Revealing the Mysteries of Acute Myeloid Leukemia: From Quantitative PCR through Next-Generation Sequencing and Systemic Metabolomic Profiling

Abstract

The efforts made in the last decade regarding the molecular landscape of acute myeloid leukemia (AML) have created the possibility of obtaining patients' personalized treatment. Indeed, the improvement of accurate diagnosis and precise assessment of minimal residual disease (MRD) increased the number of new markers suitable for novel and targeted therapies. This progress was obtained thanks to the development of molecular techniques starting with real-time quantitative PCR (Rt-qPCR) passing through digital droplet PCR (ddPCR) and next-generation sequencing (NGS) up to the new attractive metabolomic approach. The objective of this surge in technological advances is a better delineation of AML clonal heterogeneity, monitoring patients without disease-specific mutation and designing customized post-remission strategies based on MRD assessment. In this context, metabolomics, which pertains to overall small molecules profiling, emerged as relevant access for risk stratification and targeted therapies improvement. In this review, we performed a detailed overview of the most popular modern methods used in hematological laboratories, pointing out their vital importance for MRD monitoring in order to improve overall survival, early detection of possible relapses and treatment efficacy.

Keywords: MDR; RT-qPCR; acute myeloid leukemia (AML); digital droplet PCR (ddPCR); metabolomic profiling; metabolomics analysis; next-generation sequencing (NGS).

Figure 1

Workflow of diagnostic methods used in “de novo” or relapsed form of AML, as well as its MRD assessment. This scheme highlights the network of methods used in hematology laboratories and the main targets tested, divided into four levels: cytogenetic and RT-qPCR, flow cytometry, digital PCR and next generation sequencing (NGS).

Figure 2

Schematic comparison of “omic” technologies with their main advantages and disadvantages.