Bioinformatic strategies for the analysis of genomic aberrations detected by targeted NGS panels with clinical application

PeerJ. 2021 Mar 31:9:e10897. doi: 10.7717/peerj.10897. eCollection 2021.


Molecular profiling of tumor samples has acquired importance in cancer research, but currently also plays an important role in the clinical management of cancer patients. Rapid identification of genomic aberrations improves diagnosis, prognosis and effective therapy selection. This can be attributed mainly to the development of next-generation sequencing (NGS) methods, especially targeted DNA panels. Such panels enable a relatively inexpensive and rapid analysis of various aberrations with clinical impact specific to particular diagnoses. In this review, we discuss the experimental approaches and bioinformatic strategies available for the development of an NGS panel for a reliable analysis of selected biomarkers. Compliance with defined analytical steps is crucial to ensure accurate and reproducible results. In addition, a careful validation procedure has to be performed before the application of NGS targeted assays in routine clinical practice. With more focus on bioinformatics, we emphasize the need for thorough pipeline validation and management in relation to the particular experimental setting as an integral part of the NGS method establishment. A robust and reproducible bioinformatic analysis running on powerful machines is essential for proper detection of genomic variants in clinical settings since distinguishing between experimental noise and real biological variants is fundamental. This review summarizes state-of-the-art bioinformatic solutions for careful detection of the SNV/Indels and CNVs for targeted sequencing resulting in translation of sequencing data into clinically relevant information. Finally, we share our experience with the development of a custom targeted NGS panel for an integrated analysis of biomarkers in lymphoproliferative disorders.

Keywords: Bioinformatic analysis; CNV; Clinical application; Molecular markers; NGS; SNV/indel; Targeted panels.

Grants and funding

This work was supported by projects MH CR AZV NV19-03-00091, MUNI/A/1395/2019, and European Regional Development Fund-Project “A-C-G-T” (MEYS No. CZ.02.1.01/0.0/0.0/16_026/0008448). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.