Dott.ssa Elisa Giorgio

f502-c005 Dottore in Scienze Biologiche ed Oncologia
Dottorato: 26° ciclo
Matricola: 743971

Tesi di dottorato

Molecular pathogenesis of adult -onset Autosomal Dominant LeukoDystrophy (ADLD): characterization of Lamin B1 duplications, and a position effect at the LMNB1 locus.

AIM 1: to perform the analysis and the systematic molecular characterization of 20 ADLD patients with LMNB1 duplication.

Autosomal dominant leukodystrophy (ADLD) is an adult onset demyelinating disorder that is caused by duplications of the lamin B1 (LMNB1) gene. However, as only a few cases have been analyzed in detail, the mechanisms underlying LMNB1 duplications are unclear. We studied a group of twenty ADLD families by high-resolution aCGH to map the LMNB1 duplication boundaries. Two cases allowed us to define the minimum critical duplicated region required for the development of ADLD which was ∼72 kb, and extended from ∼9.9 kb upstream of the 5_-UTR, and ∼1.8 kb downstream of the 3_-UTR of LMNB1 (Fig. 1A). LMNB1 is the only gene contained in this region and no other gene is even partially duplicated. This confirms that the duplication of LMNB1 alone is sufficient to cause ADLD. Clinical features of ADLD patients were similar in all patients in whomLMNB1 was the only gene completely duplicated, and we did not notice differences associated with different duplication extents. The only possible exception is patient BR1, in whom the initial symptoms were not autonomic dysfunction. The BR1 duplication was the only one that encompassed the complete coding regions of other genes. This large inverted duplication involved three genes centromeric to LMNB1, namely GRAMD3, ALDH7A1 (MIM #107323),PHAX (MIM #604924). This may suggest that the involvement of one of these genes may play a role as modifier of the disease phenotype or that the clinical spectrum of ADLD, particularly concerning the symptoms at onset, is wider than expected. Genotyping microsatellite markers around the LMNB1 gene in patients revealed two important characteristics regarding the ADLD duplications: (1) subjects with identical junctions shared the same haplotype, suggesting the presence of a common founder. It confirms that LMNB1 duplications are indeed non recurrent and that identical duplications in different families derived from the same mutational event; (2) the duplications were the result of intra chromosomal rearrangements. Fifteen of the 16 duplications (94%) had a “simple” head to tail tandem orientation as defined by the fact that there was only a single duplication junction. One of these also showed a triplication. The only “complex” duplication was found in patient BR1, and consisted of an inverted duplicated segment. The identification, for the first time, of a large number of LMNB1duplication junction sequences has allowed us to speculate on the mechanisms that may underlie these genomic rearrangements. We propose that the genomic architecture, including the enrichment of Alu repetitive elements and higher GC%, especially in the genomic region centromeric to the LMNB1 gene may play an important role in mediating the ADLD duplications. Given the overlapping signatures of the different duplication generating mechanisms it is difficult to identify unambiguously which of these is functioning in ADLD. It is also possible that there may be more than one mechanism at play. Our results suggest that NHEJ/MMEJ and replication-based mechanisms such as FoSTeS are likely to play an important role in the formation of the duplications that cause ADLD

In patients’ fibroblasts, the expression analysis confirmed an increase of LMNB1 both at mRNA and protein levels. We demonstrated that the duplicated and normal LMNB1 alleles in ADLD patients show equal expression, suggesting regulatory regions are maintained within the rearranged segment. Given the presence of three LMNB1 alleles, the theoretical increase of its expression is1.5-fold. We demonstrated that the differences between expected and observed values for LMNB1 expression are not due to the duplicated allele alone, because the three LMNB1 alleles were always equally expressed. This may suggest that in case of duplication, the LMNB1 mRNA/protein accumulates in patients’ cells. Such expression increments and the variability among patients were also found for mRNA and protein levels in nerve biopsies from patients with duplications of the PMP22 gene. Its origin was unknown, and it did not correlate with disease severity [Katona et al., 2009].

