Kumvetsetsa matenda a airway mu JEB (2023)

Ana khumi ndi asanu ndi limodzi omwe ali ndi zizindikiro zapanjira ya mpweya anali ndi mayeso a majini omwe amasonyeza khumi ndi atatu mwa iwo anali ndi JEB, awiri anali ndi EBS ndipo mmodzi anali ndi RDEB. Jini yeniyeni ya laminin (LAMA3) inali ndi kusintha kwa majini mwa odwala khumi, kutanthauza kuti, ngakhale kuti njira yapamtunda ingakhudzidwe ndi mitundu ina ya EB chizindikiro ichi chimakhala chotheka ngati kusintha kwa chibadwa kuli mu LAMA3. Kusintha kwa majini mu majini a laminin kumalepheretsa mapuloteni a laminin kuti asapangidwe ndipo amachititsa zizindikiro za JEB chifukwa maselo a khungu sangathe kukhazikika bwino popanda izo.

Ma biopsy a Airway ochokera kwa odwala anayi adagwiritsidwa ntchito kukulitsa ma cell a 'airway EB' mu labotale. Ofufuza adawonetsa kuti akusowa mapuloteni a laminin opangidwa kuchokera ku jini ya LAMA3. Poyerekeza ndi ma cell ochokera kwa anthu opanda EB ma cell a 'airway EB' sanali abwino kumamatira ku mbale ya chikhalidwe cha ma cell. Gene therapy idapangidwa kuti iike jini yogwira ntchito ya LAMA3 m'maselo a 'airway EB'. Maselo atayamba kupanga mapuloteni kuchokera ku jini yatsopano, adakhala abwino ngati maselo omwe si a EB amamatira ku mbale ya chikhalidwe cha selo.

M'tsogolomu, maselowa adzafunika kukulitsidwa m'njira yomwe madokotala ochita opaleshoni angagwiritsire ntchito kuti alowe m'malo mwa tizilombo towonongeka kwa odwala EB. Kuti atsimikizire kuti izi ndi zotheka, ofufuza adachitapo kanthu, pogwiritsa ntchito zothandizira zosindikizidwa za 3D kuti akule ma cell a airway kukhala ma graft ndi kuwaika bwinobwino.

Chithandizo cha majinichi chimafunikira kukhathamiritsa kwina chisanathandize anthu omwe ali ndi zizindikiro za EB airway koma kafukufukuyu akuwonetsa kuti chithandizo chamtunduwu ndi chotheka.

Zotsatira za ntchitoyi zidasindikizidwa mu 2024: Lentiviral expression of wildtype LAMA3A imabwezeretsa kumamatira kwa cell m'maselo oyambira a airway kuchokera kwa ana omwe ali ndi epidermolysis bullosa. ndipo zafotokozedwa m'nkhani yofotokozera: Za LAMA3 ndi LAMB3: Njira yatsopano yochizira epidermolysis bullosa. Ntchitoyi idafotokozedwanso ndi epidermolysisbullosanews.com m'nkhani yomveka bwino: Njira yothandizira ma cell ndi majini ikuwonetsa lonjezo kwa ana a EB.

Kachilombo kamene kamasinthidwa chibadwa kwagwiritsidwa ntchito kuti agwiritse ntchito bwino majini a laminin m'maselo omwe amakula kuchokera ku biopsies odwala. Njirayi yabwerezedwa ndikuwongoleredwa kuti mupeze njira yabwino yopezera jini yatsopano m'maselo ambiri momwe mungathere. Ofufuzawa angasonyeze kuti mapuloteni a laminin amapangidwa m'maselo odwala pambuyo pa mankhwalawa komanso kuti maselo amalumikizana bwino.

Mapepala a mapuloteni apangidwa ndikuwonetsedwa kuti ndi oyenera kukulitsa maselo a khungu. Izi zitha kupanga cholumikizira chomwe chidzayesedwa kuti awone ngati maselo apulumuka. Ngati zigwira ntchito, ma cell a odwala omwe adalandira chithandizo amatha kupangidwa.

