Aβ-42 sidechain deamidation at Q15 and N27 modulate protein aggregation and microglial responses via altered cytokine production and CD68 expression
The progressive aggregation of amyloid beta (A{beta}) monomers into oligomers is a critical factor in Alzheimers disease (AD) pathogenesis. Although mutated forms of A{beta} have been shown to display altered aggregation dynamics, the specific effects of deamidated A{beta} on microglial function remain understudied. Our research group previously found that the deamidated variant A{beta}-42-N27D modified A{beta} aggregation, reduced neurotoxicity, and reduced microglial reactivity, but the impact of A{beta}-42 side chain deamidation in general on such parameters remained unclear. Here, we expanded on our prior work by investigating how two site-specific A{beta}-42 mutations (Q15E & N27D), where neutral amide side chains are replaced with negatively charged carboxylic acids, affect aggregation and microglial immune response using a mouse microglial cell line. Size exclusion chromatography revealed that A{beta}-42-Q15E and A{beta}-42-N27D exhibit distinct aggregation profiles compared to A{beta}-42 wild type (WT). Multiplexed analysis of 8 cytokines secreted into the culture medium revealed that A{beta}-42-Q15E and A{beta}-42-N27D decrease the expression of inflammatory cytokines such as IL-6, IP-10, and MIP-1 relative to A{beta}-42-WT. Immunocytochemistry revealed that A{beta}-42-Q15E and A{beta}-42-N27D decrease CD68 expression relative to A{beta}-42-WT. These findings demonstrate that deamidation significantly alters A{beta}-42 aggregation and microglial activation, suggesting structural modifications to A{beta}-42 modulate inflammatory signaling in AD. This work provides a foundation for future studies on A{beta}-42 post-translational modifications as potential therapeutic targets in AD.S