• Save
Rusu Cristina  Rm B Rare Cluj
Upcoming SlideShare
Loading in...5
×
 

Rusu Cristina Rm B Rare Cluj

on

  • 1,400 views

 

Statistics

Views

Total Views
1,400
Views on SlideShare
1,399
Embed Views
1

Actions

Likes
0
Downloads
0
Comments
0

1 Embed 1

http://www.slideshare.net 1

Accessibility

Categories

Upload Details

Uploaded via as Microsoft PowerPoint

Usage Rights

© All Rights Reserved

Report content

Flagged as inappropriate Flag as inappropriate
Flag as inappropriate

Select your reason for flagging this presentation as inappropriate.

Cancel
  • Full Name Full Name Comment goes here.
    Are you sure you want to
    Your message goes here
    Processing…
Post Comment
Edit your comment

Rusu Cristina  Rm B Rare Cluj Rusu Cristina Rm B Rare Cluj Presentation Transcript

  • DIAGNOSIS AND MANAGEMENT OF PATIENTS WITH MENTAL RETARDATION AND MULTIPLE CONGENITAL ANOMALIES – IASI MEDICAL GENETICS CENTER’S EXPERIENCE Rusu Cristina (1), Volosciuc M. (1), Braha E. (1), Butnariu L. (1), Panzaru M. (1), Popescu R. (1), Caba L (1), Ivanov Iuliu (2), Gorduza Vlad (2), Covic Mircea (1) (1) Medical Genetics Centre, “Sf. Maria” Children’s Hospital, Iasi, Romania; (2) Immunology and Genetics Lab, “Sf. Spiridon” Hospital, Iasi, Romania
  • MENTAL RETARDATION – DEFINITIONS  Mental retardation (MR) = QI < 70;  Prevalence: 3%; Mild MR - 7-10 x more frequent than moderate/ severe MR;  Evaluation supposes gathering information from specialists (educator, psychologist, physician) + parents;  Classification: ◦ Mild (IQ 50-70) – 85%; ◦ Moderate (IQ 35-50) – 10%; ◦ Severe (IQ 20-35); ◦ Profound (IQ <20); 13/07/09 2
  • MENTAL RETARDATION – DETERMINISM  Variable according to MR severity;  Mild MR: o Environment – 75-90% (social factors !); o Genetic factors – 10-25%;  Severe MR: o Environment – 50%; o Genetic factors – 50% (monogenic/ multifactorial disorders/ chromosomal abnormalities). 13/07/09 3
  • MODERATE / SEVERE M.R. – GENETIC CAUSES  CHROMOSOMAL ABNORMALITIES: ◦ Numerical: trisomy 21, 13, 18, Klinefelter → karyotype; ◦ Structural:  Deletions/ duplications: 4p-, 5p- → karyotype;  Microdeleions/ duplications: Sdr Williams, Prader Willi, Angelman → FISH;  Subtelomeric rearrangements → MLPA;  MONOGENIC DISORDERS: ◦ X-linked mental retardation: Fragile X syndrome → DNA test (PCR, Southern blot); ◦ Metabolic disorders: Lesch Nyhan, Menkes → biochemical test. ◦ POLYGENIC CAUSES ◦ MITOCHONDRIAL DISORDERS 13/07/09 4
  • MATERIAL AND METHOD  We have designed, optimized and applied an investigation protocol for mentally retarded individuals; the protocol: • Covers most genetic causes of moderate/ severe MR; • Adapted to our lab (equipments/ cost of the reagents);  We have taken 200 mentally retarded patients examined in Iasi Medical Genetics Center and analyzed the efficiency of the protocol: • If De Vries score (proposed for subtelomeric rearrangements’ identification) could be used broadly for chromosomal abnormalities detection; • If the results provided by screening methods (antiFMRP test and MLPA) are concordant with diagnostic tests (long range PCR and FISH); • If the methods chosen are reliable;
  • PROTOCOL OPTIMIZATION  Use selection scores, so that the percent of positive tests will be good enough;  Choose the cheapest, less dangerous method that provides good results;  Use a sequence of reactions: first screening tests and then diagnostic tests, so that the abnormality will be identified precisely in the end;  Reducing the cost by reducing the amount of reagents used;
  • CLINICAL PROTOCOL  PERSONAL HISTORY (pre/intra/postnatal → exclude MR produced by the environment);  Extended FAMILY HISTORY → pedigree;  ANTHROPOMETRIC MEASUREMENTS (Ht, Wt, HC, others);  Detailed PHYSICAL EXAMINATION (+ photos);  PSYCHOLOGIC EVALUATION; EVALUATION  DE VRIES SCORE for case selection;  TEST the child with MR → anomalies identified → test the parents;  GENETIC COUNSELLING (before test, followed by written consent of the parents; new session when test results were available);  Data recorded in a DATABASE specially designed;
