Endothelial eNAMPT amplifies pre-clinical acute lung injury: Efficacy of an eNAMPT-neutralising monoclonal antibody

Hector Quijada, Tadeo Bermudez, Carrie L. Kempf, Daniel G. Valera, Alexander N. Garcia, Sara M. Camp, Jin H. Song, Evelyn Franco, Jessica K. Burt, Belinda Sun, Joseph B. Mascarenhas, Kimberlie Burns, Amir Gaber, Radu C. Oita, Vivian Reyes Hernon, Christy Barber, Liliana Moreno-Vinasco, Xiaoguang Sun, Anne E. Cress, Diego MartinZhonglin Liu, Ankit Desai, Viswanathan Natarajan, Jeffrey R. Jacobson, Steven M. Dudek, Christian Bime, Saad Sammani, Joe G.N. Garcia

Research output: Contribution to journalArticlepeer-review

Abstract

Rationale: The severe acute respiratory syndrome coronavirus 2/coronavirus disease 2019 pandemic has highlighted the serious unmet need for effective therapies that reduce acute respiratory distress syndrome (ARDS) mortality. We explored whether extracellular nicotinamide phosphoribosyltransferase (eNAMPT), a ligand for Toll-like receptor (TLR)4 and a master regulator of innate immunity and inflammation, is a potential ARDS therapeutic target. Methods: Wild-type C57BL/6J or endothelial cell (EC)-cNAMPT/ knockout mice (targeted EC NAMPT deletion) were exposed to either a lipopolysaccharide (LPS)-induced (“one-hit”) or a combined LPS/ ventilator (“two-hit”)-induced acute inflammatory lung injury model. A NAMPT-specific monoclonal antibody (mAb) imaging probe (99mTc-ProNamptor) was used to detect NAMPT expression in lung tissues. Either an eNAMPT-neutralising goat polyclonal antibody (pAb) or a humanised monoclonal antibody (ALT-100 mAb) were used in vitro and in vivo. Results: Immunohistochemical, biochemical and imaging studies validated time-dependent increases in NAMPT lung tissue expression in both pre-clinical ARDS models. Intravenous delivery of either eNAMPT-neutralising pAb or mAb significantly attenuated inflammatory lung injury (haematoxylin and eosin staining, bronchoalveolar lavage (BAL) protein, BAL polymorphonuclear cells, plasma interleukin-6) in both pre-clinical models. In vitro human lung EC studies demonstrated eNAMPT-neutralising antibodies (pAb, mAb) to strongly abrogate eNAMPT-induced TLR4 pathway activation and EC barrier disruption. In vivo studies in wild-type and EC-cNAMPT/ mice confirmed a highly significant contribution of EC-derived NAMPT to the severity of inflammatory lung injury in both pre-clinical ARDS models. Conclusions: These findings highlight both the role of EC-derived eNAMPT and the potential for biologic targeting of the eNAMPT/TLR4 inflammatory pathway. In combination with predictive eNAMPT biomarker and NAMPT genotyping assays, this offers the opportunity to identify high-risk ARDS subjects for delivery of personalised medicine.

Original languageEnglish (US)
Article number2002536
JournalEuropean Respiratory Journal
Volume57
Issue number5
DOIs
StatePublished - May 1 2021

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine

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