Alzheimer's disease (AD) tsis muaj protein ntau biomarkers uas muaj kev cuam tshuam nws ntau yam hauv qab pathophysiology, cuam tshuam qhov kev nce qib ntawm kev kuaj mob thiab kev kho mob. Ntawm no, peb siv cov txheej txheem proteomics los txheeb xyuas cov kua dej cerebrospinal (CSF) biomarkers uas sawv cev rau ntau yam ntawm AD pathophysiology. Multiplex mass spectrometry tau txheeb xyuas kwv yees li 3,500 thiab kwv yees li 12,000 cov proteins hauv AD CSF thiab lub hlwb, feem. Kev tsom xam network ntawm lub hlwb proteome tau daws 44 biodiversity modules, 15 ntawm uas sib tshooj nrog cov kua cerebrospinal proteome. CSF AD cov cim nyob rau hauv cov txheej txheem sib tshooj no tau muab ntim rau hauv tsib pawg protein, sawv cev rau cov txheej txheem pathophysiological sib txawv. Cov synapses thiab metabolites hauv AD lub hlwb txo qis, tab sis CSF nce, thaum lub glial-nplua nuj myelination thiab pab pawg tiv thaiv kab mob hauv lub hlwb thiab CSF nce. Qhov sib xws thiab kab mob tshwj xeeb ntawm kev hloov pauv vaj huam sib luag tau lees paub hauv ntau dua 500 cov qauv CSF ntxiv. Cov pab pawg no kuj tau txheeb xyuas cov pab pawg lom neeg hauv asymptomatic AD. Zuag qhia tag nrho, cov txiaj ntsig no yog ib qho kev cog lus rau lub vev xaib biomarker cov cuab yeej rau kev kho mob hauv AD.
Alzheimer's disease (AD) yog qhov tshwm sim ntau tshaj plaws ntawm neurodegenerative dementia thoob ntiaj teb thiab yog tus cwj pwm los ntawm ntau yam kev ua haujlwm lom neeg, suav nrog kev sib kis ntawm synaptic, glial-mediated immunity, thiab mitochondrial metabolism (1-3). Txawm li cas los xij, nws cov biomarkers tau tsim muaj protein ntau tseem tsom rau kev kuaj pom amyloid thiab tau protein ntau, thiab yog li tsis tuaj yeem cuam tshuam txog qhov sib txawv pathophysiology. Cov "core" protein biomarkers uas ntseeg siab tshaj plaws ntsuas hauv cerebrospinal kua (CSF) suav nrog (i) amyloid beta peptide 1-42 (Aβ1-42), uas qhia txog kev tsim cov cortical amyloid plaques; (ii) tag nrho tau, lub cim ntawm axon degeneration; (iii) phospho-tau (p-tau), tus neeg sawv cev ntawm pathological tau hyperphosphorylation (4-7). Txawm hais tias cov cerebrospinal kua biomarkers tau pab txhawb peb txoj kev tshawb pom ntawm "cim" AD protein kab mob (4-7), lawv tsuas yog sawv cev rau ib feem me me ntawm cov kab mob biology tom qab tus kab mob.
Qhov tsis muaj kev sib txawv ntawm cov kab mob pathophysiological ntawm AD biomarkers tau ua rau muaj ntau yam teeb meem, suav nrog (i) tsis muaj peev xwm txheeb xyuas thiab ntsuas qhov sib txawv ntawm cov neeg mob AD, (ii) kev ntsuas tsis txaus ntawm cov kab mob hnyav thiab kev loj hlob, tshwj xeeb tshaj yog nyob rau theem preclinical, Thiab ( iii) kev txhim kho cov tshuaj kho mob uas tsis tuaj yeem daws txhua yam ntawm kev puas hlwb puas tsuaj. Peb qhov kev cia siab rau thaj chaw pathology los piav txog AD los ntawm cov kab mob cuam tshuam tsuas yog ua rau cov teeb meem no hnyav dua. Ntau thiab ntau cov pov thawj qhia tau hais tias feem ntau cov neeg laus uas muaj dementia muaj ntau tshaj ib tug pathological yam ntxwv ntawm kev txawj ntse poob (8). Ntau li 90% lossis ntau dua ntawm cov neeg uas muaj AD pathology kuj muaj kab mob vascular, TDP-43 suav nrog, lossis lwm yam kab mob degenerative (9). Cov kev faib ua feem ntau ntawm cov kab mob sib tshooj no tau cuam tshuam peb lub hauv paus kev kuaj mob tam sim no rau kev dementia, thiab xav tau kev txhais cov ntsiab lus ntawm pathophysiological ntau dua.
Raws li qhov kev xav tau ceev ceev rau ntau yam AD biomarkers, thaj chaw tau nce siv "omics" txoj kev raws li tag nrho cov txheej txheem los nrhiav biomarkers. Lub Accelerated Pharmaceutical Partnership (AMP)-AD Alliance tau tsim tawm hauv 2014 thiab yog nyob rau ntawm qhov kev pab cuam. Qhov kev sib raug zoo no los ntawm Lub Tsev Haujlwm Saib Xyuas Kev Noj Qab Haus Huv Hauv Tebchaws, kev kawm, thiab kev lag luam txhawm rau siv cov tswv yim los ntawm cov txheej txheem los txheeb xyuas cov kab mob pathophysiology ntawm AD thiab txhim kho biodiversity diagnostic tsom xam thiab cov tswv yim kho mob (10). Raws li ib feem ntawm txoj haujlwm no, network proteomics tau dhau los ua cov cuab yeej cog lus rau kev nce qib ntawm cov txheej txheem biomarkers hauv AD. Qhov kev tsis ncaj ncees ntawm cov ntaub ntawv-tsav mus kom ze tau teeb tsa cov ntaub ntawv proteomics nyuaj rau hauv cov pab pawg lossis "modules" ntawm cov proteins uas muaj feem cuam tshuam nrog qee hom cell, organelles, thiab biological functions (11-13). Yuav luag 12 cov ntaub ntawv nplua nuj network proteomics kev tshawb fawb tau ua nyob rau hauv lub hlwb AD (13-23). Zuag qhia tag nrho, cov kev tshuaj ntsuam no qhia tau hais tias AD lub hlwb network proteome tuav lub koom haum kev txuag tau zoo hauv pawg neeg ywj pheej thiab ntau thaj chaw cortical. Tsis tas li ntawd, qee qhov ntawm cov qauv no qhia txog kev hloov pauv ntawm AD-txog kev nplua nuj nyob hauv cov ntaub ntawv teev tseg, qhia txog cov kab mob ntawm ntau yam kab mob. Sib sau ua ke, cov kev tshawb pom no qhia tau hais tias muaj txiaj ntsig zoo rau qhov kev tshawb pom ntawm lub hlwb network proteome raws li qhov system-based biomarker hauv AD.
Txhawm rau hloov pauv AD hlwb network proteome rau hauv kev kho mob muaj txiaj ntsig zoo-raws li biomarkers, peb ua ke lub hlwb-derived network nrog kev tsom xam proteomic ntawm AD CSF. Qhov kev sib koom ua ke no tau coj mus rau kev txheeb xyuas ntawm tsib qhov kev cog lus ntawm CSF biomarkers uas cuam tshuam nrog ntau hom kab mob hauv lub hlwb, suav nrog cov synapses, cov hlab ntsha, myelination, o, thiab ua tsis taus pa ntawm cov kab mob metabolic. Peb tau ua tiav validated cov biomarker panels los ntawm ntau yam replication analyses, nrog rau ntau tshaj 500 CSF kuaj los ntawm ntau yam kab mob neurodegenerative. Cov kev txheeb xyuas qhov tseeb no suav nrog kev tshuaj xyuas pab pawg lub hom phiaj hauv CSF ntawm cov neeg mob asymptomatic AD (AsymAD) lossis qhia cov pov thawj ntawm qhov txawv txav ntawm amyloid tsub zuj zuj hauv ib puag ncig kev txawj ntse. Cov kev tshuaj ntsuam no qhia txog qhov tseem ceeb ntawm kev ua liaj ua teb lom neeg hauv AsymAD cov pej xeem thiab txheeb xyuas cov cim vaj huam sib luag uas tuaj yeem ua rau cov tib neeg nyob rau theem pib ntawm tus kab mob. Zuag qhia tag nrho, cov txiaj ntsig no sawv cev rau cov kauj ruam tseem ceeb hauv kev txhim kho cov cuab yeej protein biomarker raws li ntau lub tshuab uas tuaj yeem daws tau ntau yam teeb meem kev kho mob uas tau ntsib los ntawm AD.
Lub hom phiaj tseem ceeb ntawm txoj kev tshawb no yog txhawm rau txheeb xyuas cov kab mob cerebrospinal kua biomarkers tshiab uas cuam tshuam txog ntau yam kab mob hauv hlwb uas ua rau AD. Daim duab S1 qhia txog peb txoj kev tshawb fawb, uas suav nrog (i) kev tshuaj xyuas dav dav uas tau tsav los ntawm kev tshawb pom ua ntej ntawm AD CSF thiab lub network hlwb proteome los txheeb xyuas ntau lub hlwb ntsig txog CSF kab mob biomarkers, thiab (ii) tom qab rov ua dua Cov biomarkers no nyob rau hauv ntau lub hlwb cerebrospinal. cov dej sib tov. Txoj kev tshawb nrhiav pom tau pib nrog kev soj ntsuam ntawm qhov sib txawv ntawm CSF hauv 20 tus neeg paub txog ib txwm muaj thiab 20 tus neeg mob AD ntawm Emory Goizueta Alzheimer's Disease Research Center (ADRC). Kev kuaj mob ntawm AD yog txhais tau tias yog qhov tseem ceeb ntawm kev paub tsis meej nyob rau hauv qhov muaj qis Aβ1-42 thiab nce qib ntawm tag nrho tau thiab p-tau hauv cov kua cerebrospinal [Mean Montreal Cognitive Assessment (MoCA), 13.8 ± 7.0] [ELISA (ELISA) )]] (Table S1A). Kev tswj hwm (txhais tau tias MoCA, 26.7 ± 2.2) muaj cov qib CSF biomarkers ib txwm muaj.
Tib neeg CSF yog tus cwj pwm los ntawm qhov muaj zog ntau ntawm cov protein ntau, uas albumin thiab lwm cov protein ntau heev tuaj yeem tiv thaiv kom pom cov proteins txaus siab (24). Txhawm rau ua kom qhov tob ntawm kev tshawb pom cov protein, peb tshem tawm thawj 14 cov protein ntau ntau los ntawm txhua tus qauv CSF ua ntej kev soj ntsuam huab hwm coj (MS) (24). Tag nrho ntawm 39,805 peptides tau txheeb xyuas los ntawm MS, uas tau kos npe rau 3691 proteomes hauv 40 tus qauv. Protein kom muaj nuj nqis yog ua los ntawm ntau tandem mass tag (TMT) labeling (18, 25). Txhawm rau daws cov ntaub ntawv uas ploj lawm, peb suav nrog tsuas yog cov proteins uas tau suav nrog tsawg kawg 50% ntawm cov qauv hauv kev tshuaj ntsuam tom ntej, yog li thaum kawg suav txog 2875 proteomes. Vim qhov sib txawv tseem ceeb ntawm tag nrho cov protein ntau ntau, cov qauv tswj tau raug suav tias yog ib qho outlier (13) thiab tsis suav nrog hauv kev tshuaj ntsuam tom ntej. Qhov txiaj ntsig ntawm qhov seem ntawm 39 tus qauv tau hloov kho raws li hnub nyoog, poj niam txiv neej, thiab kev sib txawv ntawm cov khoom sib txawv (13-15, 17, 18, 20, 26).