AIM 2: to identify the molecular mechanism causing vADLD in a large Italian family without LMNB1 duplication.

We described a large Italian ADLD family in which the disease was in linkage with a variant LMNB1 locus but neither duplication nor point mutations were found by genetic analysis of the LMNB1 gene. An increased expression of LMNB1 in patients’ blood, fibroblasts and brain suggested to further investigate lamin B1 as disease causing gene. By array-CGH, we identified a ~660 Kb deletion upstream of LMNB1 encompassing three genes: i) GRAMD3, whose function is unknown, has a Glucosyltransferases, Rab-like GTPase activators and Myotubularins domain (GRAM) also present in the myotubularin MTMR2, whose mutations are responsible of Charcot-Marie-Tooth type 4B; ii) PHAX, identified in mouse as a component of U snRNA export complex assembly; iii) ALDH7A1, an acetaldehyde-oxidizing protein, mutated in an autosomal recessive pyridoxine-dependent epilepsy. These were not considered to be candidate disease-causing genes, and in vitro silencing experiments confirmed that they did not affect lamin B1 expression.

We hypothesized that in our family ADLD was due to the misexpression of LMNB1, and aimed at identifying LMNB1 regulatory elements using circular chromosome conformation capture (4C). This technique and its modifications have been successfully applied to discover genomic interactions and to identify the presence of regulatory elements, as for a 10,379 bp copy number variant found to contain a CDKN2B gene regulator associated with pancreatic cancer susceptibility, and a 7.4 Kb deletion found to alter long-range cis-regulatory elements of the FOXL2 gene.

Using 4C followed by dual luciferase assay we confirmed two enhancer-containing regions upstream of LMNB1: enhancer region A (Enh-A), 120-kb upstream of the LMNB1 promoter, and Enh-B, 770-kb upstream Enh-A. 4C data were also validated by nested PCR on circularized DNA, which showed the interaction of LMNB1 promoter with Enh-A both in control and patient, and with Enh-B in patient only. Furthermore, exploiting a heterozygous SNP present in patient’s LMNB1 promoter, we proved a cis-allele specific interaction between Enh-B and the allele “C” segregating with the deletion.

ENCODE data show that both the Enh-A and Enh-B elements are in a DNase I hypersensitive site, suggesting an open chromatin structure. Furthermore, several binding sites for transcription factors are predicted for the Enh-B, among which YY1 and Sp1, both active during oligodendrocytes differentiation, and BCL11A and GATA3, involved in neuron morphogenesis and sympathetic neurons survival. Both these regions were not linked to histone modifications associated with enhancer activity (e.g., H3K4me1, H3K4me2, H3K27ac), suggesting that they may be peculiar regulatory elements. Multi-species alignment showed that Enh-A and Enh-B are not evolutionary conserved, in contrast with the assumed, but not mandatory, view that a high level of conservation between species is a criterion to identify functional non-coding DNA.

In the presence of the deletion, Enh-B is re-positioned at the same genomic distance from the lamin B1 promoter as Enh-A on the wild-type allele. This suggests that the deletion brings the enhancer activity of Enh-B closer to LMNB1 promoter in patients. Because Enh-B is stronger than Enh-A, we hypothesize this causes an increase in LMNB1 level as it happens when the whole gene is duplicated.

To determine if Enh-B behaves as an enhancer in vivo, we performed a transgenic mouse enhancer assay. We generated a total of 17 transgenic embryos which were stained for LacZ reporter gene activity on embryonic day (E) 11.5 (13). Each of these founder embryos is the result of an independent oocyte injection. Transgenic embryos can therefore be expected to carry the reporter construct stably integrated into their genome at different (random) positions. In four of the transgenic embryos, Enh-B showed enhancer activity in the developing forebrain, suggesting its role as tissue specific regulatory region.