Mtsogoleri Wofufuza:

Mr Colin Butler, Pediatric ENT Fellow / Honorary Senior Lecturer, UCL Great Ormond Street Institute of Child Health, London

Colin Butler ndi Katswiri wa Ear Nose Throat Clinician pa Chipatala cha Great Ormond Street ndipo ali ndi chidwi chothandizira ana omwe ali ndi vuto lalikulu lakuyenda kwa mpweya. Iye wapanga chiyanjano cha opaleshoni m'derali ndipo ali ndi luso loika khungu munjira ya mpweya. Iye tsopano ali m'gulu la gulu lochiza ana omwe ali ndi EB omwe akukhudza njira yawo yoyendera mpweya. PhD yake yakhala mumankhwala ochiritsira ndipo adapatsidwa mwayi wa Wellcome Fellowship wofufuza njira yowonjezerera ma cell amtundu wa anthu akuluakulu kupita ku machiritso a epithelial panjira ya mpweya. Iye wasindikiza kwambiri mu gawo la airway ndipo wakhala ndi chidziwitso pakupanga zinthu kuchokera ku 'benchi kupita ku bedi'.

Ofunsira kwa Co:

Pulofesa Sam Janes, Mtsogoleri wa Dipatimenti Yofufuza Zopuma ku UCL; Pulofesa Paolo De Coppi, katswiri wa Neonatal ku UCL - Mothandizana ndi: Dr Gabriela Petrof, Dr Anna Martinez, Bambo Richard Hewitt (Paediatric Otolaryngologist Ear Nose and Throat GOSH)

“EB kukhudza njira ya mpweya ndi chinthu chosowa koma kwa omwe akhudzidwa ndi vutoli amabweretsa vuto lalikulu la kupuma chifukwa cha zipsera. Okhudzidwawo amakhala ndi machubu opumira omwe ndi aang'ono kwambiri moti angafanane ndi kupuma ndi udzu. Zosankha zachipatala ndizochepa kwambiri ndipo njira yokhayo yothetsera njira yochepetsera mpweya ndiyo kukulitsa njira yodutsa mpweya ndi baluni. Ngakhale kukulitsa kungathandize kuchepetsa kuchepa kwachangu, kuvulala kowonjezereka kumabweretsa mabala. Kafukufukuyu akufuna kumvetsetsa njirayi ndikupanga njira zochizira, makamaka popanga njira zatsopano zokulitsira khungu lapadera lanjira yapamlengalenga. Chiyembekezo ndi chakuti polimbana ndi njira ya ndege, madera ena akhoza kuyendetsedwa pogwiritsa ntchito njira zofanana. Kukhala wokhoza kuchiza zipsera za njira ya mpweya ndi minyewa yopangidwa ndi mpweya yomwe imapangidwa kuchokera ku maselo a wodwala kungasinthedi momwe tingachiritsire matendawa.

- Dr Colin Butler

Grant Title: Kuphatikiza kupuma kwa epithelial cell ndi gene therapy kuti athe kuchepetsa zizindikiro za kupuma mwa ana omwe ali ndi junctional epidermolysis bullosa (JEB)

Epidermolysis bullosa (EB) ndi vuto la majini komwe odwala amavutika ndi minyewa yosalimba kwambiri yomwe imatuluka mopweteka komanso zipsera popanda kuvulala pang'ono. Zimakhudza kwambiri khungu lakunja, komabe, bokosi la mawu ndi chitoliro cha mphepo zimatha kukhudzidwa kwambiri. Njira zothandizira pa airway EB ndizochepa kwambiri ndipo anthu omwe amakhudzidwa nthawi zambiri amavutika kumeza ndipo amatha kuvutika kwambiri kupuma chifukwa cha zipsera zam'mlengalenga. Potsirizira pake kutsekeka kwa mpweya kumapangitsa kuti pakhale kufunikira kwa tracheostomy, njira yachipatala yothandizira kutsegula njira ya mpweya. Kafukufuku m'derali wapeza kuti kulumikiza khungu mumsewu wopita kumlengalenga kungapereke mwayi wopereka maselo oyendetsa jini kuti athandize kupereka mankhwala ochiritsira matenda a EB.

Jini yomwe idzayang'ane kwambiri mu polojekitiyi ndi jini ya LAMA3, yomwe imayang'anira mapuloteni a laminin. Puloteniyi ndi yofunika kwambiri pothandiza maselo kuti azigwirizana kuti apereke mphamvu pakhungu ndi minofu ina yomwe imapezeka mumlengalenga, komanso kutenga nawo mbali pochiza mabala. Ntchito pano ikufuna kudziwa ngati chithandizo cha jini chingathandize kukonza njira yodutsa mpweya yomwe imakhudzidwa ndi EB. Maselo adzagwiritsidwa ntchito kuchokera mumsewu wa mpweya ndipo jini ya LAMA imakonzedwa kunja kwa thupi ndi kubwezeretsedwanso kuti awone ngati njirayi idzagwira ntchito kuthetsa matenda a airway.

