Avalilação PREreview de UBA1 Mutations Drive RIPK1-Mediated Cell Death and Monocyte Dysfunction in VEXAS Syndrome

Major issues

• The study does not establish the temporal and causal hierarchy between TLR signaling, TNF–NF-κB/cFLIP signaling, and RIPK1-mediated cell death. Time-resolved experiments and pathway-specific perturbations are needed to determine which pathway acts as the driver and which represents a downstream consequence.

• The study relies mainly on pharmacological inhibition to implicate RIPK1-mediated cell death but does not provide a clear rescue experiment demonstrating restoration of the UBA1-mutant phenotype. Restoring functional UBA1 or relevant downstream signaling (e.g., NF-κB/cFLIP) would provide stronger evidence for causality and reversibility.

• The monocyte population is treated largely as a homogeneous population. Single-cell analyses could determine whether all UBA1-mutant monocytes exhibit defects in both TLR and TNF responses, or whether distinct subpopulations display TLR-specific, TNF-specific, or combined/synergistic defects.

• The study would benefit from deeper molecular characterization of stimulated monocytes using scRNA-seq/ATAC-seq, followed by differential gene expression and pathway-level analyses (e.g., GSEA/KEGG), to identify transcriptional and chromatin programs underlying the impaired antimicrobial and inflammatory responses.

• The potential role of monocyte trafficking and migration is insufficiently investigated. Analysis of chemokine receptors such as CCR2, CXCR3, and CCR7, together with functional migration assays, could determine whether UBA1 mutations alter monocyte recruitment or tissue trafficking and how this relates to the inflammatory phenotype.

Minor issues

• The relationship between reduced cFLIP(L) expression, impaired NF-κB signaling, and RIPK1-mediated cell death could be clarified experimentally and temporally.

• The interaction between TLR and TNF signaling would benefit from direct comparison of TLR stimulation, TNF-α stimulation, and combined stimulation to distinguish additive from synergistic effects.

• Additional validation in primary UBA1-mutant monocytes, beyond the engineered THP-1 model, would strengthen the translational relevance of the findings.

Competing interests

The author declares that they have no competing interests.

Use of Artificial Intelligence (AI)

The author declares that they used generative AI to come up with new ideas for their review.