Steps needed to move

forward in Italy.

A professional point of view

EPIDEMIOLOGY AND PREVENTION OF

HAEMOGLOBINOPATHIES

5th EUR PEAN

SYMP SIUM

N RARE

ANAEMIAS Ferrara - Castello Estense

15-16 Novembre 2013

The Medical Genetics Unit

Dr.ssa ANNA RAVANI

Laboratorio di Genetica Molecolare

Unità Operativa di Genetica Medica

Direttore Prof.ssa Alessandra Ferlini

Azienda Ospedaliero Universitaria S.Anna

FERRARA

MEDICAL GENETIC NETWORK IN EMILIA-ROMAGNA REGION

Clinical Genetics:

The Genetic Counselling for Hemoglobinopathies

(Dr.ssa Stefania Bigoni - Hemoglobinopathies Clinic)

YEAR POSTNATAL PRENATAL

2010 160 285

2011 224 418

2012 228 434

Impact of Genetic Counselling for Hemoglobinopathies in

Clinical Genetics activities

30%

30%

Genetic counselling: Origin of the patients

MEDICAL GENETIC NETWORK IN EMILIA-ROMAGNA REGION

Impact of diagnostics for Hemoglobinopathies in the

context of the Molecular Genetics Laboratory

Molecular analysis for

hemoglobinopathies

began in the Ferrara in

1985, with indirect test

(RFLP)

In 1989 started the

direct analysis of the

b globin gene mutations

Domenica Taruscio

Istituto Superiore di Sanità

Via Giano della Bella, 34

00161 - Roma (I)

Telefono: 06 4990 4018

Fax: 06 4990 4370

taruscio@iss.it

ISS : Test Genetici : Qualità : Il controllo esterno di qualità dell'ISS

National Evaluators for b-Thalassemia External Quality Scheme:

Prof. M.Cristina Rosatelli – Medical Sciences and Byotechnologies Departement - Cagliari

Dr. Anna Ravani – Molecular Genetics Laboratory U.O Medical Genetics – Ferrara Hospital

Haemoglobinopathies 2002 Traeger-Synodinos J., Old J.M., Petrou M. EMQN

Best Practice Guidelines for Molecular Genetic Testing

Haemoglobinopathies 2012 September EMQN Workshop in Leiden

METHODS

Beta globin gene: identification of point

mutation (bthalassemia and Hemoglobin

variants) and large rearrangements

Alpha globin genes:

identification of known deletions, of point

mutation (thalassemia and Hemoglobin

variant) and large rearrangements

Delta globin gene: differential diagnosis

upon suspicion of b or thalassemia

Gamma globin genes: identification of point

mutation (modifier gene in thalassemia

intermedia) and F Hemoglobin variants

WHICH TYPES OF DIAGNOSTICS CAN FERRARA LAB OFFER?

Complete Sequence of all the genes

(promoter-3’UTR)‏

Known Mutation Detection (RDB)‏

Deletion identification in the gene

and in the cluster

Rearrangements and del/dup

identifications in control regions

(LCR and HS) by MLPA

Molecular diagnoses of haemoglobinopathies

performed in Molecular Genetics Lab of Ferrara

‏(2012 – 1989)

Beta Thalassemia 5123

Alpha thalassemia 1001

Delta globin gene

variants 119

Gamma globin genes

variants 123

Year Total b thalassemia bTh Po Delta Area bTh Italy bTh Immigrants

1989 38 22 16

1990 107 40 67

1991 209 79 128 2

1992 297 61 236

1993 291 71 220 1

1994 187 52 135 2

1995 213 59 153 1

1996 207 72 133 2

1997 180 64 104 12

1998 140 62 67 11

1999 197 66 98 33

2000 138 55 69 14

2001 167 47 101 19

2002 179 57 93 29

2003 218 62 85 71

2004 158 45 62 51

2005 164 58 47 59

2006 354 63 168 123

2007 219 47 92 80

2008 314 44 130 140

2009 237 44 84 109

2010 198 40 66 92

2011 347 88 118 141

2012 361 72(20%) 101(28%) 188(52%)

Total 5123 1370(27%) 2573(50%) 1180(23%)

Beta Thalassaemia subjects characterization in 24 years of molecular analysis

Changes during the years of the origins of patiens

undergoing molecular diagnosis of Hemoglobinopathies

1989 1995 2000 2005 2009 2012 TOTAL

21%

36%43%

10 18 18

Southern Italy/Largest Islands Immigrants Po Delta Area

Year b th prenatal

diagnoses

Origins: Po Delta

Area

Origins: Southern Italy

and major Italian Islands

Immigrating

population

Affected foetuses

1989 6 2 4 - -

1990 29 11 18 - 10

1991 62 24 38 - 11

1992 114 24 90 - 30

1993 118 21 97 - 35

1994 109 24 85 - 26

1995 101 24 77 - 29

1996 146 24 122 - 35

1997 118 31 87 - 33

1998 106 40 64 2 18

1999 82 22 56 4 16

2000 78 24 52 2 22

2001 84 14 63 7 26

2002 97 15 78 4 24

2003 81 13 51 17 17

2004 82 16 51 15 11

2005 74 10 53 11 16

2006 76 13 53 10 27

2007 93 18 50 25 22

2008 88 10 54 24 26

2009 82 14 40 28 17

2010 89 23 34 32 13

2011 98 18 52 28 22

2012 117 25 42 50(43%) 35

TOTAL 2130 460 (21.6%) 1411 (66%) 259 (12% tot) 521 (24.5 %)

b

T

H

A

L

P

R

E

N

A

T

A

L

D

I

A

G

N

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S

E

S

A‏lot‏of‏molecular‏diagnoses…….:‏ This allows us to make some researches on that material.

Many peculiar cases have been reported in national and international conferences

Two recent examples:

We identified a new Hemoglobin variant in two sisters affered to the Pediatric

Department of our Hospital (Prof.ssa Borgna). The excellent cooperation

between the two hospital units has allowed the pubblication of the case.

1

HBB: c.406G>A p.Ala135Thr

We studied a AD family with high Hemoglobin A2 but normal hematological

parameters.

We identified a deletion involving a large part of G gene, never described

before, segregating in the family with the anomalous amount of HbA2 .

We are now describing the new finding.

2

……………..AND THE FUTURE?

NEW TECHNOLOGIES ARE READY: NGS FOR HEMOGLOBINOPATHIES TESTING?

The globin genes are small, probably may not require a high

throughput methodology, but a very interesting application could be

the contestual analysis of modifier genes, in order to interpretate

peculiar phenotypes and to performe more correct prognoses.

Hypotesis of analysis: Beta globin gene AND alpha globin genes,

gamma globin genes, EKLF, polymorphsms at BCL11A locus and HBSIL-

MYB‏intergenic‏region‏…and‏so‏on..

Genetic‏modifiers‏of‏β-thalassemia and clinical severity as assessed by age at first transfusion.

Danjou F, Anni F, Perseu L, Satta S, Dessì C, Lai ME, Fortina P, Devoto M, Galanello R. - Haematologica. 2012 Jul;97(7):989-93.

but it’s also so fun!!

All this means that our work is very hard and challenging,

I thank you all for your attention