Competenze

• Biologia cellulare: colture di cellule primarie, test cellulari in vitro (adesione, proliferazione, apoptosi, co-colture, migrazione, invasione), analisi in citofluorimetria, purificazione di vescicole extracellulari da terreni di coltura e da matrici, isolamento e caratterizzazione di fibroblasti da pazienti, test di angiogenesi in 3D.
• Biologia molecolare e biochimica: silenziamento genico, SDS-PAGE, Western blotting, preparazione di campioni per analisi proteomica (incluso FASP digestion, in solution digestion, in gel digestion, high pH fractionation) e metabolomica in spettrometria di massa.
• Analisi bioinformatiche: MaxQuant, Perseus, Imaris, FlowJo, Excel, GraphPad Prism, MassHunter.

Alice Santi, PhD

Department of Experimental and Clinical Biomedical Sciences

Viale Morgagni 50 – 50134 Firenze, Italy

Email: alice.santi@unifi.it

Tel: +39 055 2751248

Citizenship: Italian

Languages: Italian mother tongue, good knowledge of spoken and written English.

Technical skills and competences:

• Cell biology: primary cells culture, cellular in vitro assays (adhesion, proliferation, apoptosis, co-culture, migration, invasion), confocal microscopy, flow cytometry, extracellular vesicle purification, isolation and characterization of primary fibroblasts from human tissue and of human monocytes, 3D angiogenesis assay.
• Molecular biology and biochemistry: gene silencing (siRNA), shRNA Technology, PCR, SDS-PAGE, Western blotting, sample preparation for mass spectrometry-based proteomics (including FASP digestion, in solution digestion, in gel digestion, high pH fractionation) and metabolomics.
• Bioinformatic tools: MaxQuant, Perseus, Imaris, FlowJo, MassHunter.

Education and training:

• From January 1^st 2011 to December 31^st 2013: PhD student in Biochemistry and Applied Biology at University of Firenze, Department of Experimental and Clinical Biomedical Sciences, section of Biochemistry, Firenze, Italy (Supervisor: Prof. Paolo Cirri). The work I have performed during my PhD demonstrated that prostate cancer associated fibroblasts boost cancer cell proliferation and that this effect is greatly dependent on stromal carbonic anhydrase IX (CA IX) activity. CA IX products, protons and bicarbonate, exert differential effects on cancer cell proliferation. While acidification of extracellular pH is detrimental for cancer cell proliferation, bicarbonate, through its organication, supplies cancer cells with intermediates that are useful to sustain their high proliferation rate. In addition, I have characterized a novel, intrinsic, property of fibroblasts, exerted by vesicle-mediated transfer of biomass, i.e. proteins and lipids, to neighbouring cells. The horizontal transfer of material from fibroblasts enhances cancer cell proliferation.
• June 30^th 2010: Master Degree in Medical Biotechnologies, 110/110 cum laude at University of Firenze, Firenze, Italy.
• From October 1^st 2009 to June 15^th 2010: Research intern at University of Firenze, Department of Experimental and Clinical Biomedical Sciences, section of Biochemistry, Firenze, Italy (Supervisor: Prof. Paolo Cirri). During the Master Degree internship, I was involved in the studies of cellular responses (i.e. proliferation and migration) of mouse fibroblast cell line NIH-3T3 at high and low oxygen concentrations, evaluating the role of hypoxia inducible factors, HIF-1α and HIF-2α, in the regulation of these cellular phenotypes.
• April 16^th 2008: Bachelor’s Degree in Biotechnologies, 110/110 cum laude at University of Firenze, Firenze, Italy.
• From October 1^st 2007 to March 31^st 2008: Research intern at University of Firenze, Department of Experimental and Clinical Biomedical Sciences, section of Pathology and Experimental Oncology, Firenze, Italy (Supervisor: Prof. Massimo Olivotto). During the Bachelor’s Degree internship, I took part in the studies on the relationship between the hypoxia-dependent (0.1-0.2% oxygen atmosphere) block of cell cycling, the NAD(P)/NAD(P)H ratio and folate utilization in purine synthesis in cancer stem cells.

Research and professional experiences:

• From May 25^th 2021 to present: Research technician at University of Firenze, Department of Experimental and Clinical Biomedical Sciences, viale Morgagni 50 – 50134 Firenze, Italy;
• From January 5^th 2016 to May 13^th 2021: Post-doctoral Fellow at CRUK Beatson Institute, Tumour Microenvironment and Proteomics Group, Glasgow, UK (Supervisor: Dr. Sara Zanivan).
• From December 1^st 2014 to November 30^th 2015: Post-doctoral Fellow at University of Firenze, Department of Experimental and Clinical Biomedical Sciences, section of Biochemistry, Firenze, Italy (Supervisor: Prof. Paolo Cirri). I participated in studies concerning the effect of microgravity in wound repair.
• From January 1^st 2014 to November 30^th 2014: Post-doctoral Fellow at University of Firenze, Department of Experimental and Clinical Biomedical Sciences, section of Biochemistry, Firenze, Italy (Supervisor: Prof. Paola Chiarugi). I took part in a collaborative study aiming at characterizing the anti-tumoural activity of antibodies directed against novel tumour markers in breast, prostate and ovarian cancers.

