耳聋研究新动向
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摘要
随着新技术和新理论不断应用于耳科学研究领域,近年来对耳聋的基础和临床研究在国内发展日新月异。研究对象更加微观化和精细化,结构生物学探寻重要功能蛋白的分子构象以及活化与失活机制,为阐释毛细胞生理功能奠定理论基础;研究对象由听觉外周向听觉中枢延伸,功能核磁探寻听觉中枢核团神经兴奋电活动,为耳鸣发病机制研究及神经生物反馈治疗耳鸣提供新思路;对耳蜗转为微环境相关蛋白质组学和小分子代谢组学的研究,明确了调控耳蜗病理生理状态的关键因子,有助于阐释耳聋机制和制定干预策略;CRISPR/Cas9技术在耳聋动物模型制备、遗传性耳聋治疗方面发挥了重要作用,与听觉早期发育相关的多种基因的联合干预为毛细胞在成年哺乳动物耳蜗实现再生提供更多的可能性。听觉医学的进一步深入研究需要有更理想的大动物模型,本课题组已经做了很好的探索,发现猪模型在听觉医学研究的独特优势,特予总结报告,以资借鉴。
With the continuous application of new technologies and theories in the field of otology, basic science and clinical research in otology has been advancing rapidly in China in recent years. Microscopical and refined, structural biology studies have shed light on the molecular conformation, activation and inactivation of important functional proteins, laying a theoretical foundation for explaining physiological functions of hair cells. In the meantime, research subjects have extended from the auditory periphery to auditory centers. Functional magnetic resonance imaging(fMRI) has been used to study neuronal excitation activities in auditory nuclei and provided new ideas in understanding pathogenesis of as well as developing neurobiofeedback treatments for tinnitus. The study of proteomics and small molecular metabolomics related to cochlea microenvironment has defined key factors in regulation of cochlear pathophysiology,which are helpful to explaining mechanisms of deafness and to formulating intervention strategies. The CRISPR/Cas9 technique has played an important role in the development of animal model of deafness and the treatment of hereditary deafness. The intervention using combined genes associated with early auditory development has provided additional possibilities for hair cell regeneration in the adult mammalian cochlea. In order to further advance hearing medicine research, better large animal models are needed. Our group has invested significant amount of efforts in this regard and identified swine as a model for hearing medicine research with unique advantages. This is summarized in this review to be shared among our colleagues.
With the continuous application of new technologies and theories in the field of otology, basic science and clinical research in otology has been advancing rapidly in China in recent years. Microscopical and refined, structural biology studies have shed light on the molecular conformation, activation and inactivation of important functional proteins, laying a theoretical foundation for explaining physiological functions of hair cells. In the meantime, research subjects have extended from the auditory periphery to auditory centers. Functional magnetic resonance imaging(fMRI) has been used to study neuronal excitation activities in auditory nuclei and provided new ideas in understanding pathogenesis of as well as developing neurobiofeedback treatments for tinnitus. The study of proteomics and small molecular metabolomics related to cochlea microenvironment has defined key factors in regulation of cochlear pathophysiology,which are helpful to explaining mechanisms of deafness and to formulating intervention strategies. The CRISPR/Cas9 technique has played an important role in the development of animal model of deafness and the treatment of hereditary deafness. The intervention using combined genes associated with early auditory development has provided additional possibilities for hair cell regeneration in the adult mammalian cochlea. In order to further advance hearing medicine research, better large animal models are needed. Our group has invested significant amount of efforts in this regard and identified swine as a model for hearing medicine research with unique advantages. This is summarized in this review to be shared among our colleagues.
引文
1 Shi,X.Cochlear Pericyte Responses to Acoustic Trauma and the Involvement of Hypoxia-Inducible Factor-1alpha and Vascular Endothelial Growth Factor.Am J Pathol,2009.174(5):1692-1704.
2 Ohlemiller,K.K.,et al.,Absence of Strial Melanin Coincides with Age-associated Marginal Cell Loss and Endocochlear Potential Decline.Hear Res,2009.249(1-2):1-14.
