I graduated in Biological Science in Italy with an experimental thesis at the Institute of Molecular Biology and Pathology of the Italian National Research Council (CNR) joining the research group of Dr. Degrassi Francesca. During my Bachelor’s degree, I explored the therapeutic potential of the kinetochore (KT) protein Hec1 (Highly Expressed in Cancer protein 1) as a molecular target to produce massive chromosome missegregation and cell death in cancer cells. Hec1 mediates KT–microtubule (MT) attachments at mitosis, as constituent of the Ndc80 complex, and is upregulated in various cancer types. I expressed Hec1 fused with an enhanced green fluorescent protein (EGFP) at its N-terminus MT-interaction domain in HeLa cells and showing that expression of this modified Hec1, localized at KTs, blocks cell proliferation and promotes apoptosis in tumor cells. 

Through western blot analysis, for apoptotic markers, and Immunofluorescence analysis, for studying microtubules-KT attachment dynamics, I have demonstrated that the overexpression of GFP-Hec1 is extremely potent in tumor cell killing and more efficient than siRNA-induced Hec1 depletion. Strikingly, normal cells showed no apparent cell proliferation defects or cell death following EGFP-Hec1 expression, while Live-cell imaging highlighted in cancer cells a massive chromosome missegregation, due to multipolar spindles, leading to cancer cell death after a prolonged mitotic arrest. I showed that EGFP-Hec1 over-expression is able to increase KT–MT attachment stability, providing a molecular explanation for the abnormal spindle architecture and the cytotoxic activity of this modified protein.

Within this project, I also had the opportunity to have my in-vitro observation translated in-vivo thanks to a collaboration with the Experimental Chemotherapy Laboratory of the Regina Elena National Cancer Institute in Rome, Italy. Consistent with my cell culture data, EGFP-Hec1 expression has been found to strongly inhibit tumor growth in a mouse xenograft model by disrupting mitosis and inducing multipolar spindles. The results of these experiments have been collected in a paper published in 2015 (Orticello et  al. N-Terminus  modified  Hec1  suppresses  tumour  growth  by  interfering  with  kinetochore-microtubule  dynamics.  Oncogene,  34:  3325-3335). 

For my Master degree in Genetics and Molecular Biology, I joined the Neuroepigenetic Lab of Andrea Fuso Ph.D. in the Department of Experimental Medicine at “La Sapienza” University in Rome, Italy. I have been working on the epigenetic alterations in the Alzheimer’s disease using mouse models and in particular, I have focused the investigation on the methylation pattern of some genes that are very well-known in this neurodegenerative disease. My Master thesis entitled "Vitamin K derivatives modulate genes involved in neuro-inflammation and neurodegeneration in SK-N-BE human neuroblastoma cells", investigates the potential role of a specific formulation of reduced vitamin K (MK7R) in SK-N-BE cells showing a remarkable modulation in the expression of specific target genes involved in amyloidogenesis (PSEN1, BACE1, APP, ADAM 10, ADAM 17) and neuroinflammation (NFkB, IL-1β, IL-6). The gene modulation strongly sustained by a coherent protein modulation suggests potential epigenetic modifications indirectly promoted by MK7R and responsible for the gene expression outcomes in the experimental conditions compared to control conditions. The overall data firmly support a "protective" role of MK7R, exerted through the mitigation of the neurodegenerative and neuroinflammatory pathways. In addition to the main thesis work, I have also been partially involved in a project on the differential expression of miRNAs in Alzheimer’s disease ( Fuso A. et al., The complex interplay between DNA methylation and miRNAs in gene expression regulation. Biochimie 2020).