Development of a p62 biodegrader for autophagy targeted degradation

ABSTRACT: Harnessing autophagy for targeted degradation of intracellular cargo is a promising extension to proteasome-based targeted protein degradation because of the capacity and versatility of lysosomes, broadening the scope of therapeutic targets. First autophagy-based degraders and glues have been identified; however, it remains unclear which component of the autophagy lysosomal pathway is most efficacious to induce the selective degradation of targets when brought into proximity. Here, we describe a platform to systematically evaluate induced proximity of autophagy candidates with mitochondria using a nanobody-based and small molecule heterobifunctional dimerizer approach. We show that induced proximity of different effectors such as autophagy cargo receptors, ATG8-like proteins or the kinases ULK1 and TBK1 are sufficient to trigger degradation of mitochondria. In contrast, self-oligomerizing autophagy cargo receptors outperform ATG8-like effectors and autophagy kinases in clearing a soluble cytosolic protein. By developing an intrabody against the autophagy cargo receptor p62, one of the strongest effectors for autophagy targeted degradation, we demonstrate that recruitment of endogenous p62 with a heterobifunctional biodegrader is sufficient to clear mitochondria. This biodegrader, however, is unable to induce degradation of soluble cytosolic proteins due to its inhibitory effect on the self-oligomerization of p62. Our study highlights the importance of avidity for autophagy targeted degradation and suggests that autophagy cargo receptors are attractive entry points for the development of heterobifunctional degraders.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hepatocyte, Cell Culture

SUBMITTER: Catrin Swantje Müller  

LAB HEAD: Catrin Swantje

PROVIDER: PXD057810 | Pride | 2025-10-16

REPOSITORIES: Pride