Siv cov ntaub ntawv txheeb xyuas t-kev tshuaj ntsuam xyuas los ntsuas qhov sib txawv ntawm cov ntaub ntawv regression teeb tsa, qhov kev tshuaj ntsuam no tau txheeb xyuas cov protein uas nws qhov kev nplua nuj tau hloov pauv ntau (P <0.05) ntawm kev tswj hwm thiab AD cov ntaub ntawv (Table S2A). Raws li pom nyob rau hauv daim duab 1A, qhov abundance ntawm tag nrho ntawm 225 proteins nyob rau hauv AD tau txo qis, thiab cov abundance ntawm 303 proteins tau loj hlob. Cov proteins uas txawv txav no suav nrog ntau yam uas tau txheeb pom yav dhau los cerebrospinal kua AD cov cim, xws li microtubule-associated protein tau (MAPT; P = 3.52 × 10−8), neurofilament (NEFL; P = 6.56 × 10−3), kev loj hlob txog Protein 43 (GAP43; P = 1.46 × 10−5), Fatty Acid Binding Protein 3 (FABP3; P = 2.00 × 10−5), Chitinase 3 like 1 (CHI3L1; P = 4.44 × 10−6), Neural Granulin (NRGN; P = 3.43 × 10−4) thiab VGF paj hlwb loj hlob (VGF; P = 4.83 × 10−3) (4-6). Txawm li cas los xij, peb kuj tau txheeb xyuas lwm lub hom phiaj tseem ceeb, xws li GDP dissociation inhibitor 1 (GDI1; P = 1.54 × 10-10) thiab SPARC-related modular calcium binding 1 (SMOC1; P = 6.93 × 10-9). Gene Ontology (GO) kev tshuaj xyuas ntawm 225 cov protein txo qis tau pom muaj kev sib raug zoo nrog cov txheej txheem hauv lub cev xws li steroid metabolism, ntshav coagulation, thiab cov tshuaj hormones (Daim duab 1B thiab Table S2B). Hauv qhov sib piv, cov protein ntau ntxiv ntawm 303 yog ze rau cov qauv ntawm tes thiab lub zog metabolism.
(A) volcano zaj duab xis qhia tau hais tias lub log2 fold hloov (x-axis) txheeb ze rau -log10 tus nqi P (y-axis) tau txais los ntawm t-test, uas yog siv los ntsuas qhov sib txawv ntawm kev tswj (CT) thiab AD cov ntaub ntawv ntawm CSF proteome Ntawm tag nrho cov proteins. Cov protein uas muaj cov qib txo qis (P <0.05) hauv AD tau pom hauv xiav, thaum cov proteins uas muaj cov kab mob ntau ntxiv tau pom liab. Cov protein uas tau xaiv yog sau npe. (B) Cov ntsiab lus GO sab saum toj ntsig txog cov protein tau txo qis (xiav) thiab nce (liab) hauv AD. Qhia peb cov ntsiab lus GO nrog cov qhab nia z siab tshaj plaws hauv thaj chaw ntawm cov txheej txheem lom neeg, kev ua haujlwm molecular, thiab cov khoom siv ntawm tes. (C) MS ntsuas MAPT qib hauv CSF qauv (sab laug) thiab nws qhov kev sib raug zoo nrog cov qauv ELISA tau qib (txoj cai). Pearson correlation coefficient nrog tus nqi P uas cuam tshuam tau tshwm sim. Vim tsis muaj ELISA cov ntaub ntawv rau ib rooj plaub AD, cov nuj nqis no suav nrog cov txiaj ntsig rau 38 ntawm 39 qhov kev soj ntsuam. (D) Kev soj ntsuam xyuas pawg (P <0.0001, Benjamini-Hochberg (BH) hloov kho P <0.01) ntawm kev tswj hwm thiab AD CSF pom cov qauv siv 65 qhov hloov pauv loj tshaj plaws hauv cov ntaub ntawv teev. Standardize, normalizes.
Cov qib proteomic ntawm MAPT yog ze ze rau qhov ntsuas ntawm tus kheej ELISA tau qib (r = 0.78, P = 7.8 × 10-9; Daim duab 1C), txhawb kev siv tau ntawm peb qhov kev ntsuas MS. Tom qab trypsin digestion ntawm qib ntawm amyloid precursor protein (APP), isoform-specific peptides mapped rau C-terminus ntawm Aβ1-40 thiab Aβ1-42 tsis tuaj yeem ionized zoo (27, 28). Yog li, APP peptides peb txheeb xyuas tsis muaj dab tsi cuam tshuam nrog ELISA Aβ1-42 qib. Txhawm rau ntsuas qhov sib txawv ntawm txhua qhov xwm txheej, peb siv cov protein sib txawv nrog P <0.0001 [false discovery rate (FDR) kho P <0.01] los ua ib qho kev soj ntsuam xyuas pawg ntawm cov qauv (Table S2A). Raws li pom nyob rau hauv daim duab 1D, cov 65 cov proteins tseem ceeb no tuaj yeem ua cov qauv kuaj kom raug raws li cov kab mob hauv lub xeev, tshwj tsis yog rau ib rooj plaub AD nrog cov yam ntxwv zoo li kev tswj hwm. Ntawm 65 cov proteins no, 63 nce hauv AD, thaum tsuas yog ob (CD74 thiab ISLR) txo. Nyob rau hauv tag nrho, cov cerebrospinal kua tshuaj ntsuam xyuas tau txheeb xyuas pua pua ntawm cov proteins hauv AD uas tuaj yeem ua tus kab mob biomarkers.
Tom qab ntawd peb tau ua qhov kev txheeb xyuas kev ywj pheej ntawm AD hlwb proteome. Lub hlwb cohort ntawm qhov kev tshawb pom no suav nrog dorsolateral prefrontal cortex (DLPFC) los ntawm kev tswj hwm (n = 10), Parkinson's disease (PD; n = 10), sib xyaw AD / PD (n = 10) thiab AD (n = 10) mob. ) Qauv. Emery Goizueta ADRC. Cov pej xeem ntawm 40 qhov xwm txheej no tau piav qhia yav dhau los (25) thiab tau sau tseg hauv Table S1B. Peb siv TMT-MS los tshuaj xyuas cov 40 lub hlwb cov ntaub so ntswg thiab cov kev sib txuas ntawm 27 tus neeg mob. Nyob rau hauv tag nrho, ob lub hlwb cov ntaub ntawv teev ua 227,121 tshwj xeeb peptides, uas tau mapped rau 12,943 proteomes (25). Tsuas yog cov proteins uas tau suav nrog tsawg kawg 50% ntawm cov neeg mob tau suav nrog hauv kev tshawb nrhiav tom ntej. Cov ntaub ntawv tshawb pom zaum kawg muaj 8817 cov protein ntau. Kho cov protein ntau ntau raws li hnub nyoog, poj niam txiv neej, thiab tom qab kev tuag (PMI). Kev tsom xam qhov sib txawv ntawm cov ntaub ntawv teeb tsa tom qab kev rov ua dua tau pom tias> 2000 cov protein ntau tau hloov pauv [P <0.05, tsom xam ntawm qhov sib txawv (ANOVA)] hauv ob lossis ntau pawg kab mob. Tom qab ntawd, peb tau ua qhov kev soj ntsuam xyuas pawg raws li qhov sib txawv ntawm cov proteins, thiab P <0.0001 hauv AD / tswj thiab / lossis AD / PD kev sib piv (Daim duab S2, A thiab B, Table S2C). Cov 165 hloov pauv cov protein no kom meej meej qhia txog cov teeb meem nrog AD pathology los ntawm kev tswj hwm thiab PD cov qauv, lees paub qhov muaj zog AD-kev hloov pauv hauv tag nrho cov proteome.
Tom qab ntawd peb tau siv ib qho algorithm hu ua Weighted Gene Co-expression Network Analysis (WGCNA) los ua kev tsom xam network ntawm lub hlwb proteome, uas npaj cov ntaub ntawv teev rau hauv cov qauv protein nrog cov qauv qhia zoo sib xws (11-13). Kev tsom xam tau txheeb xyuas 44 modules (M) co-expressed proteins, txheeb thiab suav los ntawm qhov loj tshaj (M1, n = 1821 proteins) mus rau qhov tsawg tshaj plaws (M44, n = 34 proteins) (Daim duab 2A thiab Table S2D)). Raws li tau hais los saum toj no (13) Xam cov neeg sawv cev qhia profile lossis cov yam ntxwv protein ntawm txhua qhov module, thiab sib txuas nrog cov kab mob hauv xeev thiab AD pathology, uas yog, tsim kom muaj kev sib koom tes ntawm Alzheimer's Disease Registry (CERAD) thiab Braak Score (Daim duab 2B). Zuag qhia tag nrho, 17 modules tau cuam tshuam nrog AD neuropathology (P <0.05). Ntau ntawm cov kab mob ntsig txog cov kab mob no kuj tseem muaj cov cim tshwj xeeb ntawm tes (Daim duab 2B). Raws li tau hais los saum toj no (13), cell type enrichment yog txiav txim siab los ntawm kev tshuaj xyuas cov qauv sib tshooj thiab cov npe siv ntawm cov xov tooj ntawm tes tshwj xeeb cov noob. Cov noob no tau muab los ntawm cov ntaub ntawv luam tawm hauv cov nas uas sib cais neurons, endothelial thiab glial hlwb. RNA sequencing (RNA-seq) kev sim (29).
(A) Tshawb nrhiav WGCNA ntawm lub hlwb proteome. (B) Biweight midcorrelation (BiCor) kev soj ntsuam ntawm cov qauv kos npe protein (thawj qhov tseem ceeb ntawm cov qauv qhia protein) nrog AD cov yam ntxwv neuropathological (sab saum toj), suav nrog CERAD (Aβ plaque) thiab Braak (tau tangles) cov qhab nia. Qhov kev siv zog ntawm qhov zoo (liab) thiab qhov tsis zoo (xiav) kev sib raug zoo yog qhia los ntawm ob daim duab tshav kub, thiab hnub qub qhia txog qhov tseem ceeb (P <0.05). Siv Hypergeometric Fisher's Exact Test (FET) (hauv qab) los ntsuam xyuas cov kev sib koom tes ntawm tes ntawm txhua tus qauv protein. Qhov kev siv ntawm qhov ntxoov ntxoo liab qhia txog qib ntawm cell hom enrichment, thiab lub hnub qub qhia txog qhov tseem ceeb (P <0.05). Siv txoj kev BH los kho tus nqi P uas tau los ntawm FET. (C) GO tsom xam ntawm modular proteins. Cov txheej txheem lom neeg muaj feem cuam tshuam zoo tshaj plaws yog qhia rau txhua qhov module lossis pawg module muaj feem xyuam. oligodendrocytes.
Ib txheej ntawm tsib qhov cuam tshuam nrog astrocyte thiab microglia-nplua nuj modules (M30, M29, M18, M24, thiab M5) tau pom muaj kev sib raug zoo nrog AD neuropathology (Daim duab 2B). Ontology tsom xam txuas cov glial modules nrog kev loj hlob ntawm tes, kev loj hlob, thiab kev tiv thaiv kab mob (Daim duab 2C thiab Table S2E). Ob qhov ntxiv glial modules, M8 thiab M22, kuj tseem muaj zog tiv thaiv kab mob. M8 muaj feem cuam tshuam rau txoj hauv kev hu xov tooj zoo li receptor, lub teeb liab cascade uas ua lub luag haujlwm tseem ceeb hauv kev tiv thaiv kab mob hauv lub cev (30). Nyob rau tib lub sijhawm, M22 muaj feem cuam tshuam nrog kev hloov pauv tom qab kev hloov pauv. M2, uas yog nplua nuj nyob rau hauv oligodendrocytes, qhia tau hais tias muaj zog correlation nrog AD pathology thiab ib tug ontological kev twb kev txuas nrog nucleoside synthesis thiab DNA replication, qhia txog kev txhim kho cell proliferation nyob rau hauv cov kab mob. Zuag qhia tag nrho, cov kev tshawb pom no txhawb nqa qhov nce ntawm glial modules uas peb tau pom yav dhau los hauv AD network proteome (13, 17). Tam sim no tau pom tias ntau qhov AD-txog glial modules hauv lub network qhia qis qis hauv kev tswj hwm thiab PD cov teeb meem, qhia txog lawv cov kab mob tshwj xeeb uas tau nce siab hauv AD (Daim duab S2C).
