Kurasikirwa kwekunzwa (HL) ndiyo inonyanya kuzivikanwa chirwere chekuremara muvanhu.Munyika dzakabudirira, chikamu che80 kubva muzana chekusanzwa kwevacheche muvana kunokonzerwa nemararamiro emagetsi.Inonyanya kuzivikanwa ndeye-single-gene defects (sezvinoratidzwa muFig. 1), 124 gene mutations yakawanikwa inobatanidza nekusanzwa kwekunzwa kurasikirwa kwevanhu, mamwe ose anokonzerwa nemamiriro ezvinhu ezvakatipoteredza.A cochlear implant (chigadzirwa chemagetsi chakaiswa munzeve yomukati chinopa kukurudzirwa kwemagetsi zvakananga kune tsinga yekuongorora) ndiyo nzira yakanakisisa yekurapa HL yakaoma, nepo chiyamuro chekunzwa (chigadzirwa chemagetsi chekunze chinoshandura uye kuwedzera masaisai enzwi) inogona kubatsira Varwere vane HL ine mwero.Nekudaro, parizvino hapana mishonga iripo yekurapa nhaka HL (GHL).Mumakore achangopfuura, kurapwa kwemajini kwakagamuchira kutariswa kwakawedzera senzira inovimbisa yekurapa kusashanda kwenzeve yemukati.

Fig1.Kuparadzirwa kwemhando yekusanzwa kwakabatana nematsi.[1]

Munguva pfupi yapfuura, masayendisiti kubva kuSalk Institute neYunivhesiti yeSheffield akaburitsa tsvakiridzo muMolecular Therapy - Methods & Clinical Development [2], iyo yakaratidza tarisiro yakafara yekushandiswa kwemu vivo gene kurapwa kwenhaka matsi.Uri Manor, mubatsiri purofesa paSalk Institute uye director weWaitt Center for Advanced Biophotonics, akati akaberekwa aine kurasikirwa kwakanyanya kwekunzwa uye akanzwa kuti kudzorera kunzwa chingave chipo chinoshamisa.Tsvagiridzo yake yapfuura yakawana kuti Eps8 iprotein inodzora actin ine actin inosunga uye capping zviitiko;mucochlear hair cells, protein complex inoumbwa neEps8 ine MYO15A, WHIRLIN, GPSM2 uye GNAI3 inonyanya kuwanikwa mune dzakawanda Mazano estereocilia akareba, ayo pamwe chete neMYO15A localize BAIAP2L2 pamatipi epfupi stereocilia, anodiwa pakuchengetedza masumbu ebvudzi.Nokudaro, Eps8 inogona kugadzirisa urefu hwe stereocilia yemasero emvere, iyo inokosha pakuita basa rekunzwa;Eps8 kudzima kana kuchinja kunotungamira kune pfupi stereocilia, iyo inoita kuti isakwanise kushandura zvakanaka ruzha kuita masaini emagetsi ekuona kwehuropi, izvo zvinozotungamira mukusanzwa..Panguva imwecheteyo, mushandirwi Walter Marcotti, purofesa paYunivhesiti yeSheffield, akawana kuti masero ebvudzi haagone kukura zvakajairika kana Eps8 isipo.Muchidzidzo ichi, Manor naMarcotti vakabatana kuti vaongorore kana kuwedzera Eps8 kumaseru estereociliary kunogona kudzoreredza basa ravo uye, zvakare, kunatsiridza kunzwa mumakonzo.Boka rekutsvagisa rakashandisa adeno-associated virus (AAV) vector Anc80L65 kuendesa iyo coding sequence ine wild-type EPS8 mucochlea yeEps8-/- achangozvarwa P1-P2 mbeva neraundi hwindo membrane jekiseni;mu mouse cochlear hair cells Basa re stereocilia rakagadziriswa vasati vakura;uye kugadzirisa kwekugadzirisa kwakaratidzwa ne imaging teknolojia uye kuyerwa kwe stereocilia.Mhedzisiro yacho yakaratidza kuti Eps8 yakawedzera kureba kwe stereocilia uye yakadzoreredza bvudzi sero basa mumaseru akaderera-frequency.Vakaona zvakare kuti, nekufamba kwenguva, maseru aiita kunge arasikirwa nekugona kununurwa neiyo gene therapy.Zvazvinoreva ndezvekuti kurapwa uku kungangoda kupihwa muchibereko, sezvo Eps8-/- masero ebvudzi anogona kunge akakura kana kuunganidza kukuvadzwa zvisingagone kugadziriswa mushure mekunge mbeva dzazvarwa."Eps8 ipuroteni ine mabasa mazhinji akasiyana, uye pachine zvakawanda zvekuongorora," Manor akadaro.Tsvagiridzo yeramangwana ichasanganisira kuongorora maitiro eEps8 gene therapy mukudzoreredza kunzwa pamatanho ekukura akasiyana, uye kuti zvingaite here kuwedzera mikana yekurapa.Sezvineiwo, muna Mbudzi 2020, Muzvinafundo KarenB Avraham weTel Aviv University muIsrael akaburitsa mhedzisiro yake mupepanhau EMBO Molecular Medicine [3], vachishandisa tekinoroji yekurapa kwemajini kugadzira isingakuvadze hutachiona hweadeno-inosanganiswa neAAV9-PHP.B, The gene defect mumasero emvere dzeSyne4-/- mice yakagadziridzwa nekupinza utachiona hunotakura coding sequence yeSyne4 munzeve yomukati yemakonzo, ichibvumira kupinda mumasero ebvudzi uye kusunungura zvinhu zvakatakurwa, zvichivabvumira kukura uye kushanda zvakanaka (semufananidzo 2).

