Amyloid beta 1‐40 and 1‐42 fibril ratios and maturation level cause conformational differences with minimal impact on autophagy and cytotoxicity.

Johansson, Lovisa; Sandberg, Alexander; Nyström, Sofie; Hammarström, Per; Hallbeck, Martin

The amyloid β (Aβ) peptide has a central role in Alzheimer's disease (AD) pathology. The peptide length can vary between 37 and 49 amino acids, with Aβ1‐42 being considered the most disease‐related length. However, Aβ1‐40 is also found in Aβ plaques and has shown to form intertwined fibrils with Aβ1‐42. The peptides have previously also shown to form different fibril conformations, proposed to be related to disease phenotype. To conduct more representative in vitro experiments, it is vital to uncover the impact of different fibril conformations on neurons. Hence, we fibrillized different Aβ1‐40:42 ratios in concentrations of 100:0, 90:10, 75:25, 50:50, 25:75, 10:90 and 0:100 for either 24 h (early fibrils) or 7 days (aged fibrils). These were then characterized based on fibril width, LCO‐staining and antibody‐staining. We further challenged differentiated neuronal‐like SH‐SY5Y human cells with the different fibrils and measured Aβ content, cytotoxicity and autophagy function at three different time‐points: 3, 24, and 72 h. Our results revealed that both Aβ1‐40:42 ratio and fibril maturation affect conformation of fibrils. We further show the impact of these conformation changes on the affinity to commonly used Aβ antibodies, primarily affecting Aβ1‐40 rich aggregates. In addition, we demonstrate uptake of the aggregates by neuronally differentiated human cells, where aggregates with higher Aβ1‐42 ratios generally caused higher cellular levels of Aβ. These differences in Aβ abundance did not cause changes in cytotoxicity nor in autophagy activation. Our results show the importance to consider conformational differences of Aβ fibrils, as this can have fundamental impact on Aβ antibody detection. Overall, these insights underline the need for further exploration of the impact of conformationally different fibrils and the need to reliably produce disease relevant Aβ aggregates.

PEPTIDES; ALZHEIMER'S disease; CYTOTOXINS; AMINO acids; AMYLOID

Journal of Neurochemistry, 2024, Vol 168, Issue 9, p3308

0022-3042

Academic Journal

10.1111/jnc.16201