Supplementary file 1_FDX1 is associated with CALR abundance and JAK–STAT-related inflammatory signaling in myeloproliferative neoplasms.docx

Microarray-based transcriptomic analyses identified an MPN-associated cuproptosis-related and immune-response program, and consensus clustering separated samples into exploratory transcriptomic clusters. Within this expression-derived partition, post hoc multi-algorithm feature prioritization recurrently retained FDX1 and CALR among genes associated with the Cluster 1-versus-Cluster 2 transcriptional contrast. Single-cell analyses localized Calr and Fdx1 across hematopoietic and immune populations and highlighted inferred JAK–STAT-associated communication potential.

In human MPN samples, FDX1 and total CALR protein and mRNA levels were increased and accompanied by STAT5 phosphorylation and elevated inflammatory mediators. Functionally, FDX1 knockdown reduced total CALR abundance, STAT5 phosphorylation, cytokine release, sensitivity to copper-dependent cytotoxicity, and proliferation, whereas FDX1 overexpression showed the reciprocal pattern and was partly counteracted by ruxolitinib.</p>Discussion<p>These findings support an association between FDX1 perturbation, total CALR abundance, STAT5-associated signaling, and inflammatory output in MPN, but do not establish direct regulation of mutant CALR or mutant CALR–MPL signaling.</p>