2. 1 Dr Pupak Derakhshandeh, PhD Ass Prof of Medical Science of Tehran University SMA SPINAL MUSCULAR ATROPHY

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4. 3 DELETION OF SMN AND NAIP GENES IN IRANIAN PATIENTS WITH SPINAL MUSCULAR ATROPHY

5. 4 SMA Gene: SMN (Survival Motor Neuron) Motor Neuron; Anterior Horn; Spinal Cord Second most common fatal autosomal recessive disorder after CF Second most common pediatric neuromuscular disorder after DMD Incidence : 1 in 6000-10000 live births Carrier frequency : 1 in 40-60

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7. 6 CLASSIFICATION SMA TYPE I (Werdnig-Hoffmann) SMA TYPE II (Classic) SMA TYPE III (Kugelberg-Welander)

8. 7 SMA TYPE I Severe form of SMA Onset : first 6 months Death : < 2 year Never raising the head or sitting

9. 8 SMA TYPE II Less sever Clinical appearing : < 18 months Able to sit unaid Death : about 9 years

10. 9 SMA TYPE III Mildest form of SMA Onset : > 18 months Walking without aid

11. 10 OTHER CLASSIFICATION

12. 11 DIAGNOSIS EMG Muscle Biopsy Genetic Testing/PND

13. 12 GENETICS 1990: The three types of SMA were mapped to 5q13 The SMA locus contain two inverted copies of a 500kb element The two copies named telomeric and centromeric

14. 13 GENETIC MAP Three candidate genes named SMN (Survival Motor Neuron), NAIP (Neuronal Apoptosis Inhibitory Protein) and P44 were identified in this locus Up to 95% of SMA patients (SMNI-III) are homozygously deleted for two exons (7&8) of both telomeric copy of SMN gene (SMN t )

15. 14 Deletions Up to 5% of SMA patients have frameshift mutations, gene conversions and point mutation Exons 5 and 6 of NAIP t gene are deleted in approximately 50% of type I SMA and 18% of types II and III SMA P44 t is lacked or intrrupted in 73% of SMA type I patients and 7% in types II and III

16. 15 The summery of normal alleles (N), mutant alleles (M) and deletion types (D) of SMN

17. 16 MOLECULAR DIAGNOSIS & PND PCR-SSCP or PCR-RFLP of SMN gene enables confirmation of a suspected clinical diagnosis of SMA or prenatal diagnosis These two techniques based on nucleotide differences of both exon 7 and exon 8 of telomeric and centromeric copy of SMN

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23. 22 Deletion Analysis of SMN gene Exones 7 and 8 of SMN gene were amplified and cut by Dra I and Dde I, respectively. (only centromeric copy is cutted) Absence of SMN t exone(s) 7 (and 8) confirm diagnosis of SMA

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25. 24 188 bp 164 bp 188 bp 123 bp 65 bp Exon 7, DraI Exon 8, DdeI

26. 25 SMN Deletion Analysis SMN t Exon 7 is deleted in affected child Derakhshandeh-Peykar, et al. Annal Acad Med, 2007 Derakhshandeh-Peykar, et al. Annal Acad Med, 2007 (Depart Med Gen & NRCGEB)

27. 26 NAIP Gene Deletion Analysis Exones 5 & 6 of NAIP gene were amplified with exon 13 which was the internal control Absence of exon 5 and exon 6 ( which only exist within the telomeric functional copy of NAIP) was detected in ~50% of type I SMA and 18% of types II and III SMA

28. 27 NAIP Deletion analysis Derakhshandeh-Peykar, et al. Annal Acad Med, 2007 Derakhshandeh-Peykar, et al. Annal Acad Med, 2007 (Depart Med Gen & NRCGEB)

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30. 29 The objective of this study was to genetically characterize the childhood onset spinal muscular atrophy in Iran. SMNNAIP Deletion of exon 7 & 8 Deletion of exon 5 & 6 SMA type I (n=70)70(100%)61(87%) SMA type II (n=3)2(66%)1(33%) SMA type III (n=2)1(50%)0(0%)

31. 30 Various deletion haplotypes were constructed by using genotypes of SMN and NAIP genes. Haplotype A, which has the deletions of all two involved genes, were deleted in approximately 83% of type I and II SMA but not in type III and was found predominantly in the severe group with an early onset at less than 6 month of age. we report Thirty four our experiences for prenatal diagnosis Haplotypes (%) ABC SMA type I (n=70) 871000 SMA type II (n=3) 336633 SMA type III (n=2) 050

32. 31 These studies suggested that the frequency of gene deletions of SMN1 and NAIP gene is a few higher than previous reports. It is may be due to high rate of consanguine marriage by Iranian Muslims (96 % in this families). Thus, the conformation of SMA related gene deletion will also be a useful tool for the pre and postnatal diagnostic. In addition to common PCR methods for SMN exon 7 and 8 and NAIP exons 4 and 5, we also conducted multiplex PCR of exon 5, 6 and 13 of the NAIP telomere in one reaction.

33. 32  Derakhshandeh  Esmaiili  Rahmani  Babrzadeh  Taeb  Attaran  Sajedifar  Farhud