Protease-independent action of tissue plasminogen activator in brain plasticity and neurological recovery after ischemic stroke

Hongjian Pu, Yejie Shi, Lili Zhang, Zhengyu Lu, Qing Ye, Rehana K. Leak, Fei Xu, Shubei Ma, Hongfeng Mu, Zhishuo Wei, Na Xu, Yuguo Xia, Xiaoming Hu, T. Kevin Hitchens, Michael V. L. Bennett, Jun Chen

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Emerging evidence suggests that tissue plasminogen activator (tPA), currently the only FDA-approved medication for ischemic stroke, exerts important biological actions on the CNS besides its well-known thrombolytic effect. In this study, we investigated the role of tPA on primary neurons in culture and on brain recovery and plasticity after ischemic stroke in mice. Treatment with recombinant tPA stimulated axonal growth in culture, an effect independent of its protease activity and achieved through epidermal growth factor receptor (EGFR) signaling. After permanent focal cerebral ischemia, tPA knockout mice developed more severe sensorimotor and cognitive deficits and greater axonal and myelin injury than wild-type mice, suggesting that endogenously expressed tPA promotes long-term neurological recovery after stroke. In tPA knockout mice, intranasal administration of recombinant tPA protein 6 hours poststroke and 7 more times at 2 d intervals mitigated white matter injury, improved axonal conduction, and enhanced neurological recovery. Consistent with the proaxonal growth effects observed in vitro, exogenous tPA delivery increased poststroke axonal sprouting of corticobulbar and corticospinal tracts, which might have contributed to restoration of neurological functions. Notably, recombinant mutant tPA-S478A lacking protease activity (but retaining the EGF-like domain) was as effective as wild-type tPA in rescuing neurological functions in tPA knockout stroke mice. These findings demonstrate that tPA improves long-term functional outcomes in a clinically relevant stroke model, likely by promoting brain plasticity through EGFR signaling. Therefore, treatment with the protease-dead recombinant tPA-S478A holds particular promise as a neurorestorative therapy, as the risk for triggering intracranial hemorrhage is eliminated and tPA-S478A can be delivered intranasally hours after stroke.

Original languageEnglish (US)
Pages (from-to)9115-9124
Number of pages10
JournalProceedings of the National Academy of Sciences of the United States of America
Volume116
Issue number18
DOIs
StatePublished - Apr 30 2019

Fingerprint

Tissue Plasminogen Activator
Peptide Hydrolases
Stroke
Brain
Knockout Mice
Pyramidal Tracts
Epidermal Growth Factor Receptor
Intranasal Administration
Intracranial Hemorrhages
Wounds and Injuries
Growth
Myelin Sheath
Brain Ischemia
Epidermal Growth Factor
Therapeutics

Keywords

  • Axonal sprouting
  • Diffusion tensor imaging
  • Epidermal growth factor
  • Oxygen–glucose deprivation
  • Protease-inactive tPA

ASJC Scopus subject areas

  • General

Cite this

Protease-independent action of tissue plasminogen activator in brain plasticity and neurological recovery after ischemic stroke. / Pu, Hongjian; Shi, Yejie; Zhang, Lili; Lu, Zhengyu; Ye, Qing; Leak, Rehana K.; Xu, Fei; Ma, Shubei; Mu, Hongfeng; Wei, Zhishuo; Xu, Na; Xia, Yuguo; Hu, Xiaoming; Kevin Hitchens, T.; Bennett, Michael V. L.; Chen, Jun.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, No. 18, 30.04.2019, p. 9115-9124.