Zolinga zazikulu zitatu za polojekitiyi ndikupereka zida zogwiritsira ntchito labotale kuti ziwonetsere matenda a EB airway, komanso kugwiritsa ntchito zida zosinthira ma gene kuti akonze mayendedwe apanjira okhudzidwa ndi EB.

Cholinga 1: Kumvetsetsa mozama mozama zambiri za epithelium (minofu) yomwe imakhudzidwa mumayendedwe apamlengalenga mu EB;

Cholinga 2: Kupanga njira zomwe zingatheke pakuwongolera jini ya LAMA3 pogwiritsa ntchito ma virus, (ma virus omwe amagwiritsidwa ntchito ngati njira yoperekera jini yokonzedwa);

Cholinga 3: Yesani kuti muwone momwe chithandizo chowongolera jini cha LAMA3 chimalandilidwa mumtundu wamoyo.

Zolinga zonsezi ndikuthandizira kupereka zowunikira kapena zoyambira zochiritsira zosinthidwa ndi majini za matenda a EB airway komanso kupanga chosowa cha ma cell a cell, (ma cell apadera kuti apititse patsogolo kumvetsetsa kwasayansi), kuti apange njira zamankhwala zamunthu payekha.

Kafukufukuyu atha kutseguliranso mwayi wokonza mitundu ya EB yomwe imakhudzanso minofu ina m'thupi, kuphatikizapo cornea (diso) epithelium ndi mucous nembanemba ya aerodigestive thirakiti, (mphuno, milomo, pakamwa, lilime, mmero, mawu ndi kumtunda). mbali ya mmero ndi chimphepo champhepo).

Kuthekera kopanga ma cell a matenda a airway mu EB pogwiritsa ntchito njirayi kudzatsegulanso chiyembekezo choyezetsa munthu payekhapayekha mankhwala kuti awonetsere kuchuluka kwa tinthu tating'onoting'ono ta maselo. Izi zikutanthauza kupeza chithandizo chapadera kwa munthu pogwiritsa ntchito deta yomwe yasonkhanitsidwa kuchokera mu kafukufukuyu.

Epidermolysis bullosa (EB) ndi gulu la matenda osowa majini pakhungu ndipo limakhudza kupatukana kwa minofu ndi kupanga matuza mkati mwa zigawo zosiyanasiyana za khungu. Mu mtundu wovuta kwambiri wa EB, wotchedwa junctional EB, odwala ena okhudzidwa amakhala ndi vuto la mayendedwe a mpweya kuphatikiza pa zotupa pakhungu. Kupanga matuza kutsatiridwa ndi mabala ndi kukhuthala kumachitika mumsewu wakumtunda ndipo izi zimatsekereza njira yodutsa mpweya. Ngakhale pambuyo pa kuchotsedwa kwa minyewa yopingasa, njira yowonongeka ya mpweya imapwetekanso chifukwa cha matenda obwerezabwereza ndipo pamapeto pake imatsogolera ku airway stenosis, yomwe ingakhale yosachiritsika. Chiwopsezo chachikulu cha kufa kwa odwala EB omwe ali ndi vuto la mayendedwe a mpweya watipangitsa ife kupanga chithandizo. M'gulu lathu la odwala ku Chipatala cha Great Ormond Street, tidazindikira kuti odwala a EB omwe ali ndi vuto loyendetsa ndege nthawi zambiri amanyamula masinthidwe a DNA mu jini yotchedwa LAMA3. Gulu lathu latulutsa ndikukulitsa ma cell a airway odwala mu labotale. Kapepala kogwira ntchito ka LAMA3 DNA kanalowetsedwa mu jini ya maselowa. Zotsatira zathu zasonyeza kuti chithandizo cha jinichi chikhoza kubwezeretsa maselo odwala a EB ku ntchito yachibadwa poyerekeza ndi maselo ochokera kwa anthu wamba. Gawo lotsatira la pulojekiti yathu likhala likuwonjezera mphamvu ya njira ya chithandizo cha jini ndikukonza njira yopangira opaleshoni kuti apereke maselo owongolera jini mumlengalenga. (Kuchokera ku lipoti la 2022).

Ofufuza ku University College London ndi Great Ormond Street Hospital for Children (GOSH) adalandira ndalama za DEBRA chifukwa cha ntchito yawo yopititsa patsogolo miyoyo ya odwala EB omwe amavutika ndi zovuta zapamtunda.

