Txoj Kev Kawm Pilot Ntawm Urine Proteomics hauv COVID-Associated Acute raum Injury

Hu rau: Audrey Hu Whatsapp / hp: 0086 13880143964 Email:audrey.hu@wecistanche.com

Kev Kawm Pilot ntawm Urine Proteomics hauv COVID-19-Associated Acute Kidney Injury

Ntsiab lus:tso zis, COVID-19, Mob raum raug mob,COVID

Taw qhia

Mob hnyavraumraug mobyog ib qho teeb meem loj cuam tshuam nrogCOVID-19thiab tshwm sim nyob rau hauv txog li 76 feem pua ​​​​ntawm cov neeg mob nyob rau hauv lub intensive care unit.1,2 Qhov kev tuag ntawm cov neeg mob nrogCOVID-19leej twg tsimMob raum raug mob (COVID-Mob hnyavraumraug mob) is >10 npaug siab dua cov uas tsis tau. Txawm tias tus neeg sib twMob raum raug mobCov cim muaj nyob, etiology ntawmCOVID-Mob raum raug mobyog multifactorial xav tau agnostic mus kom ze rau kev txheeb xyuas cov analytes thaum ntxov nyob rau hauv tsev kho mob chav kawm los muab kev nkag siab rau hauv biomarkers thiab mechanisms txuam nrogCOVID-Mob raum raug mobthiabCOVID-19kab mob. Tshawb nrhiav ntawmCOVID-19-Txoj kev cuam tshuam ntawm cov urinary proteome yog txwv, thiab lub raum tsis ua haujlwm tsis tau tshaj tawm.3 Kwv yees li 70 feem pua ​​​​ntawm cov proteins uas kuaj pom hauv lub raum.tso zisyog tsim nyob rau hauv lub raum nrog ib tug tseem ceeb npaum li cas lim los ntawm cov ntshav. Peb xav tias qhov kev hloov pauv ntawm cov protein ntau hauv covtso zistuaj yeem ua rau kev tshawb pom ntawm cov cim protein txuam nrogCOVID-19los yogCOVID-Mob hnyavraumraug mobthiab muab kev nkag siab txog kev siv tshuab los txhim kho kev nkag siab.

TSEEM CEEB THIAB PAUB

Peb soj ntsuamtso ziskuaj los ntawm 14 tus neeg koom (6COVID-Mob hnyavraumraug mob, 3 COVID- tsis muajMob hnyavraumraug mob,i 5 nrCOVID- tsis muajMob hnyavraumraug mob). Txhawm rau suav rau qhov sib txawv loj hauv cov ntsiab lus protein thoob plawstso zisPiv txwv li, peb siv cov bicinchoninic acid los ntsuas peptide concentration tom qab kev zom cov protein (cov khoom siv ntxiv thiab cov txheej txheem). Tag nrho cov qauv peptide tom qab ntawd normalized rau tib qhov concentration ua ntej kev soj ntsuam kom tso cai rau cov txheeb ze sib txawv ntawm qhov sib txawv ntawm cov zis proteome (Cov khoom siv ntxiv thiab cov txheej txheem).

Cistanchetuaj yeem siv rau qhovCOVID-19- koom nrogMob hnyavraumraug mob.

Covtso zisProteome of COVID-Mob hnyavraumraug mob

Tom qab lees paub qhov zoo ntawmtso zisrau kev tshawb nrhiav tshawb pom (Cov Khoom Siv Ntxiv thiab Cov Txheej Txheem, Cov Lus Ntxiv S2 thiab Daim Duab Ntxiv S1), peb tau soj ntsuam seb qhov txawv ntawm qhov txawv tuaj yeem paub qhov txawv ntawmmob hnyavraumraug mobzoo (Mob hnyavraumraug mob) (6 COVID-Mob hnyavraumraug mob) los ntawm lwm cov qauv tsis muajMob hnyavraumraug mob(8 Mob hnyavraumraug mobtsis zoo); tag nrhoMob hnyavraumraug mobkuaj tau los ntawm cov neeg mob nrogCOVID-19. Thawj 2 lub hauv paus tseem ceeb suav nrog 42.1 feem pua ​​​​ntawm qhov sib txawv thiab pom meej cais 2 pawg (Daim duab 1b), yog li qhia txog kev cuam tshuam tseem ceeb los ntawmMob hnyavraumraug mobntawmtso zisproteome ntawm cov neeg mob nrogCOVID-19. Muaj qhov txheeb cais tseem ceeb hauv 97 cov proteins thiab txo qis hauv 140 cov proteins hauv covMob hnyavraumraug mobpab pawg (hloov P < 0.05;="" daim="" duab="" 1c="" thiab="" ntxiv="" daim="" duab="" s2a).="" hierarchical="" clustering="" thiab="" pathway="" enrichment="" tsom="" xam="" qhia="" tau="" hais="" tias="" cov="" saum="" toj-up-txoj="" kev="" txoj="" kev="" yog="" complement="" activation,="" coagulation="" cascades,="" thiab="" tswj="" exocytosis="" (daim="" duab="" 1d="" thiab="">

