Distinct neuroinflammatory signatures exist across genetic and sporadic ALS cohorts

Abstract

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterised by progressive loss of upper and lower motor neurons. ALS is on a pathogenetic disease spectrum with frontotemporal dementia (FTD), with patients sometimes experiencing elements of both conditions (ALS-FTSD). For mutations associated with ALS-FTSD, such as the C9orf72 hexanucleotide repeat expansion (HRE), the factors influencing where an individual may lie on this spectrum require further characterisation. Here, using NanoString molecular barcoding with a panel of 770 neuroinflammatory genes, we interrogate inflammatory dysregulation at the level of gene expression. We identified 20 dysregulated neuroinflammatory genes in the motor cortex of deeply clinically phenotyped C9-ALS post-mortem cases, with enrichment of microglial and inflammatory response gene sets. Our analyses also revealed two distinct ALS-related neuroinflammatory panel signatures (NPS), NPS1 and NPS2, delineated by the direction of expression of proinflammatory, axonal transport and synaptic signalling pathways. Two genes with significant correlations to available clinical metrics were selected for validation: FKBP5 and BDNF. FKBP5 and its signalling partner, NF-κB, appeared to have a cell-type-specific staining distribution, with activated (i.e., nuclear) NF-κB immunoreactivity in C9-ALS. Expression of BDNF, a correlate of disease duration, was confirmed to be higher in individuals with long compared to short disease duration using BaseScope(TM) in situ hybridisation. Finally, we compared NPS between C9-ALS cases and those from deeply clinically phenotyped sporadic ALS (sALS) and SOD1-ALS cohorts, with NPS1 and NPS2 appearing across all cohorts. A subset of these signatures was also detected in publicly available RNA-sequencing data from independent C9-ALS and sALS cohorts, underscoring the relevance of these pathways across cohorts. Our findings highlight the importance of tailoring therapeutic approaches based on distinct molecular signatures that exist between and within genetic and sporadic cohorts.

Microglial CD68 and L-ferritin upregulation in response to phosphorylated-TDP-43 pathology in the amyotrophic lateral sclerosis brain

“…It remains unclear whether disease-associated changes in microglial function, and potential neuroprotective, dysfunctional, or neurotoxic effects, are consistent across ALS genotypes [42,44].…”

“…It remains unclear whether disease-associated changes in microglial function, and potential neuroprotective, dysfunctional, or neurotoxic effects, are consistent across ALS genotypes [42,44]. While pTDP-43 pathology occurs in 97% of ALS cases, mutations in SOD1, C9orf72 , or FUS genes cause unique neuropathology that may drive distinct microglial responses.…”

“…Microglial Iba1 and CD68 expression are increased in human ALS brain regions with high TDP-43 pathological load and neuronal loss [41]. Furthermore, CD68 expression in ALS is strongly correlated with motor cortex phosphorylated TDP-43 (pTDP-43) pathology load [25,42], disease progression, and upper motor neuron clinical symptoms [43,44]. However, several of these human studies lacked age-matched control cases [41,43] and the small number of microglial markers examined leave unclear whether microglial functional changes in response to TDP-43 pathology are neuroprotective or neurotoxic.…”