Selective RNAi silencing of Schwann cell Piezo1 alleviates mechanical hypersensitization following peripheral nerve injury.

IF 4.6 2区医学 Q2 MEDICINE, RESEARCH & EXPERIMENTAL

Molecular Therapy-Methods & Clinical Development Pub Date : 2025-02-12 eCollection Date: 2025-03-13 DOI:10.1016/j.omtm.2025.101433

Brandon Itson-Zoske, Uarda Gani, Alexander Mikesell, Chensheng Qiu, Fan Fan, Cheryl L Stucky, Quinn H Hogan, Seung Min Shin, Hongwei Yu

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引用次数: 0

Abstract

The present study was designed to investigate the role of Schwann cell (SC) Piezo1 in peripheral nociception. We first developed an AAV vector that has primary SC tropism after delivery into the sciatic (or tibial) nerve. This was achieved by packing AAV-GFP transcribed by a CBA promoter using a capsid AAVolig001 to generate AAVolig001-CBA-GFP. Six weeks after intraneural injection of AAVolig001-CBA-GFP in naive rats, GFP expression was detected selectively in both myelinating SCs (mSCs) and non-myelinating SCs (nmSCs). A dual promoter and bidirectional AAV encoding a U6-driven short hairpin RNA against rat Piezo1 (PZ1shRNA) and CBA-transcribed GFP was packed with capsid olig001 (AAVolig001-PZ1shRNA), and AAV was injected into unilateral sciatic (or tibial) nerve immediately after induction of common peroneal nerve injury (CPNI). Results showed that the development of mechanical hypersensitivity in the CPNI rats injected with AAVolig001-PZ1shRNA was mitigated compared to rats subjected to AAVolig001-scramble. Selective in vivo SC transduction and functional block of Piezo1 channel activity of primary cultured SCs was confirmed. These data demonstrate that (1) AAVolig001 has unique and selective primary tropism to SCs via intraneural delivery, and (2) SC Piezo1 contributes to mechanical hypersensitivity following nerve injury.

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选择性RNAi沉默雪旺细胞Piezo1可减轻周围神经损伤后的机械超敏反应。

本研究旨在探讨雪旺细胞（SC） Piezo1在外周伤害感觉中的作用。我们首先开发了一种AAV载体，该载体在进入坐骨神经（或胫神经）后具有原发性SC倾向。这是通过使用衣壳AAVolig001包装由CBA启动子转录的AAV-GFP来实现的，以生成AAVolig001-CBA- gfp。在幼稚大鼠神经内注射AAVolig001-CBA-GFP 6周后，GFP在髓鞘化SCs （mSCs）和非髓鞘化SCs （nmSCs）中都有选择性地表达。将编码u6驱动短发夹RNA的抗大鼠Piezo1 （PZ1shRNA）和cba转录GFP的双启动子和双向AAV与衣壳olig001 （AAVolig001-PZ1shRNA）包裹，在诱导腓总神经损伤（CPNI）后立即注射到单侧坐骨（或胫骨）神经。结果表明，与AAVolig001-scramble相比，注射AAVolig001-PZ1shRNA的CPNI大鼠的机械超敏反应得到了缓解。证实了原代培养SCs在体内的选择性SC转导和Piezo1通道活性的功能阻断。这些数据表明(1)AAVolig001通过神经内传递对SC具有独特和选择性的初级倾向，(2)SC Piezo1有助于神经损伤后的机械超敏反应。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Molecular Therapy-Methods & Clinical Development Biochemistry, Genetics and Molecular Biology-Molecular Biology

CiteScore

9.90

自引率

4.30%

发文量

163

审稿时长

12 weeks

期刊介绍： The aim of Molecular Therapy—Methods & Clinical Development is to build upon the success of Molecular Therapy in publishing important peer-reviewed methods and procedures, as well as translational advances in the broad array of fields under the molecular therapy umbrella. Topics of particular interest within the journal''s scope include: Gene vector engineering and production, Methods for targeted genome editing and engineering, Methods and technology development for cell reprogramming and directed differentiation of pluripotent cells, Methods for gene and cell vector delivery, Development of biomaterials and nanoparticles for applications in gene and cell therapy and regenerative medicine, Analysis of gene and cell vector biodistribution and tracking, Pharmacology/toxicology studies of new and next-generation vectors, Methods for cell isolation, engineering, culture, expansion, and transplantation, Cell processing, storage, and banking for therapeutic application, Preclinical and QC/QA assay development, Translational and clinical scale-up and Good Manufacturing procedures and process development, Clinical protocol development, Computational and bioinformatic methods for analysis, modeling, or visualization of biological data, Negotiating the regulatory approval process and obtaining such approval for clinical trials.