Optimization of nanobodies targeting pannexin1 channels for the treatment of cholestasis.

Cellular channels consisting of pannexin1 (Panx1) support paracrine communication and play key
roles in inflammation, including in liver. Compounds closing these channels are considered promising
new drugs for the treatment of inflammatory disorders. In this respect, the host laboratory has
recently developed Panx1 channel inhibitors based on nanobody technology, which counteract
inflammation accompanying acute liver disease in mice. This doctoral thesis project will build further
on this outcome by optimizing the de novo generated Panx1 nanobodies in terms of potency and
kinetic profile. First, the Panx1 nanobodies will be refined to display optimal human-like and
(thermo)stability characteristics. Subsequently, these optimized Panx1 nanobodies will be further
enhanced by generating bivalent and biparatopic nanobodies. These advanced Panx1 nanobodies will
be fused to a serum albumin-binding nanobody or Fc tail in order to increase in vivo half-life. By
combining in vitro and in vivo approaches, the resulting advanced Panx1 nanobodies will be tested
for their capacity to alleviate chronic liver disease. Focus will be hereby put on cholestasis, which
results from the accumulation of bile acids and that is associated with a prominent inflammatory
response. By doing so, this doctoral thesis project is considered of high relevance to the
pharmaceutical field, as it will provide a significant contribution to the development of new drugs for
the treatment of liver disease.