Magnetic resonance imaging (and also one autoptic analysis) in ADLD-1-TO patients showed that forebrain is affected, but cerebellum (hindbrain) is spared, a difference with families carrying LMNB1 duplication, where the whole brain white matter degenerates. This findings are in agreement with a possible forebrain specific enhancer, as supported by mouse transgenic assays.

Lamin B1 was recently suggested to be a regulator of PLP1 myelin protein through interaction with the transcription factor YY1. We may hypothesise a feedback-positive loop of lamin B1 on Enh-B, containing an YY1 binding site, which may further increase LMNB1 expression in specific cell types (e.g., the central nervous system glia) and/or developmental stages.

The type of mutation hypothesized in this work is referred to as “position effect”, i.e., the alteration of gene expression due to a modification of its genomic environment instead of its coding sequence.

We also describe a rare example of "bystander effect". The deletion upstream the LMNB1 gene involved GRAMD3, ALDH7A1 and PHAX, three genes we proved not-associated with the pathology. "Bystander effect" has only been described once in the literature for the PAX6 locus, causing Aniridia; the ELP4 gene nearby was destructed by genomic translocations but it was not the direct cause of the pathology.

Our study has shed light on the complexities of LMNB1 regulation, and unraveled a novel pathogenetic mechanism leading to alteration of lamin B1 expression and ADLD, further emphasizing that genomic alterations that change the regulatory landscapes are also important determinants of Mendelian phenotypes. Regulation mechanisms and their associated mutations leading to Mendelian disorders are among the hot topics in molecular genetics, and impact the concept of mutation screening in human disease.

Attività di ricerca

PRESENTAZIONI ORALI A CONGRESSI:

- XV Congresso SIGU 2012. Sorrento 24 novembre 2012, sessione plenaria EREDITARIETA’ NON MENDELIANA, E.Giorgio: A 660 kb deletion centromeric to the lamin B1 gene (LMNB1) mimics LMNB1 duplication and causes adult-onset autosomal dominant leukodystrophy

- European Society of Human Genetics (ESHG) 2013. Parigi 8-11 giugno 2013. Concurrent Session C13: Neurodegenerative disordiers: from gene discovery to therapy. C13.5 E.Giorgio: A large genomic deletion upstream of the lamin B1 gene (LMNB1) likely causes adult-onset autosomal dominant leukodystrophy due to alteration of the regulatory landscape of LMNB1

- Giornate Neurologiche Torinesi 2014. Torino, 28 febbraio-01 marzo 2014. E.Giorgio: Una delezione a monte del gene LMNB1 causa la leucodistrofia autosomica dominante dell’adulto attraverso un effetto posizione

PREMI VINTI:

- SIGU AWARD 2012 per la presentazione orale “A 660 kb deletion centromeric to the lamin B1 gene (LMNB1) mimics LMNB1 duplication and causes adult-onset autosomal dominant leukodystrophy”- Sorrento 24 novembre 2012

- ESHG YOUNG INVESTIGATOR AWARD 2013 to Elisa Giorgio for a presentation judged outstanding at the European Human Genetics Conference 2013- Parigi 10 June 2013

PROGETTI DI RICERCA

1) SURF I e II - Sperimentazione sull’Utilità clinica della Ricerca Farmacogenetica: progetto pilota multicentrico all’Ospedale Galliera di Genova.

Finanziato da Comitato Scientifico, Ospedali Galliera.

a) Farmacogenetica del trattamento con fluoropirimidine: tossicità acuta legata a difetti di di-idropirimidina-deidrogenasi.

b) Farmacogenetica del trattamento con anticoagulanti orali (warfarin) in soggetti a rischio tromboembolico.

2) Health Technology Assessement per gli screenin genetici: lo studio dell'appropiatezza dei test genetici di suscettibilità alla malattia tromboembolica venosa come modello di studio

PRIN 2007 finanziato dal Ministero dell’Istruzione, dell’Università e della Ricerca

3) Fattori predittivi dell’insorgenza della malattia di Alzheimer in soggetti anziani a rischio: indagine su nuovi modelli neurobiologici e sul ruolo di marcatori biologici e genetici di rischio.

