新霉素损伤哺乳动物前庭毛细胞后的转录组学变化分析
|
摘要
目的:探讨毛细胞损伤过程中的基因表达、信号通路、生物学过程变化,为毛细胞再生及前庭功能重塑提供新的思路。方法:采用RNA-seq方法研究新生小鼠椭圆囊在新霉素损伤后的基因表达变化及相关分子通路。结果:新霉素处理样品中分别有172个和232个基因表达上调和下调。这些基因的GO和KEGG通路分析揭示了许多可能与毛细胞发育相关的细胞成分、分子功能、生物过程和信号通路。结论:本研究的结果为未来的研究提供了有价值的参考和前期基础,并发现Wnt、Notch和TGFβ等信号通路可能与前庭毛细胞损伤及再生密切相关,可能为前庭毛细胞再生研究及临床眩晕防治提供重要信号通路和基因靶点。
AIM: Vestibular hair cell damage is an important cause of vestibular dysfunction. This study tried to explore the changes of gene expression, signal pathway and biological process in the process of hair cell damage, so as to provide new ideas for hair cell regeneration and vestibular function recovery. METHODS: In the present study, RNA from the newborn mouse utricles was collected for RNA-seq immediately after the damage of neomycin, in order to investigate gene expression changes. RESULTS:There were 172 and 232 genes that were found to be up-regulated and down-regulated in neomycin-treated utricles. GO and KEGG pathway analyses of these genes revealed many deregulated cellular components, molecular functions, biological processes and signaling pathways that may be related to hair cell development.CONCLUSION: These results provide valuable reference and preliminary basis for future studies, and find that Wnt, Notch TGFβ and other signaling pathways that may be closely related to vestibular hair cell injury and regeneration, which may provide important signaling pathways and gene targets for vestibular hair cell regeneration research and clinical vertigo prevention.
AIM: Vestibular hair cell damage is an important cause of vestibular dysfunction. This study tried to explore the changes of gene expression, signal pathway and biological process in the process of hair cell damage, so as to provide new ideas for hair cell regeneration and vestibular function recovery. METHODS: In the present study, RNA from the newborn mouse utricles was collected for RNA-seq immediately after the damage of neomycin, in order to investigate gene expression changes. RESULTS:There were 172 and 232 genes that were found to be up-regulated and down-regulated in neomycin-treated utricles. GO and KEGG pathway analyses of these genes revealed many deregulated cellular components, molecular functions, biological processes and signaling pathways that may be related to hair cell development.CONCLUSION: These results provide valuable reference and preliminary basis for future studies, and find that Wnt, Notch TGFβ and other signaling pathways that may be closely related to vestibular hair cell injury and regeneration, which may provide important signaling pathways and gene targets for vestibular hair cell regeneration research and clinical vertigo prevention.
引文
[1] Wu Z,Grillet N,Zhao B,et al.Mechanosensory hair cells express two molecularly distinct mechanotransduction channels[J].Nat Neurosci,2017,20(1):24-33.
[2] Bermingham-McDonogh O,Rubel EW.Hair cell regeneration:winging our way towards a sound future[J].Curr Opin Neurobiol,2003,13(1):119-126.
[3] Li W,You D,Chen Y,et al.Regeneration of hair cells in the mammalian vestibular system[J].Front Med,2016,10(2):143-151.
[4] Han J,Wu H,Hu H,et al.Characterization of the transcriptome of hair cell regeneration in the neonatal mouse utricle[J].Cell Physiol Biochem,2018,51(3):1437-1447.
[5] Kawamoto K,Sha SH,Minoda R,et al.Antioxidant gene therapy can protect hearing and hair cells from ototoxicity[J].Mol Ther,2004,9(2):173-181.
[6] Menze MA,Hutchinson K,Laborde SM,et al.Mitochondrial permeability transition in the crustacean Artemia franciscana:absence of a calcium-regulated pore in the face of profound calcium storage[J].Am J Physiol Regul Integr Comp Physiol,2005,289(1):R68-R76.
