Therapeutic signature mapping of paired direct and indirect LPS injury in an ex vivo human lung perfusion platform reveals injury-specific druggable programs

Acute Respiratory Distress Syndrome (ARDS) remains highly morbid with no approved disease-modifying therapies, partly because direct (pulmonary) and indirect (extrapulmonary) insults may drive biologically distinct early injury programs that are difficult to study in human tissue. To address this, the authors established a paired, acellular ex vivo lung perfusion (EVLP) platform using human donor lungs unsuitable for transplantation, modeling direct (endobronchial) and indirect (perfusate) lipopolysaccharide (LPS) injury within the same donor. Lung tissue proteomes were profiled at 4 hours post-insult, and therapeutic candidates were nominated by querying injury signatures against the CLUE L1000 perturbational compendium, with independent cross-platform validation.

Both injury models displayed histological injury and robust cytokine release. Direct injury preferentially enriched neutrophil degranulation, extracellular matrix remodelling, and metabolic reprogramming modules, while indirect injury showed prominent complement/coagulation perturbation and greater endothelial activation markers in perfusate. Cross-platform prioritisation converged on tractable signalling and epigenetic axes, including JAK/STAT, PI3K/AKT/mTOR, SYK, CDK, and HDAC inhibitor classes, yielding a tiered shortlist for EVLP intervention testing. This intact human lung perturbation platform enables injury-stratified mechanistic inference and therapeutic prioritisation in early lung injury relevant to ARDS, offering a translational bridge between preclinical models and clinical trials.