Finanziamento CARIGE 2008

4) AGED: Analysis of Genetic Effects in Dementia. Studio multidisciplinare integrato per l’identificazione di fattori predittivi di progressione da disturbo cognitivo lieve a malattia di Alzheimer.

Finanziamento CARIGE 2009

5) “Analisi dei risultati di 30 Anni di esperienza di Diagnosi Prenatale per le anomalie genetiche come base per lo sviluppo e validazioni di protocolli diagnostici innovativi.”

Finanziato dal Ministero della Salute

6) Clinical, neuroradiological and molecular investigtion of Adult-onset Autosomal Dominant LeukoDystrophy (ADLD): dissection of Lamin B1-mediated pathophysiological mechanisms in cellular and mouse models

Telethon-Application GGP10184

7) Lamin B1 dysregulation in Autosomal Dominant Leukodystrophy (ADLD): cellular and animal models to understand pathogenesis and move towards therapy.

Finanziato ELA Foundation 2012

ABSTRACTS

1)L’APLOTIPO H1 DEL GENE DELLA PROTEINA TAU (MAPT) È ASSOCIATO CON IL

DISTURBO COGNITIVO LIEVE, UNA CONDIZIONE PRODROMICA ALLA MALATTIA

DI ALZHEIMER (presentato al XII Congresso SIGU- Torino 08-10 novembre 2009)

E. Giorgio1, S. Cammarata2, M.I. Parodi3, R. Borghi4, L. Benussi5, M. Galli3, D. Galimberti6,

R. Ghidoni5, D. Gonella4, C. Novello2, V. Pollero2, L. Perroni3, P. Odetti4, E. Scarpini6, G. Binetti5,

M. Tabaton4, E. Di Maria1,7

2) UN CASO DI GRAVE REAZIONE AVVERSA AL 5-FLUOROURACILE ASSOCIATA

A POLIMORFISMI DEL GENE DPYD: IMPLICAZIONI PER LO SVILUPPO DI

PROTOCOLLI CLINICI BASATI SULLA FARMACOGENETICA (presentato al XII Congresso SIGU- Torino 08-10 novembre 2009).

M.I. Parodi1, C. Caroti2, G. Cassola3, E. Giorgio4, F. Faravelli5, L. Perroni1, E. Di Maria4,5

3)ESTIMATION OF WARFARIN DOSE DURING INITIAL ANTICOAGULATION: VALIDITY OF PREDICTION WITH PHARMACOGENETIC DATA

PathophysiolHaemostasisThrombosis2010-Abstract

Innocenti L [1], Schenone A [2], Giorgio E [3], Parodi MI [3], Uliana V [4], Perroni L [3], Mori M [1], Lo Pinto G [2], Di Maria E [4,5]*

4) Analisi di varianti proprie del precursore del Nerve Growth Factor (proNGF) in pazienti affetti da malattia di Alzheimer. (XIII Congresso SIGU 2010- Firenze 14-17 ottobre2010); in pubblicazione.

E. Giorgio1, C. Bonvicini2, F. Faravelli3, F. Forzano3, V. Uliana3, M.I. Parodi1, L. Perroni1, C. Bonomini4,

O. Zanetti4, M. Gennarelli2,5, E. Di Maria3,6

5) VALIDITA' DEI DATI FARMACOGENETICI NEL PREDIRE IL DOSAGGIO INIZIALE DEL WARFARIN: STUDIO PILOTA IN UN SERVIZIO OSPEDALIERO.

(XIII Congresso SIGU 2010- Firenze 14-17 ottobre2010); in pubblicazione.

Uliana V [1], Innocenti L [2], Schenone A [3], Giorgio E [4], Parodi MI [4], Forzano F [1], Perroni L [3], Mori M [2], Lo Pinto G [3], Faravelli F [1], Di Maria E [1,5]*

6) UTILITÀ CLINICA DELLA RICERCA FARMACOGENETICA: RISULTATI DI UN PROGETTO PILOTA IN UNA REALTA’ OSPEDALIERA. (XIII Congresso SIGU 2010- Firenze 14-17 ottobre2010); in pubblicazione.

