Production of nanobodies raised against transmembrane proteins that are difficult to purify still is a technical challenge. Among those are connexin43 cCx43) and pannexin1 (Panx1), which form (hemi) channels in the plasma membrane. These (hemi)channels mediate extracellular signaling and have recently emerged as key players in inflammation. This carries translational relevance, as (hemi) channel inhibition could represent a novel strategy for the treatment of a plethora of diseases.
However, clinical exploration in this direction is impeded by the lack of sound (hemi)channel inhibitors. This project will assist in overcoming this hurdle by generating nanobodies targeted against Cx43 and Panx1, thereby relying on an unprecedented protocol that combines DNA immunization with innovative nanodisc technology. By using an in vitro/ex vivo/in vivo approach, the Cx43 and Panx1 nanobodies will be tested for their potential to inhibit (hemi)channels and to alleviate pathological features in mouse models of epidemiologically relevant inflammatory disorders in the liver and the cardiovascular system. This will be accomplished by joining scientists from the bio-engineering and (bio)medical/pharmaceutical fields in a Belgian-Swiss consortium. As such, this project lays the basis for follow-up initiatives not only to scrutinize the versatile antiinflammatory therapeutic actions of the Cx43 and Panx1 nanobodies, but also to develop nanobodies raised against similar challenging targets.