Pharmacological targeting of host chaperones protects from pertussis toxin in vitro and in vivo

Ernst, K.; Mittler, A.-K.; Winkelmann, V.; Eberhardt, N.; Anastasia, A.; Sonnabend, M.; Lochbaum, R.; Wirsching, J.; Skerry, C.; Carbonetti, N. H.; Frick, M.; Barth, H.

doi:10.1101/2020.09.24.303321

bioRxiv · 10.1101/2020.09.24.303321

Pharmacological targeting of host chaperones protects from pertussis toxin in vitro and in vivo

Ernst, K.·Mittler, A.-K.·Winkelmann, V.·Eberhardt, N.·Anastasia, A.·Sonnabend, M.·Lochbaum, R.·Wirsching, J.·Skerry, C.·Carbonetti, N. H.·Frick, M.·Barth, H.

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Abstract

Whooping cough is caused by Bordetella pertussis that releases pertussis toxin (PT) which comprises enzyme A-subunit PTS1 and binding/transport B-subunit. After receptor-mediated endocytosis, PT reaches the endoplasmic reticulum from where unfolded PTS1 is transported to the cytosol. PTS1 ADP-ribosylates G-protein -subunits resulting in increased cAMP signaling. Here, the role of target cell chaperones Hsp90, Hsp70, cyclophilins and FK506-binding proteins for cytosolic PTS1-uptake is characterized in detail. PTS1 specifically and directly interacts with chaperones in vitro and in cells. Specific pharmacological chaperone inhibition protects CHO-K1, human primary airway basal cells and a fully differentiated airway epithelium from PT-intoxication by reducing cytosolic PTS1-amounts without affecting cell binding or enzyme activity. PT is internalized by human airway epithelium secretory but not ciliated cells and leads to increase of apical surface liquid. Cyclophilin-inhibitors reduced leukocytosis in infant mouse model of pertussis, indicating their promising potential for developing novel therapeutic strategies against whooping cough.

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Ernst, K., Mittler, A.-K., Winkelmann, V., Eberhardt, N., Anastasia, A., Sonnabend, M., Lochbaum, R., Wirsching, J., Skerry, C., Carbonetti, N. H., Frick, M., Barth, H.. 2020-09-25. Pharmacological targeting of host chaperones protects from pertussis toxin in vitro and in vivo. https://doi.org/10.1101/2020.09.24.303321

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