Fig2.Schematic inomiririra yemukati nzeve anatomy, ine tarisiro pane iyo organ yeCorti uye cellular basa reNesprin-4.

Zvinogona kuonekwa kuti kushandiswa kwejene therapy kuzadzisa chinangwa chekurapa zvirwere zvenhaka pazera rejini kuburikidza nekuisa, kubvisa kana kugadzirisa chero ipi zvayo yakagadziriswa yekurapa (kureva, kutonga kuchinja kwemajini muchirwere) kune kliniki yepamusoro.tarisiro yekushandisa.Ikozvino gene therapy nzira dzekusanzwa kwemajini kushaya simba dzinogona kukamurwa muzvikamu zvinotevera:

gene replacement

Kutsiva kweGene zvine nharo ndiyo yakanyanya "yakatwasuka" nzira yekurapa majini, zvichibva pakuziva uye kutsiva jena rakaremara neyakajairwa kana yemusango kopi yejini.Kutanga kubudirira kwemukati nzeve gene therapy kudzidza kwekunzwa kurasikirwa kunokonzerwa nekubviswa kwe vesicular glutamate transporter 3 (VGLUT3) gene;AAV1-mediated kuendeswa kweiyo exogenous VGLUT3 overexpression mumasero emvere dzemukati nzeve (IHCs) Inogona kuguma nekugara kwekunzwa kupora, chikamu cheribhoni synaptic morphology kupora, uye kupinduka mhinduro [4].Zvisinei, mumienzaniso inosanganisira maviri maAAV-akaunzwa magene ekutsiva anotsanangurwa munhanganyaya pamusoro, zvakakosha kuziva kuti mhando dzembeva dzinoshandiswa kune mamwe marudzi ekubvisa magene ekurasikirwa kwekunzwa kwekunzwa kwakasiyana kwenguva pfupi kubva kuvanhu, uye muP1 mbeva, nzeve yemukati iri mudanho rekukura.Kusiyana naizvozvo, vanhu vanoberekwa vaine nzeve yomukati yakakura.Musiyano uyu unodzivirira kushandiswa kwembeva zvinoguma pakurapwa kwevanhu vanogarwa nhaka yekusanzwa kunze kwekunge gene therapy yaunzwa munzeve dzembeva dzakakura.

Gene Kugadzirisa: CRISPR/Cas9

Kuenzaniswa ne "gene replacement", kuvandudzwa kweiyo gene editing tekinoroji kwaunza mambakwedza ekurapa zvirwere zvemajini kubva kumudzi.Zvinonyanya kukosha, iyo nzira yekugadzira majini inogadzirisa kukanganisa kwechinyakare overexpression gene therapy nzira dzisina kukodzera kune dzinogara nhaka dzematsi matsi, uye dambudziko rekuti nzira yekuwedzeredza haigare kwenguva refu.Mushure mokunge vatsvakurudzi veChinese vakanyatsogogodza Myo6C442Y mutant allele muMyo6WT/C442Y mice vachishandisa AAV-SaCas9-KKH-Myo6-g2 gene editing system, uye mukati memwedzi ye5 yekugogodza, mice Basa rekunzwa remuenzaniso rakadzorerwa;panguva imwecheteyo, zvakaonekwawo kuti mararamiro ekupona kwemasero emvere munzeve yemukati akavandudzwa, chimiro checilia chakava chenguva dzose, uye zviratidzo zve electrophysiological zvakagadziriswa [5].Ichi chidzidzo chekutanga pasi rose kushandisa tekinoroji yeCRISPR/Cas9 mukurapa kusanzwa kwenhaka kunokonzerwa neMyo6 gene mutation, uye ibudiriro yakakosha yetsvakiridzo yehunyanzvi hwekugadzirisa magene pakurapa kusanzwa kwenhaka.Shanduro yekiriniki yekurapa inopa hwaro hwakasimba hwesainzi.

