Hu rau: jerry.he@wecistanche.com

Wei Lin1, Jue Fan2, Long-Fei Hu2, Yan Zhang 2, Joshua D. Ooi3, Ting Meng4, Peng Jin5, Xiang Ding5, Long-Kai Peng6, Lei Song6, Rong Tang6, Zhou Xiao4, Xiang Ao4, Xiang-Cheng Xiao4, Qiao-Ling Zhou4, Ping Xiao4, Yong Zhong4

1 Department of Pathology, Xiangya Tsev Kho Mob, Central South University, Changsha, Hunan 410008, Suav teb;

2 Department of Bioinformatics thiab Data Science, Singleron Biotechnologies, Nanjing, Jiangsu 210032, Suav teb;

3Centre for Inflammatory Diseases, Monash University Department of Medicine, Monash Medical Center, Clayton, VIC 3168, Australia;

4 Department of Nephrology, Xiangya Tsev Kho Mob, Central South University, Changsha, Hunan 410008, Suav teb;

5 Department of Organ Transplantation, Xiangya Tsev Kho Mob, Central South University, Changsha, Hunan 410008, Suav teb;

6 Department ofLub raumKev hloov pauv, Lub Tsev Kho Mob Thib Ob Xiangya ntawm Central South University, Changsha, Hunan 410011, Suav.

Cistanche tuaj yeem txhim kho lub raum ua haujlwm

Abstract

Tom ntej: Txij xyoo 2019, ib qho tshiabkhaus laus nas vais lavnpe 2019 novkhaus laus nas vais lav(2019-nCoV) tau tshwm sim thoob ntiaj teb. Sib nrug los ntawm

ua npaws thiab ua pa nyuaj, mobraumkev raug mob tau pom nyob rau hauv ob peb tus neeg mob nrogkhaus laus nas vais lavKab mob 2019. Tsis tas li ntawd, raws li kev tshawb pom tsis ntev los no, tau kuaj pom tus kab mob hauv cov zis. Angiotensin-hloov enzyme II (ACE2) tau raug npaj los ua tus receptor rau kev nkag mus ntawm 2019-nCoV, uas yog tib yam li qhov ua rau mob ua pa hnyav. Txoj kev tshawb no tsom los tshawb xyuas qhov ua rau lub raum puas tsuaj thiab txoj hauv kev muaj peev xwm ntawm 2019-nCoV kab mob hauv cov zis.

Txoj kev: Peb siv ob qho tib si luam tawmraumthiab zais zis cell atlas cov ntaub ntawv thiab tshiab ywj siabraumIb leeg-cell RNA sequencing cov ntaub ntawv tsim nyob rau hauv-tsev los soj ntsuam ACE2 noob qhia nyob rau hauv tag nrho cov cell hom nyob rau hauv lub raum noj qab haus huv thiab zais zis. Pearson correlation coefficients ntawm ACE2 thiab tag nrho lwm cov noob tau tsim ua ntej. Tom qab ntawd, cov noob nrog r qhov tseem ceeb loj dua 0.1 thiab P qhov tseem ceeb me dua 0.01 tau suav tias yog qhov tseem ceeb ntawm cov noob caj noob ces nrog ACE2.

Cov txiaj ntsig: Peb cov txiaj ntsig tau pom qhov muaj txiaj ntsig zoo ntawm ACE2 hauv txhua hom subtypes ntawm proximal tubule (PT) hlwb ntawmraum. ACE2 qhia tau pom nyob rau hauv 5.12 feem pua, 5.80 feem pua, thiab 14.38 feem pua ​​ntawm cov proximal convoluted tubule hlwb, PT hlwb, thiab proximal ncaj tubule hlwb, raws li, nyob rau hauv peb luam tawm.raumcell atlas datasets. Tsis tas li ntawd, ACE2 qhia kuj tau lees paub hauv 12.05 feem pua, 6.80 feem pua, thiab 10.20 feem pua ​​​​ntawm cov hlwb ntawm cov kab mob sib kis, PT, thiab cov kab ncaj nraim ntawm peb tus kheej ob lub raum kuaj. Rau kev soj ntsuam ntawm pej xeem cov ntaub ntawv los ntawm peb lub zais zis kuaj, ACE2 qhia tau qis tab sis pom tau hauv cov zais zis epithelial. Tsuas yog 0.25 feem pua ​​​​thiab 1.28 feem pua ​​​​ntawm cov hlwb nruab nrab thiab lub kaus hlwb, feem, muaj ACE2 qhia. Xaus: Txoj kev tshawb no tau muab pov thawj bioinformatics ntawm txoj kev muaj peev xwm ntawm 2019-nCoV kab mob hauv cov zis. Keywords: 2019-nCoV; Mob raum raug mob; Angiotensin-hloov enzyme 2; Lub zais zis; COVID-19; Lub raum; Kev txheeb xyuas RNA ntu; Ib leeg-cell tsom xam

