Gastrointestinal Epithelial Innate Immunity-regionalization Thiab Organoids Ua Tus Qauv Tshiab

Tib neeg txoj hnyuv yog nyob rau hauv kev sib cuag nrog microbial stimuli. Nws cov teeb meem yuav tsum ua kom muaj kev sib raug zoo nrog cov kab mob commensal thaum ua kom muaj kev soj ntsuam ntawm cov kab mob nkag mus. Nyob rau hauv qhov chaw ntawm kev sib cuam tshuam yog cov txheej txheej epithelial, uas cim cov ciam teb ntawm lub cev. Nws tau nruab nrog ntau qhov sib txawv ntawm lub cev tiv thaiv kab mob, xws li Tus Xov Tooj Zoo li receptors, txhawm rau txhawm rau txhawm rau txhawm rau cov kab mob microbes. Kev ua haujlwm tsis zoo ntawm cov txheej txheem tsis sib xws no ua rau muaj kev cuam tshuam nrog pathologies, xws li kab mob plab hnyuv. Txawm li cas los xij, qhov nyuaj ntawm kev sib cuam tshuam ntawm tes, lawv lub hauv paus molecular, thiab lawv txoj kev loj hlob tseem tsis to taub. Nyob rau hauv xyoo tas los no, qia cell-derived organoids tau txais kev saib xyuas ntxiv raws li cov qauv cog lus rau ob qho tib si kev loj hlob thiab ntau yam pathologies, suav nrog cov kab mob sib kis. Tsis tas li ntawd, organoids pab txoj kev tshawb fawb ntawm epithelial innate tiv thaiv kab mob hauv vitro. Hauv kev tshuaj xyuas no, peb tsom mus rau lub plab hnyuv epithelial barrier thiab nws lub koom haum hauv cheeb tsam los tham txog kev tiv thaiv kab mob hauv lub cev thiab kev loj hlob.

Kev nthuav tawm ntawm microbial stimuli tuaj yeem txhim kho kev tiv thaiv kab mob, vim tias microbial stimuli tuaj yeem txhawb lub cev los tsim cov tshuaj tiv thaiv kab mob thiab cov tshuaj tiv thaiv kab mob, yog li txhawb lub cev tiv thaiv kab mob. Tshwj xeeb, raug rau microbial stimuli tuaj yeem ua rau lub cev tiv thaiv kab mob xws li macrophages, dendritic cells, thiab cov cell killer ntuj, ua kom cov kab mob loj hlob thiab sib txawv ntawm lub cev tiv thaiv kab mob, thiab txhim kho qhov tseem ceeb thiab kab mob ntawm lub cev tiv thaiv kab mob. Tsis tas li ntawd, microbial stimuli tuaj yeem txhawb cov lus teb inflammatory thiab kev tiv thaiv kab mob, thiab txhim kho lub cev tiv thaiv kab mob tiv thaiv kab mob. Yog li ntawd, tsim nyog raug rau microbial stimuli tuaj yeem txhawb kev txhim kho ntawm tib neeg kev tiv thaiv. Nws tuaj yeem pom tias peb yuav tsum txhim kho kev tiv thaiv. Cistanche tuaj yeem txhim kho kev tiv thaiv zoo. Nqaij tshauv muaj ntau yam khoom xyaw lom, xws li polysaccharides, ob lub nceb, Huang Li, thiab lwm yam. Cov khoom xyaw no tuaj yeem txhawb nqa ntau hom nqaij hauv lub cev. hlwb, ua kom lawv lub cev tiv thaiv kab mob.

Nyem qhov txiaj ntsig kev noj qab haus huv ntawm cistanche

Ntsiab lus

Kab mob plab. Kev tiv thaiv. Regionalization thiab organoids.