3 Lin,D.W.and D.R.Trune,Breakdown of Stria Vascularis Blood-labyrinth Barrier in C3H/lpr Autoimmune Disease Mice.Otolaryngol Head Neck Surg,1997.117(5):530-534.
4 Cohen-Salmon,M.,et al.,Connexin30 Deficiency Causes Instrastrial Fluid-blood Barrier Disruption within the Cochlear Stria Vascularis.Proceedings of the National Academy of Sciences,2007.104(15):6229-6234.
5 Hirose,K.,et al.,Systemic Lipopolysaccharide Compromises the Blood-labyrinth Barrier and Increases Entry of Serum Fluorescein Into the Perilymph.Journal of the Association for Research in Otolaryngology,2014.15(5):707-719.
6 Hai T,Guo W,Yao J,et al.Creation of Miniature Pig Model of Human Waardenburg Syndrome Type 2A by ENU mutagenesis.Hum Genet.201711;136(11-12):1463-1475.
7任丽丽,郭德强,陈磊等,耳前入路小型猪耳蜗内电位测定的研究.中华耳科学,2011,09(3):254-257.Ren LL,Guo DQ,Chen L,et al.A New Approach to Endocochlear Potential Measurement in Mini Pigs[J].Chinese Journal of Otology,2011,9(3):254-257.
8 Karasawa,T.and P.S.Steyger,Intracellular Mechanisms of Aminoglycoside-induced Cytotoxicity.Integrative Biology,2011.3(9):879-886.
9 Ding,D.,B.L.Allman,and R.Salvi,Ototoxic Characteristics of Platinum Antitumor Drugs.The Anatomical Record:Advances in Integrative Anatomy and Evolutionary Biology,2012.295(11):1851-1867.
10 Cai T,Seymour M.L,Zhang H,et al.Conditional Deletion of Atoh1 Reveals Distinct Critical Periods for Survival and Function of Hair Cells in the Organ of Corti[J].J.Neurosci.2013,33:10110-10122.
11郭维维,胡吟燕,翟所强等.腺病毒携带的LacZ基因在豚鼠耳蜗中的表达[J].听力学及言语疾病杂志.2001,9(1):36-37.Guo WW,Hu YY,Zhai SQ,et al.Adenoviral Mediated LacZ Gene Expression in the Guinea Pig Cochlea[J].Journal of Audiology and Speech Pathology,2001,9(1):36-37.
12 Zhai S,Shi L,Wang BE,et al.Isolation and Culture of Hair Cell Progenitors from Postnatal Rat Cochleae[J].J Neurobiol,2005,65(3):282-293.
13杨仕明,杨伟炎,韩东一等.聋病基因研究新策略-基因敲除鼠听功能和内耳形态的系统研究[J].中华耳科学杂志,2006,4(3):161-163.Yang SM,Yang WY,Han DY,et al.The General Study on Hearing Function and Inner Ear Morphology in Knockout Mice-A Novel Gene Research Strategy for Deafness[J].Chinese Journal of Otology,2006,4(3):161-163.
14 Chen W,Hu YY,Yang SM,et al.Adenoviral-mediated Hath1-EGFP Gene Transfer Into Guinea Pig Cochlear Through an Intact Round Window Membrane in Vivo[J].Journal of Otology,2008,3(1):18-23.
15陈伟,杨仕明,郭维维等.腺病毒携带EGFP基因经完整圆窗膜转导豚鼠耳蜗的实验研究[J].中华耳科学杂志,2007,5(2):231-234.Chen W,Yang SM,Guo WW,et al.Cochlear Ad-EGFP Expression by Transferred Gene through an Intact Round Window Membrane in Guinea Pigs[J].Chinese Journal of Otology,2007,5(2):231-234.
16陈伟,郭维维,胡吟燕等.Math1基因内耳导入后噪声性聋豚鼠听功能改变观察[J].中华耳科学杂志,2009,7(4):352-356.Chen W,Guo WW,Hu YY,et al.Hearing Improvement of NIHLGuinea Pig by Overexpressed Math1 Gene in the Cochlea[J].Chinese Journal of Otology,2009,7(4):352-356.