E. Di Maria [1,2], E. Giorgio [3], V. Uliana [1,2], F. Forzano [1], F. Faravelli [1], N. Sacchi [4], Sperimentatori SURF

7) SINDROMI LERI-WEILL/LANGER E MUTAZIONI NELLA “REGIONE SHOX” (XIII Congresso SIGU 2010- Firenze 14-17 ottobre2010)

Baffico M.1, Capone L.2, Baldi M.1; Bertorelli R.2, Iughetti L.3 Madeo S.3, Forzano F.4, Giorgio E.1, Vannelli S.5, Stasiowska B.5, Coviello D.A.1, Forabosco A.

8)Clinical, neuroradiologicaland molecular investigation of Adult-onset Autosomal Dominant Leukodystrophy (ADLD): dissection of Lamin B1-mediated pathophysiological mechanisms in cellular and mouse models. (XVI Telethon scientific convention -RIVA DEL GARDA_7-9 marzo 2011)

Pietro Cortelli2, Alfredo Brusco4, A. Brussino4, Sabina Capellari2, Eleonora Di Gregorio4, Daniela Lacerenza4, Elisa Giorgio4, Piero Parchi2, Rocco Liguori2, Raffele Lodi3, Giovanna Vaula5, Andrea Contestabile1, Denise Ferrera1, Claudio Canale1, Laura Gasparini1

9)Characterization of Lamin B1 duplication breakpoints and expression analysis in ADLD patients ( ICHG/ASHG 2011 Meeting Abstract_11-15 ottobre 2011)

Alessandro Brussino1, Eleonora Di Gregorio1, Elisa Giorgio1, Daniela Lacerenza1, Flavia Talarico, Giovanna Vaula, Paola Mandich, Camilo Toro, Eleonore EYMARD Pierre(a,b), Pierre Labauge (c), Sabina Capellari, Pietro Cortelli, Filippo Pinto Vairo, William Gahl, Odile Boespflug-Tanguy(b,d,e), Alfredo Brusco

10) LEUCODISTROFIA AUTOSOMICA DOMINANTE DELL’ADULTO (ADLD) ASSOCIATA A DUPLICAZIONE DEL GENE LMNB1: IDENTIFICAZIONE DEI BREAKPOINT E STUDIO DELL’ESPRESSIONE IN OTTO FAMIGLIE (XiV Congresso SIGU 2011- MILANO 13-16 NOVEMBRE 2011)

B. Alessandro1, E. Giorgio1, E. Di Gregorio1, D. Lacerenza1, F. Talarico1, G. Vaula2, P. Mandich3, S. Capellari5, P.

Cortelli5, O. Boespflug-Tanguy6, W. Gahl4, A. Brusco1

11) AUTOSOMAL DOMINANT LEUKODYSTROPHY: GENETIC CHARACTERIZATION OF EIGHT FAMILIES WITH HOMOGENEOUS PHENOTYPE (XLII Congresso NAZIONALE SOCIETA’ ITALIANA DI NEUROLOGIA, TORINO 22-25 OTTOBRE 2011)

G. VAULA, A. BRUSSINO, P. CORTELLI, E. DI GREGORIO, P. MANDICH, E. GIORGIO,

D. LACERENZA, S. CAPELLARI, F. TALARICO, W. GAHL, O. BOESPFLUG- TANGUY, E.

PIERRE, C. TORO, F. PINTO VAIRO, A. BRUSCO

12) A 660 kb deletion centromeric to the lamin B1 gene (LMNB1) mimics LMNB1 duplication and causes adult-onset autosomal dominant leukodystrophy (62° ASHG 2012- SAN FRANCISCO 6-10 novembre)

Giorgio E 1, Robyr D 2, Di Gregorio E 1,3, Lacerenza D 1, Vaula G 4, Brusco A 1,3, Antonarakis SE 2, Brussino A