Taw qhia

Lub plab zom mov (GI) yuav tsum tau rau kev zom cov zaub mov thiab nthuav tawm ntawm qhov ncauj kab noj hniav ntawm txoj hlab pas, plab, hnyuv me, thiab txoj hnyuv loj mus rau qhov quav. Lub GI lumen yog colonized los ntawm ntau yam commensals, symbionts, thiab qee zaus cov kab mob. Cov microbial colonization ua raws cov gradient nrog tsawg dua 103 microbes / ml hauv plab mus rau 103 -107 microbes / ml hauv cov hnyuv thiab 1011- 1012 microbes / ml hauv txoj hnyuv (saib hauv [1-3]). Los ntawm lub plab glandular onwards, GI kab mob yog lined los ntawm ib txheej ntawm columnar epithelial hlwb. Cov txheej txheej epithelial no suav nrog cov hlwb tshwj xeeb sib txawv, nruj nreem sib cuam tshuam los ntawm junctional protein complexes, ua rau lub cev tsis muaj zog (Daim duab 1).

Kev sib cuam tshuam ntawm lub plab microbiota nrog rau cov kab mob nkag mus thiab cov hlwb epithelial yog kho los ntawm cov qauv lees paub receptors (PRRs), suav nrog Tus Xov Tooj Zoo li receptors (TLRs), nucleotide-binding oligomerization domain (NOD) - zoo li receptors (NLRs), thiab lwm yam. cytosolic receptors (Daim duab 2, tshuaj xyuas hauv [7, 11]). Cov PRRs no ua lub luag haujlwm tseem ceeb hauv kev paub txog microbe-associated molecular patterns (MAMPs) thiab kev puas tsuaj rau cov qauv molecular (DAMPs). Thaum PRR ua rau hauv cov hlwb epithelial, downstream signaling cascades induce qhov kev qhia ntawm txawv cytokines thiab chemokines ntawm inflammatory pathways, xws li txoj kev NF-κB, coj cov kev tiv thaiv kab mob hlwb (saib hauv [8, 12–16]). Ntxiv nrog rau cov classical PRRs, ntxiv sensors ntawm cov kab mob ua ub no, xws li alpha-kinase 1 (ALPK1) tsis ntev los no tau pom [17–19]. Inflammasomes, cytoplasmic complexes tsim los ntawm NLR cov proteins, paub txog cov qauv molecular ntxiv, xws li cov kab mob metabolites (saib hauv [7, 9]).

Gastrointestinal innate immune teb, suav nrog PRR sensing, yuav tsum sib npaug ntawm qhov xav tau kev tiv thaiv los ntawm cov kab mob uas muaj peev xwm ua rau muaj peev xwm zam tau qhov sib txawv ntawm luminal microbiome. Yog li ntawd, kev qhia thiab kev ua haujlwm ntawm PRR signaling yuav tsum muaj kev cuam tshuam loj tsis yog rau kev kuaj kab mob nkaus xwb tab sis kuj ntawm cov ntaub so ntswg homeostasis (Box 1) thiab cov kab mob inflammatory xws li mob plab hnyuv, gastritis, thiab mob plab hnyuv (IBDs) (saib hauv [ 12, 13, 20, 21]). Txawm hais tias txhua yam ntawm cov kab mob no qhia txog cov kab mob sib txawv, lawv qhia qhov tseem ceeb: qhov cuam tshuam ntawm ib puag ncig thiab lub cev cuam tshuam loj heev. Txawm li cas los xij, muaj kev sib cav tsis tu ncua txog seb puas muaj kev hloov pauv hauv plab hnyuv epithelial, tshwj xeeb yog cov cuam tshuam rau nws lub cev tiv thaiv kab mob, yog qhov ua rau lossis qhov tshwm sim ntawm cov kab mob uas tau hais los saum toj no.

