High-throughput cell viability assessment using a luminescent ATP detection assay

Chronic myelomonocytic leukemia (CMML) is an aggressive hematologic malignancy for which no approved targeted therapies are available. Standard therapies as of now lack the ability to eradicate leukemic clones nor can they modify disease outcome. To address this therapeutic gap, we conducted a high-throughput chemical drug screen using a robotized platform on four genetically engineered hematopoietic murine cell lines that model recurrent CMML mutations. Our approach aimed to identify genotype-dependent therapeutic vulnerabilities through a drug repurposing strategy, leveraging the rapid clinical translation potential of pre-approved compounds. Cell viability was assessed using the CellTiter-Glo® luminescent cell viability assay, a robust method widely recognized for its accuracy and efficiency in measuring cell proliferation and cytotoxicity. The luminescent output, directly proportional to viable cell numbers, allows for high-throughput screening. The protocol detailed in this article enables the testing of a large number of compounds at multiple doses, with a 48-hour readout. This method provides a reliable, time-efficient, and flexible approach that can be adapted for different cell types and research contexts beyond oncology.