Tshwj xeeb, ntxiv cov Cheebtsam C2, C3, C5, C6, thiab C8G tau nce ntau hauvMob hnyavraumraug mobpab pawg cuam tshuam rau kev pib ua tiav (Daim duab 1d). Qhov tseem ceeb, kev lwj-accelerating factor CD55 txo qis 2.5- quav hauv qhovMob hnyavraumraug mobpab pawg. Qhov no qhia tias txo lub peev xwm los tswj kev ua kom muaj zog ntxiv, yog li ua rau qhov raug mob hnyav dua. Qhov kev tshawb pom no yog txhawb los ntawm exacerbation ntawmMob hnyavraumraug mobnyob rau hauv nas nrog tshem tawm ntawm CD55 tom qab ischemia-reperfusion raug mob.4 Ua kom cov txheej txheem ntxiv yog txuam nrog mob hnyav.COVID-19. Lub raum cell protein sC5b-9, feem ntau cuam tshuam nrog kev ua pa tsis ua haujlwm, yog tswj hwm hauv cov ntshav plasma hauv cov neeg mob.COVID-19nrog rau mob raum mob hnyav. Kev sib piv ntawmCOVID-19 cov qauv nrog thiab tsis muajMob hnyavraumraug mobkuj tau qhia txog kev txhawb nqa cov protein uas cuam tshuam nrog txoj hauv kev ntxiv (Cov Khoom Siv Ntxiv thiab Cov Txheej Txheem, Cov Lus Ntxiv S2 kab X-Z). Qhov muaj peev xwm ntawm cov tshuaj immunoglobulin complexes (pom nyob rau theem siab dua hauv cov neeg mob hnyavCOVID-19) tej zaum yuav conceivably yuav recruiting no ntxiv rau nephron yuav piav qhia txog lub mechanism rauMob hnyavraumraug mobthiab kev ua tsis tiav loj yuav tsum tau lim ntshav. Kev txheeb xyuas qhov sib txawv ntawm cov txheej txheem ntxiv hauv covtso zisntawm cov neeg mob nrogCOVID-Mob hnyavraumraug mobyog tshiab. Nws tuaj yeem ua tus qauv ntsuas los sib piv yuav ua li cas SARS-CoV-2 pathogenicity hloov zuj zus thaum sib txawv nthwv dej ntawmCOVID-19thiab nthuav txoj hauv kev rau kev ntsuas kev sib piv nrog tsis-COVID-Mob hnyavraumraug mobyav tom ntej. Yog li,tso zisyog ib qho yooj yim mus siv tau biospecimen los saib xyuas lub xeev ntawm complement system nyob rau hauv cov neeg mob nrogCOVID-19.

Tsis tas li ntawd, coagulation yam F9, F10, F13b, fibrinogen beta chain, thiab fibrinogen gamma saw tau nce ntau hauvMob hnyavraumraug mobpab pawg (Daim duab 1d). Ob peb lub cim ntawm kev tswj hwm exocytosis, suav nrog myeloperoxidase, catalase, lysozyme, leukotriene A4 hydrolase, proteinase 3, thiab matrix metallopeptidase 9, kuj tau nce hauv pawg no (Daim duab 1d). Txoj kev tswj hwm saum toj kawg nkaus suav nrog lub koom haum extracellular matrix thiab heterophilic cell-cell adhesion (Daim duab 1e), qhia txog kev puas tsuaj ntawm tubular.