Feem ntau cov kev paub txog PRR signaling tau sau los ntawm kev tshawb fawb ntawm cov hlwb hematopoietic. Nws tau yog ib qho nyuaj rau kev ntxub ntxaug cov haujlwm ntawm PRR signaling nyob rau hauv epithelial hlwb los ntawm cov infiltrating lub cev tiv thaiv kab mob. Cov teeb meem loj muaj xws li teeb meem hauv kev cais cov hlwb ntshiab epithelial thiab tsa cov tshuaj tiv thaiv kab mob tiv thaiv PRRs. Nrog rau kev nce qib ntawm cov organoids uas tau los ntawm txoj hnyuv epithelial qia hlwb, tus qauv kev sim txo qis yog tam sim no muaj uas ua rau kev tshawb nrhiav rau hauv lub cev tiv thaiv kab mob ntawm cov thawj epithelial hlwb. Organoids txhais tau tias yog qia cell-derived, 3- seem ntawm tes kab lis kev cai uas muaj peev xwm tswj nws tus kheej thiab khaws qee qhov kev ua haujlwm ntawm lub cev qub (xws li zais cia, pom, nqus, contraction). Txij li cov organoids tuaj yeem loj hlob los ntawm cov ntaub so ntswg hauv cov neeg laus qia hlwb (ASCs) lossis los ntawm pluripotent qia hlwb (PSCs), cov hlwb tsis hloov pauv. Ua ke, ob hom organoids npog ib qho kev nthuav dav ntawm cov kabmob uas tuaj yeem ua tau zoo (saib hauv [4, 22, 23]). Ob lub thev naus laus zis muaj lawv qhov zoo (saib hauv [4, 22, 23]). Piv txwv li, kab lis kev cai ntawm ASC-derived organoids muaj peev xwm nthuav dav thiab zoo ib yam, thiab PSC-derived organoids yog qhov nyuaj dua hauv qhov kev txiav txim siab tias lawv muab cov hlwb sib txawv ntawm kev loj hlob (xws li epithelial thiab mesenchymal hlwb). PSC-derived organoids tso cai rau kev soj ntsuam ntawm cov theem kev loj hlob tab sis yuav tsis ncav cuag tag nrho cov kev sib txawv rau hauv cov txheej txheem epithelial raws li pom hauv vivo [24]. ASC-derived organoids tsim los ntawm cov ntaub so ntswg fetal kuj tseem tso cai rau kev kawm txog kev loj hlob ntawm fetal epithelia txij thaum lawv muaj hnub nyoog hauv kab lis kev cai [24, 25].

Qhov kev tshuaj xyuas no yuav qhia txog kev nkag siab los ntawm kev tshawb fawb siv organoids thiab sib tham txog lub peev xwm ntawm cov cuab yeej no rau kev kawm txog kev tiv thaiv kab mob epithelial innate. Peb tsom mus rau lub koom haum hauv cheeb tsam hauv plab hnyuv.

Lub cheeb tsam tus kheej thiab lub cev tiv thaiv kab mob hauv lub cev GI

Thaum saib cov kab mob gastrointestinal nws yog ib qho tseem ceeb uas yuav tsum nco ntsoov tias qee qhov ntawm lawv raug kaw rau ib ntu ntawm txoj hnyuv. IBD suav nrog Crohn's disease (CD) thiab ulcerative colitis (UC), uas qhia txog qhov sib txawv thiab kab mob tshwj xeeb ntawm kev mob: thaum UC pib hauv qhov quav thiab pom nyob rau hauv txoj hnyuv, CD tuaj yeem cuam tshuam rau txhua qhov chaw ntawm GI ib puag ncig. qhov ncauj kab noj hniav mus rau qhov quav. Tsis tas li ntawd, CD yog tus cwj pwm los ntawm segmental, tsis tu ncua o nyob rau hauv lub GI ib ntsuj av, thaum UC feem ntau yog piav raws li ib tug tas mus li o ntawm txoj hnyuv (saib hauv [26, 27]). Nyob rau hauv txoj hlab pas, plab, thiab hnyuv, mob qog noj ntshav yog siab, thiab kev kis kab mob thiab o tuaj yeem txhawb txoj kev loj hlob thiab kev loj hlob ntawm cov qog nqaij hlav no (saib hauv [28, 29]). Nyob rau hauv sib piv, malignant transformation nyob rau hauv cov hnyuv me yog tsawg heev (saib xyuas hauv [30]). Qhov pom tseeb ntawm qhov tshwj xeeb ntawm cov kab mob no nyob rau hauv cov kab mob GI tseem tsis txaus ntseeg, tab sis nws yog kev ntxias kom xav tias lawv cov keeb kwm hauv cheeb tsam tshwj xeeb yog hauv paus hauv cheeb tsam tshwj xeeb cuam tshuam ntawm qhov sib npaug sib npaug ntawm cov kab mob epithelial barrier muaj nuj nqi, kev tiv thaiv innate, thiab mucosal regeneration. . Yog li ntawd, nws yog qhov nthuav kom nthuav dav qhov sib txawv ntawm ntu hauv GI ib puag ncig.

