ofthe

child's size. Birth

_weight

was

6,200

_g

_{(»97 centile),}

birth

length

was not

recorded,

and she had

cyanosis

and

neo-natal

_jaundice

andwastreated for4

_days

with

photother-apy. She was referred for

_genetic

evaluation because of

overgrowth,

dysmorphism,

and

_hypotonia

when 10

months old. On this occasion her

_physical

examination

showed:

_height

80cm

_{(»97 centile), weight}

9,750

_g

_(>75

centile),

OFC43 cm

(<50 centile), asymmetric

head and

face with increased size onthe

_{right; prominent}

forehead and

_metopic

suture,

_{large posteriorized}

earswith

preauric-ular

_pit

onthe

left,

bilateral

palpebral ptosis, epicanthus,

telecanthus,

blepharophimosis, high congenital myopia

and horizontal_nystagmus,

antimongoloid

slanting

of

pal-pebral

fissures,

blue

sclerae,

flat nose,

_highly

arched

pal-ate, short neck with a

_{hypochromatic}

_{spot under the chin}

region

on the

left,

umbilical

hernia,

small hands with

clinodactyly

of the fifth

_fingers

and

_{hyperflexibility}

of

fin-ger

joints, cyanosis

of

fingertips

and toes, halux of

in-creased

size,

echinovarus

_deformity

of the

feet,

global

hy-potonia,

and

_psychomotor

retardation: she started

sustain-ing

her headat8months andwasnotyet

sitting

atthetime

of examination. EEG and TAC

_findings

were normal.

Bone _age at 6 months was 9 months. Her

_paternal

aunt

hadachild that died after the first24hoursof life with

mi-tes

'

15

15

<

I

GTG

CBG

FIG. 2. Partial

_karyotypes

of

_patient

1

_showing

the transocation between chromosomes 15; the chromosome with del

15(ql2

or

_ql3)

isoriented

_upward.

GTG

_banding

technique

(4

cells)

and CBG

(1

cell).

Arrows indicate the

breakpoints

onthe schematic

representation.

crocephaly

and

_polydactyly

of hands and feet on which

further informationis notavailable.

Cytogenetic analysis

disclosedthechromosome constitu-tion

_{46,XX/46,XX,12p+,}

theextramaterial_present

_being

consistent with

_dup

12

_{(qll->ql5) (Fig. 4).}

Theabnormal constitutionwaspresent in21% of the cells

_analyzed.

Pa-rental

_karyotypes

were notavailable.

Among

the

_{overgrowth patients}

with chromosome

ab-normalities describedinthe literature thesame

_breakpoints

asfoundinour

_patients

1and2have been described intwo

patients

with Sotos

_syndrome:

_Koyama

et al.

(1985)

de-scribea

girl

with

47,XX,+inv

dup

(15)

(pter—ql2

or

_ql3;

ql2

or

ql3 pter)

and Tamaki et al.

₍₁₉₈₉₎

describe a

_girl

with

_{t(2,12) (q33.1;ql5).}

Incommonwith our

_patients

are

thechromosomal

breakpoints

in

15ql2

and

12ql5.

In addi-tion to

those,

there are twomore_reports onchromosome

alterations inSotos

_{syndrome involving}

other chromosome

regions:

the case described

_by

Nakada et al.

₍₁₉₈₂₎

with

46.XX

inv

8(p21.3;q22.1)

and the case described

_by

Schrander-Stumpel

et al.

₍₁₉₉₀₎

with

46,XY,t(3;6)

(P21;p21).

Becauseboth chromosome

_breakpoints

_15ql2

and

_12ql5

havebeen described

_previously

_amongSotos

_syndrome

pa-tients,

we reexamined those chromosomal

_regions

in all our Sotos

syndrome

patients.

We found that one ofour

patients (patient 3)

and his affected father haveadel

(15)

(ql2

or

ql3)

_(Fig.

5 and

Fig. 6).

The somatic features of this

_patient

and his father have been described

_previously

(Moretti-Ferreira

et

al,

1991).

