Project Details
Projekt
Understanding genotype impact on post entry sorting of adenoviral capsids
Subject Area
Virology
Structural Biology
Structural Biology
Term
since 2026
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 548065690
Adenoviruses (Ads) are widely used in gene therapy and vaccine development, yet their intracellular trafficking and immune activation linked to the entry process remain incompletely understood, particularly across diverse genotypes. In our proposal we integrate in vitro and in cellulo single viral particle analyses to unravel the molecular and structural mechanisms of adenoviral endosomal escape, motor binding and its consequences for host cell responses comparing different adenoviruses. We use purified viral particles from different genotypes in a reconstituted system to dissect capsid disassembly and protein VI release, identifying environmental and host-derived triggers for endosomal penetration and motor binding. These mechanistic insights are coupled with high-resolution electron cryo-microscopy (cryoEM) to capture transient structural intermediates during uncoating. In parallel, we apply advanced correlative light and electron cryo-microscopy (cryoCLEM) and electron cryo-tomography (cryoET) in infected cells to visualize virus-membrane interactions under near-native conditions. This, in cellulo approach offers critical context to the in vitro findings, allowing us to resolve virus-induced membrane disruption events, motor engagement, and recruitment of host sensing machinery at molecular resolution. The integration of these methods provides a complementary framework that links structural dynamics to biological function across multiple adenovirus genotypes, including clinically and technologically relevant types such as HAd-C5, HAd-B7, and HAd-D26. Lastly our unique approach is used to identify genotype-specific variations in post-entry sorting, autophagy regulation, and innate immune activation. By correlating these findings with capsid stability and escape timing, we provide context to the rational design of tailored adenoviral vectors for gene therapy, vaccination, and oncolytic applications. As such our proposal contributes to the broader efforts of this collaborative AdBHealth consortium investigating fundamental mechanisms, clinical implications, and vector development across adenovirus genotypes.
DFG Programme
Research Units
Subproject of
FOR 5898:
AdBHealth: Raising Knowledge & Boosting Technology - Advancing Adenovirus Biology for a Healthy Future
International Connection
France