Tsuas yog plaub lub qauv hauv peb lub network proteome (M1, M3, M10, thiab M32) muaj kev cuam tshuam tsis zoo nrog AD pathology (P <0.05) (Daim duab 2, B thiab C). Ob leeg M1 thiab M3 yog nplua nuj nyob rau hauv cov cim neuronal. M1 muaj feem xyuam rau cov teeb liab synaptic, thaum M3 muaj feem cuam tshuam nrog kev ua haujlwm mitochondrial. Tsis muaj pov thawj ntawm cell hom enrichment rau M10 thiab M32. M32 qhia txog kev sib txuas ntawm M3 thiab cell metabolism, thaum M10 muaj feem xyuam rau kev loj hlob ntawm tes thiab microtubule muaj nuj nqi. Piv nrog AD, tag nrho plaub lub modules tau nce hauv kev tswj hwm thiab PD, muab cov kab mob tshwj xeeb AD hloov pauv (Daim duab S2C). Zuag qhia tag nrho, cov txiaj ntsig no txhawb kev txo qis ntawm cov khoom siv neuron-nplua nuj uas peb tau pom yav dhau los hauv AD (13, 17). Hauv cov ntsiab lus, kev txheeb xyuas lub network ntawm lub hlwb proteome uas peb pom tau tsim cov AD-tshwj xeeb hloov cov qauv raws li peb qhov kev tshawb pom yav dhau los.
AD yog tus cwj pwm los ntawm theem asymptomatic thaum ntxov (AsymAD), uas cov tib neeg pom cov amyloid tsub zuj zuj yam tsis muaj kev paub txog kev kho mob (5, 31). Cov theem asymptomatic no sawv cev rau lub qhov rais tseem ceeb rau kev tshawb pom ntxov thiab kev cuam tshuam. Peb yav dhau los tau pom muaj zog modular preservation ntawm AsymAD thiab AD hlwb network proteome hla cov ntaub ntawv ywj pheej (13, 17). Txhawm rau kom ntseeg tau tias lub hlwb network peb tau tshawb pom tam sim no zoo ib yam nrog cov kev tshawb pom yav dhau los, peb tau txheeb xyuas qhov khaws cia ntawm 44 modules hauv cov ntaub ntawv rov ua dua los ntawm 27 DLPFC cov koom haum. Cov koom haum no suav nrog kev tswj hwm (n = 10), AsymAD (n = 8) Thiab AD (n = 9) rooj plaub. Kev tswj hwm thiab AD cov qauv tau suav nrog hauv kev txheeb xyuas ntawm peb cov kev tshawb pom lub hlwb (Table S1B), thaum AsymAD cov ntaub ntawv tsuas yog nyob rau hauv pawg neeg sib tw. Cov teeb meem AsymAD no kuj tuaj ntawm Emory Goizueta ADRC lub txhab nyiaj hlwb. Txawm hais tias kev paub zoo li qub thaum lub sijhawm tuag, cov qib amyloid tau siab txawv txav (txhais tau tias CERAD, 2.8 ± 0.5) (Table S1B).
TMT-MS tsom xam ntawm 27 lub hlwb cov ntaub so ntswg ua rau kom muaj nuj nqis ntawm 11,244 proteomes. Qhov kev suav zaum kawg no suav nrog tsuas yog cov proteins uas suav nrog tsawg kawg 50% ntawm cov qauv. Cov ntaub ntawv rov ua dua no muaj 8638 (98.0%) ntawm 8817 cov proteins uas tau kuaj pom hauv peb qhov kev tshawb pom lub hlwb, thiab muaj ze li ntawm 3000 cov protein hloov pauv loj ntawm kev tswj hwm thiab AD pawg (P <0.05, tom qab Tukey's paired t test rau kev txheeb xyuas qhov sib txawv) ( Tab S2F). Ntawm cov proteins uas txawv txav no, 910 kuj tau pom cov kev hloov pauv tseem ceeb ntawm AD thiab lub hlwb tiv thaiv kab mob (P <0.05, tom qab ANOVA Tukey paired t-test). Nws yog ib qho tsim nyog sau cia tias cov cim 910 no zoo sib xws hauv cov kev taw qhia ntawm kev hloov ntawm proteomes (r = 0.94, P <1.0 × 10-200) (Daim duab S3A). Ntawm cov protein ntau ntxiv, cov proteins uas muaj kev hloov pauv ntawm cov ntaub ntawv feem ntau yog cov tswv cuab ntawm glial-nplua nuj M5 thiab M18 modules (MDK, COL25A1, MAPT, NTN1, SMOC1, thiab GFAP). Ntawm cov proteins uas txo qis, cov kev hloov pauv zoo ib yam yog yuav luag tsuas yog cov tswv cuab ntawm M1 module (NPTX2, VGF, thiab RPH3A) cuam tshuam nrog synapse. Peb tau txheeb xyuas ntxiv txog AD-txog kev hloov pauv ntawm midkine (MDK), CD44, secreted frizzled-related protein 1 (SFRP1) thiab VGF los ntawm sab hnub poob blotting (Daim duab S3B). Module preservation tsom xam pom tias kwv yees li 80% ntawm cov protein modules (34/44) nyob rau hauv lub hlwb proteome tau txais txiaj ntsig zoo hauv cov ntaub ntawv rov ua dua (z-score> 1.96, FDR kho P <0.05) (Daim duab S3C). Kaum plaub ntawm cov qauv no tau tshwj xeeb tshwj xeeb ntawm ob lub proteomes (z-score> 10, FDR kho P <1.0 × 10−23). Zuag qhia tag nrho, kev tshawb pom thiab kev rov ua dua ntawm qib siab ntawm qhov sib xws hauv kev qhia sib txawv thiab cov qauv sib txawv ntawm lub hlwb proteome qhia txog qhov kev hloov pauv ntawm cov kev hloov pauv hauv AD frontal cortex proteins. Tsis tas li ntawd, nws kuj tau lees paub tias AsymAD thiab cov kab mob siab ntxiv muaj cov qauv paj hlwb zoo sib xws.
Ib qho kev soj ntsuam ntxaws ntxiv ntawm qhov sib txawv ntawm qhov sib txawv ntawm cov ntaub ntawv rov ua dua lub hlwb qhia txog qhov tseem ceeb ntawm AsymAD cov protein hloov pauv, suav nrog tag nrho 151 cov protein hloov pauv ntawm AsymAD thiab kev tswj hwm (P <0.05) (Daim duab S3D). Raws li kev thauj khoom amyloid, APP hauv lub hlwb ntawm AsymAD thiab AD tau nce ntau. MAPT tsuas yog hloov pauv loj hauv AD, uas zoo ib yam nrog nce qib ntawm tangles thiab nws paub txog kev sib raug zoo nrog kev txawj ntse poob (5, 7). Cov glial-nplua nuj modules (M5 thiab M18) muaj kev cuam tshuam zoo heev hauv cov protein ntau hauv AsymAD, thaum cov neuron-txog M1 modules yog tus sawv cev tshaj plaws ntawm cov protein tsawg hauv AsymAD. Ntau ntawm cov cim AsymAD no qhia txog kev hloov pauv ntau dua hauv cov tsos mob. Ntawm cov cim no yog SMOC1, glial protein nyob rau hauv M18, uas yog txuam nrog cov qog hlwb thiab kev loj hlob ntawm ob lub qhov muag thiab ceg ceg (32). MDK yog ib qho heparin-binding kev loj hlob yam cuam tshuam txog kev loj hlob ntawm tes thiab angiogenesis (33), lwm tus tswv cuab ntawm M18. Piv nrog rau pawg tswj hwm, AsymAD nce ntxiv, ua raws li kev nce ntxiv hauv AD. Hauv qhov sib piv, cov synaptic protein neuropentraxin 2 (NPTX2) tau txo qis hauv AsymAD lub hlwb. NPTX2 yav dhau los cuam tshuam nrog neurodegeneration thiab muaj lub luag haujlwm lees paub hauv kev kho kom zoo nkauj excitatory synapses (34). Zuag qhia tag nrho, cov txiaj ntsig no qhia txog ntau yam sib txawv ntawm cov kev hloov pauv hauv preclinical protein hauv AD uas zoo li kev vam meej nrog qhov mob hnyav.
Muab hais tias peb tau ua tiav qhov tob ntawm cov protein ntau hauv kev tshawb pom ntawm lub hlwb proteome, peb tab tom sim nkag siab ntau dua nws qhov sib tshooj nrog lub network-theem AD transcriptome. Yog li ntawd, peb muab piv rau lub hlwb proteome peb nrhiav tau nrog lub module peb yav tas los generated los ntawm microarray ntsuas ntawm 18,204 noob hauv AD (n = 308) thiab tswj (n = 157) DLPFC cov ntaub so ntswg (13). sib tshooj. Nyob rau hauv tag nrho, peb tau txheeb xyuas 20 qhov sib txawv RNA modules, ntau yam uas tau qhia txog kev txhawb nqa ntawm cov cell tshwj xeeb, suav nrog cov neurons, oligodendrocytes, astrocytes, thiab microglia (Daim duab 3A). Ntau qhov kev hloov pauv ntawm cov qauv no hauv AD tau pom hauv daim duab 3B. Raws li peb yav dhau los cov protein-RNA sib tshooj tsom xam siv qhov sib sib zog nqus MS proteome (txog 3000 proteins) (13), feem ntau ntawm 44 modules hauv lub hlwb proteome network peb pom muaj nyob rau hauv transcriptome network Tsis muaj qhov tseem ceeb sib tshooj hauv. peb qhov kev tshawb pom thiab rov ua dua ntawm 34 cov qauv protein ntau uas khaws cia hauv lub hlwb proteome, tsuas yog 14 (~ 40%) dhau Fisher qhov kev xeem (FET) ua pov thawj tias muaj qhov sib tshooj tseem ceeb nrog cov ntawv sau tseg (Daim duab 3A). Tau tshaj DNA kev puas tsuaj kho (P-M25 thiab P-M19), protein txhais lus (P-M7 thiab P-M20), RNA binding / splicing (P-M16 thiab P-M21) thiab protein hom phiaj (P-M13 thiab P- M23) tsis sib tshooj nrog cov qauv hauv cov ntawv sau tseg. Yog li ntawd, txawm hais tias qhov sib sib zog nqus proteome cov ntaub ntawv teev yog siv nyob rau hauv tam sim no sib tshooj tsom xam (13), feem ntau ntawm AD network proteome tsis yog mapped rau lub transcriptome network.
