LRRK2 is a negative regulator of Mycobacterium tuberculosis phagosome maturation in macrophages.

Anetta Härtlova,Heyne Lee,Sally A Cowley,Paul Davies,Antony Fearns,Ian G Ganley,Brian D Dill,Orsolya Bilkei‐Gorzo,Rowan Flynn,Matthias Trost,Alastair D Reith,Patrick A Lewis,Julien Peltier,Susanne Herbst,Dario R Alessi,Jennifer Martinez,Angela Rodgers,Maximiliano G Gutierrez

doi:10.15252/embj.201798694

Abstract

Mutations in the leucine‐rich repeat kinase 2 (LRRK2) are associated with Parkinson's disease, chronic inflammation and mycobacterial infections. Although there is evidence supporting the idea that LRRK2 has an immune function, the cellular function of this kinase is still largely unknown. By using genetic, pharmacological and proteomics approaches, we show that LRRK2 kinase activity negatively regulates phagosome maturation via the recruitment of the Class III phosphatidylinositol‐3 kinase complex and Rubicon to the phagosome in macrophages. Moreover, inhibition of LRRK2 kinase activity in mouse and human macrophages enhanced Mycobacterium tuberculosis phagosome maturation and mycobacterial control independently of autophagy. In vivo, LRRK2 deficiency in mice resulted in a significant decrease in M. tuberculosis burdens early during the infection. Collectively, our findings provide a molecular mechanism explaining genetic evidence linking LRRK2 to mycobacterial diseases and establish an LRRK2‐dependent cellular pathway that controls M. tuberculosis replication by regulating phagosome maturation.

Highlights

Mutations in the leucine-rich repeat kinase 2 (LRRK2) are associated with Parkinson’s disease, chronic inflammation and mycobacterial infections
We examined the function of LRRK2 in macrophages and show that LRRK2 negatively regulates phagosome maturation and that this contributes to mycobacterial replication and impaired innate immune responses
Given that LRRK2 is highly expressed in macrophages and several human genetic studies linked LRRK2 and mycobacterial diseases (Zhang et al, 2009; Wang et al, 2015, 2018; Fava et al, 2016), we investigated the effect of LRRK2 on Mycobacterium tuberculosis (Mtb) infection using bone marrow-derived mouse macrophages (BMDMs) from LRRK2 KO mice

Summary

Introduction

Mutations in the leucine-rich repeat kinase 2 (LRRK2) are associated with Parkinson’s disease, chronic inflammation and mycobacterial infections. Inhibition of LRRK2 kinase activity in mouse and human macrophages enhanced Mycobacterium tuberculosis phagosome maturation and mycobacterial control independently of autophagy. Our findings provide a molecular mechanism explaining genetic evidence linking LRRK2 to mycobacterial diseases and establish an LRRK2dependent cellular pathway that controls M. tuberculosis replication by regulating phagosome maturation. Mtb infects mostly macrophages and within these cells establishes a replicative niche by subverting the host cell and the normal process of phagosome maturation This cellular pathway, that represents the cornerstone of the innate immune system, targets Mtb to phagolysosomes where they are eventually eliminated (Levin et al, 2016). Using several independent experimental approaches, we show that phagosomal function is regulated by a LRRK2 kinase-dependent recruitment of the Class III phosphatidylinositol-3 kinase (PI3K) complex and its negative regulator Rubicon (RUN domain protein as Beclin-1 interacting and cysteine-rich containing). This study provides a cellular function underlying human genetic studies linking LRRK2 to mycobacterial infections and reveals an unexpected function for LRRK2 in macrophages and infectious diseases control

Results

Discussion

Materials and Methods