Engqungqutheleni Yokugoma Nezempilo, ochwepheshe bacele ukuthi "wonke umuntu anake imigomo ye-mRNA, enikeza abantu ukucabanga okungenamkhawulo."Ngakho uyini ngempela umgomo we-mRNA?Itholwe kanjani futhi liyini inani lesicelo sayo?Ingabe ingamelana ne-COVID-19 edla lubi emhlabeni wonke?Ingabe izwe lami liwusungule ngempumelelo umuthi wokugomela i-mRNA?Namuhla, ake sifunde mayelana nesikhathi esedlule nesamanje semithi yokugomela i-mRNA.

01
Iyini i-mRNA emithini yokugoma ye-mRNA?

I-mRNA (Isithunywa i-RNA), okungukuthi, i-RNA yesithunywa, iwuhlobo lwe-RNA enomucu owodwa ebhalwe ngomucu we-DNA njengesifanekiso futhi ephethe ulwazi lofuzo olungaqondisa ukuhlanganiswa kwamaprotheni.Ngokwemibandela ye-layman, i-mRNA iphindaphinda imininingwane yofuzo yochungechunge olulodwa lwe-DNA enemicu ephindwe kabili ku-nucleus, bese ishiya i-nucleus ukuze ikhiqize amaprotheni ku-cytoplasm.Ku-cytoplasm, ama-ribosomes ahamba eduze kwe-mRNA, afunde ukulandelana kwesisekelo sayo, futhi ahumushele ku-amino acid ehambisanayo, ekugcineni akhe iphrotheni (Umfanekiso 1).

Umdwebo 1 mRNA inqubo yokusebenza

02
Uyini umgomo we-mRNA futhi yini eyenza uhluke?

Imithi yokugomela i-mRNA yethula ama-antigen aqondene nezifo afaka i-mRNA emzimbeni, futhi isebenzisa indlela yokwenziwa kweprotheyini yeseli elisingethe ukukhiqiza ama-antigen, ngaleyo ndlela kubangele ukusabela komzimba.Imvamisa, ukulandelana kwe-mRNA kwama-antigen athile kungakhiwa ngokuya ngezifo ezahlukahlukene, kuhlanganiswe futhi kuthuthelwe kumaseli ngezinhlayiya zenoveli ze-lipid nanocarrier, bese ukulandelana kwe-mRNA kwama-ribosomes abantu kusetshenziselwa ukuhumusha ukulandelana kwe-mRNA ukukhiqiza amaprotheni e-antigen esifo, abonwa yi-autoimmune system ukuvikela ukusabela kwe-autoimmune ngemuva kwendima ye-2.

Umfanekiso 2. Umphumela we-in vivo womgomo we-mRNA

Ngakho-ke, yini ehlukile ngalolu hlobo lomgomo we-mRNA uma uqhathaniswa nemithi yokugoma yendabuko?Imithi yokugoma ye-mRNA iyimithi yokugomela esezingeni eliphezulu kakhulu yesizukulwane sesithathu, futhi ucwaningo olwengeziwe luyadingeka ukuze kuthuthukiswe ukuzinza, ukulawula ukuqina kwayo kokuzivikela komzimba, kanye nokuthuthukisa ubuchwepheshe obusha bokulethwa.

Isizukulwane sokuqala semithi yokugoma yendabuko ikakhulukazi ihlanganisa imithi yokugoma engasasebenzi kanye nemithi yokugoma enciphile, okuyiyona esetshenziswa kakhulu.Imithi yokugoma engasebenzi ibhekisela ekukhuliseni amagciwane noma amagciwane kuqala, bese iwavala ngokushisa noma ngamakhemikhali (imvamisa i-formalin);Imithi yokugoma enciphile ibhekisela kumagciwane ashintshashintshayo futhi enze buthaka ubuthi bawo ngemva kokwelashwa okuhlukahlukene.kodwa isagcina i-immunogenicity yayo.Ukuyijova emzimbeni ngeke kubangele ukuvela kwesifo, kodwa i-pathogen ingakhula futhi yande emzimbeni, ibangele ukusabela komzimba kwamasosha omzimba, futhi ibambe iqhaza ekutholeni ukuvikelwa kwesikhathi eside noma ukuphila konke.