Research output: Contribution to journalArticle

Pu, H, Shi, Y, Zhang, L, Lu, Z, Ye, Q, Leak, RK, Xu, F, Ma, S, Mu, H, Wei, Z, Xu, N, Xia, Y, Hu, X, Kevin Hitchens, T, Bennett, MVL & Chen, J 2019, 'Protease-independent action of tissue plasminogen activator in brain plasticity and neurological recovery after ischemic stroke', Proceedings of the National Academy of Sciences of the United States of America, vol. 116, no. 18, pp. 9115-9124. https://doi.org/10.1073/pnas.1821979116
Pu, Hongjian ; Shi, Yejie ; Zhang, Lili ; Lu, Zhengyu ; Ye, Qing ; Leak, Rehana K. ; Xu, Fei ; Ma, Shubei ; Mu, Hongfeng ; Wei, Zhishuo ; Xu, Na ; Xia, Yuguo ; Hu, Xiaoming ; Kevin Hitchens, T. ; Bennett, Michael V. L. ; Chen, Jun. / Protease-independent action of tissue plasminogen activator in brain plasticity and neurological recovery after ischemic stroke. In: Proceedings of the National Academy of Sciences of the United States of America. 2019 ; Vol. 116, No. 18. pp. 9115-9124.
@article{4dd94c66794e44e1b43583cf25d59c48,
title = "Protease-independent action of tissue plasminogen activator in brain plasticity and neurological recovery after ischemic stroke",
abstract = "Emerging evidence suggests that tissue plasminogen activator (tPA), currently the only FDA-approved medication for ischemic stroke, exerts important biological actions on the CNS besides its well-known thrombolytic effect. In this study, we investigated the role of tPA on primary neurons in culture and on brain recovery and plasticity after ischemic stroke in mice. Treatment with recombinant tPA stimulated axonal growth in culture, an effect independent of its protease activity and achieved through epidermal growth factor receptor (EGFR) signaling. After permanent focal cerebral ischemia, tPA knockout mice developed more severe sensorimotor and cognitive deficits and greater axonal and myelin injury than wild-type mice, suggesting that endogenously expressed tPA promotes long-term neurological recovery after stroke. In tPA knockout mice, intranasal administration of recombinant tPA protein 6 hours poststroke and 7 more times at 2 d intervals mitigated white matter injury, improved axonal conduction, and enhanced neurological recovery. Consistent with the proaxonal growth effects observed in vitro, exogenous tPA delivery increased poststroke axonal sprouting of corticobulbar and corticospinal tracts, which might have contributed to restoration of neurological functions. Notably, recombinant mutant tPA-S478A lacking protease activity (but retaining the EGF-like domain) was as effective as wild-type tPA in rescuing neurological functions in tPA knockout stroke mice. These findings demonstrate that tPA improves long-term functional outcomes in a clinically relevant stroke model, likely by promoting brain plasticity through EGFR signaling. Therefore, treatment with the protease-dead recombinant tPA-S478A holds particular promise as a neurorestorative therapy, as the risk for triggering intracranial hemorrhage is eliminated and tPA-S478A can be delivered intranasally hours after stroke.",
keywords = "Axonal sprouting, Diffusion tensor imaging, Epidermal growth factor, Oxygen–glucose deprivation, Protease-inactive tPA",
author = "Hongjian Pu and Yejie Shi and Lili Zhang and Zhengyu Lu and Qing Ye and Leak, {Rehana K.} and Fei Xu and Shubei Ma and Hongfeng Mu and Zhishuo Wei and Na Xu and Yuguo Xia and Xiaoming Hu and {Kevin Hitchens}, T. and Bennett, {Michael V. L.} and Jun Chen",
year = "2019",
month = "4",
day = "30",
doi = "10.1073/pnas.1821979116",
language = "English (US)",
volume = "116",
pages = "9115--9124",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "18",

}

TY - JOUR

T1 - Protease-independent action of tissue plasminogen activator in brain plasticity and neurological recovery after ischemic stroke

AU - Pu, Hongjian

AU - Shi, Yejie

AU - Zhang, Lili

AU - Lu, Zhengyu

AU - Ye, Qing

AU - Leak, Rehana K.

AU - Xu, Fei

AU - Ma, Shubei

AU - Mu, Hongfeng

AU - Wei, Zhishuo

AU - Xu, Na

AU - Xia, Yuguo

AU - Hu, Xiaoming

AU - Kevin Hitchens, T.