[7] Dean EJ,Ward T,Pinilla C,et al.A small molecule inhibitor of XIAP induces apoptosis and synergises with vinorelbine and cisplatin in NSCLC[J].Br J Cancer,2010,102(1):97-103.
[8] Friedman LM,Avraham KB.MicroRNAs and epigenetic regulation in the mammalian inner ear:implications for deafness[J].Mamm Genome,2009,20(9-10):581-603.
[9] Lewis MA,Quint E,Glazier AM,et al.An ENU-induced mutation of miR-96 associated with progressive hearing loss in mice[J].Nat Genet,2009,41(5):614-618.
[10] Yu H,Lin Q,Wang Y,et al.Inhibition of H3K9 methyltransferases G9a/GLP prevents ototoxicity and ongoing hair cell death[J].Cell Death Dis,2013,4:e506.
[11] Yoda K,Ohnuki Y,Kurosawa H.Optimization of the treatment conditions with glycogen synthase kinase-3 inhibitor towards enhancing the proliferation of human induced pluripotent stem cells while maintaining an undifferentiated state under feeder-free conditions[J].J Biosci Bioeng,2019,127(3):381-387.
[12] Campbell DP,Chrysostomou E,Doetzlhofer A.Canonical Notch signaling plays an instructive role in auditory supporting cell development[J].Sci Rep,2016,6:19484.
[13] You D,Guo L,Li W,et al.Characterization of Wnt and Notch-responsive Lgr5+ hair cell progenitors in the striolar region of the neonatal mouse utricle[J].Front Mol Neurosci,2018,11:137.
[14] Ni W,Zeng S,Li W,et al.Wnt activation followed by Notch inhibition promotes mitotic hair cell regeneration in the postnatal mouse cochlea[J].Oncotarget,2016,7(41):66754-66768.
[15] Wu J,Li W,Lin C,et al.Co-regulation of the Notch and Wnt signaling pathways promotes supporting cell proliferation and hair cell regeneration in mouse utricles[J].Sci Rep,2016,6:29418.
[16] 孔令宇,席錾,杨亚勤,等.地塞米松对小鼠急性胰腺炎胰腺组织中Notch信号及凋亡相关基因表达的影响[J].中国临床药理学与治疗学,2019,24(2):134-139.
[17] Burns J,Stone J.Development and regeneration of vestibular hair cells in mammals[J].Semin Cell Dev Biol,2017,65:96-105.
[18] Zhang S,Zhang Y,Yu P,et al.Characterization of Lgr5+ progenitor cell transcriptomes after neomycin Injury in the neonatal mouse cochlea[J].Front Mol Neurosci,2017,10:213.
[19] Mittal R,Nguyen D,Patel AP,et al.Recent advancements in the regeneration of auditory hair cells and hearing restoration[J].Front Mol Neurosci,2017,10:236.
[20] McLean WJ,Yin X,Lu L,et al.Clonal expansion of Lgr5-positive cells from mammalian cochlea and high-purity generation of sensory hair cells[J].Cell Reports,2017,18(8):1917-1929.
[21] Gu X,Chai R,Guo L,et al.Transduction of adeno-associated virus vectors targeting hair cells and supporting cells in the neonatal mouse cochlea[J].Front Cell Neurosci,2019,13(10):3389.
[22] Liu Y,Han J,Chen Z,et al.Engineering cell signaling using tunable CRISPR-Cpf1-based transcription factors[J].Nat Commun,2017,8(1):2095.
[23] Sheridan C.Interest rekindles in drug cocktails that reprogram cells[J].Nat Biotechnol,2017,35(6):489-490.
[2] Bermingham-McDonogh O,Rubel EW.Hair cell regeneration:winging our way towards a sound future[J].Curr Opin Neurobiol,2003,13(1):119-126.
[3] Li W,You D,Chen Y,et al.Regeneration of hair cells in the mammalian vestibular system[J].Front Med,2016,10(2):143-151.
[4] Han J,Wu H,Hu H,et al.Characterization of the transcriptome of hair cell regeneration in the neonatal mouse utricle[J].Cell Physiol Biochem,2018,51(3):1437-1447.
