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Transcriptomic analysis reveals essential microRNAs after peripheral nerve injury

Neural Regeneration Research 2021年第9期16卷 1865-1870页

作者： Yu Wang Shu Wang Jiang-Hong He key laboratory for neuroregeneration of jiangsu province and ministry of education Co-innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina

Studies have shown that microRNAs(miRNAs) mediate posttranscriptional regulation of target genes and participate in various physiological and pathological processes, including peripheral nerve injury. However, it is hard to select key miRNAs with essential biological functions among a large number of differentially expressed miRNAs. Previously, we collected injured sciatic nerve stumps at multiple time points after nerve crush injury, examined gene changes at different stages(acute, sub-acute, and post-acute), and obtained mRNA expression profiles. Here, we jointly analyzed mRNAs and miRNAs, and investigated upstream miRNAs of differentially expressed mRNAs using Ingenuity Pathway Analysis bioinformatic software. A total of 31, 42, 30, and 23 upstream miRNAs were identified at 1, 4, 7, and 14 days after rat sciatic nerve injury, respectively. Temporal expression patterns and biological involvement of commonly involved upstream miRNAs(miR-21, let-7, miR-223, miR-10 b, miR-132, miR-15 b, miR-127, miR-29 a, miR-29 b, and miR-9) were then determined at multiple time points. Expression levels of miR-21, miR-132, miR-29 a, and miR-29 b were robustly increased after sciatic nerve injury. Biological processes involving these miRNAs include multicellular organismal response to stress, positive regulation of the epidermal growth factor receptor signaling pathway, negative regulation of epithelial cell differentiation, and regulation of myocardial tissue growth. Moreover, we constructed mechanistic networks of let-7, miR-21, and miR-223, the most significantly involved upstream miRNAs. Our findings reveal that multiple upstream miRNAs(i.e., let-7, miR-21, and miR-223) were associated with gene expression changes in rat sciatic nerve stumps after nerve injury, and these miRNAs play an important role in peripheral nerve regeneration. This study was approved by the Experimental Animal Ethics committee of jiangsu province of China(approval No. 20190303-18) on March 3, 2019.

关键词： bioinformatic analysis Ingenuity Pathway Analysis mechanistic network microRNA peripheral nerve injury peripheral nerve regeneration RNA sequencing sciatic nerve crush

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The role of Rho GTPase family in cochlear hair cells and hearing

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Neural Regeneration Research 2023年第10期18卷 2167-2172页

作者： Yu-Bei Dai Xiang Gao Dong Liu Jie Gong School of Life Science Nantong Laboratory of Development and DiseasesKey Laboratory of Neuroregeneration of Jiangsu and Ministry of EducationCo-innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina

Rho GTPases are essential regulators of the actin cytoskeleton.They are involved in various physiological and biochemical processes such as the regulation of cytoskeleton dynamics,development,proliferation,survival,and regeneration.During the development of cochlear hair cells,Rho GTPases are activated by various extracellular signals through membrane receptors to further stimulate multiple downstream effectors.Specifically,RhoA,Cdc42,and Rac1,members of the classical subfamily of the Rho GTPase family,regulate the development and maintenance of cilia by inducing the polymerization of actin monomers and stabilizing actin filaments.In addition,they also regulate the normal morphology orientation of ciliary bundles in auditory hair cells,which is an important element of cell polarity regulation.Moreover,the actin-related pathways mediated by RhoA,Cdc42,and Rac1 also play a role in the motility of outer hair cells,indicating that the function of Rho GTPases is crucial in the highly polar auditory sensory system.In this review,we focus on the expression of RhoA,Cdc42,and Rac1 in cochlear hair cells and how these small molecules participate in ciliary bundle morphogenesis and cochlear hair cell movement.We also discuss the progress of current research investigating the use of these small molecules as drug targets for deafness treatment.

关键词： actin assembly auditory sensory neurons cell polarity cell proliferation electromotility hair cell hearing loss morphogenesis Rho GTPases stereocilia

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Robust temporal changes of cellular senescence and proliferation after sciatic nerve injury

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Neural Regeneration Research 2022年第7期17卷 1588-1595页

作者： Yin-Ying Shen Rui-Rui Zhang Qian-Yan Liu Shi-Ying Li Sheng Yi key laboratory of neuroregeneration of jiangsu and ministry of education Co-innovation Center of NeuroregenerationNMPA Key Laboratory for Research and Evaluation of Tissue Engineering Technology ProductsNantong UniversityNantongJiangsu ProvinceChina