Taw Qhia Hauv xyoo 2019, cov mob ntsws tsis paub meej tau tshwm sim thiab nthuav dav thoob plaws ntiaj teb. Kev soj ntsuam sib sib zog nqus thiab kev tshawb nrhiav etiological tom qab ntawd tau lees paub tias cov kab mob yog ib hom tshiab tau txheeb xyuas.khaus laus nas vais lavuas tau sau tias 2019 Novel coronavirus (2019-nCoV). Kev sib kis sai sai ntawm 2019-nCoV tau ua rau muaj kev kis mob ntsws thoob ntiaj teb, ua rau muaj kev hem thawj loj rau kev noj qab haus huv thoob ntiaj teb kev nyab xeeb hauv lub sijhawm luv luv.[1-3] Raws li kev tshuaj xyuas bioinformatics, nws tau pom tias tus 2019-nCoV genome yog 88 feem pua ​​​​zoo tib yam rau ob bat derived heavy acute respiratory syndrome (SARS)-like coronaviruses (bat-SL-CoVZC45 and bat-SL CoVZXC21), 79 feem pua ​​zoo tib yam rau SARS-CoV, thiab 50 feem pua. identical to Middle East respiratory syndrome coronavirus (MERS-CoV).[4] Protein structural tsom xam qhia tias

2019-nCoV muaj lub receptor-binding domain zoo ib yam li ntawm SARS-CoV, ncaj qha khi rau angiotensin-converting enzyme II (ACE2), qhia tau hais tias 2019-nCoV siv ACE2 raws li nws cov receptor, I

Feem ntau cov tsos mob ntawm 2019-nCoV kab mob yog kub taub hau thiab hnoos hauv cov neeg mob feem ntau, nrog cov tsos mob ntawm lub hauv siab tsis xis nyob, mob hnyav hnyav, lossis mob ua pa nyuaj (ARDS) hauv cov neeg mob hnyav. Mob hnyavraumKev raug mob (AKI) tau raug tshaj tawm hauv cov neeg mob tsawg uas tau lees paub tias muaj tus kab mob {{0}}nCoV. Kev kuaj pom ntawm 99 tus neeg mob kis tus kab mob 2019-nCoV pom tias peb tus neeg mob (3 feem pua) muaj qhov sib txawv ntawm cov ntshav creatinine.Pin lwm qhov kev soj ntsuam soj ntsuam, plaub (10 feem pua) ntawm 41 tus neeg mob, induding ob ntawm 13 ( 15 feem pua ​​​​) cov neeg mob hnyav (ICU) thiab ob ntawm 28 (7 feem pua ​​​​) cov neeg mob uas tsis yog ICU, tau pom cov ntshav siab creatinine.i0lA tag nrho ntawm 3.7 feem pua ​​​​ntawm cov neeg mob tuag ntawmlub raumua tsis tiav raws li qhia hauv kev tshuaj xyuas ntawm 88 tus kab mob coronavirus 2019 (COVID-19) ntsig txog kev tuag, [iil Tsis tas li ntawd, qhov kev tshawb fawb tshiab tau lees paub tias 27.06 feem pua ​​(23/85) ntawm cov neeg mob pom muaj mob hnyav.lub raumtsis ua hauj lwm, thiab mob hnyav tubular necrosis tau pom los ntawm hematoxylin-eosin staining nyob rau hauv 6 tus neeg mob tom qab tuag.Lub 2019-nCo V antigen tau kuaj pom hauv lub raum tubules los ntawm immunohistochemistry (IHC) thiab.12] Ntxiv mus, {{4 }}nCoV tau kuaj pom nyob rau hauv cov zis kuaj los ntawm qhov muaj pes tsawg leeg ntawm cov tshuaj polymer-ase chain reaction ntawm tus neeg mob uas muaj tus kab mob 2019-nCoV.134IHowever, vim li cas thiab yuav ua li cas 2019-nCoV induces AKI hauv cov zis tseem tsis paub ntau. Tsis tas li ntawd, txawm tias 2019. nCoV tuaj yeem kis tau los ntawm cov kab mob urinary tseem yog cov lus nug uas tsis muaj lus teb ntau. Raws li ACE2 ua lub luag haujlwm tseem ceeb hauv 2019-nCoVinfection, peb tsom los txheeb xyuas ACE2- qhia cov cell muaj pes tsawg leeg thiab kev faib ua feem ntawm tib neeg ib txwmlub raumthiab cov zais zis los ntawm ib leeg-cell transcriptomes los tshawb txog txoj kev kis kab mob ntawm 2019-nCoV thiab lub luag haujlwm ntawm ACE2 hauv kev kis kab mob hauv lub cev.

