Novel mitoNEET ligand NL-1 improves therapeutic outcomes in an aged rat model of cerebral ischemia/reperfusion injury

Aruvi Vijikumar; Pushkar Saralkar; Scott D. Saylor; Patrick G. Sullivan; Jason D. Huber; Werner J. Geldenhuys

doi:10.1016/j.expneurol.2022.114128

Novel mitoNEET ligand NL-1 improves therapeutic outcomes in an aged rat model of cerebral ischemia/reperfusion injury

Aruvi Vijikumar, Pushkar Saralkar, Scott D. Saylor, Patrick G. Sullivan, Jason D. Huber, Werner J. Geldenhuys

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

1 Scopus citations

Abstract

Cerebral ischemic stroke is a leading cause of mortality and disability worldwide. Currently, there are a lack of drugs capable of reducing neuronal cell loss due to ischemia/reperfusion-injury after stroke. Previously, we identified mitoNEET, a [2Fe-2S] redox mitochondrial protein, as a putative drug target for ischemic stroke. In this study, we tested NL-1, a novel mitoNEET ligand, in a preclinical model of ischemic stroke with reperfusion using aged female rats. Using a transient middle cerebral artery occlusion (tMCAO), we induced a 2 h ischemic injury and then evaluated the effects of NL-1 treatment on ischemic/reperfusion brain injury at 24 and 72 h. Test compounds were administered at time of reperfusion via intravenous dosing. Results of the study demonstrated that NL-1 (10 mg/kg) treatment markedly improved survival and reduced infarct volume and hemispheric swelling in the brain as compared aged rats treated with vehicle or a lower dose of NL-1 (0.25 mg/kg). Interestingly, the protective effect of NL-1 was significantly improved when encapsulated in PLGA nanoparticles, where a 40-fold lesser dose (0.25 mg/kg) of NL-1 produced an equivalent effect as the 10 mg/kg dose. Evaluation of changes in blood-brain barrier permeability and lipid peroxidation corroborated the protective actions of NL-1 (10 mg/kg) or NL-1 NP treatment demonstrated a reduced accumulation of parenchymal IgG, decreased levels of 4-hydroxynonenal (4-HNE) and a decreased TUNEL positive cells in the brains of aged female rats at 72 h after tMCAO with reperfusion. Our studies indicate that targeting mitoNEET following ischemia/reperfusion-injury is a novel drug target pathway that warrants further investigation.

Original language	English
Article number	114128
Journal	Experimental Neurology
Volume	355
DOIs	https://doi.org/10.1016/j.expneurol.2022.114128
State	Published - Sep 2022

Bibliographical note

Funding Information:
This project was supported by National Institutes of Health grants, U54 GM104942 and UL1 TR001998 (NIGMS), R41 NS110070 and P20 GM109098 (NIGMS to W.J.G.) and RO1 NS099918 (NINDS to J.D.H.). Content in this manuscript is solely the responsibility of the authors and does not represent the official views of the NIH. We are grateful to Deborah Corbin for technical support with tissue processing and we want to thank the WVU Microscopy Core for use of brightfield and fluorescent microscopes (P20 GM103434).

Funding Information:
This project was supported by National Institutes of Health grants, U54 GM104942 and UL1 TR001998 (NIGMS), R41 NS110070 and P20 GM109098 (NIGMS to W.J.G.) and RO1 NS099918 (NINDS to J.D.H.). Content in this manuscript is solely the responsibility of the authors and does not represent the official views of the NIH. We are grateful to Deborah Corbin for technical support with tissue processing and we want to thank the WVU Microscopy Core for use of brightfield and fluorescent microscopes (P20 GM103434).

Publisher Copyright:
© 2022 Elsevier Inc.

