Jiyya na dindindin na DEB

Mataki na farko na aiwatar da shigar da kwayar halitta mai aiki an cimma shi cikin 'sauki don amfani' sel a cikin dakin gwaje-gwaje. Masu bincike yanzu suna aiki akan amintacciyar hanya mai inganci don yin hakan a cikin ƙwayoyin fata masu haƙuri. Masu bincike sun yi amfani da nanoparticles don ɗaukar kwayar halitta mai aiki zuwa cikin sel a cikin tasa kuma yanzu suna gwada su akan fata samfurin.

An buga sakamakon a cikin 2024 a cikin Jaridar Burtaniya ta ilimin likitanci kuma a cikin Jaridar Binciken Dermatology.

Dr Jacków ya gabatar da sabuntawa game da aikin a Makon Makon 2024:

Jagoran bincike:

Dokta Joanna Jacków tana da gogewa mai yawa da ingantaccen rikodin waƙa a cikin maganin halittar bullosa na epidermolysis da aikace-aikacen gyaran kwayoyin halitta. Dokta Jacków ya nuna ingantaccen gyare-gyaren kwayoyin halitta ta amfani da kayan aikin gyaran kwayoyin halitta iri-iri, ciki har da CRISPR-Cas9, tushe da gyare-gyare na farko, a cikin DEB don gyara maye gurbi a cikin keratinocytes, fibroblasts da ƙananan ƙwayoyin cuta (iPSCs).

Masu bincike:

Farfesa John McGrath ya haɓaka hanyoyin kwantar da hankali na ƙwayoyin cuta na DEB bisa ga allurar COL7A1 da aka gyara fibroblasts, yana nuna aminci da inganci da wuri a cikin gwaji na asibiti. Zai mayar da hankali kan zaɓin samfurin tantanin halitta da halayen halayen kuma zai taimaka wajen haifar da bututu don saurin fassarar asibiti na sababbin hanyoyin maganin COL7A1 a cikin marasa lafiya.

Farfesa Stephen Hart (UCL, GOSH, London) yana da ƙwarewa mai yawa a cikin tsarin bayarwa na novel da hanyoyin maganin nucleic acid tare da nanoparticles marasa hoto da kuma aikace-aikacen su ga maganin cututtuka ciki har da cystic fibrosis, neuroblastoma, ciliary dyskinesia na farko, da melanocytic naevi. .

"Sauraron mutanen da ke da DEB, mun san cewa mafarkin "cream gene" yana da girma akan jerin abubuwan da kowa ke so… Sabon aikin binciken mu shine haɓaka nau'i na dindindin na maganin ƙwayoyin cuta na COL7A1. A wannan matakin, muna son haɓaka sabbin fasaha don saka cikakken kwafin kwayar halittar COL7A1 har abada cikin kwayar halittar wani mai DEB.”

- Dr Joanna Jacków

Sunan bayar: PASTE-Matsakaici Superexon Maye gurbin COL7A1 a matsayin Jiyya don Ciwon Cutar Epidermolysis Bullosa

A cikin shekaru 30 da suka gabata mun koyi cewa bambance-bambancen DNA ne ke haifar da DEB a cikin nau'in VII collagen gene (COL7A1). Ba tare da kwayar halittar COL7A1 mai aiki ba, fata ba za ta iya samar da isasshen nau'in furotin na VII na collagen ba wanda ke nufin cewa fata ba ta da ƙarfi ga rauni kuma blisters suna tasowa. Kalubale ga masu bincike shine yadda ake maye gurbin ko gyara kwayar halittar COL7A1. Sauraron mutanen da ke da DEB, mun san cewa mafarkin "cream gene" yana da girma akan jerin abubuwan da kowa ke so.

Har ila yau, muna farin ciki da ci gaban da aka samu a baya-bayan nan wajen samar da sabon samfurin kwayoyin halittar COL7A1 ta abokan aiki a Jami'ar Stanford tare da haɗin gwiwar Krystal Biotech, kodayake wannan tsarin yana buƙatar samun maimaita aikace-aikace don samun tasiri mai dorewa. Sabon aikin binciken mu shine game da haɓaka nau'i na dindindin na maganin kwayoyin halittar COL7A1. A wannan mataki, muna son haɓaka sabbin fasaha don saka cikakken kwafin kwayar halittar COL7A1 har abada cikin kwayar halittar wani mai DEB. Sabon tsarin jiyya na kwayoyin halitta ana kiransa PASTE, wanda ke nufin "Ƙarin Ƙarfafawa ta hanyar Abubuwan Tage-Tage-Tallafi". Don wannan aikin, za mu yi amfani da PASTE don shigar da kwayar halittar COL7A1 cikin ƙwayoyin fata na DEB. Za mu duba don ganin ko za mu iya mayar da nau'in VII collagen. Sa'an nan kuma za mu yi aiki kan inganta tsarin bayarwa na lipid don tabbatar da cewa za mu iya samun maganin cikin fata ba kawai kwayoyin halitta ba. A yanzu, ba mu yin gwajin asibiti, amma wannan shine shirin mu na gaba.