(A) Hypergeometric FET qhia txog kev txhawb nqa ntawm cov xov tooj ntawm tes-cov cim tshwj xeeb hauv RNA module ntawm AD transcriptome (sab saum toj) thiab qib sib tshooj ntawm RNA (x-axis) thiab protein (y-axis) modules ntawm lub hlwb AD (hauv qab). Qhov kev siv ntawm cov xim liab shading qhia txog qib ntawm enrichment ntawm cell hom nyob rau hauv lub sab saum toj vaj huam sib luag thiab kev siv ntawm kev sib tshooj ntawm cov modules hauv qab vaj huam sib luag. Cov hnub qub qhia txog qhov tseem ceeb (P <0.05). (B) Cov qib ntawm kev sib raug zoo ntawm cov yam ntxwv noob ntawm txhua qhov kev hloov pauv thiab AD raws li txoj cai. Cov qauv ntawm sab laug yog qhov cuam tshuam tsis zoo tshaj plaws nrog AD (xiav), thiab cov nyob sab xis yog qhov zoo tshaj plaws nrog AD (liab). Lub log-transformed BH-kho P tus nqi qhia txog qib ntawm qhov tseem ceeb ntawm txhua qhov kev sib raug zoo. (C) Cov txheej txheem sib tshooj tseem ceeb nrog kev sib koom tes hom kev txhawb nqa. (D) Kev txheeb xyuas kev sib txheeb ntawm lub log2 quav hloov ntawm cov ntawv sau npe (x-axis) thiab RNA (y-axis) hauv qhov sib tshooj. Pearson correlation coefficient nrog tus nqi P uas cuam tshuam tau tshwm sim. micro, microglia; lub cev ntawm lub cev, astrocytes. CT, tswj.
Feem ntau cov sib tshooj protein ntau thiab RNA modules qhia zoo sib xws ntawm cov xov tooj ntawm tes txhawb nqa thiab cov lus qhia hloov pauv AD (Daim duab 3, B thiab C). Hauv lwm lo lus, synapse-related M1 module ntawm lub hlwb proteome (PM1) yog mapped rau peb neuronal-nplua nuj homologous RNA modules (R-M1, R-M9 thiab R-M16), uas yog nyob rau hauv AD Ob leeg pom. txo qib. Ib yam li ntawd, glial-nplua nuj M5 thiab M18 protein modules sib tshooj nrog RNA modules nplua nuj nyob rau hauv astrocytes thiab microglial cov cim (R-M3, R-M7, thiab R-M10) thiab koom nrog cov kab mob ntau ntxiv. Cov kev sib koom modular nta nruab nrab ntawm ob cov ntaub ntawv teeb tsa ntxiv txhawb nqa hom cell enrichment thiab kab mob hloov pauv uas peb tau pom hauv lub hlwb proteome. Txawm li cas los xij, peb tau pom ntau qhov sib txawv tseem ceeb ntawm RNA thiab qib protein ntawm tus kheej cov cim hauv cov qauv sib koom no. Kev txheeb xyuas qhov sib txawv ntawm qhov sib txawv ntawm cov proteomics thiab transcriptomics ntawm cov molecules hauv cov txheej txheem sib tshooj no (Daim duab 3D) qhia txog qhov tsis sib xws. Piv txwv li, APP thiab ob peb lwm glial module proteins (NTN1, MDK, COL25A1, ICAM1, thiab SFRP1) tau pom qhov nce ntxiv hauv AD proteome, tab sis yuav luag tsis muaj qhov hloov pauv hauv AD transcriptome. Cov kev hloov pauv tshwj xeeb ntawm cov protein no tuaj yeem cuam tshuam nrog cov kab mob amyloid plaques (23, 35), qhia txog cov proteome raws li qhov chaw ntawm cov kev hloov pauv pathological, thiab cov kev hloov pauv no yuav tsis cuam tshuam rau hauv cov ntawv sau tseg.
Tom qab nws tus kheej txheeb xyuas lub hlwb thiab CSF proteomes peb nrhiav tau, peb tau ua ib qho kev ntsuam xyuas ntawm ob cov ntaub ntawv teev tseg los txheeb xyuas AD CSF biomarkers ntsig txog pathophysiology ntawm lub hlwb network. Peb yuav tsum xub txhais qhov sib tshooj ntawm ob lub proteomes. Txawm hais tias nws tau lees paub tias CSF cuam tshuam txog kev hloov pauv neurochemical hauv AD hlwb (4), qhov tseeb ntawm kev sib tshooj ntawm AD lub hlwb thiab CSF proteome tsis meej. Los ntawm kev sib piv tus naj npawb ntawm cov khoom lag luam sib koom kuaj pom hauv peb ob lub proteomes, peb pom tias ze li ntawm 70% (n = 1936) ntawm cov proteins uas pom nyob rau hauv cov kua cerebrospinal kuj muaj nyob rau hauv lub hlwb (Daim duab 4A). Feem ntau ntawm cov khoom sib tshooj no (n = 1721) yog mapped rau ib qho ntawm 44 co-expression modules los ntawm cov ntaub ntawv tshawb pom lub hlwb (Daim duab 4B). Raws li kev cia siab, rau lub hlwb loj tshaj plaws (M1 txog M6) tau nthuav tawm qhov loj tshaj plaws ntawm CSF sib tshooj. Txawm li cas los xij, muaj cov hlwb me me (piv txwv li, M15 thiab M29) uas ua tiav qhov kev xav tau siab ntawm kev sib tshooj, loj dua lub hlwb module ob zaug nws qhov loj. Qhov no txhawb peb kom tau txais cov ncauj lus kom ntxaws ntxiv, kev txheeb xyuas cov txheej txheem los xam qhov sib tshooj ntawm lub hlwb thiab cov kua cerebrospinal.
(A thiab B) Cov proteins kuaj pom hauv lub hlwb tshawb pom thiab CSF cov ntaub ntawv teeb tsa sib tshooj. Feem ntau ntawm cov protein sib tshooj no yog txuam nrog ib qho ntawm 44 co-expression modules ntawm lub hlwb co-expression network. (C) Tshawb nrhiav qhov sib tshooj ntawm cerebrospinal kua proteome thiab lub hlwb network proteome. Txhua kab ntawm daim ntawv qhia tshav kub sawv cev rau ib qho kev sib tshooj sib tshooj ntawm cov hypergeometric FET. Sab saum toj kab qhia txog qhov sib tshooj (grey / dub shading) ntawm lub hlwb module thiab tag nrho CSF proteome. Kab thib ob piav qhia tias qhov sib tshooj ntawm lub hlwb modules thiab CSF protein (shaded nyob rau hauv liab) yog ho nce-tswj nyob rau hauv AD (P <0.05). Kab thib peb qhia tau hais tias qhov sib tshooj ntawm lub hlwb modules thiab CSF protein (xiav shading) yog qhov txo qis hauv AD (P <0.05). Siv txoj kev BH los kho tus nqi P uas tau los ntawm FET. (D) Folding module vaj huam sib luag raws li kev koom tes ntawm tes thiab lwm yam GO cov ntsiab lus. Cov vaj huam sib luag no muaj tag nrho ntawm 271 lub hlwb ntsig txog cov protein, uas muaj cov ntsiab lus sib txawv hauv CSF proteome.
Siv ib leeg-tailed FETs, peb tau soj ntsuam qhov tseem ceeb ntawm cov protein sib tshooj ntawm CSF proteome thiab tib lub hlwb modules. Kev tshuaj xyuas tau qhia tias tag nrho ntawm 14 lub hlwb modules hauv CSF cov ntaub ntawv teeb tsa muaj qhov sib txawv tseem ceeb (FDR hloov kho P <0.05), thiab ib qho ntxiv module (M18) uas nws sib tshooj yog ze rau qhov tseem ceeb (FDR kho P = 0.06) (Daim duab 4C , sab saum toj kab). Peb kuj txaus siab rau cov qauv uas muaj zog sib tshooj nrog qhov sib txawv ntawm CSF cov proteins. Yog li ntawd, peb tau siv ob qhov kev tshuaj ntsuam FET ntxiv los txiav txim seb qhov twg ntawm (i) CSF protein tau nce ntau hauv AD thiab (ii) CSF protein tau txo qis hauv AD (P <0.05, paired t test AD/control) Brain modules with meaningful overlap. nruab nrab ntawm lawv. Raws li pom nyob rau hauv nruab nrab thiab hauv qab kab ntawm daim duab 4C, cov kev ntsuam xyuas ntxiv no qhia tau hais tias 8 ntawm 44 lub hlwb modules sib cuam tshuam nrog cov protein ntxiv hauv AD CSF (M12, M1, M2, M18, M5, M44, M33, thiab M38) . ), thaum tsuas yog ob lub qauv (M6 thiab M15) tau pom muaj txiaj ntsig sib tshooj nrog cov protein tsawg hauv AD CSF. Raws li kev cia siab, tag nrho 10 modules yog nyob rau hauv 15 modules nrog lub siab tshaj plaws sib tshooj nrog CSF proteome. Yog li, peb xav tias 15 cov qauv no yog cov txiaj ntsig siab ntawm AD hlwb-derived CSF biomarkers.
Peb muab cov 15 qhov sib tshooj sib tshooj no rau hauv tsib lub vaj huam sib luag loj loj raws li lawv qhov sib thooj hauv WGCNA tsob ntoo daim duab thiab lawv cov kev koom tes nrog hom cell thiab noob ontology (Daim duab 4D). Thawj lub vaj huam sib luag muaj cov modules nplua nuj nyob rau hauv cov cim neuron thiab synapse-txog cov proteins (M1 thiab M12). Lub vaj huam sib luag synaptic muaj tag nrho ntawm 94 cov proteins, thiab cov qib hauv CSF proteome tau hloov pauv ntau, ua rau nws yog qhov loj tshaj plaws ntawm lub hlwb ntsig txog CSF cov cim ntawm tsib lub vaj huam sib luag. Pab pawg thib ob (M6 thiab M15) tau qhia txog kev sib raug zoo nrog cov cim ntawm endothelial cell thiab vascular lub cev, xws li "qhov txhab kho" (M6) thiab "txoj cai ntawm kev tiv thaiv kab mob humoral" (M15). M15 kuj tseem muaj feem cuam tshuam rau lipoprotein metabolism, uas yog ze rau endothelium (36). Lub vascular vaj huam sib luag muaj 34 CSF cov cim ntsig txog lub hlwb. Pab pawg thib peb suav nrog cov qauv (M2 thiab M4) uas muaj feem cuam tshuam nrog cov cim oligodendrocytes thiab cell proliferation. Piv txwv li, cov ntsiab lus ontology saum toj kawg nkaus ntawm M2 suav nrog "kev tswj hwm zoo ntawm DNA replication" thiab "purine biosynthesis txheej txheem". Lub caij no, cov M4 suav nrog "glial cell sib txawv" thiab "chromosome cais". Lub vaj huam sib luag myelination muaj 49 CSF cov cim ntsig txog lub hlwb.
Pawg plaub muaj cov modules feem ntau (M30, M29, M18, M24, thiab M5), thiab yuav luag tag nrho cov modules yog nplua nuj nyob rau hauv microglia thiab astrocyte cov cim. Zoo ib yam li lub vaj huam sib luag myelination, lub vaj huam sib luag plaub kuj tseem muaj cov qauv (M30, M29, thiab M18) uas muaj feem cuam tshuam nrog kev loj hlob ntawm tes. Lwm cov qauv hauv pawg no muaj feem cuam tshuam nrog cov ntsiab lus tiv thaiv kab mob, xws li "txoj kev tiv thaiv kab mob" (M5) thiab "kev tiv thaiv kab mob tiv thaiv kab mob" (M24). Cov pab pawg neeg tiv thaiv kab mob glial muaj 42 CSF cov cim ntsig txog lub hlwb. Thaum kawg, lub vaj huam sib luag kawg suav nrog 52 lub hlwb ntsig txog cov cim ntawm plaub lub modules (M44, M3, M33, thiab M38), tag nrho cov ntawm lub cev muaj feem xyuam rau lub zog cia thiab cov metabolism. Qhov loj tshaj plaws ntawm cov qauv no (M3) yog ze ze rau mitochondria thiab yog nplua nuj nyob rau hauv cov cim tshwj xeeb neuron. M38 yog ib qho ntawm cov tswvcuab me me hauv cov metabolome no thiab tseem qhia txog qhov tshwj xeeb ntawm cov neuron.
