Dr Cortese on Methods for Evaluating immunologic Changes With Venetoclax in CLL

"We performed a comparative analysis, [and] we also did transcriptomics with RNA sequencing, epigenomics, and methylomics with reduced representation bisulfite sequencing. This assesses where methyl groups are sitting on portions of the genome and may regulate transcription, then eventually translation."

Matthew Cortese, MD, MPH, an assistant professor of oncology in the Department of Medicine–Lymphoma and the Department of Cancer Genetics and Genomics at Roswell Park Comprehensive Cancer Center, details the methods employed to conduct a multiomic analysis of immunologic changes in chronic lymphocytic leukemia (CLL) following treatment with venetoclax (Venclexta).

Early results from this analysis, which were presented at the 2024 ASH Annual Meeting, demonstrated that treatment with venetoclax at 400 mg daily was associated with immune synapse repair through complex immunomodulatory mechanisms beyond the cytotoxic clearance of malignant CLL cells. Cortese began by stating that this analysis incorporated a multiomic approach to assess the effect of venetoclax on immune cell function in 13 patients with CLL who had completed the study and reached the 400-mg daily target dose of venetoclax. Peripheral blood samples were collected within 24 hours prior to treatment initiation and again on day 30, immediately following the standard ramp-up to the target 400-mg daily dose, he described.

Multiomic analyses included bulk RNA sequencing, peripheral blood mononuclear cell RNA sequencing, T-cell sequencing by negative selection, epigenomic profiling, and methylomic analysis using reduced representation bisulfite sequencing, he noted. These analyses assessed transcriptomic and epigenetic changes relevant to gene regulation, Cortese explained.

Additionally, flow cytometric evaluation using an institutional immune reconstitution panel—originally developed for use in post-allogeneic stem cell transplantation—was employed to assess T-cell and natural killer (NK) cell function, macrophage activation, and stemness markers, Cortese continued. Results showed that after adjusting for treatment response at day 30, T cells, NK cells, and macrophages demonstrated evidence of repair and functional reconstitution, Cortese concluded.