Kutaya kumva (HL) ndi matenda omwe amapezeka kwambiri mwa anthu.M'mayiko otukuka, pafupifupi 80% ya milandu prelingual ugonthi ana amayamba chifukwa cha majini.Zowonjezereka ndizowonongeka kwa jini imodzi (monga momwe tawonetsera mkuyu 1), kusintha kwa 124 kwa jini kwapezeka kuti kumagwirizanitsidwa ndi kutayika kwa makutu osamva mwa anthu, zina zonse zimayambitsidwa ndi zinthu zachilengedwe.Kuyika kwa cochlear (chipangizo chamagetsi chomwe chimayikidwa mkati mwa khutu lamkati chomwe chimapereka mphamvu yamagetsi molunjika ku mitsempha yomveka) ndiyo njira yabwino kwambiri yothandizira HL kwambiri, pamene chothandizira kumva (chipangizo chamagetsi chakunja chomwe chimasintha ndi kukulitsa mafunde a phokoso) chingathandize Odwala omwe ali ndi HL yochepetsetsa.Komabe, pakali pano palibe mankhwala ochiritsira cholowa cha HL (GHL).M'zaka zaposachedwa, chithandizo cha majini chalandira chidwi chowonjezereka ngati njira yodalirika yochizira vuto lamkati la khutu.

Chithunzi 1.Kugawidwa kwa mitundu yosiyanasiyana yokhudzana ndi kusamva.[1]

Posachedwapa, asayansi ochokera ku Salk Institute ndi University of Sheffield adafalitsa zotsatira za kafukufuku mu Molecular Therapy - Methods & Clinical Development [2], zomwe zikuwonetsa chiyembekezo chogwiritsa ntchito mu vivo gene therapy ya ugonthi wobadwa nawo.Uri Manor, pulofesa wothandizira pa kafukufuku wa Salk Institute komanso mkulu wa Waitt Center for Advanced Biophotonics, adanena kuti anabadwa ndi vuto lalikulu lakumva ndipo adawona kuti kubwezeretsa kumva kungakhale mphatso yabwino kwambiri.Kafukufuku wake wam'mbuyomu adapeza kuti Eps8 ndi puloteni yowongolera actin yokhala ndi ntchito zomangirira komanso zotsekera;m'maselo a tsitsi la cochlear, mapuloteni opangidwa ndi Eps8 omwe ali ndi MYO15A, WHIRLIN, GPSM2 ndi GNAI3 makamaka amakhalapo kwambiri.Choncho, Eps8 ikhoza kulamulira kutalika kwa stereocilia ya maselo atsitsi, omwe ndi ofunikira kuti azimva bwino;Kuchotsa kapena kusintha kwa Eps8 kumayambitsa stereocilia yaifupi, zomwe zimapangitsa kuti zisathe kusintha bwino mawu kukhala zizindikiro zamagetsi kuti ubongo uzindikire, zomwe zimapangitsa kuti munthu asamve..Nthawi yomweyo, wothandizira Walter Marcotti, pulofesa ku Yunivesite ya Sheffield, adapeza kuti maselo atsitsi sangathe kukula bwino pakalibe Eps8.Mu kafukufukuyu, Manor ndi Marcotti adagwirizana kuti afufuze ngati kuwonjezera Eps8 ku maselo a stereociliary kungabwezeretse ntchito yawo, komanso kupititsa patsogolo kumva kwa mbewa.Gulu lofufuza linagwiritsa ntchito vekitala ya adeno-associated virus (AAV) Anc80L65 kuti ipereke ndondomeko yotsatizana yomwe ili ndi mtundu wa EPS8 wamtchire mu cochlea ya Eps8-/- mbewa za P1-P2 zobadwa kumene ndi jekeseni wozungulira zenera;m'maselo a tsitsi la mbewa Ntchito ya stereocilia inakonzedwa asanakhwime;ndipo kukonzanso kunkawoneka ndi luso la kulingalira ndi kuyeza kwa stereocilia.Zotsatira zake zidawonetsa kuti Eps8 idakulitsa kutalika kwa stereocilia ndikubwezeretsa ntchito ya cell ya tsitsi m'maselo otsika.Iwo adapezanso kuti, m'kupita kwa nthawi, maselo amawoneka kuti ataya mphamvu zawo zopulumutsidwa ndi mankhwala a jini.Tanthauzo lake ndilakuti mankhwalawa angafunikire kuperekedwa mu chiberekero, monga Eps8-/- ma cell atsitsi amatha kukhwima kapena kusonkhanitsa zowonongeka zomwe sizingatheke mbewa zitabadwa."Eps8 ndi mapuloteni omwe ali ndi ntchito zosiyanasiyana, ndipo pali zambiri zoti mufufuze," adatero Manor.Kafukufuku wamtsogolo adzaphatikizanso kufufuza momwe chithandizo cha majini cha Eps8 chimathandizira pakubwezeretsa kumva pakukula kosiyanasiyana, komanso ngati zingatheke kukulitsa mwayi wamankhwala.Zinangochitika kuti, mu Novembala 2020, Pulofesa KarenB Avraham wa ku Yunivesite ya Tel Aviv ku Israel adasindikiza zotsatira zake mu nyuzipepala ya EMBO Molecular Medicine [3], pogwiritsa ntchito ukadaulo waukadaulo wopangira ma gene kuti apange kachilombo kopanda vuto kogwirizana ndi adeno AAV9-PHP.B, Kuwonongeka kwa jini m'maselo atsitsi a Syne4-/- mbewa kunakonzedwa ndi jekeseni kachilombo kamene kamakhala ndi kalembedwe ka Syne4 mu khutu lamkati la mbewa, kulola kuti alowe m'maselo a tsitsi ndi kumasula ma genetic omwe amanyamula, kuwalola kuti akhwime ndikugwira ntchito bwino (monga mkuyu 2).

