Precision Medicine in Oncohematology: Integration of Cytogenomic, Molecular, and Next-Generation Sequencing Techniques in Clinical Diagnosis. A Case Report at the LIME Laboratory

Abstract

Introduction: The diagnosis of hematologic neoplasms is traditionally carried out using molecular biology, flow cytometry, and cytogenomic techniques. The integration of next-generation sequencing (NGS) is crucial to complement diagnosis, estimate prognosis, define medical management, and identify new biomarkers.

Objective: To demonstrate the importance of integrating NGS, cytogenomic, and molecular tests in the diagnosis and prognosis of hematologic neoplasms.

Methodology: Cytogenomic studies, PCR, Sanger sequencing, and NGS using the TruSight RNA Pan-Cancer panel were conducted on two patients seen at LIME.

Results: Case 1: An 83-year-old patient suspected of secondary acute myeloid leukemia (AML) due to treatment. Molecular test results for the genes MLP, JAK2E12, CALR, TP53, and BCR/ABL-P190 were negative. Cytogenetics showed a complex karyotype (monosomy 7 and trisomy 21). NGS analysis detected three pathogenic/probably pathogenic variants (IDH2, PTPN11, and RUNX1) associated with an adverse prognosis.

Case 2: An 80-year-old patient suspected of chronic myelomonocytic leukemia (CMML). Molecular test results were negative. Cytogenetics showed a normal karyotype. NGS analysis identified two pathogenic/probably pathogenic variants (RUNX1 and CSF3R) associated with an adverse prognosis. A novel mutation was found in the CEBPA gene (NM_004364.5): c.1070_1071insTGCCGG.

Conclusions: Through the implementation of NGS, pathogenic variants were detected that were not identified by classical molecular and cytogenomic methods. Additionally, prognosis was estimated, and a new variant in CEBPA was identified. These results underscore the need to integrate NGS tests in cases where classical methodologies are insufficient.