Tsis tas li ntawd, peb tau kuaj pom tias ob peb lub urinary biomarkers ntawmmob hnyavraumraug mob, suav nrog NGAL; FABP3, FABP4; MEP1A; thiab RBP4, nce hauvtso zisntawm cov neeg mob nrogCOVID-Mob hnyavraumraug mob(Daim duab 1d thiab Cov Lus Qhia Ntxiv S2). Txawm li cas los xij, kev tsom xam proteome tau nthuav tawm qhov txo qis hauv UMOD thiab EGF hauv pawg no. Urinary UMOD thiab EGF tau pom tias muaj kev sib cuam tshuam nrogMob hnyavraumraug mob, thiab lawv qhov txo qis yog txuam nrogMob hnyavraumraug mobseverity. These results further validate our proteome approach and are indicative of tubular injury affecting secretion (NGAL, UMOD, EGF) or defective absorption by proximal tubules (FABP3, FABP4). A novel observation was a >30-fold nce hauv IGFBP6 hauv cov neeg mobCOVID-Mob hnyavraumraug mob. Tsawg tsawg yog paub ntawm IGFBP6 nyob rau hauvMob hnyavraumraug mob, tab sis yav dhau los nws tau pom tias muaj feem cuam tshuam nrog txo lub raum ua haujlwm thiab lub raum tsis ua haujlwm.5 Cov kev tshawb fawb ntxiv uas siv cov pawg loj ntawmtso ziscov qauv thoob plaws tej yam kev mob yuav tsum tau kom paub meej tias cov kev hloov nyob rau hauv cov biomarkers thiab muab piv nrog rau lwm yam ua raumob hnyavraumraug mob, covariates, thiab cov txiaj ntsig.

Covtso zisProteome Associated NrogCOVID-19nyob rau hauv qhov tsis muajMob hnyavraumraug mob

VimMob hnyavraumraug mobua rau muaj kev hloov pauv tseem ceeb hauv kev nplua nuj ntawm ntau cov urinary proteins, nws nyuaj rau txheeb xyuas cov kev hloov pauv uas tsuas yog cuam tshuam nrogCOVID-19. Peb, yog li ntawd, piv covtso zisproteomes ntawmCOVID-19zoo (n ¼ 3) thiab tsis zoo (COVID) (n ¼ 5) cov neeg mob uas tsis muajMob hnyavraumraug mob(Cov duab ntxiv S2B). Qhov kev nplua nuj ntawm 57 cov protein ntau tau nce ntxiv, thiab 19 cov proteins tau txo qis hauv cov ntshav.tso zisntawmCOVID-19 patients (adjust P < 0.05, Figure 2a and Supplementary Table S2). Furthermore, a large percentage of the proteins with significantly increased levels were immunoglobulins (49.1%). The remaining proteins with increased expression were involved in the antigen presentation pathway. Proteins with significantly decreased expression were enriched in the regulated exocytosis pathway (Figure 2b). Specifically, the average abundances of major MHC-I proteins HLA-A and HLA-C increased >3tso zisvimCOVID-19. Lub MHC complex nthuav tawm antigens ntawm lub xov tooj ntawm tes mus rau lub cev tiv thaiv kab mob thaum lub hlwb kis los ntawm cov kab mob. MHC-peptide complexes tau kuaj pom hauv tib neeg lub cev sib txawv, suav nrog cov ntshav qab zib thiabtso zis.6 Cov kev cai qis ntawm exocytosis tau cim los ntawm kev txo qis ntawm DEFA1, GRN, HSPA6, SDCBP, IQGAP1, thiab CYRIB (Daim duab 2c).

Tsis tas li ntawd, muaj 4 cov proteins uas muaj ntau ntxiv hauv cov khoom nojCOVIDpawg piv nrog covCOVIDpab pawg, suav nrog MB, CA1, MANSC1, thiab ABRACL (Daim duab 2c thiab Cov Table Ntxiv S2 kab AA–AC). Tsis tas li ntawd, 5 proteins tau txo qis hauvCOVID, suav nrog GRN, CREB3L3, MUC1, CD320, thiab DLST (Daim duab 2c thiab Ntxiv Table S2 kab AA–AC). Cov proteins uas tau txheeb xyuas tshiab no tuaj yeem siv los ua biomarkers rhiab dua los ntsuas qhovCOVID-19kab mob. Ntxiv mus, cov ntaub ntawv qhia tau hais tias ntau lub cim yog extrarenal nyob rau hauvCOVIDCov neeg mob (Daim duab 2), suav nrog 5. cog lus lub hom phiaj rau noninvasivetso zisproteome profiling txhawm rau txheeb xyuas thiab saib xyuas lub raum tsis ua haujlwm ntawm lub cev.