Chithunzi 2.Kuyimira kwadongosolo lamkati mwa khutu lamkati, loyang'ana kwambiri pagulu la Corti ndi ma cell a nesprin-4.

Zitha kuwoneka kuti kugwiritsa ntchito mankhwala a jini kuti akwaniritse cholinga chochizira matenda obadwa nawo pamlingo wa jini mwa kuika, kuchotsa kapena kukonza chibadwa chilichonse chosinthika kuti chichiritsidwe (ndiko kuti, kulamulira kusintha kwa majini mu matendawa) kumakhala ndi zotsatira zachipatala.chiyembekezo chogwiritsa ntchito.Njira zamakono zochizira matenda ogontha chifukwa cha chibadwa chawo zitha kugawidwa m'magulu awa:

gene m'malo

Kusintha kwa majini mosakayikira ndi njira "yowongoka" kwambiri yochizira majini, kutengera kuzindikira ndikusintha jini yosokonekera ndi jini yabwinobwino kapena yakuthengo.Kuphunzira koyamba kopambana kwa jini yamkati yamakutu chifukwa cha kutayika kwa makutu chifukwa cha kuchotsedwa kwa jini ya vesicular glutamate transporter 3 (VGLUT3);Kupereka kwapakati kwa AAV1 kwa VGLUT3 mopitilira muyeso m'maselo atsitsi amkati (IHCs) Kutha kupangitsa kuti kumva kumveke bwino, kuchira pang'ono kwa riboni synaptic morphology, ndi mayankho okhudzika [4].Komabe, mu zitsanzo kuphatikizapo ma jini awiri operekedwa ndi AAV omwe afotokozedwa m'mawu oyamba pamwambapa, ndikofunika kuzindikira kuti zitsanzo za mbewa zomwe zimagwiritsidwa ntchito pa mitundu ina ya kuchotsedwa kwa majeremusi obadwa nawo ndi osiyana kwakanthawi ndi anthu, ndipo mu P1 mbewa, khutu lamkati liri pa msinkhu wokhwima.Mosiyana ndi zimenezi, anthu amabadwa ndi khutu lamkati lokhwima.Kusiyanaku kumalepheretsa kugwiritsa ntchito zotsatira za mbewa pochiza matenda ogontha otengera cholowa chamunthu pokhapokha ngati chithandizo cha majini chikuperekedwa ku makutu okhwima a mbewa.

Kusintha kwa Gene: CRISPR/Cas9

Poyerekeza ndi "kusintha kwa majini", chitukuko chaukadaulo wosintha ma gene kwabweretsa m'bandakucha wochiza matenda obadwa nawo kuchokera muzu.Chofunika kwambiri, njira yosinthira majini imapangitsa kuti pakhale zofooka za njira zochiritsira zamtundu wamba zomwe sizili zoyenera matenda ogontha omwe amatengera cholowa, komanso vuto loti njira yopitilira muyeso sikhala nthawi yayitali.Ofufuza a ku China atagogoda mwachindunji Myo6C442Y mutant allele mu Myo6WT / C442Y mbewa pogwiritsa ntchito AAV-SaCas9-KKH-Myo6-g2 gene editing system, ndipo mkati mwa miyezi 5 ya kugogoda, mbewa Ntchito yomvetsera ya chitsanzo inabwezeretsedwa;panthawi imodzimodziyo, adawonanso kuti kupulumuka kwa maselo atsitsi mu khutu lamkati kunakula bwino, mawonekedwe a cilia anakhala okhazikika, ndipo zizindikiro za electrophysiological zinakonzedwa [5].Uwu ndi kafukufuku woyamba padziko lonse lapansi wogwiritsa ntchito ukadaulo wa CRISPR/Cas9 pochiza kusamva kwa cholowa komwe kumachitika chifukwa cha kusintha kwa jini ya Myo6, ndipo ndikupita patsogolo kofunikira kwaukadaulo wosintha ma gene pochiza kusamva kwa cholowa.Kutanthauzira kwachipatala kwa chithandizo kumapereka maziko olimba asayansi.