Zuag qhia tag nrho, tsib lub vaj huam sib luag no qhia txog ntau hom xovtooj ntawm tes thiab kev ua haujlwm hauv AD cortex, thiab suav nrog 271 lub hlwb ntsig txog CSF cov cim (Table S2G). Txhawm rau ntsuas qhov siv tau ntawm cov txiaj ntsig MS no, peb tau siv qhov sib thooj txuas ntxiv (PEA), ib qho cuab yeej siv tshuaj tiv thaiv kab mob orthogonal nrog kev ua kom muaj peev xwm ua kom muaj peev xwm, siab rhiab heev thiab tshwj xeeb, thiab rov tshuaj xyuas cov qauv cerebrospinal kua peb pom Ib pawg ntawm cov 271 biomarkers. (nqe = 36). Cov 36 lub hom phiaj no qhia txog kev hloov pauv hauv AD ntau yam ntawm PEA, uas yog ze rau peb cov kev tshawb pom MS-raws li (r = 0.87, P = 5.6 × 10-12), uas tau txheeb xyuas cov txiaj ntsig ntawm peb cov kev tshawb fawb MS (Daim duab S4. ).
Cov ntsiab lus lom neeg tau hais los ntawm peb tsib pawg, los ntawm kev taw qhia synaptic rau lub zog metabolism, yog txhua yam cuam tshuam nrog cov kab mob ntawm AD (1-3). Yog li ntawd, tag nrho 15 modules uas muaj cov panels no muaj feem xyuam rau AD pathology nyob rau hauv lub hlwb proteome uas peb nrhiav tau (Daim duab 2B). Qhov tseem ceeb tshaj plaws yog qhov muaj txiaj ntsig zoo ntawm kev sib raug zoo ntawm peb cov glial modules thiab qhov muaj zog tsis zoo ntawm kev sib raug zoo ntawm peb cov neuronal loj tshaj plaws (M1 thiab M3). Kev tsom xam qhov sib txawv ntawm peb lub hlwb replicated proteome (Daim duab S3D) kuj tseem qhia txog M5 thiab M18-derived glial proteins. Hauv AsymAD thiab cov tsos mob AD, feem ntau nce glial proteins thiab M1-txog synapses Cov protein yog txo qis tshaj plaws. Cov kev soj ntsuam no qhia tau hais tias 271 cerebrospinal kua cim peb tau txheeb xyuas hauv tsib pawg muaj feem cuam tshuam nrog cov txheej txheem kab mob hauv AD cortex, suav nrog cov uas tshwm sim thaum ntxov asymptomatic.
Txhawm rau txhawm rau txheeb xyuas qhov kev hloov pauv ntawm lub vaj huam sib luag ntawm cov protein nyob hauv lub hlwb thiab cov leeg txha caj qaum, peb kos cov hauv qab no rau txhua qhov ntawm 15 qhov sib tshooj ntawm cov txheej txheem sib tshooj: (i) pom cov qauv kev nplua nuj nyob hauv lub hlwb cov ntaub ntawv teeb tsa thiab (ii) cov modules. protein Qhov txawv yog qhia nyob rau hauv lub cerebrospinal kua (Daim duab S5). Raws li tau hais ua ntej, WGCNA yog siv los txiav txim siab cov qauv kev nplua nuj lossis cov yam ntxwv ntawm cov protein hauv lub hlwb (13). Daim ntawv qhia roob hluav taws yog siv los piav qhia qhov sib txawv ntawm cov proteins modular hauv cov kua cerebrospinal (AD / tswj). Cov duab no qhia tau hais tias peb ntawm tsib lub vaj huam sib luag qhia cov kev qhia sib txawv hauv lub hlwb thiab cov leeg txha caj qaum. Ob lub modules ntawm lub vaj huam sib luag synapse (M1 thiab M12) qhia qhov txo qis hauv qib ntau hauv AD lub hlwb, tab sis cuam tshuam nrog cov protein ntau ntxiv hauv AD CSF (Daim duab S5A). Cov kev cuam tshuam txog neuron uas muaj cov metabolome (M3 thiab M38) tau pom zoo li lub hlwb thiab cov kua dej cerebrospinal qhia cov qauv tsis sib xws (Daim duab S5E). Lub vascular vaj huam sib luag kuj tau pom cov kev qhia sib txawv, txawm hais tias nws cov qauv (M6 thiab M15) tau nce nruab nrab hauv AD lub hlwb thiab txo qis hauv cov kab mob CSF (Daim duab S5B). Qhov seem ob lub vaj huam sib luag muaj cov glial network loj uas nws cov proteins tau tswj hwm tsis tu ncua hauv ob qhov chaw (Daim duab S5, C thiab D).
Thov nco ntsoov tias cov qauv no tsis zoo rau txhua tus cim hauv cov vaj huam sib luag no. Piv txwv li, lub vaj huam sib luag synaptic suav nrog ntau cov proteins uas txo qis hauv AD lub hlwb thiab CSF (Figure S5A). Ntawm cov kab mob cerebrospinal cov kua dej hauv qab no yog NPTX2 thiab VGF ntawm M1, thiab chromogranin B ntawm M12. Txawm li cas los xij, txawm tias cov kev zam no, feem ntau ntawm peb cov cim synaptic tau nce siab hauv AD spinal fluid. Zuag qhia tag nrho, cov kev tshuaj ntsuam no muaj peev xwm paub qhov txawv ntawm cov xwm txheej tseem ceeb hauv lub hlwb thiab cov kua cerebrospinal hauv txhua ntawm peb tsib lub vaj huam sib luag. Cov qauv no qhia txog qhov nyuaj thiab feem ntau sib txawv ntawm lub hlwb thiab CSF protein qhia hauv AD.
Tom qab ntawd, peb siv high-throughput MS replication tsom xam (CSF replication 1) kom nqaim peb 271 txheej biomarkers mus rau lub hom phiaj zoo tshaj plaws thiab rov tsim dua tshiab (Daim duab 5A). CSF daim ntawv luam 1 muaj tag nrho ntawm 96 cov qauv los ntawm Emory Goizueta ADRC, suav nrog kev tswj hwm, AsymAD, thiab AD cohort (Table S1A). Cov teeb meem AD no yog tus cwj pwm los ntawm kev paub tsis meej me ntsis (txhais tau tias MoCA, 20.0 ± 3.8), thiab kev hloov pauv hauv AD biomarkers tau lees paub hauv cov kua cerebrospinal (Table S1A). Contrary to the CSF tsom xam peb pom, qhov replication no yog ua los ntawm ib tug ntau npaum thiab high-throughput "ib zaug-shot" MS txoj kev (tsis muaj off-line fractionation), nrog rau ib tug yooj yim qauv npaj raws tu qauv uas tshem tawm qhov yuav tsum tau rau immunodepletion ntawm ib tug neeg kuaj. . Hloov chaw, ib qho kev tiv thaiv kab mob tsis txaus "txhim kho cov channel" yog siv los nthuav tawm cov teeb liab ntawm cov protein ntau tsawg (37). Txawm hais tias nws txo qis tag nrho cov kev tiv thaiv kev tiv thaiv, qhov kev txhaj tshuaj ib zaug no txo qis lub sijhawm siv lub tshuab thiab ua rau muaj cov qauv TMT-labeled uas tuaj yeem txheeb xyuas tau (17, 38). Nyob rau hauv tag nrho, kev tsom xam pom 6,487 peptides, uas tau kos npe rau 1,183 proteomes hauv 96 kis. Raws li kev soj ntsuam CSF peb pom, tsuas yog cov proteins uas muaj tsawg kawg yog 50% ntawm cov qauv tau suav nrog hauv kev suav tom qab, thiab cov ntaub ntawv tau rov qab los rau cov teebmeem ntawm hnub nyoog thiab poj niam txiv neej. Qhov no tau coj mus rau qhov kawg ntawm 792 proteomes, 95% ntawm cov uas kuj tau txheeb xyuas hauv CSF cov ntaub ntawv pom.
(A) Lub hlwb ntsig txog CSF protein cov hom phiaj tau txheeb xyuas nyob rau hauv thawj qhov rov ua dua CSF pawg thiab suav nrog hauv vaj huam sib luag zaum kawg (n = 60). (B txog E) Vaj huam sib luag biomarker qib (sib xyaw z-cov qhab nia) ntsuas hauv plaub CSF replication cohorts. Paired t-tests lossis ANOVA nrog Tukey qhov kev kho tom qab raug siv los ntsuas qhov tseem ceeb ntawm cov kev hloov pauv ntawm kev nplua nuj nyob rau hauv txhua qhov kev txheeb xyuas rov qab. CT, tswj.
Txij li thaum peb txaus siab tshwj xeeb hauv kev txheeb xyuas peb 271 lub hlwb ntsig txog CSF lub hom phiaj los ntawm kev tshuaj ntsuam xyuas, peb yuav txwv tsis pub kuaj ntxiv ntawm qhov kev hloov pauv no rau cov cim no. Ntawm cov 271 cov proteins no, 100 tau kuaj pom hauv CSF replication 1. Daim duab S6A qhia qhov sib txawv ntawm cov 100 cov cim sib tshooj ntawm kev tswj thiab AD replication qauv. Synaptic thiab metabolite histones nce ntau tshaj plaws hauv AD, thaum vascular proteins txo qis tshaj plaws hauv cov kab mob. Feem ntau ntawm 100 cov cim sib tshooj (n = 70) tuav tib txoj kev hloov pauv hauv ob cov ntaub ntawv teev tseg (Daim duab S6B). Cov 70 tau lees paub lub hlwb ntsig txog CSF cov cim (Table S2H) feem ntau cuam tshuam txog cov kev coj ua yav dhau los ntawm lub vaj huam sib luag qhia, uas yog, kev tswj hwm ntawm vascular proteins thiab kev tswj hwm ntawm tag nrho lwm cov vaj huam sib luag. Tsuas yog 10 ntawm no 70 cov proteins uas tau lees paub tau pom tias muaj kev hloov pauv hauv AD ntau ntxiv uas cuam tshuam cov qauv vaj huam sib luag no. Txhawm rau tsim kom muaj lub vaj huam sib luag uas zoo tshaj plaws qhia txog tag nrho cov qauv ntawm lub hlwb thiab cov kua dej cerebrospinal, peb tshem tawm cov 10 cov proteins los ntawm lub vaj huam sib luag uas peb tau tshawb xyuas thaum kawg (Daim duab 5A). Yog li ntawd, peb lub vaj huam sib luag thaum kawg suav nrog tag nrho 60 cov protein uas tau txheeb xyuas hauv ob pawg CSF AD ywj pheej siv cov qauv sib txawv thiab MS platform tsom xam. Z-cov qhab nia qhia cov phiaj xwm ntawm cov phiaj xwm kawg no hauv CSF daim ntawv theej 1 kev tswj hwm thiab cov xwm txheej AD tau lees paub tias lub vaj huam sib luag tau pom hauv CSF pawg peb pom (Daim duab 5B).
