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Blockade of ZFX Alleviates Hypoxia-Induced Pulmonary Vascular Remodeling by Regulating the YAP Signaling

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Abstract

High expression of the zinc finger X-chromosomal protein (ZFX) correlates with proliferation, aggressiveness, and development in many types of cancers. In the current report, we investigated the efficacy of ZFX in mouse pulmonary artery smooth muscle cells (PASMCs) proliferation during pulmonary arterial hypertension (PAH). PASMCs were cultured in hypoxic conditions. Real-time PCR and western blotting were conducted to detect the expression of ZFX. Cell proliferation, apoptosis, migration, and invasion were, respectively, measured by CCK-8, flow cytometry, wound scratchy, and transwell assays. Glycolytic ability was validated by the extracellular acidification rate and oxygen consumption rate. Transcriptome sequencing technology was used to explore the genes affected by ZFX knockdown. Luciferase and chromatin immunoprecipitation assays were utilized to verify the possible binding site of ZFX and YAP1. Mice were subjected to hypoxia for 21 days to induce PAH. The right ventricular systolic pressure (RVSP) was measured and ratio of RV/LV + S was calculated. The results show that ZFX was increased in hypoxia-induced PASMCs and mice. ZFX knockdown inhibited the proliferation, migration, and invasion of PASMC. Using RNA sequencing, we identify glycolysis and YAP as a key signaling of ZFX. ZFX knockdown inhibited Glycolytic ability. ZFX strengthened the transcription activity of YAP1, thereby regulating the YAP signaling. YAP1 overexpression reversed the effect of ZFX knockdown on hypoxia-treated PASMCs. In conclusion, ZFX knockdown protected mice from hypoxia-induced PAH injury. ZFX knockdown dramatically reduced RVSP and RV/(LV + S) in hypoxia-treated mice.

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Data Availability

The datasets used and analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

Not Applicable.

Funding

Medical and Health Science and Technology Development Plan of Shandong Province in 2020 (No. 202006011224) The Second batch of Science and Technology Plan Projects of **an Municipal Health Commission in 2020 (No. 2020-3-03).

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Authors and Affiliations

  1. Department of Pediatrics, **an Central Hospital, Shandong University, **an, 250013, Shandong, People’s Republic of China

    Ling Tang, **ao Zhou & Aili Guo

  2. Department of Pediatrics, Central Hosptial Affiliated to Shandong First Medical University, **an, 250013, Shandong, People’s Republic of China

    Ling Tang, **ao Zhou & Aili Guo

  3. Department of Neurosurgery, Qilu Hospital of Shandong University, No.107 West Wenhua Road, **an, 250012, Shandong, People’s Republic of China

    Lizhang Han

  4. Heart Center, Qingdao Women and Children’s Hospital, Shandong University, No.217 West Liaoyang Road, Qingdao, 266034, Shandong, People’s Republic of China

    Silin Pan

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  1. Ling Tang
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Contributions

LT designed the study; LT, XZ and ALG performed the research; LZH and SLP analyzed data; LT wrote the paper. All authors have read and approved the manuscript.

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Correspondence to Lizhang Han or Silin Pan.

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The authors declare that they have no conflicts of interest.

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The experimental protocol of our study was performed in accordance with the Guide for the Care and Use of Laboratory Animals and approved by **an Central Hospital Ethics committee.

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Tang, L., Zhou, X., Guo, A. et al. Blockade of ZFX Alleviates Hypoxia-Induced Pulmonary Vascular Remodeling by Regulating the YAP Signaling. Cardiovasc Toxicol 24, 158–170 (2024). https://doi.org/10.1007/s12012-023-09822-5

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