NGS and molecular profiling in-patient with Metastatic Colorectal cancer to Guide personalized Clinical Traials and Responding to Anti–PD-1/PD-L1 Therapy

Introduction: The major milestone in oncology was genomics transcription in 2008 with next-generation sequencing (NGS). In these studies, NGS used to analyze the targeted panel of protein-coding genes of the MCRC Patient tumor genome. Then preclinical cancer model therapies and phenotyping analyses performed related to patient first diagnosis and treatment lines in June 2013. There are multiple factors influencing the actual success rate when using personalized molecular diagnostics and treatments. For example, one major hurdle for precision medicine is drug access, as therapies beyond standard of care are often not available, i.e., not approved or not covered by health insurances, and access to clinical trials is limited. Methods: A tumor biopsy and preferably a matched normal control sample (blood) undergoes DNA and RNA extraction, followed by sequencing. Afterwards, the samples analyzed with regard to somatic alterations, i.e., genetic changes that appear in the tumor sample, but not in the normal tissue. Our main objective is to identify targetable aberrations and to recommend new therapy option for patient. Therefore, somatic variants such as single nucleotide variants (SNVs), small insertions and deletions (InDels), and copy number variants (CNVs) were of particular interest. Then development and validation of large molecule assays for pharmacokinetic (PK), anti-drug antibody assay and drug resistance analyses performed on preclinical patient tumor model. In addition, the overall mutational burden as an indicator of response to immunotherapy in metastatic colorectal cancer is determined based on patient NGS data. Results: In this study, we were able to identify actionable variants and corresponding therapies. We identified resistance-associated variants explaining lack of therapy response. The report indicated a high mutational load of NBPF1, MUTYH, PPIAL4G, NBPF9, and TMOD4 with amplification of the MST1L and deletion of PMS2 in the patient NGS analysis. Gene expression profile of patient programmed death ligand 1 (PD-L1) was explored in a dataset of tumor sample. PD-L1 expression was highly concordant with several genomic signatures indicative of immune-inflamed tumor microenvironment. Therefore, patients treated with immune checkpoint blockade (Pembrolizumab), which resulted in a complete response within 6 months. Which shown in supplementary data.Conclusion: Our workflow covers the way towards a more frequent use of comprehensive molecular diagnostics, which permits for the detection of genomic aberrations involved in carcinogenesis and therapy resistance. This platform objects to isolate a targetable molecular alteration in patients with metastatic colorectal cancer sponsored by the cooperative group of French Cancer Centers (UNICANCER) Gustave Roussy and funded by patient.

کلیدواژه ها:

Molecular diagnostics ، NGS ، Personalized medicine ، Metastatic colorectal Cancer diagnostics