Ntawm cov 60 cov proteins no, muaj cov molecules paub tias muaj feem xyuam nrog AD, xws li osteopontin (SPP1), uas yog ib qho pro-inflammatory cytokine uas tau txuam nrog AD hauv ntau cov kev tshawb fawb (39-41), thiab GAP43, A synaptic protein. uas yog kom meej meej txuas rau neurodegeneration (42). Cov proteins uas tau lees paub tshaj plaws yog cov cim ntsig txog lwm yam kab mob neurodegenerative, xws li amyotrophic lateral sclerosis (ALS) ntsig txog superoxide dismutase 1 (SOD1) thiab Parkinson's disease ntsig txog desaccharase (PARK7). Peb kuj tau txheeb xyuas tias ntau lwm cov cim, xws li SMOC1 thiab lub hlwb-nplua nuj daim nyias nyias txuas signaling protein 1 (BASP1), tau txwv cov kev txuas yav dhau los rau neurodegeneration. Nws yog ib nqi sau cia hais tias vim lawv tsis tshua muaj tag nrho cov abundance nyob rau hauv lub CSF proteome, nws yog ib qhov nyuaj rau peb siv no high-throughput ib leeg-shot detection txoj kev ntseeg tau mus ntes MAPT thiab lwm yam AD-related proteins (piv txwv li, NEFL thiab NRGN. (43, 44) ib.
Tom qab ntawd peb tau txheeb xyuas cov 60 qhov tseem ceeb ntawm lub vaj huam sib luag hauv peb qhov kev ntsuam xyuas ntxiv. Hauv CSF Copy 2, peb siv tib TMT-MS los tshuaj xyuas ib pawg neeg ywj pheej ntawm 297 tswj thiab AD cov qauv los ntawm Emory Goizueta ADRC (17). CSF replication 3 suav nrog kev rov ua dua tshiab ntawm TMT-MS cov ntaub ntawv los ntawm 120 tswj thiab AD cov neeg mob los ntawm Lausanne, Switzerland (45). Peb tau kuaj pom ntau tshaj li ob feem peb ntawm 60 tus cim tseem ceeb hauv txhua cov ntaub ntawv. Txawm hais tias Swiss txoj kev tshawb fawb tau siv MS platforms sib txawv thiab TMT quantification txoj kev (45, 46), peb tau rov ua dua peb lub vaj huam sib luag hauv ob qhov kev tshuaj xyuas dua (Daim duab 5, C thiab D, thiab Tables S2, I, thiab J). Txhawm rau ntsuas tus kab mob tshwj xeeb ntawm peb pab pawg, peb siv TMT-MS los tshuaj xyuas cov ntaub ntawv rov ua dua thib plaub (CSF replication 4), uas suav nrog tsis tsuas yog tswj (n = 18) thiab AD (n = 17) tus neeg mob, tab sis kuj PD ( n = 14)), ALS (n = 18) thiab frontotemporal dementia (FTD) cov qauv (n = 11) (Table S1A). Peb ua tiav qhov ntsuas ze li ob feem peb ntawm lub vaj huam sib luag cov proteins hauv pawg no (38 ntawm 60). Cov txiaj ntsig no qhia txog AD-kev hloov pauv hauv txhua tsib lub biomarker panels (Daim duab 5E thiab Table S2K). Qhov nce hauv pawg metabolite pom qhov muaj zog tshaj plaws AD tshwj xeeb, ua raws li myelination thiab glial pawg. Rau qhov tsawg dua, FTD kuj qhia tau tias muaj kev nce ntxiv ntawm cov vaj huam sib luag no, uas tuaj yeem cuam tshuam cov kev hloov pauv hauv lub network zoo sib xws (17). Hauv qhov sib piv, ALS thiab PD pom yuav luag tib yam myelination, glial, thiab metabolome profiles raws li pawg tswj hwm. Zuag qhia tag nrho, txawm tias qhov sib txawv ntawm cov qauv kev npaj, MS platform, thiab TMT cov txheej txheem kom muaj nuj nqis, cov kev soj ntsuam rov qab no qhia tau tias peb cov cim tseem ceeb ntawm pawg thawj coj muaj kev hloov pauv AD tshwj xeeb hauv ntau dua 500 tus qauv CSF tshwj xeeb.
AD neurodegeneration tau lees paub dav dav ntau xyoo ua ntej qhov pib ntawm kev paub txog cov tsos mob, yog li muaj qhov xav tau ceev ceev rau biomarkers ntawm AsymAD (5, 31). Txawm li cas los xij, ntau thiab ntau cov pov thawj qhia tau hais tias biology ntawm AsymAD nyob deb ntawm homogeneous, thiab kev sib cuam tshuam ntawm kev pheej hmoo thiab kev ua kom muaj zog ua rau muaj kev sib txawv loj hauv cov kab mob tom ntej (47). Txawm hais tias siv los txheeb xyuas cov xwm txheej AsymAD, cov theem ntawm cov tub ntxhais CSF biomarkers (Aβ1-42, tag nrho tau thiab p-tau) tsis tau muaj pov thawj kom ntseeg tau tias leej twg yuav nce mus rau dementia (4, 7), qhia ntau ntxiv. tsim nyog suav nrog holistic biomarker cov cuab yeej raws li ntau yam ntawm lub hlwb physiology kom raug stratify qhov kev pheej hmoo ntawm cov pejxeem. Yog li ntawd, peb tom qab soj ntsuam peb AD-validated biomarker vaj huam sib luag nyob rau hauv AsymAD cov pej xeem ntawm CSF daim ntawv 1. Cov 31 AsymAD cov ntaub ntawv no qhia txawv txawv core biomarker theem (Aβ1–42/tag nrho tau ELISA ratio, <5.5) thiab tiav kev txawj ntse (txhais tau tias MoCA, 27.1. ± 2.2) (Table S1A). Tsis tas li ntawd, txhua tus neeg uas muaj AsymAD muaj qhov chaw kho mob dementia qhab nia ntawm 0, qhia tias tsis muaj pov thawj ntawm kev poob qis hauv kev txawj ntse txhua hnub lossis kev ua haujlwm.
Peb thawj zaug txheeb xyuas cov qib ntawm cov vaj huam sib luag tau lees paub hauv tag nrho 96 CSF replicates 1, suav nrog AsymAD pawg. Peb pom tias ntau lub vaj huam sib luag hauv AsymAD pab pawg muaj qhov tseem ceeb AD-zoo li kev hloov pauv ntau, lub vascular vaj huam sib luag tau qhia txog kev poob qis hauv AsymAD, thaum tag nrho lwm cov vaj huam sib luag tau pom qhov nce siab (Daim duab 6A). Yog li ntawd, txhua lub vaj huam sib luag tau pom muaj kev sib raug zoo nrog ELISA Aβ1-42 thiab tag nrho tau qib (Daim duab 6B). Hauv qhov sib piv, qhov sib txheeb ntawm pab pawg thiab MoCA cov qhab nia yog qhov tsis zoo. Ib qho ntawm cov kev tshawb pom zoo tshaj plaws los ntawm cov kev tshuaj ntsuam no yog qhov loj ntawm cov vaj huam sib luag ntau hauv AsymAD pawg. Raws li pom nyob rau hauv daim duab 6A, lub vaj huam sib luag theem ntawm AsymAD pawg feem ntau hla lub vaj huam sib luag theem ntawm pawg tswj hwm thiab pawg AD, uas qhia tau tias muaj kev sib txawv heev. Txhawm rau tshawb nrhiav qhov sib txawv ntawm AsymAD no, peb tau siv qhov kev ntsuam xyuas Multidimensional Scaling (MDS) rau 96 CSF replication 1 rooj plaub. MDS tsom xam tso cai rau pom qhov zoo sib xws ntawm cov xwm txheej raws li qee qhov sib txawv hauv cov ntaub ntawv teev. Rau qhov kev tshuaj ntsuam xyuas pawg no, peb tsuas yog siv cov cim vaj huam sib luag uas tau lees paub uas muaj kev hloov pauv tseem ceeb (P <0.05, AD / tswj) hauv CSF nrhiav pom thiab rov ua dua 1 proteome (n = 29) (Table S2L) qib. Qhov kev tshuaj ntsuam no ua rau pom tseeb qhov sib txawv ntawm peb cov kev tswj hwm thiab AD cov teeb meem (Daim duab 6C). Hauv qhov sib piv, qee qhov AsymAD tau pom meej meej hauv pawg tswj hwm, thaum lwm tus nyob hauv cov xwm txheej AD. Txhawm rau tshawb nrhiav qhov AsymAD heterogeneity ntxiv, peb siv peb daim ntawv qhia MDS los txhais ob pawg ntawm cov xwm txheej AsymAD no. Thawj pab pawg suav nrog AsymAD cov teeb meem sib koom ua ke ze rau kev tswj hwm (n = 19), thaum pab pawg thib ob yog tus yam ntxwv ntawm AsymAD cov xwm txheej nrog cov cim qhia ze dua rau AD (n = 12).
(A) Qib kev qhia (z-score) ntawm pawg CSF biomarker hauv tag nrho 96 cov qauv hauv CSF replication 1 cohort, suav nrog AsymAD. Kev txheeb xyuas qhov sib txawv nrog Tukey qhov kev kho tom qab raug siv los ntsuas qhov tseem ceeb ntawm cov vaj huam sib luag hloov pauv. (B) Kev txheeb xyuas ntawm vaj huam sib luag protein ntau qib (z-score) nrog MoCA tus qhab nia thiab tag nrho tau qib hauv ELISA Aβ1-42 thiab CSF luam 1 cov qauv. Pearson correlation coefficient nrog tus nqi P uas cuam tshuam tau tshwm sim. (C) MDS ntawm 96 CSF daim ntawv luam 1 rooj plaub yog ua raws li cov qib ntau ntawm 29 cov ntawv pov thawj uas tau lees paub, uas tau hloov pauv ntau hauv ob qho kev tshawb pom thiab CSF daim ntawv luam 1 cov ntaub ntawv [P <0.05 AD/control (CT)]. Qhov kev tshuaj ntsuam no tau siv los faib pawg AsymAD rau hauv kev tswj hwm (n = 19) thiab AD (n = 12) pawg. (D) Cov phiaj xwm hluav taws xob qhia qhov sib txawv ntawm txhua CSF replication 1 proteins nrog log2 quav hloov (x-axis) txheeb ze rau -log10 tus lej P tus nqi ntawm ob pawg AsymAD. Cov vaj huam sib luag biomarkers muaj xim. (E) CSF replication 1 abundance theem ntawm pawg xaiv biomarkers yog txawv ntawm AsymAD subgroups. Tukey's post-adjusted analysis of variance yog siv los ntsuas qhov tseem ceeb ntawm kev txheeb cais.
Peb tau tshuaj xyuas cov protein sib txawv ntawm cov kev tswj thiab AD-zoo li AsymAD cov ntaub ntawv (Daim duab 6D thiab Table S2L). Daim ntawv qhia lub roob hluav taws uas tshwm sim qhia tau hais tias 14 lub vaj huam sib luag cov cim tau hloov pauv ntawm ob pawg. Feem ntau ntawm cov cim no yog cov tswv cuab ntawm synapse thiab metabolome. Txawm li cas los xij, SOD1 thiab myristoylated alanine-nplua nuj protein ntau kinase C substrate (MARCKS), uas yog cov tswv cuab ntawm myelin thiab glial immune pawg, raws li, kuj yog cov pab pawg no (Daim duab 6, D thiab E). Lub vascular vaj huam sib luag kuj tau pab txhawb ob lub cim uas tau txo qis hauv AD-zoo li AsymAD pawg, suav nrog AE binding protein 1 (AEBP1) thiab ntxiv rau cov neeg hauv tsev neeg C9. Tsis muaj qhov sib txawv tseem ceeb ntawm kev tswj hwm thiab AD-zoo li AsymAD subgroups hauv ELISA AB1-42 (P = 0.38) thiab p-tau (P = 0.28), tab sis muaj qhov sib txawv tseem ceeb hauv tag nrho cov qib tau (P = 0.0031 ) (Fig. S7). Muaj ntau lub vaj huam sib luag cov cim qhia tias qhov kev hloov pauv ntawm ob pawg AsymAD subgroups yog qhov tseem ceeb tshaj li tag nrho cov qib tau (piv txwv li, YWHAZ, SOD1, thiab MDH1) (Daim duab 6E). Zuag qhia tag nrho, cov txiaj ntsig no qhia tau tias peb lub vaj huam sib luag tau lees paub tuaj yeem muaj cov cim biomarkers uas tuaj yeem ua rau subtype thiab muaj peev xwm stratification ntawm cov neeg mob asymptomatic.