Ib leeg-cell RNA sequencing (scRNA-Seq) tau siv dav hauv 2019-nCoV kev tshawb fawb vim nws lub peev xwm los qhia cov noob caj noob ces rau txhua hom cell hauv ntau cov ntaub so ntswg tsis sib haum xeeb ntawm kev daws teeb meem siab. Qhov kev qhia ntawm ACE2 tau tshawb xyuas hauv cov hlwb alveolar hom 2 uas paub zoo hauv lub ntsws nrog rau hauv daim siab cholangiocytes, esophageal epithelial cells, thiab nqus tau[15-171enterocytes ntawm ileum thiab txoj hnyuv los ntawm scRNA-Seq. Cov txiaj ntsig no tau pom tias muaj peev xwm siv tau ntawm scRNA-Seq los tshem tawm cov phiaj xwm cell hom ntawm 2019-nCoV. Txij li thaum kis kab mob hauv cov zis thiab nws cov peev xwm tom qab tuaj yeem yog qhov tseem ceeb rau kev saib xyuas tus neeg mob thaum lub sijhawm thiab tom qab kis kab mob, peb siv ob daim ntawv teev cov ntaub ntawv scRNA-Seq transcriptome hauv kev noj qab haus huv.lub raumthiab ib cov ntaub ntawv nyob rau hauv lub zais zis noj qab haus huv los soj ntsuam cov qauv qhia ntawm cell hom nyob rau hauv lub urinary system.

Cov txheej txheem

Public dataset tau thiab ua

Gene qhia matrices ntawm scRNA-Seq cov ntaub ntawv los ntawm ib txwmlub raumntawm peb tus neeg noj qab haus huv tau rub tawm los ntawm Gene Expression Omnibus (GSE131685). Peb rov tsim dua qhov kev ntsuas qis qis siv cov cai muab los ntawm tus sau hauv daim ntawv thawj.

Peb tau txais cov ntaub ntawv scRNA-Seq ntawm cov ntaub so ntswg ntawm lub zais zis noj qab haus huv ntawm peb tus neeg mob qog noj ntshav los ntawm Gene Expression Omnibus (GSE108097). Yuav ua raws li covraumdatasets, peb siv Harmony (Lub Harmony txoj kev yog ib tug algorithm rau kev koom ua ke ntawm ib leeg-cell genomics datasets rau batch kho thiab meta-analysis, uas yog ceev, rhiab, thiab raug. los ntawm embedding.1] los ua ke cov qauv thiab ua cov kev soj ntsuam hauv qab uas siv Seurat (version 3.1, Satija Lab, https://satijalab.org/seurat/).Tag nrho cov gene qhia tau normalized thiab scaled siv NormalizeData thiab ScaleData(NormalizeData yog ib tug muaj nuj nqi. ScaleData yog ib txoj haujlwm ntawm Seurat uas tuaj yeem ntsuas thiab nruab nrab cov noob hauv cov ntaub ntawv, ua qauv qhia txog qhov muaj nuj nqis thoob plaws tag nrho cov noob.), Sab saum toj 20 00 qhov sib txawv ntawm cov noob tau raug xaiv los ntawm FindVariableFeautres rau kev tsom xam cov ntsiab lus tseem ceeb.Clustering tsom xam siv FindClusters tau ua los ntawm thawj zaug txo cov noob qhia matrix rau thawj 20 lub hauv paus ntsiab lus thiab tom qab ntawd siv qhov kev daws teeb meem ntawm 0.3 f los yog graph-based clustering.

Lub raum ua piv txwv

Lub raumcov qauv tau txais ntawm ib qhov chaw los ntawm cov npoo thiab koob biopsy ntawm cov neeg pub lub raum nyob tom qab tshem tawm ntawm tus neeg pub dawb thiab ua ntej cog rau hauv tus neeg txais.Lub raumcov qauv tau txais ntawm ib qhov chaw los ntawm cov kab thiab koob biopsy ntawm cov neeg pub nyoblub raumtom qab tshem tawm ntawm tus neeg pub dawb thiab ua ntej cog rau hauv tus neeg tau txais kev pab. Txhua cov txheej txheem cuam tshuam nrog tib neeg cov neeg koom tau ua raws li cov qauv kev coj ua ntawm Pawg Neeg Saib Xyuas Kev Ncaj Ncees ntawm Xiangya Tsev Kho Mob ntawm Central South University (IRB pom zoo tus lej 201711836) thiab nrog 1964 Tshaj Tawm ntawm Helsinki thiab nws cov kev hloov kho tom qab lossis cov qauv kev coj ncaj ncees sib piv. Lub raum biopsy kuaj tau raug tshem tawm nrog sterile phosphate buffer saline (PBS) tom qab tau txais.

Cov ntaub so ntswg dissociation

TshiabraumCov ntaub so ntswg tam sim ntawd hloov mus rau hauv GEXSCOPE "Tissue Preservation Solution (Singleton Biotechnologies, Nanjing, Jiangsu Province, Tuam Tshoj) ntawm 2 ° C mus rau 8 degree . Cov qauv tau zom hauv 2 mL ntawm GEXSCOPE "cov ntaub so ntswg dissociation solution (Singleron Biotechnologies) ntawm 37C rau 15 min nyob rau hauv ib tug 15-mL centrifuge raj nrog nruam agitation tom qab ntxuav nrog Hanks balanced ntsev tov peb zaug thiab txiav mus rau hauv kwv yees li 1 mus rau 2 hli pieces. Tom qab ntawd, cov khib nyiab ntawm tes thiab lwm yam impurities tau lim los ntawm 40-micron sterile strainer (Corning, NY, USA). Cov hlwb tau centrifuged ntawm 300 × g thiab 4 degree rau 5 min. Cell pellets raug tshem tawm hauv 1 mL ntawm PBS (HyClone, Marlborough, MA, USA). Txhawm rau tshem tawm cov qe ntshav liab.2mLGEXSCOPE Red Blood Cell Lysis Buffer (Singleton Biotechnologies) tau ntxiv rau lub cell ncua thiab incubated ntawm 25 degree rau 10 min. Qhov sib tov yog ces centrifuged ntawm 500 × g rau 5 min, thiab lub cell pellet tau resuspended hauv PBS. Cells raug suav nrog TC20 automated cell counter (Bio-Rad, Hercules, CA, USA).

