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Effect of TDP43-CTFs35 on Brain Endothelial Cell Functions in Cerebral Ischemic Injury

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Abstract

Pathological changes in the brain endothelium play an important role in the progression of ischemic stroke and the compromised BBB under ischemic stroke conditions cause neuronal damage. However, the pathophysiological mechanisms of the BBB under normal conditions and under ischemic stroke conditions have not been fully elucidated. The present study demonstrated that knockdown of TAR DNA-binding protein 43 (TDP-43) or overexpression of TDP43-CTFs35 inhibited tight junction protein expression, and mammalian sterile-20-like 1/2 (MST1/2) and YES-associated protein (YAP) phosphorylation in brain ECs and suppressed brain EC migration in vitro. The cytoplasmic TDP43-CTFs35 level was increased in brain ECs 24 h and 72 h after MCAO, but it disappeared 1 week after cerebral ischemia. The expression of tight junction proteins was also significantly deceased 24 h after MCAO and then gradually recovered at 72 h and 1 week after MCAO. The level of YAP phosphorylation was first significantly decreased 24 h after MCAO and then increased 72 h and 1 week after MCAO, accompanied by nuclear YAP translocation. The underlying mechanism is TDP43-CTFs35-mediated inhibition of Hippo signaling pathway activity through the dephosphorylation of MST1/2, which leads to the inhibition of YAP phosphorylation and the subsequent impairment of brain EC migration and tight junction protein expression. This study provides new insights into the mechanisms of brain vascular EC regulation, which may impact on BBB integrity after cerebral ischemic injury.

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Data Availability

All data generated or analyzed during this study are included in this published article.

Abbreviations

BBB:

Blood-brain barrier

ECs:

Endothelial cells

CNS:

Central nerve system

OGD:

Oxygen-glucose deprivation

LPS:

Lipopolysaccharides

TJ:

Tight junction

YAP:

YES-associated protein

TDP-43:

TAR DNA-binding protein 43

MCAO:

Middle cerebral artery occlusion

CTFs:

C-terminal TDP-43 fragments

CMV:

Cytomegalovirus immediate-early enhancer and promoter

ZOs:

Zonula occludens

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Acknowledgements

We thank Mei Xin (Division of Experimental Hematology and Cancer Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA) for providing lentivirus (YAP/TAZ shRNA) as a gift and the details of transfection experiments in the current study.

Funding

This work was supported by the National Institutes of Health (NIH)/National Institute of Neurological Disorders and Stroke (NINDS) grants, R01NS105787 (J.H.), and National Natural Science Foundation of China (grant no. 81901084).

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Authors and Affiliations

  1. Department of Neurology, The Affiliated Hospital of Yangzhou University, Jiangsu Province, Yangzhou, 225000, China

    Xiaotian Xu

  2. Department of Pharmacological & Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, 77204‐5000, USA

    Xiaotian Xu & Jiukuan Hao

  3. Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, 45229, USA

    Changwen Zhang

  4. Department of Pharmaceutical Sciences, The University of Tennessee Health Science Center, Memphis, TN, 38163, USA

    Jianxiong Jiang

  5. Division of Experimental Hematology and Cancer Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, 45229, USA

    Mei Xin

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  1. Xiaotian Xu
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Contributions

The experiments were performed by Xiaotian Xu. The manuscript was written through contributions of all authors. All authors have approved the final version of the manuscript.

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Correspondence to Jiukuan Hao.

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All animal procedures were approved by the Institutional Animal Care and Use Committees at the University of Houston and they complied with pertinent NIH guidelines for care and use of animals.

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Xu, X., Zhang, C., Jiang, J. et al. Effect of TDP43-CTFs35 on Brain Endothelial Cell Functions in Cerebral Ischemic Injury. Mol Neurobiol 59, 4593–4611 (2022). https://doi.org/10.1007/s12035-022-02869-5

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