Dupilumab rebalances immune response in pediatric type-2 disorders: impacts on t-cell clones and functionality

IntroductionDupilumab demonstrates efficacy in treating a variety of Type-2 disorders refractory to conventional therapies. Improvement in clinical outcomes establishes a need to investigate mechanistic pathways of potential immunomodulation.Methods11 patients, two with moderate-to-severe asthma and 9 with moderate-top-severe AD were recruited to give blood samples pre and three months post-Dupilumab. Longitudinal phenotyping of leukocytes using mass cytometry (CyTOF) and lineage tracing of single-cells using mitochondrial single-cell ATAC-seq (mt-scATAC-seq) was conducted in PBMC and purified T-cells. Clones were inferred using a community-based detection algorithm to cluster cells based on mitochondrial variant allele-frequency (VAF). Gene scores were computed based on open-chromatin regions in the gene and promoter. Differential expression pre- and post-dupilumab was done for each cell cluster population.ResultsCyTOF immunophenotyping showed increased Th1 and Th17 cells following dupilumab, consistent with increased activity of IL-6, TNF, IFNa, and IFNg genes in lymphocytes by mt-scATAC-Seq. Rebalancing of adaptive immunity was accompanied by upregulation of Myc, E2F, MTORC1, PI3K-AKT, and p53 signaling intermediates, and was associated with upregulation of genes controlling apoptosis, unfolded protein response, and oxidative phosphorylation. Changes in frequency of T-cell clonal lineages in vivo were detected following dupilumab.ConclusionData supports clone-specific effects of dupilumab in pediatric Type-2 disorders. This approach enables analysis of changes in gene networks in T-cell clones and populations following treatment. Upregulation of non-Type-2 signaling and changes at the clonal level suggest Dupilumab restores balance to the immune response.