AUTHOR=Gil José Vicente , Miralles Alberto , de las Heras Sandra , Such Esperanza , Avetisyan Gayane , Díaz-González Álvaro , Santiago Marta , Fuentes Carolina , Fernández José María , Lloret Pilar , Navarro Irene , Montesinos Pau , Llop Marta , Barragán Eva 

TITLE=Comprehensive detection of CRLF2 alterations in acute lymphoblastic leukemia: a rapid and accurate novel approach

JOURNAL=Frontiers in Molecular Biosciences

VOLUME=11

YEAR=2024

URL=https://www.frontiersin.org/journals/molecular-biosciences/articles/10.3389/fmolb.2024.1362081

DOI=10.3389/fmolb.2024.1362081

ISSN=2296-889X

ABSTRACT=<p><bold>Introduction:</bold> Acute lymphoblastic leukemia (ALL) is a prevalent childhood cancer with high cure rate, but poses a significant medical challenge in adults and relapsed patients. Philadelphia-like acute lymphoblastic leukemia (Ph-like ALL) is a high-risk subtype, with approximately half of cases characterized by <italic>CRLF2</italic> overexpression and frequent concomitant <italic>IKZF1</italic> deletions.</p><p><bold>Methods:</bold> To address the need for efficient, rapid, and cost-effective detection of <italic>CRLF2</italic> alterations, we developed a novel RT-qPCR technique combining SYBR Green and highresolution melting analysis on a single plate.</p><p><bold>Results:</bold> The method successfully identified <italic>CRLF2</italic> expression, <italic>P2RY8::CRLF2</italic> fusions, and <italic>CRLF2</italic> and <italic>JAK2</italic> variants, achieving a 100% sensitivity and specificity. Application of this method across 61 samples revealed that 24.59% exhibited <italic>CRLF2</italic> overexpression, predominantly driven by <italic>IGH::CRLF2</italic> (73.33%). High Resolution Melting analysis unveiled concurrent <italic>CRLF2</italic> or <italic>JAK2</italic> variants in 8.19% of samples, as well as a dynamic nature of <italic>CRLF2</italic> alterations during disease progression.</p><p><bold>Discussion:</bold> Overall, this approach provides an accurate identification of <italic>CRLF2</italic> alterations, enabling improved diagnostic and facilitating therapeutic decision-making.</p>