Njira zoperekera mankhwala a gene

Kuti chithandizo cha jini chikhale chopambana, mamolekyu amaliseche a DNA sangathe kulowa m'maselo bwino chifukwa cha hydrophilicity yawo ndi malipiro oipa a magulu a phosphate, ndikuwonetsetsa kukhulupirika kwa mamolekyu a nucleic acid, njira yotetezeka komanso yothandiza iyenera kusankhidwa.DNA yowonjezera imaperekedwa ku selo kapena minofu yomwe mukufuna.AAV imagwiritsidwa ntchito kwambiri ngati njira yoperekera chithandizo chamankhwala chifukwa champhamvu yake yopatsirana, kuchepa kwa chitetezo chamthupi, komanso kutentha kwakukulu kumitundu yosiyanasiyana ya minofu.Pakadali pano, gulu lalikulu la kafukufuku watsimikizira kufalikira kwa mitundu yosiyanasiyana ya AAV yokhudzana ndi mitundu yosiyanasiyana ya ma cell mu mbewa cochlea.Kugwiritsa ntchito mawonekedwe amtundu wa AAV ophatikizidwa ndi otsatsa omwe ali ndi ma cell amatha kukwaniritsa mawu okhudzana ndi ma cell, omwe amatha kuchepetsa zotsatira zomwe mukufuna.Kuphatikiza apo, monga m'malo mwa ma vector achikhalidwe a AAV, ma vector amtundu wa AAV atsopano akupangidwa mosalekeza ndikuwonetsa luso lapamwamba lotulutsa khutu lamkati, lomwe AAV2/Anc80L65 ndi lomwe limagwiritsidwa ntchito kwambiri.Njira zoperekera zopanda ma virus zitha kugawidwa m'njira zakuthupi (microinjection ndi electroporation) ndi njira zama mankhwala (lipid-based, polymer-based, and gold nanoparticles).Njira zonsezi zakhala zikugwiritsidwa ntchito pochiza matenda ogontha omwe timatengera kwa makolo ndipo zasonyeza ubwino ndi malire osiyanasiyana.Kuphatikiza pa galimoto yobweretsera chithandizo cha jini ngati galimoto, njira zosiyanasiyana zoyendetsera jini mu vivo zitha kugwiritsidwa ntchito kutengera mitundu yosiyanasiyana ya maselo, njira zoyendetsera, komanso chithandizo chamankhwala.Kapangidwe kake ka khutu lamkati kumapangitsa kuti zikhale zovuta kufikira ma cell omwe akukhudzidwa ndipo kugawa kwa ma genome editing agents kumachedwa.The membranous labyrinth ili mkati mwa fupa la fupa la temporal bone ndipo imaphatikizapo njira ya cochlear, semicircular duct, utricle, ndi baluni.Kudzipatula kwake, kumayenda pang'ono kwa ma lymphatic, komanso kupatukana ndi magazi ndi chotchinga chamagazi kumachepetsa njira yoperekera chithandizo kwa mbewa zakhanda zokha.Kuti mupeze ma virus oyenerera chithandizo cha majini, kubayidwa mwachindunji kwa ma virus amkati m'khutu ndikofunikira.Njira zokhazikitsidwa za jekeseni zikuphatikizapo [6]: (1) mawindo ozungulira (RWM), (2) tracheostomy, (3) endolymphatic kapena perilymphatic cochleostomy, (4) nembanemba yawindo lozungulira kuphatikizapo Tube fenestration (CF) (monga chithunzi 3).

Chithunzi 3.Kupereka makutu amkati kwa gene therapy.

Ngakhale kuti zapita patsogolo zambiri za chithandizo cha jini, malinga ndi zolinga zomasulira zachipatala, ntchito yowonjezereka iyenera kuchitidwa kuti mankhwala a jini asamakhale njira yoyamba yothandizira odwala omwe ali ndi matenda obadwa nawo, makamaka pakupanga ma vectors otetezeka komanso ogwira mtima komanso njira yobweretsera.Koma tikukhulupirira kuti posachedwapa, mitundu iyi ya chithandizo idzakhala chithandizo chamunthu payekha ndipo izikhala ndi zotsatira zabwino pamiyoyo ya anthu omwe ali ndi vuto la majini ndi mabanja awo.

Foregene yakhazikitsanso zida zowunikira zapamwamba kwambiri zama jini omwe akutsata, zomwe ndi zachangu komanso zimatha kuchita zolembera mosinthana ndi qPCR popanda kuchotsa RNA.

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