Rev. Bras. Hematol. Hemoter. vol.39 número4

rev bras hematol hemoter. 2 0 1 7;39(4):385–387

w w w . r b h h . o r g

Revista

Brasileira

de

Hematologia

e

Hemoterapia

Brazilian

Journal

of

Hematology

and

Hemotherapy

Images

in

Clinical

Hematology

Molecular

genetic

techniques

for

gains

and

losses

of

genomic

material

in

a

case

of

acute

myeloid

leukemia

Mauren

Fernanda

Moller

dos

Santos

,

Camila

Clozato

Lara

,

Elvira

Deolinda

Rodrigues

Pereira

Velloso

a_{SociedadeBeneficenteIsraelitaBrasileiraAlbertEinstein(SBIBAE),SãoPaulo,SP,Brazil}

b_{FaculdadedeMedicinadaUniversidadedeSãoPaulo(FMUSP),SãoPaulo,SP,Brazil}

a

r

t

i

c

l

e

i

n

f

o

Articlehistory:

Received30January2017 Accepted26June2017 Availableonline20July2017

A 67-year-old male presented with a four-week history of weakness.Thecomplete blood countshowed: hemoglobin: 5.1g/dL, leukocytes 182.55×103/␮L (23% blasts and 34% monocytes) and platelets: 31×109/L. A bonemarrow aspi-rateshowed50.4% ofmyeloid myeloperoxidase(MPO)-blast cellsand 42.4%ofdysplasticgranulocytic-monocyticseries, with alpha-naphthyl acetate esterase positivity in 30% of totalnucleatedcells.Flowcytometryidentifiedtwodistinct aberrantblasts (CD4-CD7-CD11c-CD13-CD34-CD117-HLA-DR-cMPO+_{) and myeloid/monocytic}

(CD14-CD33-CD35-HLA-DR-CD11b+_{)populations.Karyotypingshowedmonosomy7and}

additional material in the long arm of chromosome 2

∗ _{Correspondingauthorat:Av.AlbertEinstein,627/701,05651-901SãoPaulo,SP,Brazil.}

E-mailaddress:mauren.santos@einstein.br(M.F.Santos).

(Figure1A).Acutemyeloidleukemia(AML)-M4(FAB classifica-tion)orAMLwithmyelodysplasia-relatedchanges(WHO2008 classification)wasdiagnosed.Patientdiedtwomonthsafter withoutresponsetotherapy.

Apart from karyotyping, other molecular genetic techniques can detect gains and losses of genomic material.1–3 _{In this case, the additional material in} chro-mosome 2 was elucidated and chromosome 7 monosomy was confirmed using fluorescence in situ hybridization, multiplex ligation-dependent probe amplification and single nucleotide polymorphism-array methodologies (Figures1B,2AandB).

http://dx.doi.org/10.1016/j.bjhh.2017.06.001

386

Figure1–(A)Karyotype(G-band):45,XY,add(2)(q35),-7[20].(B)FISH(Del(7q)DeletionProbe,ref:RU-LPH025;Cytocell, Cambridge,UK):DeletionofRELNgene(chromosome7)in96%oftheanalyzednuclei.Theabsenceofagreenandared signalmayindicatethemonosomyofthechromosome.

Figure2–(A)MLPAwithSALSAMLPAprobemixP144-A2(above)andP145-A2(below)kits(MRC-Holland,Amsterdam,The Netherlands).Dosagequotientofthepatients’probesinrelationtoacontrolgroup.Probespositionedbelowthe0.65limit indicatedeletion;probespositionedabovethe1.35limitindicateduplication.Probesinredindicatemonosomy7(deletion ofallprobesofchromosome7)andprobesinblackarenormalfortheotherchromosomesstudiedinthesekits.(B)SNP-A (CytoScanHDArray;Affymetrix,SantaClara,USA):DiagramsgeneratedbyChAS(Affymetrix)forchromosome2(above)and chromosome7(below).In“Copynumberstate”,linein2.00indicatesanormalcopynumber(diploid),linein1.00indicates adeletionandlinein3.00indicatesaduplication/gain.Lineswithintermediatevaluesbetween1.00and2.00;2.00and3.00 indicatemosaicism.In“AlleleDifference”,threelinesindicateanormalgenotype(AA,ABandBBalleles),twolinesindicate deletion(AandBalleles)orlossofheterozygosity(AAandBBalleles)andfourlinesindicateduplication(AAA,AAB,ABB andBBBalleles).SNP-Arevealedtheresultarr[hg19]2p25.3p16.2(1277054276429)x3[0.8],2q35q37.3

revbrashematolhemoter.2017;39(4):385–387

387

Conflicts

of

interest

Theauthorsdeclarenoconflictsofinterest.

r

e

f

e

r

e

n

c

e

s

1.DuttaaUR.Precisioninchromosomeidentificationwithleads inmolecularcytogenetics:anillustratedreview.JPediatr Genet.2014;3(1):1–7.

2.ShenY,WuBL.Designingasimplemultiplex

ligation-dependentprobeamplification(MLPA)assayforrapid detectionofcopynumbervariantsinthegenome.JGenet Genomics.2009;36(4):257–65.

3.TiuRV,GondekLP,O’KeefeCL,HuhJ,SekeresMA,ElsonP,etal. Newlesionsdetectedbysinglenucleotidepolymorphism array-basedchromosomalanalysishaveimportantclinical impactinacutemyeloidleukemia.JClinOncol.