Cellular senescence and proliferation are essential for wound healing and tissue remodeling.However,senescence-proliferation cell fate after peripheral nerve injury has not been clearly revealed.Here,post-injury gene expression patterns in rat sciatic nerve stumps(SRP113121)and L4–5 dorsal root ganglia(SRP200823)obtained from the National center for Biotechnology Information were analyzed to decipher cellular senescence and proliferation-associated genetic changes.We first constructed a rat sciatic nerve crush model.Then,β-galactosidase activities were determined to indicate the existence of cellular senescence in the injured sciatic nerve.Ki67 and EdU immunostaining was performed to indicate cellular proliferation in the injured sciatic nerve.Both cellular senescence and proliferation were less vigorous in the dorsal root ganglia than in sciatic nerve stumps.These results reveal the dynamic changes of injury-induced cellular senescence and proliferation from both genetic and morphological aspects,and thus extend our understanding of the biological processes following peripheral nerve injury.The study was approved by the Animal Ethics committee of Nantong University,China(approval No.20190226-001)on February 26,2019.

关键词： bioinformatic analysis cellular senescence dorsal root ganglia p16 peripheral nerve regeneration peripheral nerve trauma proliferation rat sciatic nerves sciatic nerve crush β-galactosidase activities

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The combined application of stem cells and three-dimensional bioprinting scaffolds for the repair of spinal cord injury

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Neural Regeneration Research 2024年第8期19卷 1751-1758页

作者： Dingyue Ju Chuanming Dong Department of Anatomy Medical College of Nantong UniversityNantongJiangsu ProvinceChina key laboratory of neuroregeneration of jiangsu and ministry of education Co-innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina

Spinal cord injury is considered one of the most difficult injuries to repair and has one of the worst prognoses for injuries to the nervous system.Following surgery,the poor regenerative capacity of nerve cells and the generation of new scars can make it very difficult for the impaired nervous system to restore its neural functionality.Traditional treatments can only alleviate secondary injuries but cannot fundamentally repair the spinal cord.consequently,there is a critical need to develop new treatments to promote functional repair after spinal cord injury.Over recent years,there have been seve ral developments in the use of stem cell therapy for the treatment of spinal cord injury.Alongside significant developments in the field of tissue engineering,three-dimensional bioprinting technology has become a hot research topic due to its ability to accurately print complex structures.This led to the loading of three-dimensional bioprinting scaffolds which provided precise cell localization.These three-dimensional bioprinting scaffolds co uld repair damaged neural circuits and had the potential to repair the damaged spinal cord.In this review,we discuss the mechanisms underlying simple stem cell therapy,the application of different types of stem cells for the treatment of spinal cord injury,and the different manufa cturing methods for three-dimensional bioprinting scaffolds.In particular,we focus on the development of three-dimensional bioprinting scaffolds for the treatment of spinal cord injury.

关键词： biomaterials embryonic stem cells induced pluripotent stem cells mesenchymal stem cells nerve regeneration spinal cord injury stem cell therapy stem cells three-dimensional bioprinting

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Sequencing analysis of matrix metalloproteinase 7-induced genetic changes in Schwann cells

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Neural Regeneration Research 2020年第11期15卷 2116-2122页

作者： Pan-Jian Lu Gang Wang Xiao-Dong Cai Ping Zhang Hong-Kui Wang key laboratory of neuroregeneration of jiangsu and ministry of education Co-innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina

Previous research revealed the positive activity of matrix metalloproteinase 7(MMP7) on migration and myelin regeneration of Schwa nn cells(SCs). However, understanding of the molecular changes and biological activities induced by increased amounts of MMP7 in SCs remains limited. To better understand the underlying molecular events, primary SCs were isolated from the sciatic nerve stump of newborn rats and cultured with 10 nM human MMP7 for 24 hours. The results of genetic testing were analyzed at a relatively relaxed threshold value(fold change ≥ 1.5 and P-value components, diseases and functions, biosynthesis, material metabolism, cell movement, and axon guidance. The outcomes of further analysis will deepen our comprehension of MMP7-induced biological changes in SCs. This study was approved by the laboratory Animal Ethics committee of Nantong University, China(approval No. 20190225-004) on February 27, 2019.

关键词： bioinformatic analysis ingenuity pathway analysis matrix metalloproteinase 7 peripheral nerve regeneration RNA sequencing Schwann cells sciatic nerve injury

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Identification of key genes involved in axon regeneration and Wallerian degeneration by weighted gene co-expression network analysis

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Neural Regeneration Research 2022年第4期17卷 911-919页

作者： Yan Lu Qi Shan Mei Ling Xi-An Ni Su-Su Mao Bin Yu Qian-Qian Cao key laboratory of neuroregeneration of jiangsu and ministry of education Co-innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina School of Life Sciences Nantong UniversityNantongJiangsu ProvinceChina

