Single-cell and spatial profiling reveals a role for tuberculosis-induced myofibroblasts in lung immunopathology

Tuberculosis (TB) remains a global health challenge, causing ~1.3 million deaths annually. Pulmonary TB <br/>47 often leads to lung destruction and fibrosis, yet the cellular drivers of human TB immunopathology remain <br/>48 poorly defined due to limited access to relevant tissues and differences from animal models. We performed <br/>49 single-cell RNA-sequencing and spatial transcriptomics on lung tissues from TB-infected and TB-negative <br/>50 individuals in a highly endemic South African region. We identified 30 distinct immune, parenchymal, and <br/>51 stromal cell subsets, with several linked to TB pathology and corroborated through immunohistochemistry, <br/>52 flow cytometry, and an independent human lymph node granuloma cohort. Among these, we found that 53 fibroblasts were a major driver of intercellular interaction in both active TB granuloma and TB-diseased lung 54 tissue. In particular, the MMP1⁺CXCL5⁺ fibroblast subset, which expressed elevated levels of a myofibroblast55 like gene signature, was associated with severe disease, as well as higher bacterial burden in non-human 56 primate granulomas. Network analyses revealed crosstalk between MMP1⁺CXCL5⁺ fibroblasts and SPP1⁺ 57 macrophages within the granuloma cuff, which has been reported in other disease contexts, and may play an 58 important role in the immunopathology of TB. Overall, our findings highlight previously unappreciated cell 59 populations and interactions that may be targetable in host-directed TB therapies.