Biodistribution and Efficacy of Targeted Pulmonary Delivery of a Protein Kinase C-delta Inhibitory Peptide: Impact on Indirect Lung Injury

摘要

Sepsis and sepsis-induced lung injury remain a leading cause of death in intensive care units. We identified protein kinase C-delta (PKC delta) as a critical regulator of the acute inflammatory response and demonstrated that PKC delta inhibition was lung-protective in a rodent sepsis model, suggesting that targeting PKC delta is a potential strategy for preserving pulmonary function in the setting of indirect lung injury. In this study, whole-body organ biodistribution and pulmonary cellular distribution of a transactivator of transcription (TAT)-conjugated PKC delta inhibitory peptide (PKC delta-TAT) was determined following intratracheal (IT) delivery in control and septic [cecal ligation and puncture (CLP)] rats to ascertain the impact of disease pathology on biodistribution and efficacy. There was negligible lung uptake of radiolabeled peptide upon intravenous delivery [<1% initial dose (ID)], whereas IT administration resulted in lung retention of >65% ID with minimal uptake in liver or kidney (<2% ID). IT delivery of a fluorescent-tagged (tetramethylrhodamine-PKC delta-TAT) peptide demonstrated uniform spatial distribution and cellular uptake throughout the peripheral lung. IT delivery of PKC delta-TAT at the time of CLP surgery significantly reduced PKC delta activation (tyrosine phosphorylation, nuclear translocation and cleavage) and acute lung inflammation, resulting in improved lung function and gas exchange. Importantly, peptide efficacy was similar when delivered at 4 hours post-CLP, demonstrating therapeutic relevance. Conversely, spatial lung distribution and efficacy were significantly impaired at 8 hours post-CLP, which corresponded to marked histopathological progression of lung injury. These studies establish a functional connection between peptide spatial distribution, inflammatory histopathology in the lung, and efficacy of this anti-inflammatory peptide.