Stirparo, G. G.; Xu, X.; Thompson, T.; Page, K.; Harvey, J. R.; Cadwalladr, D.; Lawrence, J.; Burley, R. J.; Juvvanapudi, J.; Roberts, M.; Barker, D. F.; Mulligan, V.; Sherlock, C.; Lizio, M.; Christie, L.; Mudaliar, M.; Sheardown, S.; Margus, B.; Thompson, C.; Dickson, L.; Brice, N. L.; Carlton, M. B.; Powell, J. A.; Dawson, L. A.

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.