Identification of alternative attenuation targets of Mycobacterium tuberculosis (Mtb) is pivotal for designing new candidates for live attenuated anti-tuberculosis (TB) vaccines. In this context, the CtpF P-type ATPase of Mtb is an interesting target; specifically, this plasma membrane enzyme is involved in calcium transporting and response to oxidative stress. We found that a mutant of MtbH37Rv lacking ctpF expression (MtbΔctpF) displayed impaired proliferation in mouse alveolar macrophages (MH-S) during in vitro infection. Further, the levels of tumor necrosis factor and interferon-gamma in MH-S cells infected with MtbΔctpF were similar to those of cells infected with the parental strain, suggesting preservation of the immunogenic capacity. In addition, BALB/c mice infected with Mtb∆ctpF showed median survival times of 84 days, while mice infected with MtbH37Rv survived 59 days, suggesting reduced virulence of the mutant strain. Interestingly, the expression levels of ctpF in a mouse model of latent TB were significantly higher than in a mouse model of progressive TB, indicating that ctpF is involved in Mtb persistence in the dormancy state. Finally, the possibility of complementary mechanisms that counteract deficiencies in Ca²⁺ transport mediated by P-type ATPases is suggested. Altogether, our results demonstrate that CtpF could be a potential target for Mtb attenuation.

Mycobacterium tuberculosis is primarily a pulmonary pathogen capable of systemic dis-semination and persistence in extrapulmonary tissues, perhaps including also the brain. To investigate this potential reservoir, we analyzed human brainstem samples from 50 forensic autopsies and brain tissue from a murine model of latent tuberculosis using PCR, RT-PCR, and RNA sequencing. Mycobacterial IS6110 insertion sequence and 16S rRNA were detected in 12/50 human samples and in all murine brains, confirming persistence. Variable expression of mycobacterial latency-associated (dosR, dosT, hspX, icl) and met-abolic adaptation genes (glcB, pcaA, phoP, mprA, treS) indicated heterogeneous dorman-cy states. Differential expression analysis revealed 19 modulated genes in human brain-stems (9 upregulated, 10 downregulated) and 2,700 in murine brains (1,509 upregulated, 1,191 downregulated). Metallothioneins, USP9(Y/X), and RAB27B showed similar expres-sion in human and mice brain tissue, whereas MAPK15 and calbindin2 displayed dis-cordant patterns. Functional enrichment in mice demonstrated a shift from resident neu-ral cell identity, activating translational and mitochondrial pathways while repressing synaptic signaling. These findings provide molecular evidence that M. tuberculosis per-sists in the central nervous system in a dormancy-associated state. The observed synaptic repression overlaps with neurodegenerative processes, whereas concurrent translational activation suggests a distinct adaptive host response, highlighting the need to explore chronic neuroinflammation linked to bacterial persistence.