Kev npaj tsev qiv ntawv thiab kev txheeb xyuas cov ntaub ntawv ntawm scRNA-Seq Qhov kev ncua ntawm ib leeg tau raug rau kev npaj tsev qiv ntawv ib leeg thiab ua ntu zus, raws li tau piav qhia yav dhau los.[19] Yuav kom ua raws li cov ntaub ntawv pej xeem siv los saum toj no, peb siv qhov zoo sib xws hauv qab kev tsom xam workflflow.[18] Peb tau hloov kho tus lej ntawm cov ntsiab lus tseem ceeb rau 20 thiab siv qhov kev daws teeb meem ntawm 0.6 kom tau txais cov txiaj ntsig sib piv ntawm cov xov tooj ntawm tes nrog cov pej xeemraumcov ntaub ntawv. Cov hom cell tau sau tseg raws li cov cim canonical hauv cov ntaub ntawv.

Daim duab 1: Kev tshuaj xyuas ib leeg ntawm pej xeemraumcov ntaub ntawv los ntawm peb tus neeg pub dawb. (A) UMAP zaj duab xis nthuav tawm lub xov tooj ntawm tes annotations ntawm peb cov qauv. (B) Cov cellular muaj pes tsawg leeg ntawm peb lub raum noj qab nyob zoo kuaj. Proximal convoluted tubule hlwb suav txog 48.3 feem pua, 52.6 feem pua, thiab 38.5 feem pua; PT hlwb suav txog 26.3 feem pua, 36.9 feem pua, thiab 43.7 feem pua; thiab proximal ncaj tubule hlwb suav txog 7.6 feem pua, 2.1 feem pua, thiab 5.8 feem pua. (C) Dot zaj duab xis nthuav tawm cov cim canonical siv rau kev faib tawm ntawm tes. Lub abscissa sawv cev rau txhua hom cell, lub ordinate sawv cev rau cov noob caj noob ces, xim ntawm cov dot nyob rau hauv daim duab sawv cev rau theem ntawm kev qhia, thiab qhov luaj li cas ntawm cov dot sawv cev rau qhov feem ntawm cov noob qhia nyob rau hauv lub cell. (D) Cov qauv qhia cov noob ntawm ACE2 ntawm txhua hom kev kuaj pom ntawm tes hauvraum. Hom cell yog sawv cev kab rov tav, thiab ACE2 qhia tus nqi yog sawv cev vertically. Txhua lub ntsiab lus sawv cev rau ib lub cell. Cov hlwb uas feem ntau qhia ACE2 hauv lub raum kuaj yog PT hlwb. ACE2: Angiotensin-hloov enzyme II; PT: Proximal tubule; UMAP: Uniform manifold approximation thiab projection.

Daim duab 2: Kev tshuaj xyuas ib leeg ntawm ob lub raum biopsy kuaj. (A) Cell hom annotations pom los ntawm UMAP. Cov xov tooj sib txawv yog xim-coded, thiab cov npe ntawm tes tau ntxiv. (B) Cov qauv tshwj xeeb UMAP kev pom ntawm ob qho qauv. Cell hom yog txawv los ntawm cov xim sib txawv. (C) Heatmap ntawm canonical markers thov rau hom tes hauj lwm. Hom cell yog sawv cev kab rov tav, daim ntawv gene yog sawv cev ntsug, thiab theem qhia tau sawv cev los ntawm cov xim hauv daim duab. (D) Violin zaj duab xis qhia cov cim qhia ntawm lub raum cell hom. Hom cell yog sawv cev kab rov tav, thiab ACE2 qhia tus nqi yog sawv cev vertically. Txhua qhov scatters taw tes sawv cev rau ib lub cell. Cov hlwb uas feem ntau qhia ACE2 hauv lub raum kuaj yog PT hlwb. ACE2: Angiotensin-hloov enzyme II; PT: Proximal tubule; UMAP: Uniform manifold approximation thiab projection.

