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BMSC-Derived Exosomes Carrying lncRNA-ZFAS1 Alleviate Pulmonary Ischemia/Reperfusion Injury by UPF1-Mediated mRNA Decay of FOXD1

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Abstract

Bone marrow-derived mesenchymal stem cells (BMSCs) exert protective effects against pulmonary ischemia/reperfusion (I/R) injury; however, the potential mechanism involved in their protective ability remains unclear. Thus, this study aimed to explore the function and underlying mechanism of BMSC-derived exosomal lncRNA-ZFAS1 in pulmonary I/R injury. Pulmonary I/R injury models were established in mice and hypoxia/reoxygenation (H/R)-exposed primary mouse lung microvascular endothelial cells (LMECs). Exosomes were extracted from BMSCs. Target molecule expression was assessed by qRT-PCR and Western blotting. Pathological changes in the lungs, pulmonary edema, apoptosis, pro-inflammatory cytokine levels, SOD, MPO activities, and MDA level were measured. The proliferation, apoptosis, and migration of LMECs were detected by CCK-8, EdU staining, flow cytometry, and scratch assay. Dual-luciferase reporter assay, RNA pull-down, RIP, and ChIP assays were performed to validate the molecular interaction. In the mouse model of pulmonary I/R injury, BMSC-Exos treatment relieved lung pathological injury, reduced lung W/D weight ratio, and restrained apoptosis and inflammation, whereas exosomal ZFAS1 silencing abolished these beneficial effects. In addition, the proliferation, migration inhibition, apoptosis, and inflammation in H/R-exposed LMECs were repressed by BMSC-derived exosomal ZFAS1. Mechanistically, ZFAS1 contributed to FOXD1 mRNA decay via interaction with UPF1, thereby leading to Gal-3 inactivation. Furthermore, FOXD1 depletion strengthened the weakened protective effect of ZFAS1-silenced BMSC-Exos on pulmonary I/R injury. ZFAS1 delivered by BMSC-Exos results in FOXD1 mRNA decay and subsequent Gal-3 inactivation via direct interaction with UPF1, thereby attenuating pulmonary I/R injury.

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

Abbreviations

ANOVA:

Analysis of variance

BMSC-Exos:

BMSC-derived exosomes

BMSCs:

Bone marrow-derived MSCs

CCK-8:

Cell counting kit 8

ChIP:

Chromatin immunoprecipitation

DAPI:

4′-6-Diamidino-2-phenylindole

EdU:

5-Ethynyl-2′-deoxyuridine

FBS:

Fetal bovine serum

FOXD1:

Forkhead box D1

Gal-3:

Galectin-3

H&E:

Hematoxylin and eosin

HLA-DR:

Human leukocyte antigen DR

H/R:

Hypoxia/reoxygenation

Hsp70:

Heat shock protein

I/R:

Ischemia/reperfusion

LMECs:

Lung microvascular endothelial cells

lncRNAs:

Long non-coding RNAs

MEG3:

Maternally expressed gene 3

MSCs:

Mesenchymal stem cells

NTA:

Nanoparticle tracking analysis

PBS:

Phosphate-buffered saline

PTBP1:

Polypyrimidine tract-binding protein 1

qRT-PCR:

Quantitative real-time PCR

RBPs:

RNA-binding proteins

RIP:

RNA immunoprecipitation

shNC:

Negative control shRNA

TSG101:

Tumor susceptibility gene 101

TUNEL:

Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling

UPF1:

Up-frameshift 1

W/D:

Wet/dry

XIST:

X-inactive-specific transcript

ZFAS1:

Zinc finger antisense 1

ZFPM2-AS1:

Zinc finger protein multitype 2 antisense RNA 1

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Acknowledgements

We would like to give our sincere gratitude to the reviewers for their constructive comments.

Funding

This work was supported in part by Changzhou HighLevel Medical Talents Training Project (KY20221373 and KY20221357) and the Major Program of Science and Technology Project of Changzhou Health Commission (ZD202206).

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Authors and Affiliations

  1. Departments of Anesthesiology, The Third Affiliated Hospital of Soochow University, Changzhou, 213000, Jiangsu Province, People’s Republic of China

    Cao Gao & Zhi-xiu Meng

  2. Departments of Oncology, The Third Affiliated Hospital of Soochow University, Changzhou, 213000, Jiangsu Province, People’s Republic of China

    Yan-Jie Xu

  3. Department of Thoracic Surgery, The Third Affiliated Hospital of Soochow University, No. 185, Juqian Road, Changzhou, 213000, Jiangsu Province, People’s Republic of China

    Shuang Gu, Lei Zhang & Liang Zheng

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  1. Cao Gao
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Contributions

Cao Gao: Conceptualization, Writing-original draft preparation, Investigation, Validation, Visualization, Methodology.

Yan-Jie Xu: Software.

Zhi-xiu Meng: Methodology.

Shuang Gu: Data curation.

Lei Zhang: Visualization.

Liang Zheng: Conceptualization, Writing-original draft preparation, Supervision, Writing-reviewing and editing.

Corresponding author

Correspondence to Liang Zheng.

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The lungs were harvested after euthanasia by overdose of sodium pentobarbital. All procedures were approved by the Animal Care and Use Committee of the Third Affiliated Hospital of Soochow University.

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The informed consent obtained from study participants.

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The authors declare no competing interests.

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Gao, C., Xu, YJ., Meng, Zx. et al. BMSC-Derived Exosomes Carrying lncRNA-ZFAS1 Alleviate Pulmonary Ischemia/Reperfusion Injury by UPF1-Mediated mRNA Decay of FOXD1. Mol Neurobiol 60, 2379–2396 (2023). https://doi.org/10.1007/s12035-022-03129-2

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