Patient 1 with

_t(15;15)

and deleted

_{regions 15ql2 (or}

13)

—the

_{Prader-Willi/Angelman region}

—

presents

pheno-typic

features that resemble both

_syndromes:

_light

skin,

dup

\

12

GTG

FIG. 4. Partial

_karyotypes

from

_patient

2

_showing

dupli-cation

12(qll-ql5).

Normal

_homolog

on theleft. Arrows

indicate the

_breakpoints

on the schematic

_{representation.}

GTG

_{banding technique (three cells).}

If

15

GTG

i I

II

RBG

FIG.5. Partial

_karyotypes

of

_patient

3

_{demonstrating}

del

15

_(ql2

or

_ql3);

three cells with GTG

banding

technique

andtwocellswith RBG

_{banding technique,}

normal

homo-log

on the left. Arrows indicate the

_breakpoints

on

dia-gram.

hair,

and _eyes are observed in Prader-Willi

_patients

who

have visible

_cytogenetic

deletions of

15qll-13

(Butler,

1990);

a

tendency

to

_{obesity, though}

the fat distribution is

not

_typical

of the Prader-Willi

_syndrome;

macrostomia;

anda

_smiling

faceareobserved in

patients

with

_Angelman

syndrome.

Patient2_presentsachromosome aberrationinamosaic

form,

a

_postzygotic

_{event, and the}

_product

of the

dis-rupted

gene on chromosome

12,

possibly

a disturbed

growth

factor,

present even in _part ofthe

cells,

could be

responsible

for the

_global

overgrowth

observed in this

pa-tient.

The results ofour chromosome studies in

patients

with

overgrowth

syndromes point

toward the

hypothesis

thatin

the chromosomal

_regions

12ql5

and

_15ql2

there are loci

relatedtocellular

_growth

factors;

germinal

and/or somatic mutations in those

loci,

and

_probably

others not yet

de-tected,

mayleadto

_{overgrowth syndromes}

and

predisposi-tion to

_{tumorigenesis.}

Cytogenetic

studies of

_patients

with rare

_syndromes

are

important

forthe future

_{understanding}

and identification

ofthe molecular mechanisms involvedinthe

developmen-tal

_anomaly,

as

_{they identify}

the chromosomal

_regions

in-3p"

H

15

GTG

FIG. 6. Partial

_karyotypes

of

_patient

3's

father,

demon-strating

thesamedeletion_presentin his_son;twocells with

volved,

pointing

outnew routesfor further

_{investigations.}

This

_knowledge

will

_{certainly improve}

the

_diagnostic

and

prognostic procedures

and enhance the

_genetic

_counseling

offered the

_patients

and their families.

ACKNOWLEDGMENT

We thank Dr. P.A. Otto from the

Laboratory

of

Hu-man Genetics-IBUSP for

referring patient

1 _{to us,} and

MissLucelenidaSilva for secretarial assistance. This work

was

_{supported by}

funds from

_CNPq

and CAPES

(Brazil).

REFERENCES

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under-standingofcauseanddiagnosis.Am. J. Med. Genet. 35,

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COLE, T.R.P.,andHUGHES, H.E._(1990).Sotos_syndrome.J. Med. Genet. 27,571-576.

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HASE-GAWA, T. (1982). Acaseof cerebralgigantism with familial chromosomal aberration. Jpn. J. Human Genet. 27, 171-172.

SCHRANDER-STUMPEL, C.T.R.M., FRYNS, J.P., and

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TAMAKI, K„ HORIE, K., GO, T., OKUNO, T., MIKAWA, H., HUA, Z.Y., andABE,T. (1989). Sotossyndromewitha

balanced reciprocal translocation t(2;12) (q33.3;ql5). Ann. Genet. 32,244-246.

WAJNTAL, A., andKOIFFMANN, C.P. (1991). Letter to _the

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MORETTI-FERREIRA, D. _(1991). Chromosome

abnormali-tiesandtumorsinsyndromesleadingtoovergrowth.

Proceed-ingsof the 1991 MiamiBio/TechnologyWinterSymposium,p.

20.

Address

_reprint

_requests to:

Anita

_Wajntal

Department

of Biology

University of

SäoPaulo-USP Caixa Postal11461

Säo

Paulo,

BrazilCEP: 05499 ReceivedforpublicationDecember22, 1992;acceptedDecember