Keywords

Electron transport chain
Mitoceutical
Mitochondria
Oxidative stress
tMCAO

ASJC Scopus subject areas

Neurology
Developmental Neuroscience

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10.1016/j.expneurol.2022.114128

Cite this

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title = "Novel mitoNEET ligand NL-1 improves therapeutic outcomes in an aged rat model of cerebral ischemia/reperfusion injury",

abstract = "Cerebral ischemic stroke is a leading cause of mortality and disability worldwide. Currently, there are a lack of drugs capable of reducing neuronal cell loss due to ischemia/reperfusion-injury after stroke. Previously, we identified mitoNEET, a [2Fe-2S] redox mitochondrial protein, as a putative drug target for ischemic stroke. In this study, we tested NL-1, a novel mitoNEET ligand, in a preclinical model of ischemic stroke with reperfusion using aged female rats. Using a transient middle cerebral artery occlusion (tMCAO), we induced a 2 h ischemic injury and then evaluated the effects of NL-1 treatment on ischemic/reperfusion brain injury at 24 and 72 h. Test compounds were administered at time of reperfusion via intravenous dosing. Results of the study demonstrated that NL-1 (10 mg/kg) treatment markedly improved survival and reduced infarct volume and hemispheric swelling in the brain as compared aged rats treated with vehicle or a lower dose of NL-1 (0.25 mg/kg). Interestingly, the protective effect of NL-1 was significantly improved when encapsulated in PLGA nanoparticles, where a 40-fold lesser dose (0.25 mg/kg) of NL-1 produced an equivalent effect as the 10 mg/kg dose. Evaluation of changes in blood-brain barrier permeability and lipid peroxidation corroborated the protective actions of NL-1 (10 mg/kg) or NL-1 NP treatment demonstrated a reduced accumulation of parenchymal IgG, decreased levels of 4-hydroxynonenal (4-HNE) and a decreased TUNEL positive cells in the brains of aged female rats at 72 h after tMCAO with reperfusion. Our studies indicate that targeting mitoNEET following ischemia/reperfusion-injury is a novel drug target pathway that warrants further investigation.",

keywords = "Electron transport chain, Mitoceutical, Mitochondria, Oxidative stress, tMCAO",

author = "Aruvi Vijikumar and Pushkar Saralkar and Saylor, {Scott D.} and Sullivan, {Patrick G.} and Huber, {Jason D.} and Geldenhuys, {Werner J.}",

note = "Funding Information: This project was supported by National Institutes of Health grants, U54 GM104942 and UL1 TR001998 (NIGMS), R41 NS110070 and P20 GM109098 (NIGMS to W.J.G.) and RO1 NS099918 (NINDS to J.D.H.). Content in this manuscript is solely the responsibility of the authors and does not represent the official views of the NIH. We are grateful to Deborah Corbin for technical support with tissue processing and we want to thank the WVU Microscopy Core for use of brightfield and fluorescent microscopes (P20 GM103434). Funding Information: This project was supported by National Institutes of Health grants, U54 GM104942 and UL1 TR001998 (NIGMS), R41 NS110070 and P20 GM109098 (NIGMS to W.J.G.) and RO1 NS099918 (NINDS to J.D.H.). Content in this manuscript is solely the responsibility of the authors and does not represent the official views of the NIH. We are grateful to Deborah Corbin for technical support with tissue processing and we want to thank the WVU Microscopy Core for use of brightfield and fluorescent microscopes (P20 GM103434). Publisher Copyright: {\textcopyright} 2022 Elsevier Inc.",

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doi = "10.1016/j.expneurol.2022.114128",

language = "English",

volume = "355",

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T1 - Novel mitoNEET ligand NL-1 improves therapeutic outcomes in an aged rat model of cerebral ischemia/reperfusion injury

AU - Vijikumar, Aruvi

AU - Saralkar, Pushkar

AU - Saylor, Scott D.

AU - Sullivan, Patrick G.

AU - Huber, Jason D.

AU - Geldenhuys, Werner J.