Mu gawo loyamba la kafukufuku wawo, asayansi adayang'ana kumvetsetsa zosowa zenizeni za odwala EB omwe amatchulidwa ku dipatimenti ya Ear, Nose, ndi Throat ku GOSH. Kafukufukuyu anaphatikizapo odwala 15, amuna ndi akazi, omwe ali ndi zaka zopitirira miyezi 9 panthawi yotumizidwa. Pakati pa odwalawa, mitundu yosiyanasiyana ya EB idadziwika, ndipo Junctional Epidermolysis Bullosa-Simplex (JEB-S) ndiyofala kwambiri. Kufufuza kwa majini a gulu la odwalawa kunavumbula kuti odwala asanu ndi anayi mwa 14 anali ndi mitundu yosiyanasiyana ya tizilombo toyambitsa matenda mu jini imodzi, LAMA3, kutanthauza kuti jini iyi imakhala ndi chizolowezi choyambitsa mavuto akuyenda.

Gulu lofufuza linakhazikitsa bwino chikhalidwe cha maselo kuchokera ku ma biopsies a airway kuchokera kwa odwala anayi omwe ali ndi 'airway EB', kuwalola kuti aphunzire maselo mu labotale. Iwo adapeza kuti panali kusowa kwa LAMA3 jini ndi kufotokoza kwa mapuloteni m'maselo a chikhalidwe, komanso kuti maselo analephera kutsata mbale za chikhalidwe cha pulasitiki komanso maselo ochokera kwa opereka omwe si a EB. Izi zikusonyeza kuti chitsanzo chochokera m'maselo ndichothandiza popitiriza kuphunzira njira ya EB, ndipo chitha kugwiritsidwa ntchito kuyesa mankhwala omwe angakhalepo.

Kutengera zidziwitso zomwe zidapezedwa kuchokera ku Aim 1, gulu lofufuza lidapanga ma lentiviral vectors kuti apereke LAMA3 ku ma cell a epithelial a airway omwe adakulira mu cell culture. Kugwiritsa ntchito vekitala ya LAMA3 ku ma cell odwala a EB kunapangitsa kuti LAMA3 RNA ichuluke kwambiri komanso mawu a protein. Chofunika kwambiri, ma cell a airway EB odwala omwe amawongolera amawonetsa kulumikizidwa kwa cell, ndipo anali ofanana ndi ma cell omwe si a EB. Kupeza uku kukuwonetsa kuti kuwongolera kwachidziwitso kwa mawu a LAMA3 kumatha kuthana ndi zovuta zomata ma cell kwa odwala ena a EB.

Kuti kafukufukuyu apitirire patsogolo, asayansi amayenera kutenga ma cell okonzedwa kuchokera ku chikhalidwe ndikuwaikanso kumayendedwe a mpweya. Komabe, mosiyana ndi kulumikiza khungu, palibe njira yopangira opaleshoni yomwe ingalole kuti izi zichitike pamene mukusunga mpweya wovomerezeka. Momwemo, gululo linayesa kuyesa kwa epithelial cell engraftment pogwiritsa ntchito chitsanzo cha opaleshoni. Analekanitsa ma cell a basal cell ndikuwakulitsa m'mikhalidwe yama cell yofanana ndi yomwe idagwiritsidwa ntchito kwa odwala a EB m'mbuyomu, kenako adapanga masinthidwe osindikizidwa a 3D opangidwa kuti azithandizira kuphatikizika kwa ma cell a tracheal epithelial cell. Kusanthula pambuyo pa opaleshoni kunawonetsa kulowetsedwa bwino kwa maselo otukuka. Chofunika kwambiri, izi zimatsegula chitseko cha kuthekera kwa ma cell a autologous opangidwa ndi ex vivo kuti azichita bwino pambuyo pomuika m'malo oyenera azachipatala.

Zotsatira za phunziroli zimapereka chiyembekezo kuti m'tsogolomu zidzakhala zotheka kuphatikiza chithandizo cha maselo ndi majini kuti apange njira yatsopano kwa odwala omwe ali ndi mawonetseredwe a EB. Kupita patsogolo komwe kunachitika pakumvetsetsa maziko a chibadwa cha EB, kupanga njira zowongolera ma jini, ndikulemba bwino maselo otukuka mumtundu wa nyama zomwe zimakhala ngati opaleshoni yofanana ndi anthu zimatsimikizira kuthekera kwa njira zochiritsira zosintha. Njira zotsatila mu phunziroli zikhala kukonza bwino ma lentiviral vectors kuti akhale oyenera kugwiritsidwa ntchito kwa odwala, komanso kupititsa patsogolo bwino komanso kuberekana kwa transplantation ya cell.