Worm product-based therapies for prevention of bloodbrain barrier breakdown

Background:
Blood-brain barrier (BBB) dysfunction has a major role in small vessel disease (SVD) due to the infiltration of blood-borne substances and inflammatory cells. Studies show that synthetic small molecule analogues (SMAs) 11a and 12b, derived from ES-62, an anti-inflammatory molecule secreted by the parasitic worm Acanthocheiloneama viteae, have potent barrier protecting and anti-inflammatory properties at peripheral sites.

Aims:
1. To determine if SMA11a and SMA12b reduce BBB breakdown and pro-inflammatory responses in vivo using Col4a1+/Svc mutant mice, a relevant mouse model for SVD.
2. To elucidate SMA11a- and SMA12b-mechanism(s) of action in vitro using Col4 gene-edited human brain microvascular endothelial cells (Col4-HBMECs) and mouse microglia (SIM-A9s).

Methods:
Treatments incorporated administration of SMA11a, SMA12b versus vehicle, with and without exacerbation by interleukin-1β (IL-1β), a cytokine associated with cerebral SVD. Randomisation and blinding were implemented.
1. Microbleeds, BBB proteins, inflammatory cells and cytokines will be measured in female adult Col4a1+/Svc and Col4a2 mutant mice using quantitative histopathology and immunofluorescence, immunoblotting and ELISAs.
2. Migration, angiogenesis, proliferation, BBB proteins and cytokines will be measured in Col4-HBMECs and, where appropriate SIM-A9s, using in vitro wound healing assay, tube formation assay, cell proliferation assay, immunofluorescence, and cytokine arrays.

Results/Conclusions:
This is preliminary work, in progress. We envisage BBB components (occludin, claudin-5, collagen IV, and laminin) to be increased and inflammatory responses to be decreased by ES-62 SMAs versus vehicle in vivo and in vitro. For the first time, we will determine the efficacy of parasitic worm-related therapies in ameliorating SVD and reveal urgently required novel strategies that reverse or repair BBB breakdown