Marques, M. P. | bioRxiv Science

bioRxiv2025

In-situ glial cell-surface proteomics identifies pro-longevity factors in Drosophila

Much focus has shifted towards understanding how glial dysfunction contributes to age-related neurodegeneration due to the critical roles glial cells play in maintaining healthy brain function. Cell-cell interactions, which are largely mediated by cell-surface proteins, control many critical aspects of development and physiology; as such, dysregulation of glial cell-surface proteins in particular is hypothesized to play an important role in age-related neurodegeneration. However, it remains technically difficult to profile glial cell-surface proteins in intact brains. Here, we applied a cell-surface proteomic profiling method to glial cells from intact brains in Drosophila, which enabled us to fully profile cell-surface proteomes in-situ, preserving native cell-cell interactions that would otherwise be omitted using traditional proteomics methods. Applying this platform to young and old flies, we investigated how glial cell-surface proteomes change during aging. We identified candidate genes predicted to be involved in brain aging, including several associated with neural development and synapse wiring molecules not previously thought to be particularly active in glia. Through a functional genetic screen, we identified one surface protein, DIP-{beta}, which is down-regulated in old flies and can increase fly lifespan when overexpressed in adult glial cells. We further performed whole-head single-nucleus RNA-seq and revealed that DIP-{beta} overexpression mainly impacts glial and fat cells. We also found that glial DIP-{beta} overexpression was associated with improved cell-cell communication, which may contribute to the observed lifespan extension. Our study is the first to apply in-situ cell-surface proteomics to glial cells in Drosophila, and to identify DIP-{beta} as a potential glial regulator of brain aging.

neuroscience↗

bioRxiv2025

miR-34c-5p is a novel regulator of T cell differentiation that targets FOXP3

MicroRNAs regulate key genes and pathways essential for T cell differentiation and function; however, many remain poorly characterized. We previously identified miR-34c-5p as a T cell receptor-inducible microRNA in naive CD4 T cells, but its role in immune regulation remained largely unexplored. In this study, we mapped miR-34c expression across multiple sort-purified CD4 T cell subsets and found that its induction is restricted to FOXP3 cells, both natural regulatory T cells (Tregs) and cells undergoing inducible Treg (iTreg) differentiation trajectory. miR-34c expression correlates with high FOXP3 levels and is absent in conventional memory effector subsets. Functional studies using miR-34c antagomiRs reveal that miR-34c limits iTreg differentiation by restraining FOXP3 expression. Mechanistically, we show that FOXP3 activates miR-34c transcription through direct binding to its promoter, while miR-34c targets the FOXP3 3'UTR, establishing a negative feedback loop. Together, our findings identify miR-34c as a FOXP3-responsive miRNA that fine-tunes iTreg development via post-transcriptional repression of FOXP3, uncovering a novel layer of regulatory control in T cell responses.

immunology↗