N1 - Funding Information: This project was supported by National Institutes of Health grants, U54 GM104942 and UL1 TR001998 (NIGMS), R41 NS110070 and P20 GM109098 (NIGMS to W.J.G.) and RO1 NS099918 (NINDS to J.D.H.). Content in this manuscript is solely the responsibility of the authors and does not represent the official views of the NIH. We are grateful to Deborah Corbin for technical support with tissue processing and we want to thank the WVU Microscopy Core for use of brightfield and fluorescent microscopes (P20 GM103434). Funding Information: This project was supported by National Institutes of Health grants, U54 GM104942 and UL1 TR001998 (NIGMS), R41 NS110070 and P20 GM109098 (NIGMS to W.J.G.) and RO1 NS099918 (NINDS to J.D.H.). Content in this manuscript is solely the responsibility of the authors and does not represent the official views of the NIH. We are grateful to Deborah Corbin for technical support with tissue processing and we want to thank the WVU Microscopy Core for use of brightfield and fluorescent microscopes (P20 GM103434). Publisher Copyright: © 2022 Elsevier Inc.

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N2 - Cerebral ischemic stroke is a leading cause of mortality and disability worldwide. Currently, there are a lack of drugs capable of reducing neuronal cell loss due to ischemia/reperfusion-injury after stroke. Previously, we identified mitoNEET, a [2Fe-2S] redox mitochondrial protein, as a putative drug target for ischemic stroke. In this study, we tested NL-1, a novel mitoNEET ligand, in a preclinical model of ischemic stroke with reperfusion using aged female rats. Using a transient middle cerebral artery occlusion (tMCAO), we induced a 2 h ischemic injury and then evaluated the effects of NL-1 treatment on ischemic/reperfusion brain injury at 24 and 72 h. Test compounds were administered at time of reperfusion via intravenous dosing. Results of the study demonstrated that NL-1 (10 mg/kg) treatment markedly improved survival and reduced infarct volume and hemispheric swelling in the brain as compared aged rats treated with vehicle or a lower dose of NL-1 (0.25 mg/kg). Interestingly, the protective effect of NL-1 was significantly improved when encapsulated in PLGA nanoparticles, where a 40-fold lesser dose (0.25 mg/kg) of NL-1 produced an equivalent effect as the 10 mg/kg dose. Evaluation of changes in blood-brain barrier permeability and lipid peroxidation corroborated the protective actions of NL-1 (10 mg/kg) or NL-1 NP treatment demonstrated a reduced accumulation of parenchymal IgG, decreased levels of 4-hydroxynonenal (4-HNE) and a decreased TUNEL positive cells in the brains of aged female rats at 72 h after tMCAO with reperfusion. Our studies indicate that targeting mitoNEET following ischemia/reperfusion-injury is a novel drug target pathway that warrants further investigation.

AB - Cerebral ischemic stroke is a leading cause of mortality and disability worldwide. Currently, there are a lack of drugs capable of reducing neuronal cell loss due to ischemia/reperfusion-injury after stroke. Previously, we identified mitoNEET, a [2Fe-2S] redox mitochondrial protein, as a putative drug target for ischemic stroke. In this study, we tested NL-1, a novel mitoNEET ligand, in a preclinical model of ischemic stroke with reperfusion using aged female rats. Using a transient middle cerebral artery occlusion (tMCAO), we induced a 2 h ischemic injury and then evaluated the effects of NL-1 treatment on ischemic/reperfusion brain injury at 24 and 72 h. Test compounds were administered at time of reperfusion via intravenous dosing. Results of the study demonstrated that NL-1 (10 mg/kg) treatment markedly improved survival and reduced infarct volume and hemispheric swelling in the brain as compared aged rats treated with vehicle or a lower dose of NL-1 (0.25 mg/kg). Interestingly, the protective effect of NL-1 was significantly improved when encapsulated in PLGA nanoparticles, where a 40-fold lesser dose (0.25 mg/kg) of NL-1 produced an equivalent effect as the 10 mg/kg dose. Evaluation of changes in blood-brain barrier permeability and lipid peroxidation corroborated the protective actions of NL-1 (10 mg/kg) or NL-1 NP treatment demonstrated a reduced accumulation of parenchymal IgG, decreased levels of 4-hydroxynonenal (4-HNE) and a decreased TUNEL positive cells in the brains of aged female rats at 72 h after tMCAO with reperfusion. Our studies indicate that targeting mitoNEET following ischemia/reperfusion-injury is a novel drug target pathway that warrants further investigation.

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Novel mitoNEET ligand NL-1 improves therapeutic outcomes in an aged rat model of cerebral ischemia/reperfusion injury

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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