[5] Kawamoto K,Sha SH,Minoda R,et al.Antioxidant gene therapy can protect hearing and hair cells from ototoxicity[J].Mol Ther,2004,9(2):173-181.
[6] Menze MA,Hutchinson K,Laborde SM,et al.Mitochondrial permeability transition in the crustacean Artemia franciscana:absence of a calcium-regulated pore in the face of profound calcium storage[J].Am J Physiol Regul Integr Comp Physiol,2005,289(1):R68-R76.
[7] Dean EJ,Ward T,Pinilla C,et al.A small molecule inhibitor of XIAP induces apoptosis and synergises with vinorelbine and cisplatin in NSCLC[J].Br J Cancer,2010,102(1):97-103.
[8] Friedman LM,Avraham KB.MicroRNAs and epigenetic regulation in the mammalian inner ear:implications for deafness[J].Mamm Genome,2009,20(9-10):581-603.
[9] Lewis MA,Quint E,Glazier AM,et al.An ENU-induced mutation of miR-96 associated with progressive hearing loss in mice[J].Nat Genet,2009,41(5):614-618.
[10] Yu H,Lin Q,Wang Y,et al.Inhibition of H3K9 methyltransferases G9a/GLP prevents ototoxicity and ongoing hair cell death[J].Cell Death Dis,2013,4:e506.
[11] Yoda K,Ohnuki Y,Kurosawa H.Optimization of the treatment conditions with glycogen synthase kinase-3 inhibitor towards enhancing the proliferation of human induced pluripotent stem cells while maintaining an undifferentiated state under feeder-free conditions[J].J Biosci Bioeng,2019,127(3):381-387.
[12] Campbell DP,Chrysostomou E,Doetzlhofer A.Canonical Notch signaling plays an instructive role in auditory supporting cell development[J].Sci Rep,2016,6:19484.
[13] You D,Guo L,Li W,et al.Characterization of Wnt and Notch-responsive Lgr5+ hair cell progenitors in the striolar region of the neonatal mouse utricle[J].Front Mol Neurosci,2018,11:137.
[14] Ni W,Zeng S,Li W,et al.Wnt activation followed by Notch inhibition promotes mitotic hair cell regeneration in the postnatal mouse cochlea[J].Oncotarget,2016,7(41):66754-66768.
[15] Wu J,Li W,Lin C,et al.Co-regulation of the Notch and Wnt signaling pathways promotes supporting cell proliferation and hair cell regeneration in mouse utricles[J].Sci Rep,2016,6:29418.
[16] 孔令宇,席錾,杨亚勤,等.地塞米松对小鼠急性胰腺炎胰腺组织中Notch信号及凋亡相关基因表达的影响[J].中国临床药理学与治疗学,2019,24(2):134-139.
[17] Burns J,Stone J.Development and regeneration of vestibular hair cells in mammals[J].Semin Cell Dev Biol,2017,65:96-105.
[18] Zhang S,Zhang Y,Yu P,et al.Characterization of Lgr5+ progenitor cell transcriptomes after neomycin Injury in the neonatal mouse cochlea[J].Front Mol Neurosci,2017,10:213.
[19] Mittal R,Nguyen D,Patel AP,et al.Recent advancements in the regeneration of auditory hair cells and hearing restoration[J].Front Mol Neurosci,2017,10:236.
[20] McLean WJ,Yin X,Lu L,et al.Clonal expansion of Lgr5-positive cells from mammalian cochlea and high-purity generation of sensory hair cells[J].Cell Reports,2017,18(8):1917-1929.
[21] Gu X,Chai R,Guo L,et al.Transduction of adeno-associated virus vectors targeting hair cells and supporting cells in the neonatal mouse cochlea[J].Front Cell Neurosci,2019,13(10):3389.
[22] Liu Y,Han J,Chen Z,et al.Engineering cell signaling using tunable CRISPR-Cpf1-based transcription factors[J].Nat Commun,2017,8(1):2095.
[23] Sheridan C.Interest rekindles in drug cocktails that reprogram cells[J].Nat Biotechnol,2017,35(6):489-490.