proximal tubule [PT] hlwb rau lub raum thiab lub kaus mom rau lub zais zis). Pearson correlation coefficients ntawm ACE2 thiab tag nrho lwm cov noob tau tsim ua ntej. Tom qab ntawd, cov noob uas muaj r qhov tseem ceeb loj dua 0.1 thiab P qhov tseem ceeb me dua 0.01 tau suav tias yog cov noob caj ces tseem ceeb nrog ACE2. Peb tshem tawm cov noob mitochondrial rau cov hauv qab no txoj kev txhawb nqa thiab kev sib cuam tshuam ntawm cov protein-protein (PPI) analyses. Rau cov ntaub ntawv rau lub raum, peb siv cov noob sib koom ua ke hauv tsawg kawg yog ob ntawm peb lub xov tooj ntawm tes ntawm PT, cov kab ncaj nraim, thiab cov kab sib txuas sib txuas ntawm cov tubule hauv daim duab Venn. Tag nrho ntawm 126 thiab 65 cov noob tseem nyob rau cov ntaub ntawv rau pej xeem thiab ntiag tug raum, raws li. Rau cov ntaub ntawv zais zis, cov txheej txheem kawg muaj 372 cov noob. Gene Ontology (GO) thiab Kyoto Encyclopedia of Genes thiab Genomes (KEGG) txoj kev txhim kho kev txheeb xyuas tau raug siv los ntawm R pob ClusterProfiller (version 3.8.1, Bioconductor, https:// bioconductor.org/packages/release/bioc/html/clusterPro filer.html). Txoj hauv kev nrog P_adj tus nqi tsawg dua 0.05 raug suav tias yog qhov muaj txiaj ntsig zoo. Rau kev soj ntsuam PPI, ob qhov kawg ntawm cov noob los ntawm ob lub raum cov ntaub ntawv tau muab tso ua ke los ua ib daim ntawv teev npe noob. PPI tsom xam tau kwv yees raws li paub txog kev sib cuam tshuam ntawm cov noob nrog cov ntsiab lus GO cuam tshuam hauv R pob STRINGdb (version 1.20.0, https://string-db.org/). Raws li qhov sib txawv ntawm qhov sib txawv ntawm cov noob protein sib cuam tshuam network cov ntaub ntawv, PPI network tau raug xa tawm siv Cytoscape software (version 3.4,https://cytoscape.org/).

Daim duab 3: Public zais zis ib leeg-cell dataset. (A) Ob-dimensional visualization ntawm lub zais zis cell atlas los ntawm UMAP. Cov xov tooj sib txawv yog xim-coded, thiab cov npe ntawm tes tau ntxiv. (B) Kev nthuav qhia tus qauv ntawm cov cim canonical siv los txheeb xyuas cov zais zis ntawm tes. X-axis sawv cev rau txhua hom ntawm tes, Y-axis sawv cev rau cov noob caj noob ces, cov xim ntawm cov dot hauv daim duab sawv cev rau theem ntawm kev qhia, thiab qhov luaj li cas ntawm cov dot sawv cev rau qhov sib npaug ntawm cov noob qhia hauv lub cell. (C) ACE2 qhia thoob plaws 12 cell subgroups hauv lub zais zis dataset. Hom cell yog nyob rau kab rov tav axis, thiab ACE2 qhia tus nqi yog sawv cev vertically. Txhua qhov scatters taw tes sawv cev rau ib lub cell. ACE2 yog qhia nyob rau hauv lub kaus hlwb nyob rau hauv lub zais zis kuaj.

ACE2: Angiotensin-hloov enzyme II; UMAP: Uniform manifold approximation thiab projection.

Cistanche tuaj yeem txhim kho lub raum ua haujlwm

Kev txheeb cais

Marker genes for each cluster were identified with the Wilcox likelihood-ratio test with default parameters via the FindAllMarkers function in Seurat v.3.1.2. Marker genes that are expressed in >10% of the cells in a cluster and average log (fold change) of >{{0}}}.25 tau xaiv. Pearson correlation tau siv los txheeb xyuas cov noob caj noob ces nrog ACE2, thiab P-tus nqi qis dua 0.01 tau siv los txheeb xyuas cov noob caj noob ces nrog ACE2. Rau cov ntaub ntawv scRNA-Seq, statistical analysis thiab cov duab tau ua tiav los ntawm R (version 3.5.1, https://www.r-project.org/).

Cov txiaj ntsig

Kev nthuav qhia cov qauv ntawm ACE2 hauv ob lub raum Los ntawm kev tshuaj xyuas cov pej xeem ib leeg-cell transcriptome dataset ntawm tib neeg lub raum hlwb los ntawm peb tus neeg pub dawb, peb pom tias ACE2 qhia tau muab faib thoob plaws ntau hom cell. Tshwj xeeb tshaj yog, ACE2 feem ntau enriched hauv PT hlwb, nrog rau ob leeg convoluted tubules thiab ncaj tubules [Daim duab 1A–1D]. Lwm yam nephron subtypes, xws li khaws cov duct thiab distal tubule nrog rau lub cev tiv thaiv kab mob, txhua tus pom cov noob qes qis heev.