Wannan aikin yana mai da hankali kan haɓaka niyya don warkar da jiyya don dystrophic epidermolysis bullosa (DEB). An sami ci gaba mai yawa don haɓaka maganin ƙwayoyin cuta don DEB ta amfani da ƙwayoyin cuta marasa haɗaka don isar da COL7A1 ga fata da ta ji rauni kodayake maimaita gudanarwa yana da mahimmanci don dorewar fa'ida.

A matsayin ƙarin hanya, muna nufin yin amfani da haɗin kai na dindindin na cikakken tsayin COL7A1 gene ta amfani da ƙarin shirye-shirye ta hanyar abubuwan da aka yi niyya ta musamman (PASTE). PASTE ya haɗu da ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun kayan aiki tare da babban ɗaukar nauyi na haɗin gwiwar serine don gabatar da har zuwa 36kb na kayan a wuraren da aka ƙera musamman a cikin genome. Haɗin kai na dindindin na COL7A1 mai cikakken tsayi zai ba da izinin "girma ɗaya ya dace da duka" tsarin gyaran DNA.

Da farko, za mu zaɓa da kuma kwatanta keratinocytes da / ko fibroblasts daga fata mai haƙuri na DEB. Daga nan za mu inganta ƙirar ƙirar PASTE ta amfani da electroporation na plasmids na musamman. COL7A1 gene hadewa za a tabbatar ta amfani da PCR- da Sanger sequencing. Kwayoyin da aka gyara cikin nasara za a keɓe su kuma a sake fasalin su don bincika ceton nau'in VII collagen. Bayan ingantacciyar ƙirar gini, a madadin DNA na plasmid, PASTE enzymes za a isar da su ta hanyar in-vitro haɗakarwar mRNA. Na gaba, za mu mai da hankali kan hanyoyin isarwa, haɓaka masu karɓar nanoparticles na tushen lipid (LNPs) don sadar da injin PASTE zuwa sel masu niyya. Nunin faifai zai gano jerin peptide waɗanda ke ɗaure da ƙarfi da fifiko ga fibroblasts da keratinocytes. Za a bincika abubuwan da aka gyara na lipid don dacewarsu don aikace-aikacen yanayi a cikin ƙirar fata na 3D. A takaice, wannan aikin yana nufin haɓaka sabon nau'in maganin maye gurbin kwayoyin halitta don DEB.

Aikin namu yana da manyan manufofi guda biyu:

Da fari dai, muna so mu ɗauki cikakken kwafin kwayar halittar da ba ta canza ba mu saka shi cikin DNA na marasa lafiya domin ƙwayoyin su su fara yin furotin da ake buƙata don lafiyayyen fata. Wannan zai ba mu damar ƙirƙirar genotype-agnostic (watau yana aiki komai maye gurbin da majiyyaci ya samu) da kuma magani na dindindin. A cikin watanni shida da suka gabata, mun sami matakin farko na gyaran DNA yana aiki sosai a cikin sel masu sauƙin canzawa, amma ƙwayoyin fata masu haƙuri suna nuna taurin kai. Yanzu muna aiki don tabbatar da cewa muna da amintacciyar yarjejeniya mai inganci don canza DNA a cikin sel waɗanda ke da mahimmanci.

Abu na biyu, muna son ƙirƙirar nanoparticles waɗanda za su iya ɗaukar wannan injin ɗin gyaran DNA zuwa cikin ƙwayoyin fata tare da tsari mai sauƙin amfani da kayan shafa. Muna gwaji da dabaru daban-daban na waɗannan nanoparticles. Mun sami wasu sakamako masu ban sha'awa da ake amfani da nanoparticles zuwa sel a cikin tasa, kuma yanzu muna gwada su akan fata samfurin. Wannan zai zama mafi wahala (fata ta samo asali musamman don kiyaye abubuwa), amma a cikin marasa lafiya na EB, shingen fata ya riga ya raunana kuma ya rushe. Muna binciko hanyoyin da za a bi da fata samfurin don yin kwafin fatar marasa lafiya na EB, kuma da fatan hakan zai bar nanoparticles su shiga zurfi cikin nama.

A halin yanzu, muna tattaunawa da masanin tattalin arziki don ganin 'yar jarida ta zo ta duba aikinmu don ta rubuta labarin ci gabanmu.