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Dissecting the regulation, structural dynamics, and physiological role of human Ninj1 activity during Mtb-induced cell death on a molecular scale.

Objective

Mycobacterium tuberculosis (Mtb) causes tuberculosis a leading infectious killer worldwide. Mtb primarily infects macrophages, leading to the death of some of these cells via distinct regulated cell death (RCD) mechanisms. While apoptosis facilitates Mtb elimination, necrotic cell deaths, like pyroptosis, necroptosis, and ferroptosis may enhance bacterial replication and spread. Mtb effectors, including the type VII secretion system ESX-1 and the cell wall lipid PDIM, induce host membrane damage, inducing necrotic cell death. Notably, the host lab has shown that inhibiting all RCD pathways does not prevent Mtb-induced cell lysis. However, depleting Ninjurin-1 (Ninj1), a membrane protein vital for plasma membrane (PM) rupture during various RCD pathways, mitigates Mtb-induced necrosis. Ninj1 oligomerization appears to occur after initial PM integrity disruption, yet the precise mechanisms governing Ninj1 activation and its role in Mtb-induced cell lysis remain unclear. I hypothesize that Ninj1 plays a crucial role in Mtb-induced lysis and that Mtb exploits this process through effector-mediated PM permeabilization. To investigate how Ninj1 oligomerization influences cell fate and the inflammatory response, I will leverage my expertise in super-resolution microscopy of pore-forming proteins alongside the host lab’s skills in Mtb infection biology. First, I will identify PM chemical and physical changes during RCD that correlate with Ninj1 oligomerization to elucidate the activation mechanism. Next, I will analyze Ninj1 dynamics and the functional impact of its macromolecular structures using DNA-PAINT super-resolution microscopy and long-term single-molecule tracking. Finally, I will determine whether Mtb activates Ninj1 via effector-induced PM damage and quantify the impact of this activation on the inflammatory response. This project aims to provide groundbreaking insights into cellular responses to Mtb and molecular inflammation research.

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Topic(s)

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HORIZON-TMA-MSCA-PF-EF - HORIZON TMA MSCA Postdoctoral Fellowships - European Fellowships

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Call for proposal

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(opens in new window) HORIZON-MSCA-2024-PF-01

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Coordinator

NORGES TEKNISK-NATURVITENSKAPELIGE UNIVERSITET NTNU
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 267 418,56
Address
HOGSKOLERINGEN 1
7491 TRONDHEIM
Norway

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Region
Norge Trøndelag Trøndelag
Activity type
Higher or Secondary Education Establishments
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Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

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