. 2022 Mar 23;14(7):1628. doi: 10.3390/cancers14071628

Table 2.

Challenges associated with routine clinical testing of KRAS mutations in NSCLC for a pathology laboratory. NGS, next generation sequencing; IHC, immunohistochemistry; IVDR, in vitro drug regulation.

Challenges
To obtain a sufficient quantity and quality of extracted RNA/DNA from formalin fixed tissue biopsies, cytological samples, or liquid biopsies To select the best molecular biology methods (targeted sequencing versus NGS) To integrate the evaluation of genomic alterations of interest (at least on TP53, STK11, and KEAP1) associated with the KRAS assessment status To assess the pathogenicity and the functionality of somatic variants of KRAS mutations in NSCLC (oncogenicity) To be able to evaluate the PD-L1 status by IHC and the KRAS status at the same time to provide in the future immune treatment and/or targeted therapy against a KRAS G12C mutation at baseline To deal with possible tumor heterogeneity (according to the size of the sample) To assess the gene KRAS status in non-adenocarcinoma lung carcinoma To handle liquid biopsies at baseline and at progression in daily practice To select an optimal gene panel To master the turnaround time required to obtain all molecular biology and IHC results To integrate the costs and the reimbursement according to molecular testing To obtain accreditation according to the ISO 15,189 norm for KRAS molecular testing To deal with the next IVDR in Europe To be able to look for mechanisms of resistance at baseline and at progression

Challenges

To obtain a sufficient quantity and quality of extracted RNA/DNA from formalin fixed tissue biopsies, cytological samples, or liquid biopsies
To select the best molecular biology methods (targeted sequencing versus NGS)
To integrate the evaluation of genomic alterations of interest (at least on TP53, STK11, and KEAP1) associated with the KRAS assessment status
To assess the pathogenicity and the functionality of somatic variants of KRAS mutations in NSCLC (oncogenicity)
To be able to evaluate the PD-L1 status by IHC and the KRAS status at the same time to provide in the future immune treatment and/or targeted therapy against a KRAS G12C mutation at baseline
To deal with possible tumor heterogeneity (according to the size of the sample)
To assess the gene KRAS status in non-adenocarcinoma lung carcinoma
To handle liquid biopsies at baseline and at progression in daily practice
To select an optimal gene panel
To master the turnaround time required to obtain all molecular biology and IHC results
To integrate the costs and the reimbursement according to molecular testing
To obtain accreditation according to the ISO 15,189 norm for KRAS molecular testing
To deal with the next IVDR in Europe
To be able to look for mechanisms of resistance at baseline and at progression