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Alma universitas studiorum parmensis A.D. 962 - Università di Parma Alma universitas studiorum parmensis A.D. 962 - Università di Parma
Department of
Medicine and Surgery
Università degli Studi di Parma

Laboratory of Medical Oncology

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Who we are

Scientific Head: Prof. Marcello Tiseo, Full Professor of Medical Oncology

For info and contacts: marcello.tiseo@unipr.it, 0521-702669 direct; Lab number. 0521-702313

Affiliating Personnel:

  • Prof. Marcello Tiseo
  • Dr. Roberta Minari
  • Dr. Benedetta Pellegrino
  • Dr. Giulia Mazzaschi
  • Dr. Gianmarco Martines
  • Dr. Chiara Tornali

.

Where we are

Hall 24 - UOC Medical Oncology, New Center for Oncology and Radiotherapy, floor 0

What we do

Main lines of research :

  • Research of predictive factors of response and study of mechanisms of resistance to molecular targeted therapies: application of next-generation methods, digital droplet PCR and Next Generation Sequencing, for the study of liquid biopsy (cell free DNA extracted from plasma) and tumor tissue in order to identify predictive biomarkers of response and mechanisms of resistance to molecularly targeted therapies in patients with solid neoplasms with specific molecular driver.
  • Research for predictive factors of response to immunotherapy: Characterization of the immunoprofile, by study of circulating lymphocyte subpopulations (flow cytofluorimetry) and tumor microenvironment (immunohistochemistry), genomic and transcriptional profiling (NGS, RNA-sequencing and nanostring), and radiomic profiling of solid neoplasms treated with immunotherapy.
  • Research for biomarkers of outcome in early disease:Use of liquid biopsy in operable disease for the study of minimal residual molecular disease.
  • Study of the prognostic and predictive role of functional homologous recombination deficiency (HRD)in triple-negative breast cancer or with germline mutation in BRCA1/2 genes or with early onset. Study of the association between HRD and immune system activation in breast and ovarian cancer patients. Study of the prognostic role of HRD in breast and/or ovarian cancer patients carrying germline Variant of Unknown Significance (VUS) in BRCA1/2 genes. Study of the prognostic and predictive role of HRD in ovarian carcinoma.

Main methodologies applied:

  • Next-generation sequencing:Parallel massive fluorescence sequencing exploiting sequencing Chemistry by Illumina synthesis. Employment of targeted panels (50 to 500 genes) for analysis of molecular alterations (SNVs, indels, CNVs, and gene fusions) in tumor tissue and/or liquid biopsy.
  • Digital droplet PCR:Digital droplet PCR based on water-oil emulsification. The sample is fractionated into approximately 20,000 droplets within which, in each droplet, the template amplification reaction takes place. Massive partitioning of the sample is one of the key aspects of this technique that gives it a high sensitivity useful for somatic variant analysis, minimal residual disease, expression assays, and analysis of gene copy number alterations.
  • ELISA assays: immuno-enzymatic assays.
  • RAD51 immunofluorescence assay.
  • Characterization of tumor immune infiltrate by immunohistochemistry.

Main instrumentation provided:

  • Miseq (Illumina) The MiSeq system is a next-generation sequencing (NGS) platform that integrates cluster generation, amplification, sequencing and data analysis into a single instrument. It is ideal for targeted resequencing and sequencing of small genomes.
  • NextSeq 550Dx (Illumina) The NextSeq™ 550Dx Illumina® instrument is a CE-IVD-marked next-generation sequencing (NGS) platform with dual modes of use: Diagnostic (Dx Mode) and Research (Research Mode). NextSeq™ 550 enables sequencing of exomes, whole genomes, transcriptomes, and provides a transition between high-processivity sequencing and array scanning.
  • Droplet Digital™ PCR (ddPCR™, BIO-RAD) Droplet Digital PCR (ddPCR) employs advanced microfluidics technology to generate thousands of highly uniform droplets of a few nanoliters in volume, within which a PCR reaction takes place. Once generated, the droplets can be amplified, read and analyzed. This highly sensitive technology can provide absolute quantification of the target sample, and detect rare genomic variants for clinical and research purposes.
  • 4200 TapeStation System (Agilent)The Agilent 4200 TapeStation system is an automated electrophoresis instrument for quality control of nucleic acids (DNA and RNA) by analyzing sample size, concentration and integrity. The TapeStation system finds application within any next-generation sequencing (NGS) workflow.
  • Maxwell® 16 Instrument (Promega)is used to perform automated purification of nucleic acids (DNA, RNA, circulating free DNA and miRNA) from whole blood, tissue, plasma and buffy-coat. Up to 16 samples can be processed simultaneously in about 30 minutes. The extracted nucleic acids are of high quality and concentration and can be used for various types of clinical and research applications.
  • Fluorescence microscope Nikon Eclipse Ni-U equipped with 100-W mercury lamp with Nikon Plan Fluor 20X and 60X objectives.
  • VENTANA DP 200 slide scanner (CE-IVD).

Keywords for Laboratory description: biomarkers, liquid biopsy, resistance mechanisms, homologous recombination (HRD), immune-profiling.

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