citogenetica, citogenomica citogenetica medica (anomalie
TRANSCRIPT
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TANDEM 2
GENETICA MEDICA
A.A.2019/2020
Alberto Turco
27 gennaio 2020
CITOGENETICA, CITOGENOMICA
CITOGENETICA MEDICA
(anomalie cromosomiche)
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Ecografia ostetrica: aumento dello spessore della plica nucale
(NT Nucal Translucency)
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S.di Edwards - Trisomia 18
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Candy Chromosomes……
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1956: IDENTIFICAZIONE DEL NUMERO CORRETTO DI CROMOSOMI
NELL’UOMO (46 non 48)
Un po’ di storia………….
PS Nel 1953 (tre anni prima) identificata la struttura
molecolare a doppia elica del DNA….
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Avvolgimento e condensazione cromatina
50:1
5000:1
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PREPARAZIONE CARIOTIPO STANDARD(Citogenetica convenzionale)
-Cc somatiche nucleate (es.linfociti sangue periferico, fibroblasti)
-Mitogeno (PHA fitoemoagglutinina)
-Colchicina (blocco in metafase - inibizione del fuso)
-Soluzione ipotonica
-Fissazione
-Colorazione (bandeggio)
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Regione
subtelomerica
Regione
subtelomerica
Anatomia di un cromosoma umano
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Cariotipo umano
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Cariotipo:esempi di colorazione G (Giemsa) a 550 bande
(Thompson & Thompson, 2016)
Esempio di referto di analisi citogenetica
standard per infertilità
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«Bandeggio
Cromosomico»
(standard)
Braccio
Regione
Banda
Sottobanda
Sotto-sottobanda
Es 12q24.32
p = petit
q = …..
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CARIOTIPO STANDARD UMANO(G-banding pattern)
CROMOSOMI ACROCENTRICI
Satelliti = geni in tandem per rRNA
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Morfologia dei cromosomi umani (dimensioni e posizione centromero)
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Hello everybody……Dove andiamo oggi? Australia?…un’amica, Nancy, abita a Sydney, in Watkins Street
….Nancy!
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= Citogenetica standard («tradizionale»)
= Citogenetica molecolare («citogenomica»,«Molecular Karyotyping»)
«Evoluzione» (progressi) della Citogenetica
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La Citogenetica Classica
(convenzionale, standard)
permette di identificare riarrangiamenti
cromosomici fino a circa 5-10 Mb
La Citogenetica Molecolare
(Citogenomica)
permette di identificare
riarrangiamenti fino a poche Kb
EVOLUZIONE DELLA CITOGENETICA
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CITOGENETICA MOLECOLARE
(Citogenomica, «chromonomics»)Livello di risoluzione< 4-5Mb (4-5 milioni di basi)
F.I.S.H.(Fluorescent In Situ Hybridization)
Array – CGH (aCGH)(array-based Comparative Genomic Hybridization)
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F.I.S.H.
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Diagnosi PGD di sesso «embrionale» per
gravi malattie X-linked (FISH multicolor)
X
Y
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S.da microdelezione: es. S.Wolf - Hirschhorn
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NB In alcuni casi la delezione nella s.di Wolf in 4p è
talmente estesa da essere visibile al cariotipo standard……
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Microarray CGH (vd oltre) (WHS = Wolf Hirnschorn Syndrome)
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ArrayCGH: aCGH)Screening dell’intero genoma
Cianine, sostanze
fluorescenti
Ibridazione Genomica Comparatica (CGH)
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…o CMA (Chromosomal MicroArray)
Dimensioni di un array
(DNA chip)
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Microchip di DNA (Microarray)
Permettono in tempi brevi e a bassi costi
l’analisi dell’intero genoma – Possono contenere fino
a 1 milione di «sonde» (brevi sequenze di DNA)
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aCGH: alcuni esempi applicativi
Identificazione di una microdelezione nel cromosoma 11
del(11)(q13.2-q13.4) mediante aCGH
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ANOMALIE CROMOSOMICHE(Chromosome Disorders)
- Abortività (reproductive wastage)
- Malformazioni congenite
- Ritardo mentale («ID, Intellectual Disability»)
- Le a.c. sono presenti in quasi l’1% dei neoanti (1/150-1/200)…
- ….e in >50% degli aborti spontanei del I trimestre
- La maggior parte sono diagnosticabili alla nascita
(Thompson & Thompson, 8th Ed., Elsevier, 2016)
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(Thompson & Thompson, 2016)
Incidenza delle anomalie cromosomiche alla nascita
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S.di Edwards (trisomia 18)
PGD-FISH interfasica
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Caso clinico….Cariotipo fetale da L.A.(liquido amniotico)
46,XY, t(2;5)(q3.4;q2.4)
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….cariotipo materno….
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«5p- syndrome» – Cri du chat
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