Multimodal spatial transcriptomics redefines cortical pathology in Huntington's disease

Huntington's disease (HD) is caused by CAG expansion in the huntingtin gene (HTT), yet how somatic repeat instability and HTT inclusions relate to selective cell loss in the human brain remains unclear. We have developed a multimodal spatial transcriptomics approach that enables the definition of transcriptional programs with subcellular resolution, somatic CAG repeat lengths, and eight other pathology markers, including HTT inclusions, in every cell of intact brain sections. Imaging 428,173 cells in the control and HD cortex revealed the selective vulnerability of layer 5-6 (L5-6) near-projecting (NP) neurons and L6b deep-layer excitatory neurons, which undergo a >50% loss that is closely linked to large somatic repeat expansions (>450 ± 55 CAGs). Intranuclear inclusions were most prevalent at intermediate somatic repeat expansions (250-350 CAGs) and were associated with broader transcriptional changes than somatic repeat expansion alone. These data provide a comprehensive resource and establish a general framework for connecting repeat expansion and protein pathology across diverse cell types.