Through genomic and transcriptomic sequencing techniques (whole exome sequencing, WES; whole transcriptome sequencing, WTS) patients with enrichment for KEAPness and specific gene interactions associated with it.
Age >18 years;
Histological diagnosis of NSCLC
metastatic disease
Availability, at the time of enrollment, of adequate biological material to be able to perform molecular analyses, taken (surgical or by biopsy) before administration of any anti-tumor treatment (chemotherapy and/or radiotherapy);
Previous systemic therapy for metastatic disease;
Comorbidities not controlled with adequate medical therapy.
Age >18 years;
Histological diagnosis of NSCLC
Patients with metastatic NSCLC who are candidates for and/or undergoing treatment first-line chemotherapy-immunotherapy combination treatment or immunotherapy from alone (if PD-L1\>50% as per clinical practice) that meet the following criteria.
Other
Cross-sectional
Not listed
Innovative molecular tool
To generate an innovative molecular tool for prediction DNA/RNA will be extracted from 5μm FFPE tissue sections using the AllPrep DNA/RNA FFPE kit (Qiagen). The quality of the RNA will be evaluated with the Bioanalyzer, the integrity of genomic DNA through the Agilent NGS FFPE qPCR QC Kit (Agilent). The libraries for RNA-Seq will be prepared using the RNA TruSeq Exome Kit (Illumina). The quality of the libraries resulting will be checked using Bioanalyzer (high sensitivity DNA kit). The intermediate library before exon enrichment will be quantified with Qubit, the final library with qPCR. The samples will be sequenced in paired-end mode, sequencing 76 bp on each side. The Exome DNA sequencing will be performed using SureSelectXT Low Input Kit reagents for the initial preparation of the library followed by enrichment with Clinical Research Exome Kit (Agilent).
Tracing the evolutionary path
Tracing the evolutionary path of NRF-2 dependent tumors, in his association with the microenvironment, and in particular the related immune subtypes, and characterize the interactions between pairs of genes (epistatic) underlying the KEAPness.i phenotype, the multi-region sequencing technology that will be used involves the use of surgical samples, taken in a single session. This allows you to isolate different regions of the tumor, corresponding to different time points (central region, R1; expansion zone, R2; locoregional metastases, R3). Through this methodology it is possible to trace the evolution of the tumor over time, and in particular the interactions between pairs of mutant genes linked to the KEAPness phenotype.
Identification of therapeutic vulnerabilities specific
Identification of therapeutic vulnerabilities specific to biological processes related to KEAPness. Cell lines derived from patients affected by NSCLC and GC, already available in the laboratories of Promoter/IFO, will initially be characterized by genomic sequencing and transcriptomic (WES and WTS) to identify the enrichment in the KEAPness phenotype and previously identified epistatic interactions.
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