Murine Functional Lung Imaging Using X-Ray Velocimetry for Longitudinal Noninvasive Quantitative Spatial Assessment of Pulmonary Airflow

<b>Background/Objectives:</b> The recent development of four-dimensional X-ray velocimetry (4DXV) technology (three-dimensional space and time) provides a unique opportunity to obtain preclinical quantitative functional lung images. Only single-scan measurements in non-survival studies have been obtained to date; thus, methodologies enabling animal survival for repeated imaging to be accomplished over weeks or months from the same animal would establish new opportunities for the assessment of pathophysiology drivers and treatment response in advanced preclinical drug-screening efforts. <b>Methods:</b> An anesthesia protocol developed for animal recovery to allow for repetitive, longitudinal scanning of individual animals over time. Test-retest imaging scans from the lungs of healthy mice were performed over 8 weeks to assess the repeatability of scanner-derived quantitative imaging metrics and variability. <b>Results:</b> Using a murine model of fibroproliferative lung disease, this longitudinal scanning approach captured heterogeneous progressive changes in pulmonary function, enabling the visualization and quantitative measurement of averaged whole lung metrics and spatial/regional change. Radiation dosimetry studies evaluated the effects of imaging acquisition protocols on X-ray dosage to further adapt protocols for the minimization of radiation exposure during repeat imaging sessions using these newly developed image acquisition protocols. <b>Conclusions:</b> Overall, we have demonstrated that the 4DXV advanced imaging scanner allows for repeat measurements from the same animal over time to enable the high-resolution, noninvasive mapping of quantitative lung airflow dysfunction in mouse models with heterogeneous pulmonary disease. The animal anesthesia and image acquisition protocols described will serve as the foundation on which further applications of the 4DXV technology can be used to study a diverse array of murine pulmonary disease models. Together, 4DXV provides a novel and significant advancement for the longitudinal, noninvasive interrogation of pulmonary disease to assess spatial/regional disease initiation, progression, and response to therapeutic interventions.