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 Martin; Zhonglin Liu; Ankit A. Desai; Viswanathan Natarajan; Jeffrey R. Jacobson; Steven M. Dudek; Christian Bime; Saad Sammani; Joe G.N. Garcia

doi:10.1183/13993003.02536-2020

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 A. Desai, Viswanathan Natarajan, Jeffrey R. Jacobson, Steven M. Dudek, Christian Bime, Saad Sammani, Joe G.N. Garcia

Research output: Contribution to journal › Article › peer-review

65 Scopus citations

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 language	English (US)
Article number	2002536
Journal	European Respiratory Journal
Volume	57
Issue number	5
DOIs	https://doi.org/10.1183/13993003.02536-2020
State	Published - May 1 2021

Keywords

Acute Lung Injury
Animals
Antibodies, Monoclonal
COVID-19
Humans
Mice
Mice, Inbred C57BL
SARS-CoV-2

ASJC Scopus subject areas

Pulmonary and Respiratory Medicine

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10.1183/13993003.02536-2020

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Quijada, H., Bermudez, T., Kempf, C. L., Valera, D. G., Garcia, A. N., Camp, S. M., Song, J. H., Franco, E., Burt, J. K., Sun, B., Mascarenhas, J. B., Burns, K., Gaber, A., Oita, R. C., Hernon, V. R., Barber, C., Moreno-Vinasco, L., Sun, X., Cress, A. E., ... Garcia, J. G. N. (2021). Endothelial eNAMPT amplifies pre-clinical acute lung injury: Efficacy of an eNAMPT-neutralising monoclonal antibody. European Respiratory Journal, 57(5), Article 2002536. https://doi.org/10.1183/13993003.02536-2020

Quijada, H, Bermudez, T, Kempf, CL, Valera, DG, Garcia, AN, Camp, SM, Song, JH, Franco, E, Burt, JK, Sun, B, Mascarenhas, JB, Burns, K, Gaber, A, Oita, RC, Hernon, VR, Barber, C, Moreno-Vinasco, L, Sun, X, Cress, AE, Martin, D, Liu, Z, Desai, AA, Natarajan, V, Jacobson, JR, Dudek, SM, Bime, C, Sammani, S & Garcia, JGN 2021, 'Endothelial eNAMPT amplifies pre-clinical acute lung injury: Efficacy of an eNAMPT-neutralising monoclonal antibody', European Respiratory Journal, vol. 57, no. 5, 2002536. https://doi.org/10.1183/13993003.02536-2020

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

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.",

keywords = "Acute Lung Injury, Animals, Antibodies, Monoclonal, COVID-19, Humans, Mice, Mice, Inbred C57BL, SARS-CoV-2",

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

note = "Publisher Copyright: Copyright {\textcopyright} ERS 2021.",

year = "2021",

month = may,

day = "1",

doi = "10.1183/13993003.02536-2020",

language = "English (US)",

volume = "57",

journal = "European Respiratory Journal",

issn = "0903-1936",

publisher = "European Respiratory Society",

number = "5",

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TY - JOUR

T1 - Endothelial eNAMPT amplifies pre-clinical acute lung injury

T2 - Efficacy of an eNAMPT-neutralising monoclonal antibody

AU - Quijada, Hector

AU - Bermudez, Tadeo

AU - Kempf, Carrie L.

AU - Valera, Daniel G.

AU - Garcia, Alexander N.

AU - Camp, Sara M.

AU - Song, Jin H.

AU - Franco, Evelyn

AU - Burt, Jessica K.

AU - Sun, Belinda

AU - Mascarenhas, Joseph B.

AU - Burns, Kimberlie

AU - Gaber, Amir

AU - Oita, Radu C.

AU - Hernon, Vivian Reyes

AU - Barber, Christy

AU - Moreno-Vinasco, Liliana

AU - Sun, Xiaoguang

AU - Cress, Anne E.

AU - Martin, Diego

AU - Liu, Zhonglin

AU - Desai, Ankit A.

AU - Natarajan, Viswanathan

AU - Jacobson, Jeffrey R.

AU - Dudek, Steven M.

AU - Bime, Christian

AU - Sammani, Saad

AU - Garcia, Joe G.N.

PY - 2021/5/1

Y1 - 2021/5/1

N2 - 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.

AB - 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.

KW - Acute Lung Injury

KW - Animals

KW - Antibodies, Monoclonal

KW - COVID-19

KW - Humans

KW - Mice

KW - Mice, Inbred C57BL

KW - SARS-CoV-2

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

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