Differential Expression Profiles and Functional Prediction of tRNA-Derived Small RNAs in Rats After Traumatic Spinal Cord Injury
Differential Expression Profiles and Functional Prediction of tRNA-Derived Small RNAs in Rats After Traumatic Spinal Cord Injury
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脊髓创伤后大鼠中 tRNA 衍生小 RNA 的差异表达谱和功能预测
DOI：
10.3389/fnmol.2019.00326
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发表时间：
2020-01-10
影响因子：
4.8
通讯作者：
Li, Jian-Jun
中科院分区：
文献类型：
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作者：
Qin, Chuan;Feng, Hao;Li, Jian-Jun
脊髓损伤（Spinal cord injury，SCI）是一种常见的脊髓损伤。由于原发性机械性损伤是不可避免的，因此有必要关注SCI诱导继发性损伤的潜在分子机制，以开发有希望的SCI患者治疗方法。转移RNA衍生的小RNA（transferRNA-derivedsmallRNA，tsRNA）是一类新型的非编码短RNA，在多种疾病中具有潜在的调控功能。然而，tsRNA在创伤性SCI中的功能作用尚未确定。我们使用测序，定量逆转录-聚合酶链反应（qRT-PCR），生物信息学和荧光素酶报告基因分析的组合，以筛选表达谱，并确定脊髓损伤后的功能作用的tsRNA。结果，在大鼠脊髓挫伤后1天，鉴定出297个差异表达的tsRNA。其中，155个tsRNA有显著差异表达：91个在SCI后显著上调，而64个在SCI后显著下调（倍数变化> 1.5; P < 0.05）。生物信息学分析显示候选的tsRNA（tiRNA-Gly-GCC-001、tRF-Gly-GCC-012、tRF-Gly-GCC-013和tRF-Gly-GCC-016）可能通过靶向脑源性神经营养因子（BDNF）通过丝裂原活化蛋白激酶（MAPK）和神经营养因子信号通路发挥调节作用。我们验证了候选的tsRNA，并发现SCI后tsRNA和BDNF的表达水平呈相反的趋势。最后，使用荧光素酶报告基因测定鉴定了tiRNA-Gly-GCC-001靶向BDNF。总之，我们发现了一种改变的tsRNA表达模式，并预测tiRNA-Gly-GCC-001可能通过靶向BDNF参与MAPK和神经营养因子通路，从而调节SCI后的病理生理过程。本研究为进一步探讨脊髓损伤的发病机制和治疗靶点提供了新的思路。
Spinal cord injury (SCI) is mostly caused by trauma. As the primary mechanical injury is unavoidable, a focus on the underlying molecular mechanisms of the SCI-induced secondary injury is necessary to develop promising treatments for patients with SCI. Transfer RNA-derived small RNA (tsRNA) is a novel class of short, non-coding RNA, possessing potential regulatory functions in various diseases. However, the functional roles of tsRNAs in traumatic SCI have not been determined yet. We used a combination of sequencing, quantitative reverse transcription-polymerase chain reaction (qRT-PCR), bioinformatics, and luciferase reporter assay to screen the expression profiles and identify the functional roles of tsRNAs after SCI. As a result, 297 differentially expressed tsRNAs were identified in rats' spinal cord 1 day after contusion. Of those, 155 tsRNAs were significantly differentially expressed: 91 were significantly up-regulated, whereas 64 were significantly down-regulated after SCI (fold change > 1.5; P < 0.05). Bioinformatics analyses revealed candidate tsRNAs (tiRNA-Gly-GCC-001, tRF-Gly-GCC-012, tRF-Gly-GCC-013, and tRF-Gly-GCC-016) that might play regulatory roles through the mitogen-activated protein kinase (MAPK) and neurotrophin signaling pathways by targeting brain-derived neurotrophic factor (BDNF). We validated the candidate tsRNAs and found opposite trends in the expression levels of the tsRNAs and BDNF after SCI. Finally, tiRNA-Gly-GCC-001 was identified to target BDNF using the luciferase reporter assay. In summary, we found an altered tsRNA expression pattern and predicted tiRNA-Gly-GCC-001 might be involved in the MAPK and neurotrophin pathways by targeting the BDNF, thus regulating the post-SCI pathophysiologic processes. This study provides novel insights for future investigations to explore the mechanisms and therapeutic targets for SCI.