Gene therapy yekuendesa nzira

Kuti kurapa kwemajini kubudirire, mamorekuru eDNA akashama haakwanise kupinda maseru zvinobudirira nekuda kwehydrophilicity yavo uye mhosva yakaipa yemapoka ephosphate, uye kuve nechokwadi chekutendeseka kweakawedzera nucleic acid mamorekuru, nzira yakachengeteka uye inoshanda inofanirwa kusarudzwa.Iyo yakawedzerwa DNA inounzwa kune yakanangwa sero kana tishu.AAV inoshandiswa zvakanyanya semotokari yekuendesa kurapwa kwechirwere nekuda kwehupamhi hwekutapukira, yakaderera immunogenicity, uye yakafara tropism kune akasiyana matishu marudzi.Parizvino, boka rakakura rebasa rekutsvagisa rakateya tropism yeakasiyana subtypes eAAV zvine chekuita nemhando dzakasiyana dzesero mugonzo cochlea.Kushandisa maAAV ekutumira maitiro akasanganiswa nesero-chaiwo vanosimudzira vanogona kuwana sero-chaiyo kutaura, izvo zvinogona kuderedza kure-kunangwa mhedzisiro.Pamusoro pezvo, seimwe nzira kune echinyakare maAAV vectors, matsva ekugadzira AAV mavheta ari kugara achigadzirwa uye anoratidza hukuru hwekufambisa munzeve yemukati, iyo AAV2/Anc80L65 ndiyo inonyanya kushandiswa.Nzira dzisiri dzehutachiona dzekutumira dzinogona kukamurwazve kuita nzira dzemuviri (microinjection uye electroporation) uye nzira dzemakemikari (lipid-based, polymer-based, uye goridhe nanoparticles).Maitiro ese ari maviri akashandiswa mukurapa kwekugara nhaka kusanzwa uye aratidza zvakanakira zvakasiyana uye zvisingakwanisi.Pamusoro pemotokari yekutakura yejene kurapwa semotokari, nzira dzakasiyana dzemu vivo gene manejimendi dzinogona kushandiswa zvichienderana nemhando dzakasiyana dzemasero, nzira dzekutonga, uye kurapa kwakanaka.Chimiro chakaomesesa chenzeve yemukati chinoita kuti zviome kusvika kumaseru anonangwa uye kugoverwa kwemajenome editing agents kunononoka.Membranous labyrinth iri mukati mebhonzo rebhonzo repfupa renguva uye inosanganisira cochlear duct, semicircular duct, utricle, uye balloon.Kusarudzika kwayo, kutenderera kuri kuita lymphatic kushoma, uye kupatsanurwa kubva muropa neropa-maze barriers inodzikamisa iyo inoshanda systemic kuendesa kwekurapa kune mbeva dzinongozvarwa chete.Kuti uwane hutachiona hwehutachiona hunokodzera kurapwa kwemajini, jekiseni rakananga remunharaunda remavhairasi munzeve yemukati inodiwa.Nzira dzakasimbiswa dzejekiseni dzinosanganisira [6]: (1) round window membrane (RWM), (2) tracheostomy, (3) endolymphatic kana perilymphatic cochleostomy, (4) round window membrane pamwe neTube fenestration (CF) (semufananidzo 3).

Fig3.Inner nzeve kuendeswa kwejene therapy.

Kunyange zvazvo kufambira mberi kwakawanda kwakaitwa mukurapa kwemajini, zvichibva pazvinangwa zvekushandura kwekiriniki, basa rakawanda rinoda kuitwa kusati kwaitwa gene therapy inogona kuva nzira yekutanga yekurapa kune varwere vane zvirwere zvemajini, kunyanya mukugadzirwa kwemavector akachengeteka uye anobudirira uye nzira yekuendesa.Asi isu tinotenda kuti munguva pfupi iri kutevera, marudzi aya ekurapa anozove musimboti wekurapa kwemunhu uye achave nemhedzisiro yakanaka pahupenyu hwevanhu vane genetic disorder nemhuri dzavo.

Foregene yatanga zvakare yakakwira-kuburikidza yekuongorora kit kune yakanangwa majini, inokurumidza uye inogona kuita reverse transcription uye qPCR maitiro pasina RNA kutorwa.

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