Non-Small Cell Lung cancer is the most frequently diagnosed lung cancer type (80-90% of all cases) and lung adenocarcinoma (LUAD) is the major subtype. Early diagnosis by low-dose computed tomography was recently shown to be effective in reducing lung cancer mortality (~20% at 5 years). However, the management of lung cancer patients can be difficult due to pathological and genetic heterogeneity of the disease. We, recently, discovered a new molecular subtype of lung adenocarcinoma using a 10-genes signature, namely C1- LUAD, which correlates with the worst patients prognosis and has peculiar features of mesenchymal and stem-like cells. Yet, we also identified an intracellular signature of 7 microRNAs as a surrogate of the 10-genes to distinguish the C1-LUAD subtype. Of note, analyzing intracellular differentially expressed miRNAs in C1 patients, we found a targeted network of genes as hubs of mechanisms for lung cancer progression. Importantly, miRNAs can be secreted in body fluids (e.g., Urine, breast milk, blood) via extracellular vesicles (i.e. exosomes). In the last years, tumor exosomes have also drawn increasing attention due to their stability and accessibility as diagnostic and prognostic biomarkers. For these reasons, we selected 22-miRNAs which target genes involved in cancer progression and we analyzed their expression in isolated exosomes derived from C1 and nonC1 serum samples. Performing a differential expression analysis we found 5 miRNAs up-regulated and 6 miRNAs down-regulated in C1 compared with nonC1 patients. Next, using differential centrifugation protocol, we purified and characterized exosomes from a panel of LUAD cell lines classified by gene expression and phenotypic behaviors in C1-like and nonC1-like. Notably, we evaluated the heterogenic exosomes kinetic up-take in C1-like and nonC1-like cells, highlighting that C1-like cells are more prone to internalized labeled-exosomes, in a time and dose dependent manner. Overall, we identified for the first time a set of exosomal-miRNAs linked to C1-LUAD subtype, which pave the way to make an early diagnosis for this new aggressive subtype of lung cancer. Moreover, we found that C1-like cells internalize more efficiently exosomes, suggesting a role of these extracellular vesicles in the maintenance of the metastatic features characteristic of the C1 tumors.
Analysis of extracellular-vesicle miRNAs in a new aggressive molecular subtype of lung cancer / Mazzarelli, Francesco; Colangelo, Tommaso; Cuttano, Roberto; Melocchi, Valentina; Dama, Elisa; Kiptiu, Patricia; Maria Perrone, Rosa; Afanga, Kuku; Bianchi, Fabrizio. - (2022). (Intervento presentato al convegno 62nd Annual Meeting of the Italian Cancer Society The exciting path from preclinical research to clinical application tenutosi a Hotel NH Venezia Laguna Palace Viale Ancona, 2 30172 – Venezia, Italy).
Analysis of extracellular-vesicle miRNAs in a new aggressive molecular subtype of lung cancer
Francesco Mazzarelli†Primo
Writing – Original Draft Preparation
;Fabrizio Bianchi
Supervision
2022
Abstract
Non-Small Cell Lung cancer is the most frequently diagnosed lung cancer type (80-90% of all cases) and lung adenocarcinoma (LUAD) is the major subtype. Early diagnosis by low-dose computed tomography was recently shown to be effective in reducing lung cancer mortality (~20% at 5 years). However, the management of lung cancer patients can be difficult due to pathological and genetic heterogeneity of the disease. We, recently, discovered a new molecular subtype of lung adenocarcinoma using a 10-genes signature, namely C1- LUAD, which correlates with the worst patients prognosis and has peculiar features of mesenchymal and stem-like cells. Yet, we also identified an intracellular signature of 7 microRNAs as a surrogate of the 10-genes to distinguish the C1-LUAD subtype. Of note, analyzing intracellular differentially expressed miRNAs in C1 patients, we found a targeted network of genes as hubs of mechanisms for lung cancer progression. Importantly, miRNAs can be secreted in body fluids (e.g., Urine, breast milk, blood) via extracellular vesicles (i.e. exosomes). In the last years, tumor exosomes have also drawn increasing attention due to their stability and accessibility as diagnostic and prognostic biomarkers. For these reasons, we selected 22-miRNAs which target genes involved in cancer progression and we analyzed their expression in isolated exosomes derived from C1 and nonC1 serum samples. Performing a differential expression analysis we found 5 miRNAs up-regulated and 6 miRNAs down-regulated in C1 compared with nonC1 patients. Next, using differential centrifugation protocol, we purified and characterized exosomes from a panel of LUAD cell lines classified by gene expression and phenotypic behaviors in C1-like and nonC1-like. Notably, we evaluated the heterogenic exosomes kinetic up-take in C1-like and nonC1-like cells, highlighting that C1-like cells are more prone to internalized labeled-exosomes, in a time and dose dependent manner. Overall, we identified for the first time a set of exosomal-miRNAs linked to C1-LUAD subtype, which pave the way to make an early diagnosis for this new aggressive subtype of lung cancer. Moreover, we found that C1-like cells internalize more efficiently exosomes, suggesting a role of these extracellular vesicles in the maintenance of the metastatic features characteristic of the C1 tumors.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
