LONKE I-GENOME RESEQUENING
Ukuqapha kwe-Genomics kwe-SARS-CoV-2 kwembula ukwahluka kokususwa kwe-Nsp1 okushintsha impendulo yohlobo lwe-interferon
Nanopore |Ilumina |Ukulandelanisa kabusha kwe-genome |i-metagenomics |I-RNA-Seq |Sanger
I-Biomarker Technologies inikeze ukwesekwa kobuchwepheshe ekulandeleni kwesampula kulolu cwaningo.
Amaphuzu avelele
I-1.SARS-CoV-2 i-genome sequencing kanye nokuhlaziywa kwe-phylognetic kuhlonza izinguquko eziphindaphindayo ezingama-35 ezihlanganisa ama-SNP angama-31 kanye nama-Indels angu-4.
2.Ukuhlotshaniswa nama-phenotypes emtholampilo we-117 kuveza okungenzeka
izinguquko ezibalulekile.
∆500-532 esifundeni sekhodi se-Nsp1 sihlotshaniswa negciwane eliphansi
3.umthwalo kanye neserum IFN-β.
4.Ama-Viral isolate anokuguqulwa kuka-∆500-532 adala i-IFN-I ephansi
impendulo kumaseli angenwe yileli gciwane.
Idizayini Yokuhlola
Izimpumelelo
1. I-COVID-19 epidemiological and genomic surveillance
Idatha yomtholampilo yaqoqwa esifundazweni saseSichuan, e-China phakathi nesikhathi sokuqubuka kukaJan. 22, 2020 kuya ku-Feb. 20, 2020. Amacala angama-538 e-COVID-19 aqinisekiswe ukuhlolwa kwe-qPCR eSichuan, ama-28.8% awo abevela esifundazweni. capital.Amacala aqinisekisiwe e-Sichuan akhuphuke kakhulu, afinyelela umvuthwandaba ngomhla ka-Jan. 30.Futhi, idatha esekelwa ukuthi ukuqhelelana komphakathi kungaba yisici esibalulekile ekuvimbeleni ukusabalala kwegciwane.
Umfanekiso 1. Ucwaningo lwe-Epidemiological lwe-COVID-19 esifundazweni sase-Sichuan, e-China
2. Ukwakhiwa kofuzo lwe-SARS-CoV-2 kanye nokuhlonza okuhlukile
Ngokukhuliswa kwe-PCR ye-multiplex elandelwa ukulandelana kwe-nanopore, inani lama-genomes acishe abe ngu-310 aseduze noma ingxenye avela ezigulini ezingama-248 akhiqizwa cishe.U-80% wama-genomes ambozwe ngokufundwa okungu-10(Ukujula okushiwo: 0.39 M okufundwayo ngesampula ngayinye).
Umfanekiso 2. Ukuvama kokuhluka ngakunye eqenjini le-Sichuan
Isamba sama-SNP angu-104 kanye nama-Indel angu-18 ahlonzwe ku-SARS-CoV-2 genomes, lapho ama-SNP angu-31 nama-Indel angu-4 ahlonzwa njengezinhlobonhlobo zofuzo eziphindaphindayo.Ngokuwaqhathanisa namasampula angu-169 avela e-Wuhan kanye nama-81,391 asezingeni eliphezulu lokulandelana kwe-genome yomphakathi ku-GISAID, okuhlukile okungu-29 kwezingu-35 okutholwe kwethulwa kwamanye amazwekazi.Ngokuphawulekayo, izinhlobo ezine ezihlanganisa ∆500-532, ACC18108AT, ∆729-737 kanye ne-T13243C, zitholakale kuphela e-Sichuan nase-Wuhan futhi zingekho kudatha ye-GISAID, okubonisa ukuthi lezi zinhlobonhlobo kungenzeka kakhulu zifakwe e-Wuhan, ehlangabezana ne- amarekhodi okuhamba eziguli.
Ukuhlaziywa kokuziphendukela kwemvelo okunethuba eliphezulu(ML) indlela kanye nezindlela zewashi lemolekyuli ye-Bayesian kwacutshungulwa kumagciwane amasha angama-88 asuka eSichuan kanye nama-genome akhethiwe angama-250 asuka kwezinye izifunda.Ama-Genome ano-∆500-532 (Ukususwa endaweni yekhodi ye-Nsp1) atholwe asatshalaliswa kancane esihlahleni se-phylogenetic.Ukuhlaziywa kwe-Haplotype kokuhlukile kwe-Nsp1 kukhombe ezi-5 zazo emadolobheni amaningi.Le miphumela iphakamise ukuthi i-∆500-532 yenzeka emadolobheni amaningi futhi ingase ingeniswe izikhathi eziningi ivela e-Wuhan.