13) Una delezione di 660 Kb a monte del gene lamina B1 (LMNB1) ha un effetto analogo alla sua duplicazione genica e causa leucodistrofia autosomica dominante dell’adulto (ADLD) (15° congreSSO SIGU 2012, SORRENTO 21-24 NOVEMBRE 2012)

E. Giorgio1, D. Robyr2, E. Di Gregorio 1, D. Lacerenza3, G. Vaula4, A. Brusco1, S. Antonarakis2, A. Brussino1

14) Clinical, neuroradiological and molecular investigation of Adult-onset Autosomal Dominant Leukodystrophy (ADLD): dissection of Lamin B1-mediated pathophysiological mechanisms in cellular and mouse models (XVII CONVENTION TELETHON RIVA DEL GARDA 11-13 MARZO 2013)

Denise Ferrera1, Caterina Giacomini1, Claudio Canale2, Elisa Giorgio5, A. Brussino5, Sabina Capellari4, Roberto Marotta3, Eleonora Di Gregorio5, Giovanna Vaula7, Robyr D.8, Antonarakis S.E.8, Mandich P.9, Alfredo Brusco5,6, Pietro Cortelli4, Laura Gasparini1

15) A large genomic deletion upstream of the lamin B1 gene (LMNB1) likely causes adult-onset autosomal dominant leukodystrophy due to

alteration of the regulatory landscape of LMNB1.(ESHG 2013- PARIGI 8-11 GIUGNO 2013)

E. Giorgio, D. Robyr, E. Di Gregorio, D. Lacerenza, G. Vaula, D. Imperiale, C. Atzori, A. Brusco, S. Antonarakis, A. Brussino

16) A novel gene for Spinocerebellar Ataxia (SCA) linked to chromosome 6 and fatty acid metabolism (SPATAX 2013, PARIGI 11-13 GIUGNO)

Eleonora Di Gregorio1,2, Barbara Borroni3, Elisa Giorgio1, Daniela Lacerenza1, Cecilia Mancini1, Alessandro Calcia1, Isabella Mura4, Domenico Coviello4, Nico Mitro5, Marion Gaussen6, Nicola Lo Buono1, Ada Funaro1, Giovanna Vaula7, Isabelle Lagroua6, Laura Orsi7, Alexandra Durr6,8, Chiara Costanzi3, Alessandro Padovani3, Alexis Brice6,8, Loredana Boccone9, Eriola Hoxha10, Filippo Tempia10, Donatella Caruso5, Giovanni Stevanin6,8, Alfredo Brusco1,2

17) A novel gene for Spinocerebellar Ataxia (SCA) linked to chromosome 6 and involved in fatty acid metabolism (16° congresso sigu, roma 25-28 settembre 2013)

18) Exome sequencing reveals an atypical case of SCAR1 with myoclonic movements at onset. (16° congresso sigu, roma 25-28 settembre 2013)

Cecilia Mancini1, Laura Orsi2, Simona Cavalieri1,3, Eleonora Di Gregorio1,3, Elisa Giorgio1, Alessandro Calcia1, Daniela Lacerenza1, Elisa Pozzi1, Thomas Langer4,5, Quasar Saleem Padiath6, Alfredo Brusco1,3

19) A novel gene for Spinocerebellar Ataxia (SCA) linked to chromosome 6 and involved in fatty acid metabolism (ASHG 2013, BOSTON 22-26 OTTOBRE)

20) Lamin B1 dysregulation in autosomal dominant leukodystrophy (ADLD): genetics and biological mechanisms (SINS 2013, Roma 3-5 OTTOBRE)

Giorgio E, Ferrero M, , Boda E, Eleonora Di Gregorio,Vaula G, Mandich P,Gahl W,Pinto Vairo F, Boespflug-Tanguy O, Cortelli P, Brussino A, Buffo A, Brusco A

