NINJ1 is activated by Mycobacterium tuberculosis ESX-1 secreted effector EsxA and mediates necrosis of infected human macrophages
Lytic cell death contributes to tissue damage and dissemination during Mycobacterium tuberculosis (Mtb) infection, yet the mechanisms governing plasma membrane rupture (PMR) remain incompletely defined. Here, we identify Ninjurin-1 (NINJ1), a mediator of PMR, as a central effector of macrophage lysis during Mtb infection. Mtb infection induced NINJ1 oligomerization, and genetic deletion or pharmacological inhibition of NINJ1 markedly reduced lactate dehydrogenase (LDH) release. Cytokine secretion was largely preserved in NINJ1-deficient macrophages, with the exception of CXCL10. Unexpectedly, inhibition of pyroptosis, apoptosis, necroptosis, and ferroptosis - individually or in combination - did not prevent NINJ1 activation or PMR, indicating that Mtb-induced membrane rupture proceeds independently of canonical regulated cell death pathways. However, NINJ1 oligomerization and PMR required the Mtb ESX-1 secretion system, identifying a bacterial virulence determinant as a critical upstream trigger. Although osmoprotective PEG partially reduced LDH release, neither calcium signaling nor cell swelling accounted for NINJ1-dependent PMR in Mtb-infected macrophages. Together, these findings establish NINJ1 as a key executioner of Mtb-induced lytic cell death and reveal an ESX-1-dependent pathway of PMR that is uncoupled from canonical host cell death programs. TeaserNINJ1 mediates plasma membrane rupture of macrophages infected with Mycobacterium tuberculosis