17 Chen W,Jongkamonwiwat N,Abbas L,et al.Restoration of Auditory Evoked Responses by Human ES-Cell-Derived Otic Progenitors[J].Nature,2012,490:278-282.
18 Akil O,Seal RP,Burke K,et al.Restoration of Hearing in the VGLUT3 Knockout Mouse Using Virally Mediated Gene Therapy[J].Neuron 2012(75):283-293.
19 Xue Gao,Yong Tao,Veronica Lamas,et al.Treatment of Autosomal Dominant Hearing Loss By in Vivo Delivery of Genome Editing Agents.Nature[J].2018,11;553(7687):217-221.
20 Pan B,Askew C,Galvin A,et al.Gene Therapy Restores Auditory and Vestibular Function 1 in a Mouse Model of Usher Syndrome,Type 1c[J].Nat Biotechnol.2017,35(3):264-272.
21 Iizuka T1,Kamiya K1,Gotoh S,et al.Perinatal Gjb2 Gene Transfer Rescues Hearing in Amousemodel of Hereditary Deafness[J].Hum Mol Genet.2015,1;24(13):3651-3661.
22杨仕明.小型猪动物模型在耳科学领域的应用[J].中华耳科学杂志,2016,14(1):1-5.Yang SM.The Miniature Pig as an Animal Model in Otological Research.Chinese Journal of Otology,2016,14(1):1-5.
23 Pracy JP,White A,Mustafa Y.The Comparative Anatomy of the Pig Middle Ear Cavity.J Anat 1998;192:359-368.
24 Gurr A,Stark T,Probst G,et al.The Temporal Bone of Lamb and Pig as an Alternative in ENT-Education[J].Laryngorhinootologie.2010,89(1):17-24.
25 AndréGurr·K,Kevenh Rster·T,Stark·M.The Common Pig:a Possible Model for Teaching Ear Surgery[J].Eur Arch Otorhinolaryngol,2010,267:213-217.
26 Yi HJ,Guo W,Wu N,et al.The Temporal Bone Microdissection of Miniature Pigs as a Useful Large Animal Model for Otologic Research.Acta Otolaryngol.2013 Oct 9.[Epub ahead of print]
27 Guo WW,Chen L,Yang SM.Hearing and Cochlear Structures in a Swine Model.Journal of Otology 2010,5(2):97-100.
28 Guo W,Yi HJ,Ren LL,Chen L,Li DZ,Wei S,Yang SM.The Morphology and Electrophysiology of the Cochlea of the Miniature Pig.THE ANATOMICAL RECORD 298:494-500(2015).
29 Wei Chen,Haijin Yi,Liang Zhang,et al.Shuolong Yuan,Yue Zhang,Lili Ren,Jianan Li,Lei Chen,Weiwei Guo&Shiming Yang(2017):Establishing the Standard Method of Cochlear Implant in Rongchang pig,Acta Oto-Laryngologica,DOI:10.1080/00016489.2016.1267406
30郝青青.小型猪ENU诱变建立内耳Mondini畸形耳聋模型的初步研究[D].解放军医学院,2014.Hao QQ.Mondini Dysplasia in the ENU-Mutagenised Miniature Pig as an Animal Model[D].Chinese PLA Medical School,2014.
31陈志婷.基于mPEG_PLGA纳米载体蛋白质内耳递送研究[D].解放军医学院,2014.Chen ZT.Research of Inner Ear Delivery Based on m PEG-PLGANanocarrier for Protein[D].Chinese PLA Medical School,2014.
32 Hai T,Guo W,Yao J,et al.Creation of Miniature Pig Model of Human Waardenburg Syndrome Type 2A by ENU Mutagenesis[J].Human Genetics,2017(Pt 3):1-13.