Honours and awards:

• ‘IMaSS’ Mass Spectrometry School winner to attend the Shimadzu Mass Spectrometry training course, Kyoto, Japan (2015);
• Selected for oral communication at IX Italian Proteomic Association (ItPA) Annul Congress (2014);
• FEBS bursary winners to attend at 38^th FEBS Congress, Saint Petersburg, Russia (2013);
• Selected for oral communication at 56^th National Meeting of Italian Society of Biochemistry and Molecular Biology (2012).

Teaching Experience:

• I was responsible for training and tutoring Biology MSc under-graduated students.
• Teaching of modules within the Course of Biochemistry (Degree in Applied Pharmaceutical Sciences, University of Firenze) and within the Course of Biochemical Methods (Degree in Biology, University of Firenze).

Training Courses:

• Q Exactive HF Biotech training course, Glasgow, UK, 2016;
• MaxQuant Summer School, Oxford, UK, 2016;
• Shimadzu Mass Spectrometry training course, Kyoto, Japan, 2015;
• ‘IMaSS’ Mass Spectrometry training course, Italy, 2015.

Publications:

• Toti A, Santi A, Pardella E, Nesi I, Tomasini R, Mello T, Paoli P, Caselli A, Cirri P. Activated fibroblasts enhance cancer cell migration by microvesicles-mediated transfer of Galectin-1. J Cell Commun Signal. 2021 Sep;15(3):405-419. doi: 10.1007/s12079-021-00624-4. Epub 2021 May 22.
• Santi A, Kugeratski FG, Zanivan S. Cancer associated fibroblasts: the architects of stroma remodelling. Proteomics. 2017 Dec 27. doi: 10.1002/pmic.201700167.
• Reid SE, Kay EJ, Neilson LJ, Henze AT, Serneels J, McGhee EJ, Dhayade S, Nixon C, Mackey JB, Santi A, Swaminathan K, Athineos D, Papalazarou V, Patella F, Román-Fernández Á, ElMaghloob Y, Hernandez-Fernaud JR, Adams RH, Ismail S, Bryant DM, Salmeron-Sanchez M, Machesky LM, Carlin LM, Blyth K, Mazzone M, Zanivan S. Tumor matrix stiffness promotes metastatic cancer cell interaction with the endothelium. EMBO J. 2017 Aug 15;36(16):2373-2389. doi: 10.15252/embj.201694912. Epub 2017 Jul 10.
• Hernandez-Fernaud JR, Ruengeler E, Casazza A, Neilson LJ, Pulleine E, Santi A, Ismail S, Lilla S, Dhayade S, MacPherson IR, McNeish I, Ennis D, Ali H, Kugeratski FG, Al Khamici H, van den Biggelaar M, van den Berghe PV, Cloix C, McDonald L, Millan D, Hoyle A, Kuchnio A, Carmeliet P, Valenzuela SM, Blyth K, Yin H, Mazzone M, Norman JC, Zanivan S. Secreted CLIC3 drives cancer progression through its glutathione-dependent oxidoreductase activity. Nat Commun. 2017 Feb 15;8:14206. doi: 10.1038/ncomms14206.
• Grandi A, Santi A, Campagnoli S, Parri M, De Camilli E, Song C, Jin B, Lacombe A, Castori-Eppenberger S, Sarmientos P, Grandi G, Viale G, Terracciano L, Chiarugi P, Pileri P, Grifantini R. ERMP1, a novel potential oncogene involved in UPR and oxidative stress defense, is highly expressed in human cancer. Oncotarget. 2016 Aug 23. doi: 10.18632/oncotarget.11550. [Epub ahead of print];
• Caselli A, Paoli P, Santi A, Mugnaioni C, Toti A, Camici G, Cirri P. Low molecular weight protein tyrosine phosphatase: Multifaceted functions of an evolutionarily conserved enzyme. Biochim Biophys Acta. 2016 Oct; 1864(10):1339-55. doi:10.1016/j.bbapap.2016.07.001
• Caselli A, Cirri P, Santi A, Paoli P. Morin: a promising natural drug. Curr Med Chem 2016; 23(8):774-791.
• Santi A, Caselli A, Ranaldi F, Paoli P, Mugnaioni C, Michelucci E, Cirri P. Cancer associated fibroblasts transfer lipids and proteins to cancer cells through cargo vesicles supporting tumor growth. Biochim Biophys Acta. 2015 Dec;1853(12):3211-23. doi:10.1016/j.bbamcr.2015.09.013
• Santi A, Caselli A, Paoli P, Corti D, Camici G, Pieraccini G, Taddei ML, Serni S, Chiarugi P, Cirri P. The effect of CA IX catalysis products within tumor microenvironment. Cell Commun Signal. 2013 Oct 29;11:81. doi:101186/1478-811X-11-81.
• Paoli P, Cirri P, Caselli A, Ranaldi F, Bruschi G, Santi A, Camici G. The insulin-mimetic effect of Morin: a promising molecule in diabetes treatment. Biochim Biophys Acta. 2013 Apr;1830(4):310211. doi:10.1016/j.bbagen.2013.01.017.