-Master’s Degree in Molecular and Cell Biology and Biomedical Sciences at University ofRome “Tor Vergata” – Final grade: 110/110 cum laude (14/06/2021)

 Thesis: “Deregolazione del pathway del TGF-Beta in pazienti affetti da sindrome mielodisplastica con mutazione del gene SF3B1”.

Supervisors: Prof. Bianca Maria Ciminelli – Prof. Maria Teresa Voso PhD Candidate in Immunology, Molecular Medicine and Applied Biotechnology at University of Rome “Tor Vergata” – Oncohematology laboratory (01/11/2021 – ongoing) Supervisors: Prof. Maria Teresa Voso, Tiziana Ottone, PhD

-PhD Candidate in Immunology, Molecular Medicine and Applied Biotechnology at University of Rome “Tor Vergata” – Oncohematology laboratory (01/11/2021 – ongoing)

Supervisors: Prof. Maria Teresa Voso, Tiziana Ottone, PhD

-I am currently doing my research at the Laboratory of Oncohematology (Department of Biomedicine and Prevention), directed by Prof. Maria Teresa Voso, University of Rome “Tor Vergata,” under the supervision of Tiziana Ottone, PhD. In agreement with the project, I delved into the molecular characterization of extramedullary acute myeloid leukemia (EM-AML) employing a high-throughput approach. In detail, I focused on genomic an transcriptomic analysis of patients who developed an extramedullary localization at the time of diagnosis or disease relapse, of whom we had paired samples of bone marrow and extramedullary site, mainly skin lesion. Notably, transcriptome analysis showed a significant difference in gene expression between the two sample types. This result provided the rationale for a functional enrichment analysis, which revealed an up-regulation of extracellular receptor-matrix interaction and focal adhesion pathways in extramedullary localization samples. These are signal transduction pathways known to be involved in the processes of tumorigenesis and metastasis, thereby promoting cancer progression. Subsequently, I validated by SYBR Green Real-Time PCR (qRT-PCR) the differential expression of some genes of interest belonging to these pathways, showing that there is a strong up-regulation in secondary localizations of genes such as collagen, fibronectin, integrins, laminins, which make up the extracellular matrix (ECM). Indeed, tumor progression depends on the ability of neoplastic cells to cross ECM and establish metastasis at sites distinct from the primary tumor mass, a process that in solid tumors is known as epithelial-mesenchymal transition (EMT), but in the case of hematologic malignancies is still under investigation. Among the genes on which I have focused my attention is TWIST1, one of the transcription factors involved in EMT. To assess its effective role in the process of carcinogenesis, I set up gene silencing by RNA interference and I treated in vitro the OCI-AML3 cell line with metformin, a drug capable of inhibiting this process. The study showed that TWIST1 is actually involved in leukemia cell migration and invasion, as confirmed by a functional transmigration assay. The results of the study led to the paper entitled “Expression profiling of extramedullary acute myeloid leukemia suggests involvement of epithelial-mesenchymal transition pathways” published on Leukemia, DOI: 10.1038/s41375-023-02054-0. Still remaining in the field of acute myeloid leukemias, I have another research project in progress focused on core binding factor leukemias, with particular reference to inv(16). The aim of this work was to deeply analyze the genomic and transcriptomic profile of this subset of AML in order to develop more effective therapies for all those patients who experience disease relapse after treatment. The evidence gathered allowed me to channel our findings into a paper that is currently under submission.

–Cell Biology

• Adherent cell cultures (HEK 293, SH-SYSY, MSC) and suspension cell cultures (leukemia cell lines)

• Isolation of mononuclear cells from peripheral blood and bone marrow by density gradient

• Magnetic beads cell isolation

• Cell viability assay (MTS)

• Trypan-Blue dye exclusion assay

• Transient transfection of HEK293 and SH-SYSY cells

• Gene silencing by RNA interference (RNAi)

• DNA-jetPRIME transfection reagent

•  In vitro cell treatments

• Preparation of slides with May-Grunwald Giemsa staining

• Electroporation

• Sonication

Molecular Biology

• Nucleic acid quantification (Nanodrop and Qubit)

• DNA and RNA extraction (Phenol-chloroform extraction, spin-column extraction, from TRIzol and automatic with Maxwell, from FFPE samples) and cryopreservation

• PCR, RT-PCR, Real-Time PCR, Q-

• Sanger sequencing and Next Generation Sequencing (NGS)

• Capillary electrophoresis/Fragment analysis

• SDS-PAGE electrophoresis/Agarose native gel electrophoresis

• Western Blot

• Gene cloning in competent cells

Systems Biology and Bioinformatics

• Excellent knowledge of Cytoscape, CellDesigner, DAVID, SIGNOR 0 (The SIGnaling Network Open Resource)

• Gene expression analysis

• Good knowledge of GraphPad Prism software

• Good knowledge of R software

In vivo models

 Authorized to use animals for experimental and scientific purposes (species: Mus musculus).