Muaj qhov xav tau ceev ceev rau cov cuab yeej siv biomarker los ntsuas qhov zoo dua thiab tsom mus rau ntau yam pathophysiology tom qab AD. Cov cuab yeej no yuav tsum tsis tsuas yog hloov pauv peb cov txheej txheem kuaj mob AD, tab sis kuj txhawb kev siv cov tswv yim kho mob tshwj xeeb (1, 2). Txog rau qhov kawg no, peb tau siv qhov kev tsis sib haum xeeb ntawm kev ua haujlwm rau AD hlwb thiab CSF txhawm rau txheeb xyuas qhov web-based CSF biomarkers uas muaj kev cuam tshuam ntau yam ntawm lub hlwb-raws li pathophysiology. Peb qhov kev soj ntsuam tau tsim tsib CSF biomarker panels, uas (i) cuam tshuam cov synapses, cov hlab ntsha, myelin, tiv thaiv kab mob thiab metabolic tsis ua haujlwm; (ii) ua kom pom muaj zog rov tsim dua tshiab ntawm ntau lub MS platforms; (iii) Qhia txog kev hloov pauv ntawm kab mob tshwj xeeb thoob plaws rau theem pib thiab lig ntawm AD. Zuag qhia tag nrho, cov kev tshawb pom no sawv cev ib kauj ruam tseem ceeb rau kev txhim kho ntau haiv neeg, txhim khu kev qha, web-oriented biomarker cov cuab yeej rau AD tshawb fawb thiab kev siv tshuaj kho mob.
Peb cov txiaj ntsig tau qhia txog lub koom haum tau txais txiaj ntsig zoo ntawm AD lub hlwb network proteome thiab txhawb nqa nws siv los ua cov thauj tog rau nkoj rau kev txhim kho biomarker. Peb qhov kev ntsuam xyuas qhia tau hais tias ob qho kev ywj pheej TMT-MS datasets uas muaj AD thiab AsymAD lub hlwb muaj zog modularity. Cov kev tshawb pom no txuas ntxiv peb txoj haujlwm yav dhau los, qhia txog kev khaws cia ntawm cov qauv muaj zog ntawm ntau dua 2,000 lub hlwb cov ntaub so ntswg los ntawm ntau pawg neeg ywj pheej nyob rau hauv pem hauv ntej, parietal, thiab sab cev nqaij daim tawv (17). Qhov kev pom zoo network no qhia txog ntau yam kev hloov pauv ntawm cov kab mob uas tau pom hauv kev tshawb fawb tam sim no, suav nrog kev nce ntawm glial-nplua nuj inflammatory modules thiab txo qis ntawm neuron-nplua nuj modules. Zoo li kev tshawb fawb tam sim no, lub network loj loj no tseem muaj cov kev hloov pauv tseem ceeb hauv AsymAD, qhia txog ntau yam sib txawv ntawm cov kab mob preclinical (17).
Txawm li cas los xij, nyob rau hauv cov txheej txheem kev saib xyuas zoo heev no, muaj ntau qhov zoo-grained biological heterogeneity, tshwj xeeb tshaj yog cov tib neeg nyob rau theem pib ntawm AD. Peb lub vaj huam sib luag biomarker tuaj yeem piav qhia ob pawg hauv AsymAD, uas ua rau pom qhov sib txawv tseem ceeb ntawm ntau CSF cov cim. Peb pab pawg muaj peev xwm hais txog qhov sib txawv ntawm kev lom neeg ntawm ob pawg sub, uas tsis pom tseeb ntawm qib ntawm AD biomarkers. Piv nrog rau pawg tswj hwm, Aβ1-42 / tag nrho tau qhov sib piv ntawm cov neeg AsymAD no txawv txav tsawg. Txawm li cas los xij, tsuas yog tag nrho cov qib tau sib txawv ntawm ob pawg AsymAD pawg, thaum qib Aβ1-42 thiab p-tau tseem sib piv. Txij li thaum siab CSF tau zoo li yog qhov kev kwv yees zoo dua ntawm kev paub txog cov tsos mob dua li Aβ1-42 qib (7), peb xav tias ob pawg AsymAD tuaj yeem muaj kev pheej hmoo sib txawv ntawm kev kis kab mob. Muab qhov piv txwv loj ntawm peb AsymAD thiab tsis muaj cov ntaub ntawv ntev, xav tau kev tshawb fawb ntxiv kom ntseeg tau cov lus xaus no. Txawm li cas los xij, cov txiaj ntsig no qhia tau hais tias qhov system-based CSF vaj huam sib luag tuaj yeem txhim kho peb lub peev xwm los txhim kho cov tib neeg zoo thaum lub sijhawm asymptomatic ntawm tus kab mob.
Zuag qhia tag nrho, peb qhov kev tshawb pom txhawb nqa lub luag haujlwm ntawm ntau yam kev ua haujlwm lom neeg hauv cov kab mob ntawm AD. Txawm li cas los xij, dysregulated lub zog metabolism tau dhau los ua lub ntsiab lus tseem ceeb ntawm tag nrho peb tsib daim ntawv pov thawj validated. Metabolic proteins, xws li hypoxanthine-guanine phosphoribosyltransferase 1 (HPRT1) thiab lactate dehydrogenase A (LDHA), yog cov feem ntau validated synaptic biomarkers, qhia tias qhov nce hauv AD CSF yog kev sib deev heev. Peb cov hlab ntsha thiab glial panels kuj muaj ob peb lub cim kev koom tes hauv cov metabolism ntawm cov tshuaj oxidative. Cov kev tshawb pom no tau ua raws li lub luag haujlwm tseem ceeb uas cov txheej txheem metabolic ua si hauv tag nrho lub hlwb, tsis tsuas yog ua kom tau raws li lub zog xav tau ntawm cov neurons, tab sis kuj ua kom tau raws li qhov xav tau ntawm lub zog siab ntawm astrocytes thiab lwm cov hlwb glial (17, 48). Peb cov txiaj ntsig tau txhawb nqa cov pov thawj loj hlob uas hloov pauv hauv redox muaj peev xwm thiab cuam tshuam ntawm lub zog txoj hauv kev tuaj yeem yog qhov tseem ceeb ntawm kev sib txuas ntawm ntau cov txheej txheem tseem ceeb koom nrog hauv pathogenesis ntawm AD, suav nrog cov kab mob mitochondrial, glial-mediated o, thiab vascular puas (49). Tsis tas li ntawd, cov metabolic cerebrospinal kua biomarkers muaj cov protein ntau sib txawv ntawm peb cov kev tswj hwm thiab AD-zoo li AsymAD subgroups, qhia tias kev cuam tshuam ntawm lub zog thiab redox txoj hauv kev yuav yog ib qho tseem ceeb nyob rau hauv preclinical theem ntawm tus kab mob.
Qhov sib txawv ntawm lub hlwb thiab cerebrospinal kua vaj huam sib luag tiam sis peb tau pom kuj muaj qhov cuam tshuam txog kev lom neeg lom zem. Synapses thiab metabolomes nplua nuj nyob rau hauv neurons qhia txo qis hauv AD lub hlwb thiab nce ntau hauv cov kua cerebrospinal. Muab hais tias neurons yog nplua nuj nyob rau hauv lub zog-tsim mitochondria ntawm synapses los muab lub zog rau lawv cov cim tshwj xeeb heev heev (50), qhov zoo sib xws ntawm kev qhia profiles ntawm ob pawg neuron yuav tsum. Kev poob ntawm cov neurons thiab extrusion ntawm cov hlwb puas tuaj yeem piav qhia cov hlwb thiab CSF vaj huam sib luag hauv cov kab mob tom qab, tab sis lawv tsis tuaj yeem piav qhia txog kev hloov pauv ntawm lub vaj huam sib luag thaum ntxov peb saib (13). Ib qho kev piav qhia ua tau rau cov kev tshawb pom no nyob rau hauv cov kab mob asymptomatic thaum ntxov yog qhov txawv txav synaptic pruning. Cov ntaub ntawv pov thawj tshiab hauv cov qauv nas qhia tias microglia-mediated synaptic phagocytosis tuaj yeem ua haujlwm txawv txav hauv AD thiab ua rau lub hlwb poob qis (51). Cov khoom siv hluavtaws pov tseg no tuaj yeem khaws cia hauv CSF, uas yog vim li cas peb thiaj li pom qhov nce hauv CSF hauv lub vaj huam sib luag neuron. Immune-mediated synaptic pruning kuj tseem tuaj yeem piav qhia txog qhov nce ntawm glial proteins uas peb saib hauv lub hlwb thiab cov kua cerebrospinal thoob plaws hauv cov txheej txheem kab mob. Ntxiv nrog rau synaptic pruning, tag nrho cov kev txawv txav ntawm txoj kev exocytic kuj tseem tuaj yeem ua rau lub hlwb sib txawv thiab CSF kab lus ntawm cov cim neuronal. Ntau cov kev tshawb fawb tau pom tias cov ntsiab lus ntawm exosomes hauv pathogenesis ntawm AD hlwb tau hloov pauv (52). Txoj kev extracellular kuj koom nrog hauv kev loj hlob ntawm Aβ (53, 54). Nws yog tsim nyog sau cia tias kev tawm tsam ntawm exosomal secretion yuav txo tau AD-zoo li pathology hauv AD transgenic nas qauv (55).
Nyob rau tib lub sijhawm, cov protein nyob rau hauv lub vascular vaj huam sib luag tau pom muaj qhov nce nruab nrab hauv AD lub hlwb, tab sis txo qis hauv CSF. Ntshav-brain barrier (BBB) tsis ua haujlwm tuaj yeem piav qhia cov kev tshawb pom no. Ntau qhov kev tshawb fawb ywj pheej ntawm tib neeg tau ua pov thawj BBB kev puas tsuaj hauv AD (56, 57). Cov kev tshawb fawb no tau lees paub ntau yam kev txawv txav ntawm qhov kev sib tw endothelial, suav nrog cov tshuaj tiv thaiv hlwb ntawm cov ntshav ntawm lub hlwb (57). Qhov no tuaj yeem muab cov lus piav qhia yooj yim rau kev nce siab vascular proteins hauv lub hlwb, tab sis nws tsis tuaj yeem piav qhia tag nrho cov txiaj ntsig ntawm cov protein no hauv cov kua cerebrospinal. Ib qho ua tau yog tias lub hauv nruab nrab lub paj hlwb yog nquag cais cov molecules los daws cov teeb meem ntawm kev mob o thiab oxidative kev nyuaj siab. Qhov txo qis hauv qee qhov CSF cov protein ntau tshaj plaws hauv lub vaj huam sib luag no, tshwj xeeb tshaj yog cov neeg koom nrog hauv kev tswj hwm lipoprotein, muaj feem cuam tshuam nrog kev txwv tsis pub muaj kev phom sij ntawm qhov mob thiab cov txheej txheem neuroprotective ntawm cov pa oxygen reactive. Qhov no muaj tseeb rau Paroxonase 1 (PON1), lipoprotein binding enzyme lub luag haujlwm rau txo qis oxidative kev ntxhov siab hauv kev ncig (58, 59). Alpha-1-microglobulin/bikunin precursor (AMBP) yog ib qho tseem ceeb txo qis-tswj cim ntawm pawg vascular. Nws yog qhov ua ntej ntawm lipid transporter bikunin, uas tseem koom nrog hauv kev tiv thaiv kab mob thiab kev tiv thaiv paj hlwb (60, 61).
