Investigation of chromosomal abnormalities and microdeletions in 50 patients with multiple congenital anomalies

Investigation of chromosomal abnormalities and microdeletions in 50 patients with multiple congenital anomalies Akbar Mohammadzadeh MD, PhD candidate in Medical Genetics Genetics Research Center University of Social Welfare and Rehabilitation Sciences

Congenital Anomalies A congenital anomaly is defined as a structural defect, present at birth. Congenital anomalies may result in long-term disability, which may have significant impacts on individuals, families, health-care systems and societies.

Birth defects can be further divided into two categories: 1. Major anomalies a. May be lethal. b. Clearly disturb life of the individual. c. Require medical and surgical care. 2. Minor anomalies a. Never lethal. b. Problems may be mainly cosmetic and do not have medical significance. c. Some anomalies may mildly disturb life. d. These mild anomalies may, however, be a sign of more severe congenital anomalies or may be associated with a larger entity of multiple anomalies or a syndrome

Congenital Anomalies The other classifcation of congenital anomalies: I.Single congenital anomalies a. Multifactorial inheritance b. Correctable with surgery c. Good prognosis(commonly) II. Multiple congenital anomalies a. Two or more unrelated major structural malformations b. High mortality & morbidity rate c. Poor prognosis

Causes of Multiple Congenital Anomalies

Causes of 2.7 million neonatal deaths in 193 countries in 2010 Source: Adapted from WHO. Born too soon. The global action report on preterm birth. Geneva, World Health Organization, 2012

Incidence of Congenital Structural Abnormalities Incidence % according to birth record a year in Iran Major abnormality apparent at birth 2-3 32,000-48,000 MCAs in newborns (IRAN) 1 16,000 Mortality Rate <5years in IRAN 15.5/1000 24,800 Mortality Rate <5years in IRAN due to CAs(WHO, 2015) 18 4,464

Patients selection Inclusion Criteria Multiple congenital anomalies 2 Major Congenital Anomalies in 2 different organs. Parents should be alive. Parents should be unrelated((preferably ). Sporadic /single affected.

46,XY,der(4)t(4;12)(q33;q15)mat 50 Karyotypes 3 Abnormal 46,XY, der(18)[12]/46,xy,der(18),+mar[8]de novo 46,XY, del(3)(?p25p26) 47 Normal

Patient 1: A 16-month-year old boy Non-consanguineous parents A 24-year-old G1P1 mother at term CHD(PDA+ASD2 & heart murmur ) Severe DD Hypotonia Infantile spasms Microcephaly Hypertelorism Long face Wide & depressed nasal bridge frontal bossing Simian crease Epicanthal fold Low set & large & anteverted & posteriorly rotated ears Bilateral 2nd and 3rd toes syndactyly Imperforated anus (repaired) Rocker bottom & flat feet Curly hair High arched palate SNHL Brain MRI: Normal & EEG: Abnormal Brain Sono: Thin corpus callosum

Proband s karyotype result 46,XY,der(4)t(4;12)(q33;q15)mat

Mother s karyotype result 46,XX,t(4;12)(q33;q15)

Signs 12q dup 4q del Our patient Growth retardation + + + Intellectual disability + + + Microcephaly + + + Hypoplasia of corpus callosum + - + Hypotonia + + + Heart defects + + + Short neck + - + Hypertelorism + - + Epicanthal folds + - + Downslanting-palpebral fissures +? + Low-set ears + + + Micrognathia + + + Long philtrum + - + High-arched palate + + + Clinodactyly + + +

Signs 12q dup 4q del Our patient Nose prominent tip + + + Anteverted nares + + + Genitourinary malformtion + - - Simian creases + N/A + Skeletal anomalies + + + Foot deformity + + + Anal abnormality - - + Hearing loss - +(Conductive) +(SN) Infantile spasms +(rare) - + Recurrent rhinitis - + +

Patient 2: A 10-year-old boy Non-consanguineous parents A 24-year-old G1P1 mother at term Moderate to severe ID Microcephaly Autistic behavior seizure Microcephaly Polydactyly(4 limbs) Ptosis Arched eyebrows & Synophrys Hypertelorism Triangular face Low hairline frontal bossing Epicanthal folds Low set ears Retrognathia High arched palate Bilateral moderate SNHL Brain MRI: Normal & EEG: Abnormal

46,XY,del(3)(?p25p26)

46,XY,del(3)(?p25p26)

Signs 3p25 del Our patient Growth retardation + + Intellectual disability + + Microcephaly + + Hypotonia + + Hypertelorism - + Epicanthal folds + + Ptosis + + Downslanting-palpebral fissures + + Low-set ears + + Retrognathia + + Long philtrum + + High-arched palate + +

Signs 3P25 del Our patient Polydactyly + + GI abnormalities + - Seizures or abnormal EEG + + Hearing loss + + Arched eyebrows + + Synophrys + + Triangular face + + Low hairline + + frontal bossing + + Renal abnormalities + - Kyphosis or scoliosis + - Cardiovascular abnormalities + -