Txhawm rau kom ntseeg tau qhov txiaj ntsig no, peb tau txheeb xyuas ntxiv cov qauv rau lub raum los ntawm ob tus neeg pub noj qab haus huv. Lub 4736 hlwb tau muab faib ua cuaj hom cell, uas tau sib tshooj nrog cov ntaub ntawv pej xeem, suav nrog xya nephron-specific subtypes raws li canonical marker genes [Daim duab 2A]. Lub PT (CUBN, SLC13A3, thiab SLC22A8) tau muab faib ua cov tubules sib thooj thiab cov tubule ncaj nraim raws li cov cim qhia sib txawv. (ATP6V0D2), thiab glomerular parietal epithelial hlwb (KRT8 thiab KRT18) kuj tau sau tseg, thiab cov txiaj ntsig no tau ua raws li qhov tshwm sim dhau los. Tsis tas li ntawd, peb tau txheeb xyuas ib pawg ntawm cov voj voog ntawm Henle hlwb (UMOD, SLC12A1, thiab CLDN16), uas tsis nyob hauv cov ntaub ntawv pej xeem. Tsis tas li ntawd, cov kab mob stroma thiab lub cev tiv thaiv kab mob, tsim los ntawm cov kab mob endothelial tsawg (CD34 thiab PECAM1) thiab monocytes (CD14, FCN1, thiab VCAN), tau pom [Daim duab 2B]. Tus neeg sawv cev cim cov noob rau tag nrho cov subpopulations yog npaj rau hauv daim duab 2C.

Raws li tau pom nyob rau hauv cov ntaub ntawv pej xeem, peb tau soj ntsuam ACE2 qhia tawm hauv txhua lub hlwb PT piv nrog lwm hom cell [Daim duab 2D]. Quantitatively, ACE2 qhia kuj tau lees paub hauv 12.05 feem pua, 6.79 feem pua, thiab 10.20 feem pua ​​​​ntawm cov hlwb ntawm cov kab mob sib kis, PT, thiab cov kab ncaj nraim, raws li, hauv peb tus kheej ob lub raum zoo ib txwm kuaj. Yav dhau los IHC kev tshuaj xyuas [21] tau qhia txog qhov sib txawv ntawm qhov sib txawv ntawm ACE2 cov protein qhia pom nyob rau hauv txhuam ciam teb ntawm PTs.

Kev nthuav qhia tus qauv ntawm ACE2 hauv lub zais zis

Raws li cov ntaub ntawv zais zais rau pej xeem ntawm 12 hom cell [Daim duab 3A thiab 3B], peb pom qhov qhia qis ntawm ACE2 hauv txhua hom cell epithelial. Qhov concentration ntawm ACE2 zoo li muaj qhov txo qis los ntawm txheej txheej ntawm lub zais zis epithelium (kaum hlwb) mus rau txheej sab hauv (basal hlwb) nrog cov hlwb nruab nrab hauv nruab nrab. Lwm hom cell, xws li endothelial thiab lub cev tiv thaiv kab mob, feem ntau tsis zoo rau ACE2 [Daim duab 3C]. Qhov feem pua ​​​​ntawm cov hlwb hauv lub zais zis qhia ACE2 (1.3 feem pua) qis dua li ntawm lub raum epithelium. Kev txheeb xyuas yav dhau los ntawm SARS cov neeg mob tau qhia tias tus kab mob SARS (SARS-CoV) tuaj yeem muaj sia nyob hauv cov zis ntawm qib kuaj pom tau. , uas yog nyob rau hauv kev pom zoo nrog peb tsom xam saum toj no.

Kev soj ntsuam ua haujlwm ntawm cov noob caj noob ces nrog ACE2

Nyob rau hauv ob lub raum datasets, peb pom ib tug loj tus naj npawb ntawm noob co-expressed nrog ACE2, tshwj xeeb tshaj yog nyob rau hauv proximal ncaj tubule hlwb [Daim duab 4A thiab 4D]. Cov noob caj noob ces sib koom ua ke tau ua kom muaj ntau yam ntawm cov kab mob ntsig txog cellular (CC) [Daim duab 4B thiab 4E]. Qhov tseem ceeb, txhuam ciam teb thiab txhuam ciam teb daim nyias nyias tau ua kom muaj txiaj ntsig zoo ntawm CC cov ntsiab lus, uas zoo sib xws nrog cov chaw cellular ntawm ACE2 kuaj pom los ntawm kev tshuaj ntsuam immunohistochemical.[21] Cov ntsiab lus ntxiv KEGG suav nrog cov haujlwm ib txwm muaj ntawm ACE2, xws li renin-angiotensin system, PT.

Daim duab 4: Kev soj ntsuam ua haujlwm ntawm ACE-txog cov noob hauv lub raum. (A, D) Venn daim duab ntawm ACE2 co-expression noob los ntawm peb PT hlwb hauv cov pej xeemraumdataset (Data 1) and the private kidney dataset (Data 2). (B, E) Analysis of ACE2 co-expression genes in each cell (expression related to >2 hom cell) rau GO CC term enrichment nyob rau hauv cov ntaub ntawv 1 thiab cov ntaub ntawv 2. X-axis sawv cev rau log-transformed P qhov tseem ceeb. (C, F) Kev soj ntsuam ntawm ACE2 co-expression genes rau KEGG lub sij hawm enrichment nyob rau hauv cov ntaub ntawv 1 thiab cov ntaub ntawv 2. Lub sij hawm nyob rau hauv liab yog txoj kev ntawm cov ntaub ntawv teev. ACE2: Angiotensin-hloov enzyme II; CC: Cellular Cheebtsam; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes thiab Genomes; PT: Proximal tubule.