Txawm hais tias muaj ntau qhov kev xav zoo siab, qhov tsis muaj peev xwm txheeb xyuas cov kab mob biochemical ncaj qha yog qhov paub zoo txog kev txwv ntawm kev tshawb pom-tsav proteomics tsom xam. Yog li ntawd, kev tshawb fawb ntxiv yog tsim nyog los ntseeg siab txhais cov txheej txheem hauv qab no biomarker panels. Txhawm rau txav mus rau kev txhim kho ntawm MS-raws li kev soj ntsuam kev soj ntsuam, cov kev taw qhia yav tom ntej tseem yuav tsum tau siv cov txheej txheem tsom xam ntau rau kev txheeb xyuas qhov loj ntawm biomarker, xws li xaiv los yog sib piv cov tshuaj tiv thaiv (62). Peb tsis ntev los no tau siv cov tshuaj tiv thaiv kab mob sib luag (63) kom siv tau ntau ntawm CSF protein hloov tau piav qhia ntawm no. Ob peb lub hom phiaj tseem ceeb ntawm lub vaj huam sib luag tau suav nrog qhov tseeb qhov tseeb, suav nrog YWHAZ, ALDOA, thiab SMOC1, uas qhia txog peb cov synapse, metabolism, thiab cov kab mob o, raws li (63). Independent Data Acquisition (DIA) thiab lwm cov tswv yim raws li MS kuj tseem yuav pab tau rau kev txheeb xyuas lub hom phiaj. Bud et al. (64) Nws tsis ntev los no tau pom tias muaj qhov sib tshooj tseem ceeb ntawm AD biomarkers tau txheeb xyuas hauv peb cov ntaub ntawv tshawb pom CSF thiab cov ntaub ntawv ywj pheej DIA-MS, uas muaj ze li ntawm 200 CSF cov qauv los ntawm peb pawg European sib txawv. Cov kev tshawb fawb tsis ntev los no txhawb nqa lub peev xwm ntawm peb cov vaj huam sib luag los hloov mus rau kev txhim kho MS-raws li kev tshawb nrhiav. Ib txwm siv tshuaj tiv thaiv kab mob thiab kev tshawb nrhiav aptamer kuj tseem ceeb rau kev txhim kho ntxiv ntawm AD biomarkers tseem ceeb. Vim tias tsis tshua muaj ntau ntawm CSF, nws nyuaj dua los kuaj xyuas cov biomarkers siv cov txheej txheem MS siab. NEFL thiab NRGN yog ob qho piv txwv ntawm cov khoom siv qis qis CSF biomarkers, uas tau kos npe rau hauv lub vaj huam sib luag hauv peb cov kev txheeb xyuas dav dav, tab sis tsis tuaj yeem tshawb pom kev ntseeg siab siv peb lub tswv yim MS nkaus xwb. Kev tsom xam cov tswv yim raws li ntau yam tshuaj tiv thaiv kab mob, xws li PEA, tuaj yeem txhawb kev hloov kho ntawm cov cim no.
Zuag qhia tag nrho, txoj kev tshawb no muab ib qho tshwj xeeb proteomics mus kom ze rau kev txheeb xyuas thiab txheeb xyuas ntawm CSF AD biomarkers raws li cov kab ke sib txawv. Kev ua kom zoo dua cov cim npe no hla cov pawg AD ntxiv thiab MS platforms tuaj yeem ua pov thawj cog lus tias yuav ua rau AD pheej hmoo stratification thiab kho. Cov kev tshawb fawb uas ntsuas qhov ntev ntawm cov panels nyob rau lub sij hawm kuj tseem yog ib qho tseem ceeb los txiav txim seb qhov sib xyaw ua ke ntawm cov cim twg zoo tshaj plaws stratifies qhov kev pheej hmoo ntawm cov kab mob ntxov thiab kev hloov ntawm cov kab mob hnyav.
Tsuas yog 3 cov qauv luam tawm los ntawm CSF, tag nrho cov qauv CSF siv hauv txoj kev tshawb fawb no tau sau los ntawm kev txhawb nqa ntawm Emory ADRC lossis cov koom haum tshawb fawb zoo sib xws. Tag nrho ntawm plaub pawg ntawm Emory CSF cov qauv tau siv hauv cov kev tshawb fawb proteomics. CSF pawg tau pom tias muaj cov qauv los ntawm 20 kev tswj hwm kev noj qab haus huv thiab 20 tus neeg mob AD. CSF daim ntawv luam 1 suav nrog cov qauv ntawm 32 kev tswj hwm kev noj qab haus huv, 31 AsymAD cov tib neeg, thiab 33 tus neeg AD. CSF daim ntawv luam 2 muaj 147 tswj thiab 150 AD qauv. Cov kab mob ntau yam CSF replication 4 cohort suav nrog 18 tswj, 17 AD, 19 ALS, 13 PD, thiab 11 FTD cov qauv. Raws li kev pom zoo pom zoo los ntawm Emory University Institutional Review Board, txhua tus neeg koom nrog Emory tau txais kev pom zoo. Raws li 2014 National Institute of Aging Practice Guidelines for Alzheimer's Centers (https://alz.washington.edu/BiospecimenTaskForce.html), cerebrospinal kua tau sau thiab khaws cia los ntawm lumbar puncture. Kev tswj hwm thiab AsymAD thiab AD cov neeg mob tau txais kev ntsuas kev txawj ntse ntawm Emory Cognitive Neurology Clinic lossis Goizueta ADRC. Lawv cov cerebrospinal kua kuaj tau kuaj los ntawm INNO-BIA AlzBio3 Luminex rau ELISA Aβ1-42, tag nrho tau thiab p-tau tsom xam (65). ELISA qhov tseem ceeb yog siv los txhawb kev kuaj mob ntawm cov ncauj lus raws li tsim los ntawm AD biomarker txiav tawm cov txheej txheem (66, 67). Cov ntaub ntawv tseem ceeb thiab kev kuaj mob rau lwm yam kev kuaj mob CSF (FTD, ALS, thiab PD) kuj tau txais los ntawm Emory ADRC lossis cov koom haum tshawb fawb koom nrog. Cov ntaub ntawv ntsiab lus metadata rau cov xwm txheej Emory CSF tuaj yeem pom hauv Table S1A. Cov yam ntxwv ntawm Swiss CSF replication 3 cohort tau luam tawm yav dhau los (45).
CSF pom tus qauv. Txhawm rau nce qhov tob ntawm peb qhov kev tshawb pom ntawm CSF cov ntaub ntawv teeb tsa, kev tiv thaiv kev noj qab haus huv ntawm cov protein ntau ntau tau ua ua ntej trypsinization. Hauv luv luv, 130 μl ntawm CSF los ntawm 40 tus neeg CSF cov qauv thiab qhov sib npaug (130 μl) ntawm High Select Top14 Abundance Protein Depletion Resin (Thermo Fisher Scientific, A36372) tau muab tso rau hauv kab rov tav (Thermo Fisher Scientific, A89868) ntawm chav. kub Incubate). Tom qab spinning rau 15 feeb, centrifuge tus qauv ntawm 1000g rau 2 feeb. Lub 3K ultracentrifugal lim ntaus ntawv (Millipore, UFC500396) tau siv los mloog cov qauv effluent los ntawm centrifuging ntawm 14,000g rau 30 feeb. Dilute tag nrho cov qauv ntim rau 75 μl nrog phosphate buffered saline. Cov protein concentration tau soj ntsuam los ntawm txoj kev bicinchoninic acid (BCA) raws li cov chaw tsim khoom raws tu qauv (Thermo Fisher Scientific). Lub immunodepleted CSF (60 μl) los ntawm tag nrho 40 cov qauv tau zom nrog lysyl endopeptidase (LysC) thiab trypsin. Hauv luv luv, cov qauv raug txo thiab alkylated nrog 1.2 μl 0.5 M tris-2 (-carboxyethyl)-phosphine thiab 3 μl 0.8 M chloroacetamide ntawm 90 ° C rau 10 feeb, thiab tom qab ntawd sonicated hauv da dej rau 15 feeb. Cov qauv no tau diluted nrog 193 μl 8 M urea buffer [8 M urea thiab 100 mM NaHPO4 (pH 8.5)] mus rau qhov kawg ntawm 6 M urea. LysC (4.5 μg; Wako) yog siv rau kev zom zaub mov thaum hmo ntuj ntawm chav tsev kub. Tom qab ntawd cov qauv yog diluted rau 1 M urea nrog 50 mM ammonium bicarbonate (ABC) (68). Ntxiv qhov sib npaug ntawm qhov sib npaug (4.5 μg) ntawm trypsin (Promega), thiab tom qab ntawd incubate tus qauv rau 12 teev. Acidify lub digested peptide tov rau qhov kawg concentration ntawm 1% formic acid (FA) thiab 0.1% trifluoroacetic acid (TFA) (66), thiab ces desalt nrog ib tug 50 mg Sep-Pak C18 kem (Dej) raws li tau piav saum toj no (25) . Tom qab ntawd cov peptide tau eluted hauv 1 ml ntawm 50% acetonitrile (ACN). Txhawm rau kom muaj nuj nqis protein ntau thoob plaws batch (25), 100 μl aliquots los ntawm tag nrho 40 CSF cov qauv tau muab tso ua ke los tsim cov qauv sib xyaw, uas tau muab faib ua tsib tus qauv thoob ntiaj teb sab hauv (GIS) (48) cov qauv. Txhua tus qauv thiab cov qauv sib xyaw ua ke yog qhuav los ntawm lub tshuab nqus tsev siab ceev (Labconco).
CSF luam cov qauv. Dayon thiab cov npoj yaig yav dhau los tau piav qhia txog kev tiv thaiv kev tiv thaiv kab mob thiab kev zom zaub mov ntawm CSF luam 3 cov qauv (45, 46). Cov qauv rov ua dua uas tseem tshuav tsis tau txhaj tshuaj tiv thaiv tus kheej. Digest cov qauv uas tsis tau tshem tawm hauv trypsin raws li tau piav qhia dhau los (17). Rau txhua qhov kev soj ntsuam rov qab, 120 μl aliquots ntawm eluted peptide los ntawm txhua tus qauv tau muab sib xyaw ua ke thiab muab faib ua qhov sib npaug ntawm cov ntim aliquots los siv raws li TMT-labeled thoob ntiaj teb sab hauv tus qauv (48). Txhua tus qauv thiab cov qauv sib xyaw ua ke yog qhuav los ntawm lub tshuab nqus tsev siab ceev (Labconco). Txhawm rau txhim kho cov teeb liab ntawm cov protein ntau CSF tsawg, los ntawm kev sib txuas ntawm 125 μl los ntawm txhua tus qauv, ib qho "hloov" qauv tau npaj rau txhua qhov kev soj ntsuam rov ua dua [piv txwv li, cov qauv lom neeg uas ua raws li cov qauv kev tshawb fawb, tab sis tus nqi muaj nyob. loj dua (37, 69)] muab tso rau hauv ib qho kev sib xyaw CSF qauv (17). Cov qauv sib xyaw yog tom qab ntawd tshem tawm cov tshuaj tiv thaiv kab mob siv 12 ml ntawm High Select Top14 Abundance Protein Removal Resin (Thermo Fisher Scientific, A36372), zom raws li tau piav qhia saum toj no, thiab suav nrog ntau daim ntawv TMT tom ntej.
Post lub sij hawm: Aug-27-2021