REPORT SULLE ATTIVITÀ SVOLTE
Corso di dottorato Scienze Cliniche e Traslazionali
Cognome Rofi Nome Eleonora
Titolo della tesi
Liquid biopsy to monitor genetic alterations in plasma as non-invasive pharmacogenetic-based approach to evaluate predictive biomarkers during treatment in cancer patients
Progetto di ricerca
At present, in oncology the therapeutic strategies are defined according to the molecular landscape assessed with tissue biopsies and, thus, using DNA and/or RNA obtained from a fragment of the primary tumour or a single metastatic lesion. Anyway, mutational analysis in a single-site biopsy is encumbered by several limitations.
A huge step forward in oncology to characterize tumor heterogeneity and plasticity at diagnosis and during the course of treatment was done with the use of a promising minimally invasive tool., i.e. via a blood draw. All cells can release in human blood their materials, including cell-free DNA (cfDNA), RNA, proteins and vesicles (such as exosomes). This approach is the expression of the overall spatial heterogeneity from the overall sites of disease and it is able to identify emerging sub-clones responsible for treatment resistance.
This thesis was aimed at evaluating if a pharmacogenetic approach based on circulating predictive
biomarkers (i.e. circulating cell-free tumor DNA and RNA extracted from exosomes) may be a good
and useful tool to monitor treatment outcome in solid tumors.
Patients affected by lung, breast, prostate, pancreatic cancer and melanoma have been enrolled and monitored during different treatment, i.e. targeted therapy, hormonal therapy, chemotherapy and immunotherapy by circulating tumor nucleic acids. Nucleic acids were analysed by digital droplet PCR for targetable drivers, for acquired mutations and gene expression selected during therapy. Circulating tumor nucleic acids were find to correlate with tumor burden, disease status and response to treatment, beeing a good candidate to monitor tumor response and support routine clinical practice.
Attività formative previste dal corso di Dottorato
- Inglese accademico I “C1” aa 2016/2017 (votazione esame finale: very good) - Inglese accademico II “C1+” aa 2017/2018 (votazione esame finale: very good)
- Statistica (votazione finale: idonea)
Partecipazione a convegni di società scientifiche nazionali ed internazionali e seminari tenuti nel triennio
- ESMO 2018 – Munich (Germany) – 19-23 Ottobre 2018 - Poster Presenter . Title: “Selective induction of PD-L1 expression in plasma-derived exosomes by gemcitabine-nab-paclitaxel vs. FOLFIRINOX in pancreas cancer”;
- I Congresso Monotematico SIF “Research progress in Pharmacogenomics: personalised medicine and clinical translation”, Bologna, 5-6 Luglio 2018 - Oral Communication. Title: “Therapeutic monitoring in NSCLC by liquid biopsy”;
- Astrazeneca ONCOLOGY SCIENCE CONFERENCE - Milano, 21-22 Maggio 2018 – Poster Presenter. Title: “An in vitro study to assess the COoperation of EGFR Mutations in Promoting Actionable REsistance in NSCLC and explore new therapeutic Strategies”;
- ESMO Preceptorship on Immuno-Oncology, Lugano (Svizzera), 4-5 Maggio 2018; “PHARMACOGENETICS: The way to find individual therapy solutions”, Milano, 10 aprile 2018; - Workshop IMI Immuno-oncologia, Pisa, 22-23 Marzo 2018;
- III Convegno monotematico SIF "New and old challenges for clinical pharmacology: from therapeutic drug monitoring to pharmacogenetics and pharmacovigilance", Napoli, 15-16 Marzo 2018 – Oral Communication. Title: “L’associazione gemcitabina e nab-paclitaxel incrementa l’espressione del PD-L1 nel tumore del pancreas;
- 38° Congresso Nazionale SIF, Rimini – 25-28 Ottobre 2017 – Poster Presenter. Title: “Early variations of KRAS mutations in circulating cell-free tumor DNA: a promising biomarker to monitor chemotherapy resistance in pancreatic cancer”;