SARS-CoV-2 virion tsis feem ntau tso rau hauvtso zis, tab sis nws muaj tropism rau lub raum cell hom.7–9 Peb soj ntsuam sebtso zisproteomics tuaj yeem ntes SARS-CoV-2 proteins. Interestingly, peptides los ntawm SARS-CoV-2 nucleoprotein tau kuaj pom hauvtso ziscov qauv sau los ntawm 2 cov neeg mob hnyav nrogCOVID-Mob hnyavraumraug mobleej twg tuag (Cov Khoom Siv Ntxiv thiab Cov Txheej Txheem, Daim duab 1a thiab Cov Lus Ntxiv S1). Kev txheeb xyuas peptide tau lees paub los ntawm kev tshawb nrhiav nrog MaxQuant thiab MS-GFþ software. Ze-ua tiav qhov kev pab them nqi ntawm RNA binding domain tau ua tiav los ntawm ib qhov dej chromatography ua ke rau tandem loj spectrometry tsom xam. Sequences ze N-terminus of SARS CoV-2 nucleoprotein kuj tau pom (Daim duab 2d). Raws li qhov tsawg-suav peptide-spectrum matches ntawm SARS-CoV{10}} peptides pom, qhov concentration ntawm SARS-CoV-2 nucleoprotein tau kwv yees kom tsawg hauv cov no.tso ziscov qauv (Table Ntxiv S3).

Txawm hais tias peb txoj kev tshawb fawb yog ib qho pov thawj ntawm kev tshawb nrhiav nrog cov neeg mob tsawg tsawg, peb tuaj yeem tsim txoj hauv kev rau cov khoom zoo.tso zisproteomics ntawm cov neeg mob nrogCOVID-19nyob rau hauv qhov chaw kho mob. Peb tshaj tawm tej zaum biomarkers ntawmCOVID-19kab mob thiabCOVID-Mob hnyavraumraug mobuas yuav pab tau rau kev kuaj xyuas thiab saib xyuas kev txhim kho ntawmCOVID-19kab mob los yogMob hnyavraumraug mob. Hauv particular, txoj hauv kev ntxiv thiab coagulation cascade thiab txoj hauv kev exocytosis tau raug tswj hwm zoo rau cov neeg mobCOVID-Mob hnyavraumraug mob. Tsis tas li ntawd, cov protein cuam tshuam nrog kev nthuav qhia antigen tau cuam tshuam nrogCOVID-19kab mob. Peb nthuav tawm cov npe ntawm cov muaj peev xwm tshiab biomarker proteins (ie, MB, CA1, MANSC1, ABRACL, GRN, CREB3L3, DEFA1, MUC1, DLST, thiab IGFBP6) thiab ntawm yav dhau los qhiaMob hnyavraumraug mobbiomarker proteins (piv txwv li, NGAL, FABP3, FABP4, MEP1A, thiab RBP4) kom paub tseeb ntxiv ntawm lawv qhov tshwj xeeb, rhiab heev, thiab hnyav. Ib subset ntawm peb cov neeg mob nrogCOVID-Mob hnyavraumraug mobtsim tawmMob hnyavraumraug mobntawm preexisting mob raum kab mob, uas kuj yuav cuam tshuam rautso zisproteins pom enriched nyob rau hauv pawg no. Parsing cov nyhuv ntawm cov kab mob raum ntev nyob rau hauvCOVID-19yuav xav tau cov pab pawg loj dua; Txawm li cas los xij, peb pom tias qhov ua kom muaj txiaj ntsig ntawm kev sib xyaw ua ke thiab coagulation txoj hauv kev proteins tau pom nyob hauv cov neeg mobCOVID-Mob hnyavraumraug mob nrog thiab tsis muaj kab mob raum ntev (Daim duab 1d thiab Cov Lus Qhia Ntxiv S1). Ua ke, cov txiaj ntsig zoo siab ua ntej no qhia meej meej qhia qhov xav tau rau kev tshawb nrhiav ntxiv hauv kev nthuav dav kev kho mob uas suav nrog cov neeg mob uas tsis yog-COVID-19- koom nrogMob hnyavraumraug mob.

Qhia tawm

CRP ceeb toom yog ib tug tswv cuab ntawm pawg thawj coj saib xyuas thiab muaj kev ncaj ncees hauv Renalytix AI thiab ua tus kws pab tswv yim rau Genfifit. AV ceeb toom ua tus kws pab tswv yim rau NxStage Medical Inc. thiab Astute Medical Inc. Cov koom haum no tsis txhawb kev tshawb fawb hauv qhov kev tshawb fawb no. Tag nrho lwm tus kws sau ntawv tshaj tawm tsis muaj kev sib tw txaus siab.