Daim duab 5: Kev soj ntsuam ua haujlwm ntawm ACE2-cov noob muaj feem xyuam hauv lub zais zis. (A) Kev soj ntsuam ntawm ACE2 co-expression noob nyob rau hauv lub kaus hlwb ntawm lub zais zis rau GO CC term enrichment. X-axis sawv cev rau lub log-transformed Pvalues. (B) Kev soj ntsuam ntawm ACE2 co-expression noob nyob rau hauv lub kaus hlwb ntawm lub zais zis rau KEGG lub sij hawm enrichment. (C, D) PPI tes hauj lwm qhia txog kev sib raug zoo ntawm ACE2 thiab nws cov noob muaj feem xyuam hauv ob lub raum thiab zais zis. Qhov luaj li cas ntawm cov node (gene) sawv cev rau tus naj npawb ntawm cov neeg koom tes (cov neeg nyob sib ze). Cov kab dav thiab xim (qis mus rau dav sib haum rau xiav rau liab) yog proportional rau lub zog sib cuam tshuam hauv STRING database. ACE2: Angiotensin-hloov enzyme II; CC: Cellular Cheebtsam; GO: Gene Ontology;

KEGG: Kyoto Encyclopedia of Genes thiab Genomes; PPI: Protein-protein sib cuam tshuam.

bicarbonate reclamation, mineral absorption, thiab ntau yam metabolism-txog txoj hauv kev [Daim duab 4C thiab 4F]. Cov txiaj ntsig tau lees paub tias ACE2 thiab nws cov noob muaj feem xyuam yog qhov tseem ceeb los tswj kev ua haujlwm ntawm lub raum.[24] Rau cov ntaub ntawv zais zis, CC thiab KEGG cov txiaj ntsig tau txais txiaj ntsig feem ntau zoo ib yam nrog rau lub raum, qhov uas cov proteins thiab lub raum ua haujlwm xws li kev nqus thiab cov metabolism tau koom nrog [Daim duab 5A thiab 5B]. Peb tau ua PPI tsom xam rau ACE2 thiab nws cov kev sib koom ua ke hauv ob qho tib siraumthiab zais zis, raws li qhia hauv daim duab 5C thiab 5D. Txawm hais tias cov protein sib cuam tshuam no cuam tshuam nrog kev hloov pauv pathogenic ntawm PT thiab kab mob kab mob hauv lub plab tsis paub meej thiab yuav tsum tau tshawb xyuas ntxiv. Discusion AKI in 2019-nCoV and SARS-CoV infection COVID-19 is a new infectious disease with a formidable speed of transmission. Txawm hais tias feem ntau ntawm cov neeg mob no pom cov tsos mob ua pa, ntau yam ntawm cov tsos mob tsis ua pa kuj tau tshaj tawm, qhia tias lwm yam kabmob raug cuam tshuam thaum muaj tus kabmob. Hauv kev tshuaj ntsuam xyuas cov yam ntxwv ntawm cov neeg mob uas muaj tus kab mob 2019- nCoV, lub raum kev ua haujlwm puas tau raug lees paub, [9,10] thiab txuas ntxiv mus.raumKev kho mob hloov pauv tau xav tau nyob hauv kwv yees li 9 feem pua ​​​​ntawm tag nrho 99 tus neeg mob uas muaj tus kab mob.[9] Gabarre et al[25] tau tshuaj xyuas AKI hauv cov neeg mob hnyav nrog COVID-19. Tsis tas li ntawd, lub raum raug mob thiab lub raum tsis ua haujlwm tau pom nyob rau hauv lwm yamcov khaus laus nas vais lavcuam ​​tshuam nrog cov kab mob ua pa, tshwj xeeb tshaj yog ob betacov khaus laus nas vais lav(SARS-CoV and MERS-CoV),[26] which are genetically similar to 2019-nCoV.[4,8] Ib txoj kev tshawb fawb dhau los qhia tias 36 (6.7 feem pua) ntawm 536 tus neeg mob SARS tsim AKI, thiab 33 ntawm tag nrho 36 tus neeg mob (91.7 feem pua) nrog rau lub raum tsis ua haujlwm tuag.[27] Qhov tseem ceeb, SARS-CoV fragments tau kuaj pom los ntawm cov tshuaj tiv thaiv polymerase hauv cov zis ntawm qee tus neeg mob.[28] Tsis tas li ntawd, kev kis tus kab mob SARS-CoV tsis tu ncua thiab rov ua dua tau pom nyob hauvraumtshwj xeeb tshaj yog nyob rau hauv tubular epithelial hlwb nyob rau hauv vitro, [29] qhia tias lub raum puas nyob rau hauv cov neeg mob uas muaj SARS yuav yog vim cov kab mob ncaj qha nyob rau hauv lub hom phiaj hlwb tshwj tsis yog rau lwm yam triggers, xws li tus tswv tsev tiv thaiv kab mob, mob ntsws loj, thiab ARDS. Txawm hais tias muaj cov kab mob zoo sib xws rau SARS-CoV, cov ncauj lus kom ntxaws txog kev koom tes ntawm lub raum ntawm 2019-nCoV tseem tsis paub meej.