- ESPT Congress – Catania – 4-7 ottobre 2017 - Poster Presenter . Title: “Patients with NSCLC may display a low ratio of p.T790M vs. activating EGFR mutations in plasma at disease progression: implications for personalised treatment.;
- EPMA World Congress – Malta, 14-17 Settembre 2017 – Oral Communication. Title: “Early changes in plasma DNA levels of mutant KRAS as a sensitive marker of response to chemotherapy in pancreatic cancer.”;
- PROGRESSI NEL NSCLC ALK TRASLOCATO, Milano, 10 maggio 2017;
- Convegno SIF "Il ruolo della Farmacologia Clinica in Italia: prospettive future nell'ambito del SSN", Napoli, 15-16 Dicembre 2016 – Poster Presenter. Title: “Monitoring KRAS mutations from plasma circulating free tumor DNA as non-invasive approach to evaluate pancreatic cancer dynamics.”; - XVIII CONGRESSO Nazionale AIOM, Roma, 28-30 Ottobre 2016;
- XIX Seminario SIF Dottorandi e Assegnisti - Rimini, 20-22 settembre 2016 – Poster Presenter. Title: “Monitoring KRAS mutations from plasma circulating free tumor DNA as non-invasive approach to evaluate pancreatic cancer dynamics.”;
- The Young Biomarkers Day – The Golden Helix symposium on Circulating Nucleic Acids - Lucca 15-16 giugno 2015-16
- II corso annuale - Tumori gastrointestinali: dalla pratica clinica alla biologia. Highlights from ASCO GI 2016 - Prato 19 Febbraio 2016
Riconoscimenti e premi ottenuti nel triennio
2018: Vincitrice del travel grant ESO per partecipare alla conferenza “ESMO Preceptorship on Immuno-Oncology, Lugano (Svizzera), 4-5 Maggio 2018
Lavori pubblicati nel triennio
1. Rofi E, Del Re M, Arrigoni E, Rizzo M, Fontanelli L, Crucitta S, Gianfilippo G, Restante G, Fogli S, Porta C, Danesi R, Schmidinger M. Clinical pharmacology of monoclonal antibodies targeting anti PD-1 axis in urothelial cancers. Critical Reviews in Oncology/Hematology. Volume 144, 2019, 102812. doi: 10.1016/j.critrevonc.2019.09.004.
2. Crucitta S, Restante G, Del Re M, Bertolini I, Bona E, Rofi E, Fontanelli L, Gianfilippo G, Fogli S, Stasi I, Ghilli M, Fontana A, Danesi R. Endothelial nitric oxide synthase c.-813C>T predicts for proteinuria in metastatic breast cancer patients treated with bevacizumab-based chemotherapy. Cancer Chemother Pharmacol. 2019 Sep 16. doi: 10.1007/s00280-019-03933-z.
3. Pasqualetti F, Restante G, Gonnelli A, Rofi E, Molinari A, Crucitta S, Paiar F, Rudà R, Danesi R, Soffietti R, Del Re M. Dabrafenib treatment in a patient with BRAF V600E ganglioglioma: circulating exosome-derived cancer RNA supports treatment choice and clinical monitoring. Neuro Oncol. 2019 Aug 24. pii: noz157. doi: 10.1093/neuonc/noz157.
4. Del Re M, Bertolini I, Crucitta S, Fontanelli L, Rofi E, De Angelis C, Diodati L, Cavallero D, Gianfilippo G, Salvadori B, Fogli S, Falcone A, Scatena C, Naccarato AG, Roncella M, Ghilli M, Morganti R, Fontana A, Danesi R. Overexpression of TK1 and CDK9 in plasma-derived exosomes is associated with clinical resistance to CDK4/6 inhibitors in metastatic breast cancer patients. Breast Cancer Res Treat. 2019 Nov;178(1):57-62. doi: 10.1007/s10549-019-05365-y.
5. Del Re M, Crucitta S, Sbrana A, Rofi E, Paolieri F, Gianfilippo G, Galli L, Falcone A, Morganti R, Porta C, Efstathiou E, van Schaik R, Jenster G, Danesi R. AR-V7 and AR-FL expression is associated with clinical outcome: a translational study in patients with castrate resistant prostate cancer. BJU Int. 2019 May 4. doi: 10.1111/bju.14792.
6. Rofi E, Del Re M, Cappelli C, Puppo G, Crucitta S, Valeggi S, Chella A, Danesi R, Petrini I. The increase in activating EGFR mutation in plasma is an early biomarker to monitor response to osimertinib: a case report. BMC Cancer. 2019 Apr 30;19(1):410. doi: 10.1186/s12885-019-5604-6.