2 Ohlemiller,K.K.,et al.,Absence of Strial Melanin Coincides with Age-associated Marginal Cell Loss and Endocochlear Potential Decline.Hear Res,2009.249(1-2):1-14.
3 Lin,D.W.and D.R.Trune,Breakdown of Stria Vascularis Blood-labyrinth Barrier in C3H/lpr Autoimmune Disease Mice.Otolaryngol Head Neck Surg,1997.117(5):530-534.
4 Cohen-Salmon,M.,et al.,Connexin30 Deficiency Causes Instrastrial Fluid-blood Barrier Disruption within the Cochlear Stria Vascularis.Proceedings of the National Academy of Sciences,2007.104(15):6229-6234.
5 Hirose,K.,et al.,Systemic Lipopolysaccharide Compromises the Blood-labyrinth Barrier and Increases Entry of Serum Fluorescein Into the Perilymph.Journal of the Association for Research in Otolaryngology,2014.15(5):707-719.
6 Hai T,Guo W,Yao J,et al.Creation of Miniature Pig Model of Human Waardenburg Syndrome Type 2A by ENU mutagenesis.Hum Genet.201711;136(11-12):1463-1475.
7任丽丽,郭德强,陈磊等,耳前入路小型猪耳蜗内电位测定的研究.中华耳科学,2011,09(3):254-257.Ren LL,Guo DQ,Chen L,et al.A New Approach to Endocochlear Potential Measurement in Mini Pigs[J].Chinese Journal of Otology,2011,9(3):254-257.
8 Karasawa,T.and P.S.Steyger,Intracellular Mechanisms of Aminoglycoside-induced Cytotoxicity.Integrative Biology,2011.3(9):879-886.
9 Ding,D.,B.L.Allman,and R.Salvi,Ototoxic Characteristics of Platinum Antitumor Drugs.The Anatomical Record:Advances in Integrative Anatomy and Evolutionary Biology,2012.295(11):1851-1867.
10 Cai T,Seymour M.L,Zhang H,et al.Conditional Deletion of Atoh1 Reveals Distinct Critical Periods for Survival and Function of Hair Cells in the Organ of Corti[J].J.Neurosci.2013,33:10110-10122.
11郭维维,胡吟燕,翟所强等.腺病毒携带的LacZ基因在豚鼠耳蜗中的表达[J].听力学及言语疾病杂志.2001,9(1):36-37.Guo WW,Hu YY,Zhai SQ,et al.Adenoviral Mediated LacZ Gene Expression in the Guinea Pig Cochlea[J].Journal of Audiology and Speech Pathology,2001,9(1):36-37.
12 Zhai S,Shi L,Wang BE,et al.Isolation and Culture of Hair Cell Progenitors from Postnatal Rat Cochleae[J].J Neurobiol,2005,65(3):282-293.
13杨仕明,杨伟炎,韩东一等.聋病基因研究新策略-基因敲除鼠听功能和内耳形态的系统研究[J].中华耳科学杂志,2006,4(3):161-163.Yang SM,Yang WY,Han DY,et al.The General Study on Hearing Function and Inner Ear Morphology in Knockout Mice-A Novel Gene Research Strategy for Deafness[J].Chinese Journal of Otology,2006,4(3):161-163.
14 Chen W,Hu YY,Yang SM,et al.Adenoviral-mediated Hath1-EGFP Gene Transfer Into Guinea Pig Cochlear Through an Intact Round Window Membrane in Vivo[J].Journal of Otology,2008,3(1):18-23.
15陈伟,杨仕明,郭维维等.腺病毒携带EGFP基因经完整圆窗膜转导豚鼠耳蜗的实验研究[J].中华耳科学杂志,2007,5(2):231-234.Chen W,Yang SM,Guo WW,et al.Cochlear Ad-EGFP Expression by Transferred Gene through an Intact Round Window Membrane in Guinea Pigs[J].Chinese Journal of Otology,2007,5(2):231-234.
