Il ruolo del next generation sequencing nelle leucemie acute 12 aprile 2013 Bologna Ilaria Iacobucci Dipartimento di Medicina Specialistica, Diagnostica e Sperimentale, Università di Bologna, Bologna

Genetic characterization of BCR-ABL1 like ALL Sample IDRearrangementKey Lesions PAKTALSTRN3-JAK2IKZF1 del and p.Leu117fs PAKKCAEBF1-PDGFRBIKZF1 (IK6); EBF1 del; PAX5 inv; CDKN2A/CDKN2B del PAKVKKNUP214-ABL1IKZF1 p.Ser402fs; PAX5 del; CDKN2A/CDKN2B del e1-5 del; CDKN2A/CDKN2B del PAKYEBCR-JAK2IKZF1 (IK6); EBF1 del; PAX5 del and p.Gly24Arg; CDKN2A/CDKN2B del p.Ile682_Arg683insGlyPro; IKZF1 del e1-e6; EBF1 del; PAX5 p.Val319fs; CDKN2A/CDKN2B del (IK6); CDKN2A/CDKN2B del; FLT3 p.Asn609ins23aa p.Arg867Gln; CDKN2A/CDKN2B del PANNGLPAX5-JAK2IKZF1 del PANSFDETV6-ABL1IKZF1 (IK6); PAX5 del; CDKN2A/CDKN2B del PANEHFRCSD1-ABL1N/A SJBALL085NUP214-ABL1IKZF1 (IK6) and p.Ala79fs SJBALL010RANBP2-ABL1PAX5 del Novel rearrangements, copy number alterations, and sequence mutations constitutively activating kinase signaling and dysregulating cytokine receptor signaling. Roberts KG et al. Cancer Cell, 2012

Genetic characterization of hypodiploid ALL Near haploid Receptor tyrosine kinase signaling and Ras signaling (71%) Focal deletions of a histone gene cluster at 6p22 (19.1%) IKZF3 encoding AIOLOS (13%) Low hypodiploidy TP53 (91.2%) IKZF2 encoding HELIOS (53%) RB1 (41%) Nature Genetics 45, 242–252 (2013) Li- Fraumeni associated mutations

5 validated novel missense mutations in BCR-ABL1-positive ALL Genetic characterization of BCR-ABL1 positive ALL

Genetic characterization of T-ALL Next Generation Sequencing in T-ALL Whole genomeX chromosome targeted sequencing 12 male T-ALL cases 12 ETP ALL Inactivating mutations and deletions in the X- linked plant homeodomain finger 6 (PHF6) gene in 16% of pediatric and 38% of adult primary T-ALL sample. Vlierberghe P, Nat Genet Activating mutations in genes regulating cytokine receptor and RAS signalling (67% ); inactivating lesions disrupting haematopoietic development (58%); and histone-modifying genes (48%). Zhang G, et al. Nature 2012 Whole exome 67 T-ALL Mutations of CNOT3 7.9% adult T-ALLs; mutations affecting the ribosomal proteins RPL5 and RPL10 in 9.8% pediatric T-ALLs. De Keersmaecker K. Nature Genetics, 2013 Whole exome 5 T-ALL Mutations in NT5C2, which encodes a 5′- nucleotidase enzyme that is responsible for the inactivation of nucleoside-analog chemotherapy drugs. Tsoneva G. Nature Medicne, 2013

Minimal residual disease monitoring and NGS in ALL Deep-sequencing approach for minimal residual disease detection in acute lymphoblastic leukemia Faham M et al. Blood 2012;120: High-Throughput Sequencing Detects Minimal Residual Disease in Acute T Lymphoblastic Leukemia Wu D et al. Sci Transl Med 2012 Minimal residual disease monitoring and NGS in ALL