Di Maggio, Federica (2021) New dimension of cell culture in physiopathology: development of patient-derived organoid structure and analysis in colorectal cancer. [Tesi di dottorato]

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Item Type: Tesi di dottorato
Resource language: English
Title: New dimension of cell culture in physiopathology: development of patient-derived organoid structure and analysis in colorectal cancer
Creators:
CreatorsEmail
Di Maggio, Federicadimaggio@ceinge.unina.it
Date: 13 December 2021
Number of Pages: 86
Institution: Università degli Studi di Napoli Federico II
Department: Medicina Molecolare e Biotecnologie Mediche
Dottorato: Medicina molecolare e biotecnologie mediche
Ciclo di dottorato: 34
Coordinatore del Corso di dottorato:
nomeemail
Santoro, Massimomasantor@unina.it
Tutor:
nomeemail
Pastore, LucioUNSPECIFIED
Salvatore, FrancescoUNSPECIFIED
Date: 13 December 2021
Number of Pages: 86
Keywords: Colorectal cancer, Patient derived organoids, Next Generation Sequencing, Third-generation sequencing.
Settori scientifico-disciplinari del MIUR: Area 05 - Scienze biologiche > BIO/12 - Biochimica clinica e biologia molecolare clinica
Date Deposited: 17 Dec 2021 09:09
Last Modified: 28 Feb 2024 11:44
URI: http://www.fedoa.unina.it/id/eprint/14281

Collection description

Colorectal cancer (CRC) is the fourth most frequent malignancy and the third leading cause of cancer death worldwide. About 5-10% of patients affected by CRC carry pathogenic germline variants in genes associated with cancer risk development. This suggests that the mutational status in predisposing genes may be a tool to identify at risk subjects; these can be considered good candidates for targeted prevention and/or treatment. In this context, the establishment of patient-derived organoids (PDOs) represent a milestone toward a successful precision medicine. The aim of this study was to establish, and then to determine, whether PDOs reflect the genes molecular pattern of tumoral tissue of CRC patients and so identify suitable candidates for treatment. We enrolled eighty patients affected by CRC who underwent surgery and, from these we stabilized 11 PDOs. They were monitored during the various phases of growth using an advanced microscopy instrumentation (Cell Discoverer-7, Zeiss). Moreover, we carried out molecular analyses, first using customized multi-gene panel, and then using whole genome sequencing (WGS) with Oxford Nanopore Technology (ONT) to evaluate the gene pattern of four genomes derived from the same patient (blood, PDOs, tumor-derived tissue and paired healthy tissue). The established model system showed genomic sequencing concordance with paired tumor in the same patient. Indeed, we confirmed the presence of 37 pathogenic mutations at somatic level in the PDOs. With these experiments we indicate a robust validity of the strategy used, also to pursue with further experimentation toward the precision medicine goal. In conclusion, the full spectrum of mutations to the aim of precision medicine studies, as also demonstrated by this work, includes the possibility of subsequent screening of cancer drug libraries, then leading to targeting specific mutations.

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