Umfanekiso 2. Izinhlobonhlobo zofuzo eziphindelelayo kanye nokuhlaziywa kwe-phylogenetic ku-SARS-CoV-2 genomes.
3. Ukuhlanganiswa kwezinhlobonhlobo zofuzo eziphindelelayo nemithelela yomtholampilo
Ama-phenotypes angu-117 emtholampilo ahlotshaniswa nokuqina kwe-COVID-19, lapho ama-phenotypes ahlobene nokuqina ayi-19 ahlukaniswa abe yizici ezinzima nezingezona ezinzima.Ubudlelwano phakathi kwalezi zici kanye nokwehluka kofuzo okungu-35 kwabonwa kumephu yokushisa ye-bi-cluster.Ukuhlaziywa kokucebisa okulinganiselwe okufana ne-GSEA kuboniswe ukuthi ∆500-532 ihlobene kabi ne-ESR, i-serum IFN-β kanye ne-CD3+CD8+ T cell count counts egazini.Ngaphezu kwalokho, ukuhlolwa kwe-qPCR kubonise ukuthi iziguli ezingenwe igciwane eligcina ∆500-532 zinenani eliphakeme kakhulu le-Ct, okungukuthi inani legciwane egazini eliphansi kakhulu.
Umfanekiso 3. Izinhlangano zezinhlobo zofuzo ezingama-35 eziphindaphindayo ezinama-phenotypes omtholampilo
4. Ukuqinisekiswa kokuguquguquka kwegciwane okuhambisana nama-phenotypes omtholampilo
Ukuze uqonde ukuthintana kuka-∆500-532 emisebenzini ye-Nsp1, amaseli e-HEK239T adluliswa ngama-plasmids aveza ubude obugcwele, i-WT Nsp1 namafomu aguquguqukayo asuswe.Amaphrofayili e-Transcriptome wamaseli e-HEK239T ngamunye aphathwe acutshungulwa ukuze kuhlaziywe i-PCA, okubonisa ukuthi okuguquguqukayo kokususa kwakuhlangene ngokuqhathaniswa futhi kwakuhluke kakhulu ku-WT Nsp1.Izakhi zofuzo ezazilawulwa kakhulu eziguquguqukayo zazinothiswe ngokuyinhloko “kunqubo ye-peptide biosynthetic/metabolic”, “ribonucleoprotein complex biogenesis”, “amaprotheni aqondiswe ku-membrane/ER”, njll. Ngaphezu kwalokho, ukususwa okubili kubonise iphethini yokuqhuma ehlukile evela ku-WT.
Umfanekiso 4. Ukuhlaziywa kwe-Transcriptome kumaseli e-HEK239T adluliswa yi-WT Nsp1 kanye nalokho ngokususwa.
Imithelela yokususwa ekuphenduleni kwe-IFN-1 nayo ihlolwe esifundweni esine-overexpressed.Konke ukususwa okuhloliwe kuboniswe ukunciphisa impendulo ye-IFN-1 kumaseli adlulisiwe we-HEK239T kanye ne-A549 kuwo womabili izinga le-transcriptome kanye neleveli yamaprotheni.Kuyathakazelisa ukuthi izakhi zofuzo ezilawulwa phansi kakhulu ekususweni zathuthukiswa "ekuphenduleni kokuzivikela kugciwane", "i-viral genome replication", "ukulawulwa kokubhala nge-RNA polymerase II" kanye "nempendulo yohlobo lwe-interferon I".
Umfanekiso 5. Ukulawulwa phansi kwezindlela zokusayina ze-interferon ku-∆500-532 mutant
Kulolu cwaningo, umthelela walokhu kususwa kugciwane kwaqinisekiswa ngokwengeziwe izifundo zokutheleleka ngegciwane.Amagciwane anama-mutant athile ahlukanisiwe kumasampula omtholampilo futhi atheleleka kumaseli e-Calu-3.Imiphumela enemininingwane yocwaningo lokutheleleka ngegciwane ingafundwa ephepheni.
doi:10.1016/j.chom.2021.01.015
Ireferensi
U-Lin J, uTang C, u-Wei H, et al.Ukuqapha kwe-genomic kwe-SARS-CoV-2 kwembula ukwahluka kokususwa kwe-Nsp1 okushintsha impendulo yohlobo lwe-interferon yohlobo lwe-I[J].I-cell host & microbe, 2021.
Izindaba Namaphuzu Avelele ihlose ukwabelana ngamacala akamuva ayimpumelelo ne-Biomarker Technologies, ithwebula impumelelo yesayensi yenoveli kanye namasu avelele asetshenziswa phakathi nocwaningo.
Isikhathi sokuthumela: Jan-06-2022