  • DE VRIES SCORE (2001): 3/> points necessary Criteria Score Mental retardation family history • Compatible with monogenic inheritance 1 • Incompatible with monogenic inheritance (including discordant phenotypes) 2 Prenatal onset growth retardation 2 Abnormal postnatal growth (1 point each, maximum 2 points) • Microcephaly 1 • Short stature 1 • Macrocephaly 1 • Tall stature 1 2/> facial anomalies (mostly hypertelorism, nasal and auricular defects) 2 Extrafacial anomalies (1 point each, maximum 2 points), especially: • Hand anomalies 1 • Heart defects 1 • Hypospadias +/- criptorchydism 1
  • INVESTIGATION PROTOCOL  KARYOTYPE (→ numerical & structural chromosomal abnormalities);  BARR TEST – for cases that associate abnormal sexual development;  Fragile X screening and diagnosis – done for cases with speech delay/ autism (early signs of Fragile X); positive & negative samples included; ◦ ANTI-FMRP TEST (immunohistochemical test done on hair root) → screening; ◦ Long range PCR (identifies premutations and complete mutations) → diagnostic;  MLPA (2 separate kits P036 and P070) → subtelomeric rearrangements (if the results in both kits are abnormal) and polymorphisms (if the result is abnormal in a single kit);  FISH → microdeletions / confirm subtelomeric rearrangements;  DNA stored for subsequent tests (after parental consent);
  • RESULTS  53% of patients (106) have been selected for genetic testing; the rest were due to social causes/ perinatal events/ fetopathies etc;  4 patients had specific monogenic disorders and a DNA sample has been taken for further confirmation;  15 patients had a Barr test → 3 cases confirmed;  99 patients had a karyotype → 43 cases identified; many of them were Down syndrome;  7 patients had a FISH test for microdeletion → 1 case confirmed;  55 patients had a MLPA test → 4 cases identified; FISH confirmation is in due course;  8 patients had FraX testing → all normal;  Structural chromosomal abnormalities identified by karyotype have been confirmed by MLPA.
  • TEST EFFICIENCY 100% 90% 80% 70% 60% 50% Abnormal Normal 40% 30% 20% 10% 0% Barr Kar FISH MLPA FraX
  • 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 01-001.2 SCNN1D 01-001.0 TNFRSF18 01-247.1 SH3BP5L 02-000.3 ACP1 02-000.3 ACP1 02-241.2 CAPN10 03-000.3 CHL1 03-000.3 CHL1 03-198.8 BDH1 04-000.5 GPI7 04-000.3 ZNF141 04-191.1 FRG1 05-000.3 PDCD6 05-000.3 LOC133957 05-180.6 GNB2L1 06-000.3 IRF4 06-000.3 IRF4 06-170.7 PSMB1 07-000.9 CENTA1 07-000.9 UNC84A 07-158.6 VIPR2 08-000.4 FBXO25 08-000.4 FBXO25 08-144.6 ZC3H3 09-000.9 DMRT1 09-000.4 DOCK8 09-139.8 EHMT1 10-000.5 DIP2C 10-000.2 ZMYND11 10-135.0 PAOX 11-000.2 RIC8A 11-000.2 BET1L 11-133.6 hCAP-D3 12-000.12 SLC6A12 12-000.3 JARID1A 12-132.2 ZNF10 Mapview 13-019.2 PSPC1 Mapview 13-019.2 PSPC1 13-112.8 F7 14-019.9 CCNB1IP1 14-019.9 PARP2 po36-88.D04_08121819RL^CEQsCSV.csv 14-105.0 MTA1 15-021.5 NDN 15-021.4 MKRN3 15-099.3 ALDH1A3 P070.88.G01_09032408H9^CEQsCSV.csv 16-000.4 DECR2 16-000.04 POLR3K 16-088.6 GAS8 17-000.1 RPH3AL 17-000.1 RPH3AL 17-078.4 TBCD 18-000.2 THOC1 18-000.2 USP14 18-075.9 C18orf22 19-000.2 PPAP2C 19-000.5 CDC34 19-063.8 CHMP2A 20-000.2 ZCCHC3 20-000.3 SOX12 20-062.2 OPRL1 21-014.7 STCH 21-014.5 RBM11 21-046.9 HRMT1L1 DUPLICATION 9p 22-016.0 IL17R 22-016.6 BID 22-049.6 RABL2B X-000.5 SHOX X/Y-000.5 SHOX (PAR region) X/Y-154.8 SYBL1 (PAR region)
  • Ratio 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 01-001.0 TNFRSF18 02-000.3 ACP1 03-000.3 CHL1 04-000.3 ZNF141 5-000.3 LOC133957 06-000.3 IRF4 07-000.9 UNC84A 08-000.4 FBXO25 09-000.4 DOCK8 10-000.2 ZMYND11 11-000.2 BET1L 12-000.3 JARID1A 13-019.2 PSPC1 14-019.9 PARP2 15-021.5 NDN P070.111.H01_09032408HA^CEQsCSV.csv 16-000.4 DECR2 17-000.1 RPH3AL 18-000.2 THOC1 19-000.2 PPAP2C 20-000.2 ZCCHC3 21-014.7 STCH 22-016.0 IL17R dup(21)(q22.3;q11.2) X-000.5 SHOX
  • CONCLUSIONS  De Vries score - useful for case selection (both chromosomal abnormalities and subtelomeric rearrangements);  MLPA - reliable, fast and unexpensive method that can be used as a screening method for subtelomeric rearrangements;  MLPA may be used as a complementary method to identify marker chromosomes/ complex chromosomal abnormalities;  The protocol could be further optimized by extending MLPA for other applications;  Cases with normal results/ polymorphisms - reevaluated in order to get a final diagnosis;  Cases identified - followed in order to prevent complications; clinical description of subtelomeric rearrangements is an ongoing process.
  • 13/07/09 18