AUTHOR=Stirparo Giuliano G. , Xu Xiao , Thompson Toni , Page Keith , Harvey Jenna R. M. , Cadwalladr David , Lawrence Jason , Burley Russell J. , Juvvanapudi Joel , Roberts Megan , Barker Daniel F. , Mulligan Victoria , Sherlock Chloe , Lizio Marina , Christie Louisa , Mudaliar Mani , Sheardown Steven , Margus Brad , Thompson Craig , Dickson Louise , Brice Nicola L. , Carlton Mark B. , Powell Justin A. C. , Dawson Lee A. 

TITLE=NETSseq reveals inflammatory and aging mechanisms in distinct cell types, driving cerebellar decline in ataxia telangiectasia

JOURNAL=Frontiers in Neuroscience

VOLUME=Volume 19 - 2025

YEAR=2025

URL=https://www.frontiersin.org/journals/neuroscience/articles/10.3389/fnins.2025.1636787

DOI=10.3389/fnins.2025.1636787

ISSN=1662-453X

ABSTRACT=Ataxia–telangiectasia (A–T) is a rare, autosomal recessive, multisystem disorder caused by mutations in the Ataxia–Telangiectasia Mutated (ATM) gene and is characterized by a devastating and progressive neurological pathology. The cellular and molecular changes driving the neurological abnormalities associated with A-T are not well understood. Here, we applied our proprietary Nuclear Enriched Transcript Sort sequencing (NETSseq) platform to investigate changes in cell type composition and gene expression in human cerebellar post-mortem tissue from A-T and control donors. We found dysregulation in neurotransmitter signaling in granule neurons, potentially underlying the impaired motor coordination in A-T. Astrocytes and microglia have evidence of accelerated aging, with astrocytes being characterized by neurotoxic signatures, while microglia showed activation of DNA damage response pathways. Compared to single-nuclei technologies, NETSseq provided a more robust detection of genes with low abundance, a higher cell type specific expression pattern, and significantly lower levels of cross-contamination. These findings highlight the importance of NETSseq as a resource for investigating mechanisms and biological processes associated with disease, providing high-sensitivity, cell-specific insights to advance targeted therapies for neurodegenerative diseases.