21) Lamin B1 expression is affected by EBV infection in lymphoblasts of patients with Autosomal Dominant Leukodystrophy through mir-23 deregulation (ESHG 2014, MILANO 31 maggio-3 giugno 2014)

Elisa Giorgio1, Livio Favaro 2, Nicola Lo Buono1, Cecilia Mancini1, Giovanna Vaula 3, Pietro Cortelli 4, Sabina Capellari 4 ,Paola Mandich 5, Niklas Dahl 6, Atle Melberg 7, Elisa Pozzi1, Eleonora Di Gregorio8, Simona Cavalieri8, Pierre Labauge 9, Eleonore Eymard Pierre 9, Harshvardhan Rolyan 10, Odile Boespflug-Tanguy 9,11-13, Laura Gasparini 14, Quasar Saleem Padiath 10, Alessandro Brussino 1 and Alfredo Brusco 1,8

22) TWO NOVEL MISSENSE COL4A1 MUTATIONS AND GENETICS HETEROGENEITY IN PORENCEPHALY (ESHG 2014, MILANO 31 maggio-3 giugno 2014).

Giovanna Vaula1, Elisa Giorgio2, Giovanni Bosco1, Martina Conterno1, Davide Quartana1, Alessandro Calcia2, Cecilia Mancini2, Eleonora Di Gregorio3, Simona Cavalieri2, Elisa Pozzi2, Alessandro Brussino2, Alfredo Brusco2,3.

23) AN ATYPICAL FORM OF PROGRESSIVE EXTREME HETEROTOPIC CALCIFICATION IN A PATIENT WITH A DE NOVO INSERTIONAL TRANSLOCATION DER(X)INS(X;2)(Q26.1;P13.3) (ESHG 2014, MILANO 31 maggio-3 giugno 2014).

Elisabetta Flex1; Elisa Giorgio2; Margherita Silengo3; Claudio Defilippi4; Elga Belligni3; Elisa Biamino3; Laura Bernardini5, Eleonora Di Gregorio6; Cecilia Bracco2; Giovanni Battista Ferrero3; Raoul C. Hennekam7; Marco Tartaglia1*; Alfredo Brusco2*.

24) LARGE CRYPTIC GENOMIC REARRANGEMENTS WITH APPARENTLY NORMAL KARYOTYPES DETECTED BY ARRAY-CGH (ESHG 2014, MILANO 31 maggio-3 giugno 2014).

Elisa Savin1, Eleonora Di Gregorio 1,2, Franco Fiocchi 1, Valeria Giorgia Naretto 1, Elisa Biamino 3, Elga Belligni 3, Cecilia Mancini 2, Simona Cavalieri2, Elisa Pozzi2, Elisa Giorgio 2, Eva Colombo 1, Flavia Talarico 1, Patrizia Pappi 1, Enrico Grosso 1, Margherita Cirillo Silengo 3, Giovanni Battista Ferrero 3, Alfredo Brusco 1,2

25) Exome sequencing reveals a new CLN5 mutation in an adult form of cerebellar ataxia (ESHG 2014, MILANO 31 maggio-3 giugno 2014).

C. Mancini1, S. Nassani2, Y. Guo3, E. Giorgio1, A. Calcia1, X. Liu4, E. Di Gregorio5, S. Cavalieri5, E. Pozzi1, A. Brussino1, Y. Xie4, F. Wang3, L. Tian3, W. Chen4, B. Nmezi6, Q. S. Padiath6, H. Jiang4,7, A. Kyttala8, N. R. Pizio2, H. Hakonarson3,9,10, A. Brusco1,5 .

26) CHALLENGES IN MEDICAL GENETICS: EXOME SEQUENCING UNCOVERS RECESSIVE MUTATIONS IN TWO CASES WITH DE NOVO CNV. (SIGU 2014, BOLOGNA 30-31 ottobre 2014).