Isizukulwane sesibili semithi yokugoma emisha sihlanganisa imithi yokugoma engaphansi kweyunithi kanye nemithi yokugoma yamaphrotheni.Umuthi wokugoma we-Subunit uwumuthi wokugoma owenziwe ngezingxenye eziyinhloko ze-immunogen ezivikelayo zamabhaktheriya e-pathogenic, okungukuthi, ngokubola kwamakhemikhali noma i-proteolysis elawulwayo, ukwakheka okukhethekile kwamaprotheni amabhaktheriya namagciwane kuyakhishwa futhi kuhlolwe.Imithi yokugoma eyenziwe ngezingcezu ze-immunologically asebenzayo;Imithi yokugoma yamaphrotheni enziwe kabusha ingamaprotheni e-antigen recombinant akhiqizwa ezinhlelweni ezihlukene zokuchazwa kwamaseli.

Isizukulwane sesithathu semithi yokugoma esezingeni eliphezulu ihlanganisa imigomo ye-DNA kanye nemithi yokugoma ye-mRNA.Iwukwethula ngokuqondile ucezu lofuzo lwegciwane (i-DNA noma i-RNA) efaka ikhodi ethile ye-antigenic kumaseli esilwane somatic (umjovo wokugoma emzimbeni womuntu), futhi ukhiqize iphrotheni ye-antigenic ngohlelo lwe-protein synthesis yeseli eliphethe, okuyenga umsingathi ukuthi akhiqize ukungatheleleki ekuphenduleni kwephrotheni ye-antigenic ukuze kuzuzwe inhloso yokuvimbela nokwelashwa kwezifo.Umehluko phakathi kwalokhu okubili ukuthi i-DNA iqala ukubhalwa ku-mRNA bese kuba amaprotheni ahlanganiswa, kuyilapho i-mRNA ihlanganiswa ngokuqondile.

03
Umlando wokutholwa kanye nenani lokusetshenziswa komgomo we-mRNA

Uma kukhulunywa ngemigomo ye-mRNA, kufanele sikhulume ngososayensi wesifazane ovelele, uKati Kariko, obeke isisekelo esiqinile socwaningo lwesayensi ngokufika kwemigomo ye-mRNA.Wayegcwele intshisekelo yocwaningo ku-mRNA ngenkathi efunda.Eminyakeni engaphezu kwengu-40 yomsebenzi wakhe wokucwaninga ngesayensi, wabhekana nezingqinamba eziphindaphindiwe, akazange afake isicelo sezimali zocwaningo lwesayensi, futhi akazange abe nesimo esizinzile socwaningo lwesayensi, kodwa ubelokhu ephikelela ocwaningweni lwe-mRNA.

Kati Karito

Kunama-node amathathu abalulekile ekufikeni kwemigomo ye-mRNA.

Esinyathelweni sokuqala, uphumelele ukukhiqiza i-molecule ye-mRNA ayeyifuna ngokusebenzisa isiko lamaseli, kodwa wahlangabezana nenkinga ekwenzeni i-mRNA isebenze emzimbeni: ngemva kokujova i-mRNA egundaneni, yayizogwinywa amasosha omzimba egundane.Wabe esehlangana no-Weissman.Basebenzise i-molecule ku-tRNA ebizwa nge-pseudouridine ukwenza i-mRNA igweme impendulo yokuzivikela komzimba.[2].
Esinyathelweni sesibili, cishe ngo-2000, uProf. Pieter Cullis wafunda i-lipid nanotechnology LNPs ekulethweni kwe-vivo ye-siRNA yezinhlelo zokusebenza zokuthulisa izakhi zofuzo [3][4].Inhlangano ye-Weissman uKariko et al.ithole ukuthi i-LNP ingumthwali ofanelekayo we-mRNA ku-vivo, futhi ingase ibe ithuluzi elibalulekile lokuletha i-mRNA encoding amaprotheni okwelapha, futhi ngemva kwalokho iqinisekiswe ekuvimbeleni igciwane leZika, i-HIV nezimila [5] [6][7][8].

Esinyathelweni sesithathu, ngo-2010 nango-2013, iModerna neBioNTech zathola amalayisense elungelo lobunikazi ngokulandelana okuhlobene nokuhlanganiswa kwe-mRNA eNyuvesi yasePennsylvania ukuze kuthuthukiswe.UKatalin uphinde waba iphini likamongameli omkhulu we-BioNTech ngo-2013 ukuze aqhubeke nokuthuthukisa imigomo ye-mRNA.

Namuhla, imigomo ye-mRNA ingasetshenziswa ezifweni ezithathelwanayo, izimila, kanye nesifuba somoya.Esimeni sokudlondlobala kwe-COVID-19 emhlabeni wonke, imigomo ye-mRNA ingase idlale indima njenge-vanguard.