Cistanche tuaj yeem txhim kho lub raum ua haujlwm

Kev soj ntsuam ntawm ACE2 qhia hauv cov zis

Lub etiology ntawm AKI nyob rau hauv SARS zoo nkaus li yog multifactorial thiab tseem tsis paub meej, uas tau xav tias muaj ib qho kev sib raug zoo nrog rau cov kev qhia siab ntawm ACE2 nyob rau hauv lub raum.[29,30] scRNA-seq, ib tug tshiab txoj kev uas yog nce ingly siv nyob rau hauv. lub fifield ntawm biology thiab tshuaj, tuaj yeem txheeb xyuas hom cell thiab qhia cov noob ntawm qib cellular. Raws li peb cov kev txheeb xyuas ib leeg hauv ob qho tib silub raum(ob qho tib si pej xeem thiab peb cov ntaub ntawv) thiab zais zis, peb pom muaj qib ntawm ACE2 hauv ob lub raum thiab zais zis. Zoo ib yam li peb cov ntaub ntawv, Deng thiab cov neeg ua haujlwm sib koom ua ke [31] txheeb xyuas pej xeemraumcov ntaub ntawv thiab qhia tias lub raum muaj kev pheej hmoo siabkhaus laus nas vais lavkab mob. Tsis tas li ntawd, hauv peb cov ntaub ntawv, peb pom tias PT hlwb muaj feem pua ​​​​ntawm cov lus qhia ntau dua li lub zais zis epithelial hlwb, uas yuav qhia tau tias lub raum muaj kev cuam tshuam rau 2019-nCoV ntau dua li lub zais zis. Piv nrog rau cov theem qhia ntawm ACE2 hauv kev zom zaub mov, [15] qhov qis dua ntawm ACE2 hauv cov zis tuaj yeem qhia cov neeg mob tsawg dua nrog rau cov tsos mob ntawm lub raum, uas cuam tshuam nrog kev soj ntsuam rau cov neeg mob SARS. Kev sib koom ua ke thiab kev tsom xam tau qhia txog kev ua haujlwm tseem ceeb ntawm ACE2 hauv cov zis, uas tuaj yeem piav qhia txog kev ua haujlwm tsis zoo ntawm lub raum los ntawm ACE2- khi cov kab mob. Ntxiv mus, ACE2 qhia nyob rau sab nrauv hlwb (PT hlwb nyob rau hauv lub raum tubules thiab umbrella cells nyob rau hauv lub zais zis epithelium) kuj yog ib qho ntawm cov ua rau muaj cov kab mob nyob rau hauv cov zis.

Qhov tseem ceeb ntawm txoj kev kawm

AKI, uas tau raug pov thawj los ua tus kwv yees ntawm kev tuag siab hauv cov neeg mob SARS, [27] kuj tseem tuaj yeem ua rau muaj kev nyuaj ntawm kev kho mob, kev mob hnyav zuj zus, thiab txawm tias yog qhov ntsuas tsis zoo rau kev muaj sia nyob ntawm 2019-nCoV cov neeg mob. . Txoj kev tshawb fawb tam sim no tau muab pov thawj ncaj qha rau qhov muaj peev xwm kis tau ntawm lub raum epithelial hlwb nrog 2019-nCoV, uas tau thov kom muaj kev saib xyuas ntau ntxiv rau COVID-19 cov neeg mob AKI, tshwj xeeb tshaj yog thaum muaj kev sib kis loj thiab kev kis thoob ntiaj teb txuas ntxiv mus ntxiv. . Tsis tas li ntawd, kev faib tawm ntawm ACE2 hauv cov zis thiab nws qhov kev kuaj pom [32] hauv cov zis kuaj pom pom qhov tsim nyog ntawm kev kuaj kab mob hauv cov zis kuaj ntawm COVID-19 cov neeg mob, uas tuaj yeem txo qhov kev pheej hmoo kis mus rau qee qhov. Peb cov txiaj ntsig yuav tsum tau saib raws li lawv cov kev txwv. Ua ntej, peb tau tshuaj xyuas ACE2 cov lus qhia tsuas yog hauv cov qauv kev noj qab haus huv ib leeg xwb, thiab qhov tsis muaj cov qauv ntawm cov neeg mob nrog COVID-19 yog qhov kev txwv loj. Tsis tas li ntawd, peb tsis tau suav nrog pawg pov thawj kev lees paub los lees paub peb qhov kev tshawb pom. Ntau cov pov thawj ncaj qha los ntawm kev sim hauv vitro thiab cov qauv neeg mob xav tau.

Kev Pab Nyiaj Txoj Haujlwm no tau txais kev txhawb nqa los ntawm Txoj Haujlwm Saib Xyuas Kev Noj Qab Haus Huv ntawm Hunan Xeev (No. C2019184) thiab National Natural Science Foundation ntawm Tuam Tshoj (No. 81800641).

Kev tsis sib haum xeeb Tsis muaj.