Elisa Giorgio1, Viviana Caputo2, Andrea Ciolfi3,4, Eleonora Di Gregorio5, Alessandro Calcia1, Cecilia Mancini1, Simona Cavalieri5, Elga Belligni6, Elisa Biamino6, Cristina Molinatto6, Margherita Cirillo6, Giovanni Battista Ferrero6, Marco Tartaglia3, Alfredo Brusco1,5.

PAPERS

1) Severe fluoropyrimidine-related toxicity: clinical implications of DPYD analysis and UH2/U ratio evaluation.

E. Giorgio¹ , M. I. Parodi¹, C. Caroti², Mauro D’Amico², G. Cassola³, F. Faravelli⁴, F. Forzano, V. Uliana⁴,F. Mattioli⁵, C. Fucile⁵, V. Marini⁵, A. Martelli⁵, E. Di Maria^4,5 Cancer Chemother Pharmacol. 2011 Aug 11

2) Possible Influence of a Non-Synonymous Polymorphism Located in the NGF Precursor on Susceptibility to Late-Onset Alzheimer's Disease and Mild Cognitive Impairment.

Di Maria E, Giorgio E, Uliana V, Bonvicini C, Faravelli F, Cammarata S, Novello MC, Galimberti D, Scarpini E, Zanetti O, Gennarelli M, Tabaton M. J Alzheimers Dis. 2012 Jan 1;29(3):699-705.

3) health technology assessment of genetic testing for susceptibility to venous thromboembolism in italy-Chapter 2.2: Provision of genetic testing for inherited thrombophilia in Italy.

Elisa Giorgio, Vera Uliana, The Project Unit Investigators, Emilio Di Maria IJPH 2012, Vol9, Number 2, Supp1

4) Analysis of LMNB1 Duplications in Autosomal Dominant Leukodystrophy Provides Insights into Duplication Mechanisms and Allele-Specific Expression

Giorgio E, Rolyan H, Kropp L, Chakka AB, Yatsenko S, Gregorio ED, Lacerenza D, Vaula G, Talarico F, Mandich P, Toro C, Pierre EE, Labauge P, Capellari S, Cortelli P, Vairo FP, Miguel D, Stubbolo D, Marques LC, Gahl W, Boespflug-Tanguy O, Melberg A, Hassin-Baer S, Cohen OS, Pjontek R, Grau A, Klopstock T, Fogel B, Meijer I, Rouleau G, Bouchard JP, Ganapathiraju M, Vanderver A, Dahl N, Hobson G, Brusco A, Brussino A, Padiath QS.

Hum Mutat. 2013 Aug;34(8):1160-71

5) ELOVL5 Mutations Cause Spinocerebellar Ataxia 38.

Di Gregorio E, Borroni B, Giorgio E, Lacerenza D, Ferrero M, Lo Buono N, Ragusa N, Mancini C, Gaussen M, Calcia A, Mitro N, Hoxha E, Mura I, Coviello DA, Moon YA, Tesson C, Vaula G, Couarch P, Orsi L, Duregon E, Papotti MG, Deleuze JF, Imbert J, Costanzi C, Padovani A, Giunti P, Maillet-Vioud M, Durr A, Brice A, Tempia F, Funaro A, Boccone L, Caruso D, Stevanin G, Brusco A. Am J Hum Genet. 2014 Aug 7;95(2):209-17

6) Novel mutation of SLC20A2 in an Italian patient presenting with migraine.

Rubino E, Giorgio E, Gallone S, Pinessi L, Orsi L, Gentile S, Duca S, Brusco A.

J Neurol. 2014 Oct;261(10):2019-21. doi: 10.1007/s00415-014-7475-8.

7) Adult-onset autosomal recessive ataxia associated with neuronal ceroid lipofuscinosis type 5 gene (CLN5) mutations.

Mancini C, Nassani S, Guo Y, Chen Y, Giorgio E, Brussino A, Di Gregorio E, Cavalieri S, Lo Buono N, Funaro A, Pizio NR, Nmezi B, Kyttala A, Santorelli FM, Padiath QS, Hakonarson H, Zhang H, Brusco A. J Neurol. 2014 Oct 31.