7. Bordi P, Del Re M, Minari R, Rofi E, Buti S, Restante G, Squadrilli A, Crucitta S, Casartelli C, Gnetti L, Azzoni C, Bottarelli L, Petrini I, Cosenza A, Ferri L, Rapacchi E, Danesi R, Tiseo M. From the beginning to resistance: Study of plasma monitoring and resistance mechanisms in a cohort of patients treated with osimertinib for advanced T790M-positive NSCLC. Lung Cancer. 2019 May;131:78-85. doi: 10.1016/j.lungcan.2019.03.017.
8. Macera L, Spezia PG, Medici C, Rofi E, Del Re M, Focosi D, Mazzetti P, Navarro D, Antonelli G, Danesi R, Pistello M, Maggi F. Comparative evaluation of molecular methods for the quantitative measure of torquetenovirus viremia, the new surrogate marker of immune competence. J Med Virol. 2019 Apr 19. doi: 10.1002/jmv.25488.
9. Rofi E, Vivaldi C, Del Re M, Arrigoni E, Crucitta S, Funel N, Fogli S, Vasile E, Musettini G, Fornaro L, Falcone A, Danesi R. The emerging role of liquid biopsy in diagnosis, prognosis and treatment monitoring of pancreatic cancer. Pharmacogenomics. 2019 Jan;20(1):49-68. doi: 10.2217/pgs-2018-0149.
10. Cremolini C, Rossini D, Dell'Aquila E, Lonardi S, Conca E, Del Re M, Busico A, Pietrantonio F, Danesi R, Aprile G, Tamburini E, Barone C, Masi G, Pantano F, Pucci F, Corsi DC, Pella N, Bergamo F, Rofi E, Barbara C, Falcone A, Santini D. Rechallenge for Patients With RAS and BRAF Wild-Type Metastatic Colorectal Cancer With Acquired Resistance to First-line Cetuximab and Irinotecan: A Phase 2 Single Arm Clinical Trial. JAMA Oncol. 2019 Mar 1;5(3):343-350. doi: 10.1001/jamaoncol.2018.5080.
11. Del Re M, Bordi P, Rofi E, Restante G, Valleggi S, Minari R, Crucitta S, Arrigoni E, Chella A, Morganti R, Tiseo M, Petrini I, Danesi R. The amount of activating EGFR mutations in circulating cell-free DNA is a marker to monitor osimertinib response. Br J Cancer. 2018 Nov;119(10):1252-1258. doi: 10.1038/s41416-018-0238-z.
12. Fogli S, Polini B, Del Re M, Petrini I, Passaro A, Crucitta S, Rofi E, Danesi R. EGFR-TKIs in non-small cell lung cancer: focus on clinical pharmacology and mechanisms of resistance. Pharmacogenomics. 2018 Jun 1;19(8):727-740. doi: 10.2217/pgs-2018-0038.
13. Del Re M, Crucitta S, Restante G, Rofi E, Arrigoni E, Biasco E, Sbrana A, Coppi E, Galli L, Bracarda S, Santini D, Danesi R. Pharmacogenetics of androgen signaling in prostate cancer: Focus on castration resistance and predictive biomarkers of response to treatment. Crit Rev Oncol Hematol. 2018 May;125:51-59. doi: 10.1016/j.critrevonc.2018.03.002.
14. Del Re M, Marconcini R, Pasquini G, Rofi E, Vivaldi C, Bloise F, Restante G, Arrigoni E, Caparello C, Bianco MG, Crucitta S, Petrini I, Vasile E, Falcone A, Danesi R. PD-L1 mRNA expression in plasma-derived exosomes is associated with response to anti-PD-1 antibodies in melanoma and NSCLC. Br J Cancer. 2018 Mar 20;118(6):820-824. doi: 10.1038/bjc.2018.9.
15. Arrigoni E, Del Re M, Galimberti S, Restante G, Rofi E, Crucitta S, Baratè C, Petrini M, Danesi R, Di Paolo A. Concise Review: Chronic Myeloid Leukemia: Stem Cell Niche and Response to Pharmacologic Treatment. Stem Cells Transl Med. 2018 Mar;7(3):305-314. doi: 10.1002/sctm.17-0175.
