Optineurin Deficiency Collapses the Host Endolysosomal Network and Impairs Xenophagy to Accelerate Mycobacterium tuberculosis Growth

Selective autophagy is a host defense mechanism against Mycobacterium tuberculosis (Mtb) that restricts bacterial growth by targeting ubiquitin-coated bacilli for lysosomal degradation via autophagy receptors. Optineurin is a selective autophagy receptor that targets pathogens and modulates immune signaling; however, its precise structural mechanism during Mtb infection remains poorly defined. Here, we show that while Optineurin deficiency spares the global host transcriptomic response to infection, it collapses the host endolysosomal network, reducing LAMP1+ and LysoTracker+ reserves by half. Multi-dose bafilomycin A1 flux assays demonstrated that this structural depletion selectively blocks the dynamic, directional trafficking and functional delivery of autophagosomes to the pathogen, significantly reducing Mtb-DQ-BSA colocalization. Genetic complementation restored bacterial restriction in a manner dependent on three phosphosites (Ser187, Ser530, and the uncharacterized Ser556). In the context of reduced autophagic containment and increased Mtb replication, Optineurin deficiency accelerated necrotic-like host cell death. In vivo, Optineurin deficiency enhanced bacterial replication and impaired the Type I interferon response during acute Mtb infection but did not affect long-term survival. Together, these findings identify Optineurin as a critical regulator of autophagic flux, host cell death, and Type I interferon responses that limit early Mtb pathogenesis.