Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS 10 articles published in JoVE Cancer Research Evaluation of Colorectal Cancer Risk and Prevalence by Stool DNA Integrity Detection Claudia Rengucci1, Giulia De Maio1, Maura Menghi2, Francesca Benzi2, Daniele Calistri1 1Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 2Diatech Pharmacogenetics srl The presented diagnostic FL-DNA kit is a time-saving and user-friendly method to determine the reliable probability of the presence of colorectal cancer lesions. Medicine A Whole Body Dosimetry Protocol for Peptide-Receptor Radionuclide Therapy (PRRT): 2D Planar Image and Hybrid 2D+3D SPECT/CT Image Methods Maria Luisa Belli1, Emilio Mezzenga1, Valentina Di Iorio2, Monica Celli3, Paola Caroli3, Elisabeth Canali3, Federica Matteucci3, Elisa Tardelli3, Ilaria Grassi3, Maddalena Sansovini3, Silvia Nicolini3, Stefano Severi3, Marta Cremonesi4, Mahila Ferrari5, Giovanni Paganelli3, Anna Sarnelli1 1Medical Physics Unit, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 2Oncology Pharmacy, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 3Nuclear Medicine Unit, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 4Radiation Research Unit, Istituto Europeo di Oncologia (IEO) IRCCS, Milano, Italy, 5Medical Physics Unit, Istituto Europeo di Oncologia (IEO) IRCCS, Milano, Italy This method estimates the absorbed dose of different structures for peptide-receptor-radionuclide-therapy (PRRT) with the possibility of avoiding organ overlap on 2D-projections. Serial whole-body planar images permit estimation of mean absorbed doses along the whole body, while the hybrid approach, combining planar images and 3D-SPECT/CT image, overcomes the limitations of structure overlapping. Cancer Research Detection of a Circulating MicroRNA Custom Panel in Patients with Metastatic Colorectal Cancer Matteo Canale1, Giorgia Marisi1, Alessandro Passardi2, Emanuela Scarpi3, Paola Ulivi1 1Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Department of Medical Oncology, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 3Unit of Biostatistics and Clinical Trials, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS We present a protocol to evaluate the expression levels of circulating microRNA (miRNAs) in plasma samples from cancer patients. In particular, we used a commercially available kit for circulating miRNA extraction and reverse transcription. Finally, we analyzed a panel of 24 selected miRNAs using real-time pre-spotted probe custom plates. Cancer Research Dried Blood and Serum Spots As A Useful Tool for Sample Storage to Evaluate Cancer Biomarkers Laura Mercatali1, Patrizia Serra2, Giacomo Miserocchi1, Chiara Spadazzi1, Chiara Liverani1, Alessandro De Vita1, Giorgia Marisi3, Alberto Bongiovanni1, Federica Recine1, Akwilina Pangan4, Nestory Masalu4, Toni Ibrahim1, Dino Amadori1 1Osteoncology and Rare Tumors Center, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 2Unit of Biostatistics and Clinical Trials, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 3Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 4Department of Oncology, Bugando Medical Center, Mwanza, Tanzania This protocol describes a simple and useful method to store peripheral blood and serum/plasma for downstream analyses such as single nucleotide polymorphism (SNP) evaluation and ELISA assay. Cancer Research Establishment of a Primary Culture of Patient-derived Soft Tissue Sarcoma Alessandro De Vita1, Laura Mercatali1, Giacomo Miserocchi1, Chiara Liverani1, Chiara Spadazzi1, Federica Recine1, Alberto Bongiovanni1, Federica Pieri2, Davide Cavaliere3, Valentina Fausti1, Dino Amadori1, Toni Ibrahim1 1Osteoncology and Rare Tumors Center, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Pathology Unit, Morgagni-Pierantoni Hospital, 3Unit of Surgery and Advanced Oncologic Therapies, Morgagni-Pierantoni Hospital The following protocol focuses on the establishment of a primary culture of patient-derived soft tissue sarcoma (STS). This model could help us to better understand the molecular background and pharmacological profile of these rare malignancies and could represent a starting point for further research aimed at improving STS management. Cancer Research Detection of a CDH1 Rare Transcript Variant in Fresh-frozen Gastric Cancer Tissues by Chip-based Digital PCR Chiara Molinari1, Raefa Abou Khouzam2, Samanta Salvi1, Tania Rossi1, Guglielmina Nadia Ranzani2, Daniele Calistri1 1Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Department of Biology and Biotechnology, University of Pavia The manuscript describes a chip-based digital PCR assay to detect a rare CDH1 transcript variant (CDH1a) in fresh-frozen normal and tumor tissues obtained from patients with gastric cancer. Cancer Research Characterization of Tumor Cells Using a Medical Wire for Capturing Circulating Tumor Cells: A 3D Approach Based on Immunofluorescence and DNA FISH Giulia Gallerani1, Claudia Cocchi2, Martine Bocchini3, Filippo Piccinini1, Francesco Fabbri1 1Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Associazione Annastaccatolisa Onlus, 3Nuclear Medicine Unit, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS We present a new approach to characterize tumor cells. We combined immunofluorescence with DNA fluorescent-in-situ-hybridization to evaluate cells captured by a functionalized medical wire capable of in vivo enriching CTCs directly from patient blood. Cancer Research Serum and Plasma Copy Number Detection Using Real-time PCR Samanta Salvi1, Vincenza Conteduca1, Filippo Martignano1, Giorgia Gurioli1, Daniele Calistri1, Valentina Casadio1 1Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS This manuscript describes the copy number variation analysis performed in serum or plasma DNA using real-time PCR approach. This method is suitable for the prediction of drug resistance in castration resistant prostate cancer patients, but it could be informative also for other diseases. Cancer Research Looking for Driver Pathways of Acquired Resistance to Targeted Therapy: Drug Resistant Subclone Generation and Sensitivity Restoring by Gene Knock-down Chiara Arienti1, Sara Pignatta1, Michele Zanoni1, Michela Cortesi1, Alice Zamagni1, Filippo Piccinini2, Anna Tesei1 1Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS This is a time- and cost-effective in vitro protocol investigating the mechanisms of acquired resistance to targeted therapeutic agents, which is a highly unmet medical need in cancer management. Cancer Research Cell-Free DNA Integrity Analysis in Urine Samples Valentina Casadio1, Samanta Salvi1, Filippo Martignano1, Roberta Gunelli2, Sara Ravaioli1, Daniele Calistri1 1Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Department of Urology, Morgagni-Pierantoni Hospital A method for analyzing DNA integrity in the cell-free supernatant fraction of urine samples is proposed. The method is suitable for early detection of urological malignancies and has proven accurate for the early diagnosis of bladder cancer.