Selected clinical strains of Mycobacterium avium unlock in vivo models for NTM drug discovery programs

Mycobacterium avium pulmonary disease is an emerging global health challenge for which drug development remains limited by preclinical models that rely on laboratory strains and invasive endpoint analyses. Here, we compared recent clinical M. avium isolates with the reference strain ATCC 700898 across macrophage, Galleria mellonella , and murine infection models and evaluated longitudinal micro-computed tomography (µCT) as a non-invasive tool to monitor disease progression and treatment response. While extracellular growth rates were comparable, clinical isolates demonstrated enhanced host-associated fitness and induced higher bacterial burdens and more severe pulmonary pathology in mice than the reference strain. These strain-dependent differences were detected by quantitative µCT imaging. Using the hypervirulent isolate MYC_0069, we further show that clarithromycin monotherapy and standard-of-care triple therapy significantly reduced bacterial burden and lung pathology. Together, these findings establish a clinically relevant chronic M. avium model that combines clinical isolates with longitudinal imaging to enable preclinical anti-mycobacterial drug evaluation in vivo .