04
Ithemba lokufaka isicelo somgomo we-mRNA ku-COVID-19

Ngobhubhane lomhlaba wonke lwe-COVID-19, amazwe asebenza kanzima ukwenza umuthi wokugomela ukunqanda lolu bhubhane.Njengohlobo olusha lokugoma, umuthi wokugomela i-mRNA ubambe iqhaza eliphambili ekusungulweni kobhubhane olusha lomqhele.Amajenali amaningi aphezulu abike indima ye-mRNA ku-SARS-CoV-2 i-coronavirus entsha (Umfanekiso 3).

Umfanekiso wesi-3 Umbiko ngemithi yokugomela i-mRNA ukuvimbela i-coronavirus entsha (kusuka ku-NCBI)

Okokuqala, ososayensi abaningi babike ucwaningo lomgomo we-mRNA (SARS-CoV-2 mRNA) ngokumelene ne-coronavirus entsha kumagundane.Isibonelo: umgomo we-lipid nanoparticle-encapsulated-nucleoside-modified mRNA (mRNA-LNP), umjovo womthamo owodwa udala izimpendulo zamaseli e-CD4+ T kanye ne-CD8+ T eqinile, i-plasma ehlala isikhathi eside kanye nezimpendulo ze-memory B zamaseli, kanye nokuphendula okuqinile nokuqhubekayo kwe-antibody okungathathi hlangothi.Lokhu kubonisa ukuthi umuthi wokugomela i-mRNA-LNP uyikhandidethi elithembisayo ngokumelene ne-COVID-19[9][10].

Okwesibili, abanye ososayensi baqhathanisa imiphumela ye-SARS-CoV-2 mRNA kanye nemithi yokugoma yendabuko.Uma kuqhathaniswa nemithi yokugoma ye-protein recombinant: Imithi yokugoma ye-mRNA iphakeme kakhulu kunemithi yokugoma yamaprotheni ekuphenduleni kwesikhungo se-germinal, ukusebenza kwe-Tfh, ukukhiqizwa kwe-antibody okungathathi hlangothi, inkumbulo ethile yamaseli B, namaseli e-plasma ahlala isikhathi eside [11].

Kwathi lapho abantu abazogomela i-SARS-CoV-2 mRNA bengena ezivivinyweni zomtholampilo, kwaphakama ukukhathazeka mayelana nesikhathi esifushane sokuvikela umgomo.Ososayensi benze uhlobo lwe-lipid-encapsulated lomuthi wokugomela i-nucleoside-modified mRNA obizwa nge-mRNA-RBD.Umjovo owodwa ungakhiqiza amasosha omzimba aqinile nezimpendulo zamaselula, futhi ungacishe uvikele ngokuphelele amagundane ayimodeli atheleleke nge-2019-nCoV, enamazinga aphezulu amasosha omzimba agcina egcinwe okungenani izinyanga eziyi-6.5.Le datha iphakamisa ukuthi umthamo owodwa we-mRNA-RBD unikeza isivikelo sesikhathi eside ngokumelene nenselelo ye-SARS-CoV-2 [12].
Kukhona nososayensi abasebenzela ukwakha imithi emisha ephephile nesebenzayo yokugomela i-COVID-19, njengomgomo we-BNT162b.Ama-macaque avikelwe kwa-SARS-CoV-2, avikela umgudu wokuphefumula ophansi ku-RNA ebangelwa yigciwane, akhiqiza amasosha omzimba anamandla, futhi awazange abonise zimpawu zokuthuthukisa isifo.Amakhandidethi amabili okwamanje angaphansi kokuhlolwa ezivivinyweni zesigaba I, futhi ukuhlolwa ezivivinyweni zesigaba II/III zomhlaba jikelele nakho kuyaqhubeka, futhi isicelo siseduze kakhulu [13].

05
Isimo somgomo we-mRNA emhlabeni

Njengamanje, i-BioNTech, iModerna kanye neCureVac baziwa njengabaholi abathathu abaphambili bezokwelapha be-mRNA emhlabeni.Phakathi kwabo, iBioNTech kanye neModerna bahamba phambili ocwaningweni nasekuthuthukisweni komgomo omusha wokugomela umqhele.IModerna ibigxile ocwaningweni nasekuthuthukisweni kwemithi nemijovo yokugoma ehlobene ne-mRNA.Umuthi wokugomela i-COVID-19 isigaba III i-mRNA-1273 uyiphrojekthi yenkampani ekhula ngokushesha kakhulu.I-BioNTech iphinde ibe yinkampani ehamba phambili emhlabeni yocwaningo lwe-mRNA yocwaningo lwemithi nokugoma, enesamba semithi/imithi yokugoma engu-19 mRNA, engu-7 yayo engene esigabeni somtholampilo.I-CureVac ibigxile ocwaningweni nasekuthuthukisweni kwemithi/imithi yokugoma ye-mRNA, futhi iyinkampani yokuqala emhlabeni ukusungula ulayini wokukhiqiza we-RNA othobela i-GMP, egxile emathumba, izifo ezithathelwanayo nezifo ezingandile.

