Human Ex Vivo Lung Perfusion: A Model System to study Lung diseases and direct targeted therapies

In the present study, we demonstrate the feasibility of the EVLP technique to perfuse lungs from deceased patients with a broad spectrum of chronic lung diseases. We were able to assess gene expression changes over the course of EVLP and between end-stage diseases by RNA sequencing transcriptomics from tissue samples collected at different time points, revealing fundamental genetic information which has important avenues for modulation in the future. Thus, EVLP can serve as a clinically relevant experimental model to derive mechanistic insights into pulmonary pathophysiology and various human lung diseases. Overall design: Method: Tissues from chronic lung diseases and normal lungs were collected at time 0 and end of EVLP (6hr), and quality RNA libraries were generated using Illumina TruSeq RNA Access library preparation kit. The coding regions of the transcriptome were then captured from this library using sequence-specific probes to create the final library. The cDNA libraries were validated using KAPA Biosystems primer premix kit with Illumina-compatible DNA primers and Qubit 2.0 fluorometer and quality examined using Agilent Tapestation 2200. Cluster generation and 75 bp Paired-read dual-indexed sequencing was performed on Illumina NextSeq 500's.