16陈伟,郭维维,胡吟燕等.Math1基因内耳导入后噪声性聋豚鼠听功能改变观察[J].中华耳科学杂志,2009,7(4):352-356.Chen W,Guo WW,Hu YY,et al.Hearing Improvement of NIHLGuinea Pig by Overexpressed Math1 Gene in the Cochlea[J].Chinese Journal of Otology,2009,7(4):352-356.
17 Chen W,Jongkamonwiwat N,Abbas L,et al.Restoration of Auditory Evoked Responses by Human ES-Cell-Derived Otic Progenitors[J].Nature,2012,490:278-282.
18 Akil O,Seal RP,Burke K,et al.Restoration of Hearing in the VGLUT3 Knockout Mouse Using Virally Mediated Gene Therapy[J].Neuron 2012(75):283-293.
19 Xue Gao,Yong Tao,Veronica Lamas,et al.Treatment of Autosomal Dominant Hearing Loss By in Vivo Delivery of Genome Editing Agents.Nature[J].2018,11;553(7687):217-221.
20 Pan B,Askew C,Galvin A,et al.Gene Therapy Restores Auditory and Vestibular Function 1 in a Mouse Model of Usher Syndrome,Type 1c[J].Nat Biotechnol.2017,35(3):264-272.
21 Iizuka T1,Kamiya K1,Gotoh S,et al.Perinatal Gjb2 Gene Transfer Rescues Hearing in Amousemodel of Hereditary Deafness[J].Hum Mol Genet.2015,1;24(13):3651-3661.
22杨仕明.小型猪动物模型在耳科学领域的应用[J].中华耳科学杂志,2016,14(1):1-5.Yang SM.The Miniature Pig as an Animal Model in Otological Research.Chinese Journal of Otology,2016,14(1):1-5.
23 Pracy JP,White A,Mustafa Y.The Comparative Anatomy of the Pig Middle Ear Cavity.J Anat 1998;192:359-368.
24 Gurr A,Stark T,Probst G,et al.The Temporal Bone of Lamb and Pig as an Alternative in ENT-Education[J].Laryngorhinootologie.2010,89(1):17-24.
25 AndréGurr·K,Kevenh Rster·T,Stark·M.The Common Pig:a Possible Model for Teaching Ear Surgery[J].Eur Arch Otorhinolaryngol,2010,267:213-217.
26 Yi HJ,Guo W,Wu N,et al.The Temporal Bone Microdissection of Miniature Pigs as a Useful Large Animal Model for Otologic Research.Acta Otolaryngol.2013 Oct 9.[Epub ahead of print]
27 Guo WW,Chen L,Yang SM.Hearing and Cochlear Structures in a Swine Model.Journal of Otology 2010,5(2):97-100.
28 Guo W,Yi HJ,Ren LL,Chen L,Li DZ,Wei S,Yang SM.The Morphology and Electrophysiology of the Cochlea of the Miniature Pig.THE ANATOMICAL RECORD 298:494-500(2015).
29 Wei Chen,Haijin Yi,Liang Zhang,et al.Shuolong Yuan,Yue Zhang,Lili Ren,Jianan Li,Lei Chen,Weiwei Guo&Shiming Yang(2017):Establishing the Standard Method of Cochlear Implant in Rongchang pig,Acta Oto-Laryngologica,DOI:10.1080/00016489.2016.1267406
30郝青青.小型猪ENU诱变建立内耳Mondini畸形耳聋模型的初步研究[D].解放军医学院,2014.Hao QQ.Mondini Dysplasia in the ENU-Mutagenised Miniature Pig as an Animal Model[D].Chinese PLA Medical School,2014.
31陈志婷.基于mPEG_PLGA纳米载体蛋白质内耳递送研究[D].解放军医学院,2014.Chen ZT.Research of Inner Ear Delivery Based on m PEG-PLGANanocarrier for Protein[D].Chinese PLA Medical School,2014.
32 Hai T,Guo W,Yao J,et al.Creation of Miniature Pig Model of Human Waardenburg Syndrome Type 2A by ENU Mutagenesis[J].Human Genetics,2017(Pt 3):1-13.