Peripheral nerve injury repair requires a certain degree of cooperation between axon regeneration and Wallerian degeneration.Therefore,investigating how axon regeneration and degeneration work together to repair peripheral nerve injury may uncover the molecular mechanisms and signal cascades underlying peripheral nerve repair and provide potential strategies for improving the low axon regeneration capacity of the central nervous system.In this study,we applied weighted gene co-expression network analysis to identify differentially expressed genes in proximal and distal sciatic nerve segments from rats with sciatic nerve injury.We identified 31 and 15 co-expression modules from the proximal and distal sciatic nerve segments,respectively.Functional enrichment analysis revealed that the differentially expressed genes in proximal modules promoted regeneration,while the differentially expressed genes in distal modules promoted neurodegeneration.Next,we constructed hub gene networks for selected modules and identified a key hub gene,Kif22,which was up-regulated in both nerve segments.In vitro experiments confirmed that Kif22 knockdown inhibited proliferation and migration of Schwann cells by modulating the activity of the extracellular signal-regulated kinase signaling pathway.collectively,our findings provide a comparative framework of gene modules that are co-expressed in injured proximal and distal sciatic nerve segments,and identify Kif22 as a potential therapeutic target for promoting peripheral nerve injury repair via Schwann cell proliferation and migration.All animal experiments were approved by the Institutional Animal Ethics committee of Nantong University,China(approval No.S20210322-008)on March 22,2021.

关键词： axon regeneration extracellular signal-regulated kinase signaling pathway hub genes Kif22 peripheral nerve injury protein kinase Cα Schwann cells Wallerian degeneration weighted gene co-expression network analysis

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Peripheral nerve fibroblasts secrete neurotrophic factors to promote axon growth of motoneurons

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Neural Regeneration Research 2022年第8期17卷 1833-1840页

作者： Qian-Ru He Meng cong Fan-Hui Yu Yu-Hua Ji Shu Yu Hai-Yan Shi Fei Ding key laboratory of neuroregeneration of jiangsu and ministry of education Co-Innovation Center of NeuroregenerationJiangsu Clinical Medicine Center of Tissue Engineering and Nerve Injury RepairNantong UniversityNantongJiangsu ProvinceChina

Peripheral nerve fibroblasts play a critical role in nerve development and regeneration.Our previous study found that peripheral nerve fibroblasts have different sensory and motor phenotypes.Fibroblasts of different phenotypes can guide the migration of Schwann cells to the same sensory or motor phenotype.In this study,we analyzed the different effects of peripheral nerve-derived fibroblasts and cardiac fibroblasts on motoneurons.compared with cardiac fibroblasts,peripheral nerve fibroblasts greatly promoted motoneuron neurite outgrowth.Transcriptome analysis results identified 491 genes that were differentially expressed in peripheral nerve fibroblasts and cardiac fibroblasts.Among these,130 were significantly upregulated in peripheral nerve fibroblasts compared with cardiac fibroblasts.These genes may be involved in axon guidance and neuron projection.Three days after sciatic nerve transection in rats,peripheral nerve fibroblasts accumulated in the proximal and distal nerve stumps,and most expressed brain-derived neurotrophic factor.In vitro,brain-derived neurotrophic factor secreted from peripheral nerve fibroblasts increased the expression ofβ-actin and F-actin through the extracellular regulated protein kinase and serine/threonine kinase pathways,and enhanced motoneuron neurite outgrowth.These findings suggest that peripheral nerve fibroblasts and cardiac fibroblasts exhibit different patterns of gene expression.Peripheral nerve fibroblasts can promote motoneuron neurite outgrowth.

关键词： brain-derived neurotrophic factor differential gene expression F-actin fibroblasts motoneurons mRNA sequencing neurite outgrowth peripheral nervous system β-actin

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A comparative analysis of differentially expressed genes in rostral and caudal regions after spinal cord injury in rats

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Neural Regeneration Research 2022年第10期17卷 2267-2271页

作者： Xue-Min Cao Sheng-Long Li Yu-Qi Cao Ye-Hua Lv Ya-Xian Wang Bin Yu Chun Yao key laboratory of neuroregeneration of jiangsu and ministry of education NMPA Key Laboratory for Research and Evaluation of Tissue Engineering Technology ProductsCo-innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina

The initial mechanical damage of a spinal cord injury(SCI)triggers a progressive secondary injury cascade,which is a complicated process integrating multiple systems and cells.It is crucial to explore the molecular and biological process alterations that occur after SCI for therapy development.The differences between the rostral and caudal regions around an SCI lesion have received little attention.Here,we analyzed the differentially expressed genes between rostral and caudal sites after injury to determine the biological processes in these two segments after SCI.We identified a set of differentially expressed genes,including col3a1,col1a1,Dcn,Fn1,Kcnk3,and Nrg1,between rostral and caudal regions at different time points following SCI.Functional enrichment analysis indicated that these genes were involved in response to mechanical stimulus,blood vessel development,and brain development.We then chose col3a1,col1a1,Dcn,Fn1,Kcnk3,and Nrg1 for quantitative real-time PCR and Fn1 for immunostaining validation.Our results indicate alterations in different biological events enriched in the rostral and caudal lesion areas,providing new insights into the pathology of SCI.