Imikhiqizo ehlobene:I-RNase Inhibitor
Amagama angukhiye: umgomo we-miRNA, i-RNA Isolation, i-RNA extraction, i-RNase Inhibitor

Izikhombo:1.K Karikó, Buckstein M , Ni H , et al.Ukucindezelwa Kokuqashelwa Kwe-RNA Ngama-Toll-like Receptors: Umthelela Wokuguqulwa Kwe-Nucleoside kanye Ne-Evolutionary Origin ye-RNA[J].Ukungavikeleki, 2005, 23(2):165-175.
2. K Karikó, Muramatsu H, Welsh FA, et al.Ukufakwa Kwe-Pseudouridine Ku-mRNA Kukhiqiza IVector Ephakeme Engenayo Immunogenic Enamandla Angeziwe Okuhumusha Nokuqina Kwebhayoloji[J].I-Molecular Therapy, 2008.3.Chonn A , Cullis PR .Intuthuko yakamuva kubuchwepheshe be-liposome kanye nezicelo zabo zokulethwa kwezakhi zofuzo zesistimu[J].Ukubuyekezwa Okuthuthukisiwe Kokulethwa Kwezidakamizwa, 1998, 30(1-3):73.4.Kulkarni JA , Witzigmann D , Chen S , et al.I-Lipid Nanoparticle Technology yokuhumusha komtholampilo kwe-siRNA Therapeutics[J].Ama-akhawunti Ocwaningo Lwekhemikhali, 2019, 52(9).5.UKariko, Katalin, Madden, et al.I-Expression kinetics ye-nucleoside-modified mRNA ilethwe ngama-lipid nanoparticles kumagundane ngemizila ehlukahlukene[J].Ijenali Yokukhishwa Okulawulwayo Ijenali Esemthethweni Yenhlangano Yokukhishwa Okulawulwayo, 2015.6.Ukuvikelwa kwegciwane le-Zika ngomuthi wokugomela we-mRNA womthamo owodwa ophansi we-nucleoside-modified[J].Imvelo, 2017, 543(7644):248-251.7.Pardi N , Secreto AJ , Shan X , et al.Ukuphathwa kwe-nucleoside-modified mRNA efaka ama-antibody e-nucleoside-modified widely neutralizing kuvikela amagundane enziwe abantu enseleleni ye-HIV-1[J].Ukuxhumana Kwemvelo, 2017, 8:14630.8.Stadler CR , B?Hr-Mahmud H , Celik L , et al.Ukuqedwa kwezimila ezinkulu kumagundane ngama-mRNA-encoded bispecific antibodies[J].Imithi Yemvelo, 2017.9.NN Zhang, Li XF , Deng YQ , et al.Umuthi wokugomela i-mRNA we-Thermostable ngokumelene ne-COVID-19[J].Iseli, 2020.10.D Laczkó, Hogan MJ , Toulmin SA , et al.Ukugonywa Okukodwa NgeMijovo ye-mRNA eshintshiwe ye-Nucleoside Kuletha Izimpendulo Eziqinile Zeselula kanye Nezimpendulo Zamasosha omzimba aHumoral ngokumelene ne-SARS-CoV-2 kumagundane - ScienceDirect[J].2020.11.Lederer K , Castao D , Atria DG , et al.I-SARS-CoV-2 mRNA Vaccines Foster Potent Antigen-Specific Germinal Center Izimpendulo Ezihlotshaniswa Ne-Neutralizing Antibody Generation[J].Ukungavikeleki, 2020, 53(6):1281-1295.e5.12.Huang Q , Ji K , Tian S , et al.Umuthi wokugomela umthamo owodwa we-mRNA uhlinzeka ngokuvikeleka kwesikhathi eside kwamagundane e-hACE2 ashintshashintshayo avela kwa-SARS-CoV-2[J].Ezokuxhumana Ngemvelo.13.Vogel AB , Kanevsky I , Ye C , et al.Imithi yokugomela i-BNT162b ivikela ama-rhesus macaques kwa-SARS-CoV-2[J].Imvelo, 2021:1-10.