16. Fogli S, Del Re M, Rofi E, Posarelli C, Figus M, Danesi R. Clinical pharmacology of intravitreal anti-VEGF drugs. Eye (Lond). 2018 Jun;32(6):1010-1020. doi: 10.1038/s41433-018-0021-7. 17. Del Re M, Arrigoni E, Restante G, Passaro A, Rofi E, Crucitta S, De Marinis F, Di Paolo A,
Danesi R. Concise Review: Resistance to Tyrosine Kinase Inhibitors in Non-Small Cell Lung Cancer: The Role of Cancer Stem Cells. Stem Cells. 2018 May;36(5):633-640. doi: 10.1002/stem.2787.
18. Del Re M, Rofi E, Restante G, Crucitta S, Arrigoni E, Fogli S, Di Maio M, Petrini I, Danesi R. Implications of KRAS mutations in acquired resistance to treatment in NSCLC. Oncotarget. 2017 Dec 21;9(5):6630-6643. doi: 10.18632/oncotarget.23553.
19. Bordi P, Tiseo M, Rofi E, Petrini I, Restante G, Danesi R, Del Re M. Detection of ALK and KRAS Mutations in Circulating Tumor DNA of Patients With Advanced ALK-Positive NSCLC With Disease Progression During Crizotinib Treatment. Clin Lung Cancer. 2017 Nov;18(6):692-697. doi: 10.1016/j.cllc.2017.04.013.
20. Del Re M, Bordi P, Petrini I, Rofi E, Mazzoni F, Belluomini L, Vasile E, Restante G, Di Costanzo F, Falcone A, Frassoldati A, van Schaik RHN, Steendam CMJ, Chella A, Tiseo M, Morganti R, Danesi R. Patients with NSCLC may display a low ratio of p.T790M vs. activating EGFR mutations in plasma at disease progression: implications for personalised treatment. Oncotarget. 2017 Sep15;8(49):86056-86065. doi: 10.18632/oncotarget.20947.
21. Di Paolo A, Del Re M, Arrigoni E, Di Desidero T, Rofi E, Danesi R, Bocci G. Pharmacokinetic Markers of 5-Fluorouracil Toxicity in Clinical Trials and Real World. Clinical Cancer Drugs 2017. doi: 10.2174/2212697X04666170817124625.
22. Del Re M, Vivaldi C, Rofi E, Vasile E, Miccoli M, Caparello C, d'Arienzo PD, Fornaro L, Falcone A, Danesi R. Early changes in plasma DNA levels of mutant KRAS as a sensitive marker of response to chemotherapy in pancreatic cancer. Sci Rep. 2017 Aug 11;7(1):7931. doi: 10.1038/s41598-017-08297-z.
23. Del Re M, Restante G, Di Paolo A, Crucitta S, Rofi E, Danesi R. Pharmacogenetics and Metabolism from Science to Implementation in Clinical Practice: The Example of Dihydropyrimidine Dehydrogenase. Curr Pharm Des. 2017;23(14):2028-2034. doi: 10.2174/1381612823666170125155530.
24. Del Re M, Rofi E, Citi V, Fidilio L, Danesi R. Should CYP2D6 be genotyped when treating with tamoxifen? Pharmacogenomics. 2017 Jun;18(8):755-756. doi: 10.2217/pgs-2017-0065. 25. Del Re M, Biasco E, Crucitta S, Derosa L, Rofi E, Orlandini C, Miccoli M, Galli L, Falcone A,
Jenster GW, van Schaik RH, Danesi R. The Detection of Androgen Receptor Splice Variant 7 in Plasma-derived Exosomal RNA Strongly Predicts Resistance to Hormonal Therapy in Metastatic Prostate Cancer Patients. Eur Urol. 2017 Apr;71(4):680-687. doi: 10.1016/j.eururo.2016.08.012.
26. Del Re M, Citi V, Crucitta S, Rofi E, Belcari F, van Schaik RH, Danesi R. Pharmacogenetics of CYP2D6 and tamoxifen therapy: Light at the end of the tunnel? Pharmacol Res. 2016 May;107:398-406. doi:10.1016/j.phrs.2016.03.025.
Attività di tutoraggio
- Tutoraggio a studenti per tesi di laurea in Medicina e Chirurgia, Farmacia, Tecnico di Laboratorio, per la realizzazione della parte sperimentale e scrittura della tesi.