AU - Bennett, Michael V. L.

AU - Chen, Jun

PY - 2019/4/30

Y1 - 2019/4/30

N2 - Emerging evidence suggests that tissue plasminogen activator (tPA), currently the only FDA-approved medication for ischemic stroke, exerts important biological actions on the CNS besides its well-known thrombolytic effect. In this study, we investigated the role of tPA on primary neurons in culture and on brain recovery and plasticity after ischemic stroke in mice. Treatment with recombinant tPA stimulated axonal growth in culture, an effect independent of its protease activity and achieved through epidermal growth factor receptor (EGFR) signaling. After permanent focal cerebral ischemia, tPA knockout mice developed more severe sensorimotor and cognitive deficits and greater axonal and myelin injury than wild-type mice, suggesting that endogenously expressed tPA promotes long-term neurological recovery after stroke. In tPA knockout mice, intranasal administration of recombinant tPA protein 6 hours poststroke and 7 more times at 2 d intervals mitigated white matter injury, improved axonal conduction, and enhanced neurological recovery. Consistent with the proaxonal growth effects observed in vitro, exogenous tPA delivery increased poststroke axonal sprouting of corticobulbar and corticospinal tracts, which might have contributed to restoration of neurological functions. Notably, recombinant mutant tPA-S478A lacking protease activity (but retaining the EGF-like domain) was as effective as wild-type tPA in rescuing neurological functions in tPA knockout stroke mice. These findings demonstrate that tPA improves long-term functional outcomes in a clinically relevant stroke model, likely by promoting brain plasticity through EGFR signaling. Therefore, treatment with the protease-dead recombinant tPA-S478A holds particular promise as a neurorestorative therapy, as the risk for triggering intracranial hemorrhage is eliminated and tPA-S478A can be delivered intranasally hours after stroke.

AB - Emerging evidence suggests that tissue plasminogen activator (tPA), currently the only FDA-approved medication for ischemic stroke, exerts important biological actions on the CNS besides its well-known thrombolytic effect. In this study, we investigated the role of tPA on primary neurons in culture and on brain recovery and plasticity after ischemic stroke in mice. Treatment with recombinant tPA stimulated axonal growth in culture, an effect independent of its protease activity and achieved through epidermal growth factor receptor (EGFR) signaling. After permanent focal cerebral ischemia, tPA knockout mice developed more severe sensorimotor and cognitive deficits and greater axonal and myelin injury than wild-type mice, suggesting that endogenously expressed tPA promotes long-term neurological recovery after stroke. In tPA knockout mice, intranasal administration of recombinant tPA protein 6 hours poststroke and 7 more times at 2 d intervals mitigated white matter injury, improved axonal conduction, and enhanced neurological recovery. Consistent with the proaxonal growth effects observed in vitro, exogenous tPA delivery increased poststroke axonal sprouting of corticobulbar and corticospinal tracts, which might have contributed to restoration of neurological functions. Notably, recombinant mutant tPA-S478A lacking protease activity (but retaining the EGF-like domain) was as effective as wild-type tPA in rescuing neurological functions in tPA knockout stroke mice. These findings demonstrate that tPA improves long-term functional outcomes in a clinically relevant stroke model, likely by promoting brain plasticity through EGFR signaling. Therefore, treatment with the protease-dead recombinant tPA-S478A holds particular promise as a neurorestorative therapy, as the risk for triggering intracranial hemorrhage is eliminated and tPA-S478A can be delivered intranasally hours after stroke.

KW - Axonal sprouting

KW - Diffusion tensor imaging

KW - Epidermal growth factor

KW - Oxygen–glucose deprivation

KW - Protease-inactive tPA

UR - http://www.scopus.com/inward/record.url?scp=85065511979&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85065511979&partnerID=8YFLogxK

U2 - 10.1073/pnas.1821979116

DO - 10.1073/pnas.1821979116

M3 - Article

C2 - 30996120

AN - SCOPUS:85065511979

VL - 116

SP - 9115

EP - 9124

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 18

ER -