TXOJ CAI

Qhov kev tshawb nrhiav no tau siv cov qauv uas tau los ntawm covCOVID-19biorepository ntawm Washington University Tsev Kawm Ntawv Tshuaj Kho Mob, uas tau txais kev txhawb nqa los ntawm cov hauv qab no: Barnes Jewish Hospital Foundation; lub Siteman Cancer Center muab P30 CA091842 los ntawm National Cancer Institute ntawm National Institutes of Health; thiab Washington University Institute of Clinical and Translational Sciences muab UL1TR002345 los ntawm National Center for Advancing Translational Sciences ntawm National Institutes of Health. Peb ua tsaug rau George M. O'Brien Center ntawm Johns Hopkins University rau kev ua yeeb yamraumraug mobbiomarker kev soj ntsuam (pab P30DK079310). Ib feem ntawm qhov kev tshawb fawb no tau ua tiav siv EMSL, lub chaw siv kev tshawb fawb hauv tebchaws tau txhawb nqa los ntawm Lub Tsev Haujlwm Saib Xyuas Lub Zog Lub Chaw Haujlwm ntawm Kev Tshawb Fawb Txog Kev Tshawb Fawb thiab Ib puag ncig thiab nyob ntawm PNNL. Txoj haujlwm proteomics tau txais kev txhawb nqa los ntawm NIH Common Fund, los ntawm Lub Chaw Haujlwm Saib Xyuas Kev Ua Haujlwm / Lub Chaw Haujlwm ntawm NIH Tus Thawj Coj raws li khoom plig UG3HL145593, thiab NIH pab nyiaj U01DK124020. Cov ntsiab lus tsuas yog lub luag haujlwm ntawm cov kws sau ntawv thiab tsis tas yuav sawv cev rau qhov pom ntawm National Institutes of Health. Peb ua tsaug rau lub raum Translational Research Center (KTRC) ntawm Division of Nephrology ntawm Washington University hauv St. Louis rau kev txhawb nqa kev ua, khaws cia, thiab khaws cia.tso ziscov qauv. Peb ua tsaug rau Charles Goss, Adriana Rauseo Acevedo, thiab Rachel Presti rau lawv cov kev siv zog hauv kev tswj cov ntaub ntawv thiab kev nrhiav neeg mob. Peb kuj ua tsaug rau Jana Chiang rau nws txoj kev koom tes hauv kev kho, formatting, thiab tshuaj xyuas cov ntsiab lus.

SUPPLEMENTARY KHOOM

Supplementary File (PDF) Supplementary Materials and Methods. Xws li cov ntaub ntawv hais txog cov txheej txheem siv rau kev nrhiav tau tus qauv thiab cov neeg mob suav nrog,tso zisKev ua thiab khaws cia, kev npaj cov qauv proteomics, LC-MS/MS-based proteomic tsom xam, proteomics data processing, correlation analysis of patient with COV-Mob hnyavraumraug mobpiv nrog cov tsis muajmob hnyavraumraug mob, thiab qhov twg cov ntaub ntawv muaj mus download tau. Daim duab S1. Pearson correlation ntawm urinary proteomics thoob plaws cov qauv kawm. Daim duab S2. Kev faib tawm ntawm P qhov tseem ceeb ua ntej hloov kho. (A) Cov qhabnias ntawm qhov kev xeem limma uas piv cov kev siv ntawmtso zisproteins raws li txoj cai ntawmmob hnyavraumraug mob. (B) Cov txiaj ntsig ntawm kev ntsuas limma uas piv cov kev siv ntawmtso zisproteins raws li covCOVID-19kab mob hauv lubMob hnyavraumraug mob- pab pawg tsis zoo. Tab S1. Master daim ntawv ntawm kev tshawb fawb cov neeg koom. Tab S2. Kev txheeb xyuas cov txiaj ntsig ntawm urinary proteomics. Tab S3. SARS-CoV-2 peptides tau kuaj pom nyob rau hauv 2 cov qauv tso zis los ntawm LC-MS/MS. Cov ntaub ntawv proteomics tau tshawb xyuas nrog MSGFþ software. STROBE Cov Lus Qhia (PDF).

CEEB TOOM

Lub hauv paus yog los ntawm Yinyin Ye, Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA, thiab lwm yam.