Patient3: A 27-month-old boy Consanguineous parents (half first cousins) A 27-year-old G1P1 mother at term CHD(PDA) NDD Facial dysmorphic features Hypotonia Aphagia Optic hypoplasia Recurrent Lower respiratory tract infections & Otitis media Brachydactyly Left ingunal hernia & UDT Umbilical hernia 2nd and 3rd toes syndactyly Rocker bottom feet 5th toe clinodactyly SGOT (AST) was 99 and SGPT (ALT) was 93 Thyroid function tests: normal Axial spiral CT scan of the brain without contrast: mild ventriculomegaly in frontal horns Brain MRI at age of 8 months: Extra axial hydrocephalus & brain atrophy metabolic screening: normal Immune deficiency

Karyotype Result 46,XY, der(18)[12]/46,xy,der(18),+mar[8], nuc ish(d18z1 3)[37]/(D18Z1 2)[63] de novo

nuc ish(d18z1 3)[37]/(D18Z1 2)[63] de novo FISH Result

nuc ish(d18z1 3)[37]/(D18Z1 2)[63] de novo FISH Result

nuc ish(d18z1 3)[37]/(D18Z1 2)[63] de novo FISH Result

Array- CGH Result

Array- CGH Result

Array- CGH Result

Array- CGH Result Gene content Start Cyto End Cyto Type Start End Size (bp) Gene content 18p11.32 18p11.21 LOSS 148,992 13,448,995 13,300,004 44 OMIM genes 18q21.31 18q23 GAIN 54,532,626 78,012,800 23,480,175 60 OMIM genes

Symptoms 18p del 18q dup 18q21.31-q23 dup Full trisomy 18 Our patient Growth retardation + + - ++ ++ Intellectual disability + + + Mild/moderate ++ ++ Microcephaly + +? ++ + Defects of CNS + -? + + Hypotonia + - - + + Heart defects + +? ++ +(PDA) Optic nerve hypoplasia + - - - + Strabismus + - +? - Hypertelorism + - - + + Epicanthal folds + - - ++ + Downslanting-palpebral fissures +? - + + Low-set ears + + - ++ + Micrognathia + + - ++ + Long philtrum - - - + + High-arched palate + + - ++ + Clinodactyly + + - ++ + IgA, IgG, or IgM deficiency + - -? +,+,+ Prominent occiput - +? ++ - Genital malformtion + - - ++ +

Sotos syndrome 5q35.3 deletion syndrome 47 Normal 17 MLPA Williams-Beuren syndrome DiGeorge syndrome 22q11.21 deletion syndrome Rubinstein-Taybi syndrome

SALSA MLPA P245 Microdeletion Syndromes-1 probemix 1p36 deletion syndrome, 2p16 microdeletion, 2q23 microdeletion/mbd5, 2q33 microdeletion/satb2, 3q29 microdeletion, 9q22.3 microdeletion, 15q24 deletion syndrome, 17q21 microdeletion, 22q13/Phelan-McDermid, Cri du Chat syndrome, 5p15, DiGeorge syndrome 22q11, Distal 22q11 region, DiGeorge region 2, 10p14, Langer-Giedion syndrome, 8q, Miller-Dieker syndrome, 17p, NF1 microdeletion syndrome, Prader-Willi / Angelman, MECP2 / Xq28 duplication, Rubinstein-Taybi syndrome, Smith-Magenis syndrome, Sotos syndrome 5q35.3, Wolf-Hirschhorn 4p16.3, Williams syndrome,

MLPA results of four patients with MCAs Phenotype MLPA result 5y, CHD+ NDD+ Autism+ Renal reflux+ Seizure+ Frontal bossing+ Sparse hair+ Vesicoureteral reflux+ Constipation 2y, CHD(supravalvular aortic stenosis)+ NDD+ ADHD+ Left visual defect+ Strabismus+ Hypotonia+ Without hypercalcemia Sotos syndrome 5q35.3 deletion syndrome Williams -Beuren syndrome 3y, CHD+ NDD+ Bilateral UDT+ Facial dysmorphic feature+ Simian crease+ low set & posteriorly rotated & malformed ears+ Fifth finger clinodactyly DiGeorge syndrome 22q11.21 deletion syndrome 7m, CHD+ NDD+ Bilateral cataract+ Hypotonia+ Micrognathia+ Low set ears+ Depressed & broad nasal bridge+ Long philtrum Rubinstein -Taybi syndrome MLPA kit: SALSA MLPA P245 Microdeletion Syndromes-1 probemix

Acknowledgement susan akbaroghli 2., Ehsan Aghaei-Moghadam 5., Nejat Mahdieh 6, Reza Shervin Badv 7, Peyman Jamali 8, Hossein Najmabadi 1,3, Yousef Shafaghati 4,1. Farkhondeh Behjati 1* 1. Genetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran. 2. Clinical Genetics Division, Mofid Children s Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran Pediatric Neurology Research Center, Mofid Children's Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran 3. Kariminejad-Najmabadi Pathology and Genetics Center, Tehran, Iran 4. Sarem Cell Research Center(SCRC), Sarem women s hospital, Tehran, Iran. 5. Children Medical Center, Tehran Medical University, Tehran, Iran 6. Cardiogenetic Research Laboratory, Rajaie Cardiovascular Medical and Research Center, Iran University of Medical Sciences 7. Children's Medical Center, Pediatrics Center of Excellence, Tehran University of Medical Sciences, Tehran, Iran 8. Shahroud Welfare Organization, Shahroud, Iran * Correspondent: Professor of Medical Genetics, Genetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran. AND patients families

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