关键词： biological process caudal differentially expressed genes Gene Ontology hemisection immunostaining Rattus norvegicus RNA-sequencing rostral spinal cord injury

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Heat shock factor 1 promotes neurite outgrowth and suppresses inflammation in the severed spinal cord of geckos

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Neural Regeneration Research 2023年第9期18卷 2011-2018页

作者： Bing-Qiang He Ai-Cheng Li Yu-Xuan Hou Hui Li Xing-Yuan Zhang Hui-Fei Hao Hong-Hua Song Ri-Xin Cai Ying-Jie Wang Yue Zhou Yong-Jun Wang key laboratory of neuroregeneration of jiangsu and ministry of education Co-innovation Center of NeuroregenerationMedical School of Nantong UniversityNantongJiangsu ProvinceChina Department of Rehabilitation Medicine Affiliated Hospital of Nantong UniversityNantongJiangsu ProvinceChina

The low intrinsic growth capacity of neurons and an injury-induced inhibitory milieu are major contributo rs to the failure of sensory and motor functional recovery following spinal cord injury.Heat shock transcription factor 1(HSF1),a master regulator of the heat shock response,plays neurogenetic and neuroprotective roles in the damaged or diseased central nervous system.However,the underlying mechanism has not been fully elucidated.In the present study,we used a gecko model of spontaneous nerve regeneration to investigate the potential roles of gecko HSF1(gHSF1) in the regulation of neurite outgrowth and inflammatory inhibition of macrophages following spinal cord injury.gHSF1 expression in neurons and microglia at the lesion site increased dramatically immediately after tail amputation.gHSF1 ove rexpression in gecko primary neuro ns significantly promoted axonal growth by suppressing the expression of suppressor of cytokine signaling-3,and fa cilitated neuro nal survival via activation of the mitogen-activated extracellular signal-regulated kinase/extracellular regulated protein kinases and phosphatidylinositol 3-kinase/protein kinase B pathways.Furthermore,gHSF1 efficiently inhibited the macrophagemediated inflammatory response by inactivating 1kappa B-alpha/NF-kappaB signaling.Our findings show that HSF1 plays dual roles in promoting axonal regrowth and inhibiting leukocyte inflammation,and provide new avenues of investigation for promoting spinal co rd injury repair in mammals.

关键词： apoptosis gecko heat shock factor 1 inflammation neuron regeneration spinal cord suppressor of cytokine signaling-3

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miR-30c promotes Schwann cell remyelination following peripheral nerve injury

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Neural Regeneration Research 2017年第10期12卷 1708-1715页

作者： Sheng Yi Qi-hui Wang Li-li Zhao Jing Qin Ya-xian Wang Bin Yu Song-lin Zhou key laboratory of neuroregeneration of jiangsu and ministry of education Co-innovation Center of NeuroregenerationNantong UniversityNantongJiangsu ProvinceChina

Differential expression of mi RNAs occurs in injured proximal nerve stumps and includes mi RNAs that are firstly down-regulated and then gradually up-regulated following nerve injury.These mi RNAs might be related to a Schwann cell phenotypic switch.mi R-30 c,as a member of this group,was further investigated in the current study.Sprague-Dawley rats underwent sciatic nerve transection and proximal nerve stumps were collected at 1,4,7,14,21,and 28 days post injury for analysis.Following sciatic nerve injury,mi R-30 c was down-regulated,reaching a minimum on day 4,and was then upregulated to normal levels.Schwann cells were isolated from neonatal rat sciatic nerve stumps,then transfected with mi R-30 c agomir and co-cultured in vitro with dorsal root ganglia.The enhanced expression of mi R-30 c robustly increased the amount of myelin-associated protein in the co-cultured dorsal root ganglia and Schwann cells.We then modeled sciatic nerve crush injury in vivo in Sprague-Dawley rats and tested the effect of perineural injection of mi R-30 c agomir on myelin sheath regeneration.Fourteen days after surgery,sciatic nerve stumps were harvested and subjected to immunohistochemistry,western blot analysis,and transmission electron microscopy.The direct injection of mi R-30 c stimulated the formation of myelin sheath,thus contributing to peripheral nerve regeneration.Overall,our findings indicate that mi R-30 c can promote Schwann cell myelination following peripheral nerve injury.The functional study of mi R-30 c will benefit the discovery of new therapeutic targets and the development of new treatment strategies for peripheral nerve regeneration.

关键词： nerve regeneration peripheral nerve regeneration peripheral nerve injury sciatic nerve mi RNAs mi R-30c dedifferentiation Schwann cells myelination in vivo in vitro neural regeneration

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