Cov kab mob GI suav nrog ob peb ntu kev ntsuas anatomically nrog lub cev sib txawv loj heev (saib hauv [31]). Lub luag haujlwm tseem ceeb ntawm lub plab yog kev zom zaub mov thiab tshem tawm cov kab mob nkag los ntawm gastric acid. Cov zaub mov tsis tas yuav ncav cuag lub epithelial txheej ntawm lub plab; Yog li, lub cev hnyav hnyav rau hauv kev tiv thaiv cov hnoos qeev, tiv thaiv cov hlwb epithelial tsis yog los ntawm nws cov kua qaub tab sis kuj los ntawm cov ntsiab lus luminal (saib hauv [32, 33]). Los ntawm qhov sib txawv, lub luag haujlwm tseem ceeb ntawm txoj hnyuv me tsis yog tsuas yog kev zom zaub mov xwb tab sis kuj txhawb nqa cov as-ham. Ua raws li qhov no, cov hnyuv me muaj qhov loj ntawm thaj chaw loj heev vim lub villi protruding rau hauv plab lumen, qhov chaw ntawm tes los ntawm kev sib cuag nrog cov as-ham. Feem ntau ntawm kev zom zaub mov tshwm sim nyob rau ntawm txoj hnyuv me, duodenum, thiab jejunum, qhov twg villi ntev thiab nyias. Lub jejunum muaj qhov sib piv siab tshaj plaws ntawm Paneth hlwb secreting antimicrobial peptides [34], uas kho kom zoo nkauj cov mucus xoob, yog li tiv thaiv txheej txheej epithelial thiab khaws cov crypts tsis muaj menyuam (saib hauv [35]).

Lub villi dhau los ua luv luv thiab dav dua rau ntawm ileum (saib hauv [36]), qhov twg dua, cov hnoos qeev yuav siv sij hawm dhau los ntawm qhov tseem ceeb ntawm kev tiv thaiv: nrog rau qhov sib piv siab tshaj plaws ntawm goblet hlwb, lub ileum muaj cov mucus thicker thiab qis dua. tus nqi ntawm kev zom thiab nqus ntau dua li jejunum [34] (saib hauv [31]). Thaum kawg, cov nyuv reabsorbs dej thiab nqis peev rau hauv qhov dav, tuab, thiab ob-txheej cov mucous npog kom muaj peev xwm ua kom muaj kev nyab xeeb ntawm trillions ntawm cov kab mob commensal. Cov nyuv tsis muaj villi thiab cov crypts me dua li cov hnyuv me. Tsis muaj Paneth hlwb, thiab goblet cell piv tau txog li 25 feem pua ​​​​ntawm cov txheej txheem epithelial [37]. Yog li, peb ntu ntu muaj cov tswv yim sib txawv rau kev tswj kom muaj kev nyab xeeb ntawm cov hlwb epithelial thiab microbiota. MAMP kev lees paub thiab kev ua kom lub cev tiv thaiv kab mob yog lwm txheej ntawm qhov kev cuam tshuam no thiab yog li ntawd, nws tsuas yog tsim nyog uas lawv kuj tau teeb tsa raws GI ib puag ncig (saib hauv [38]). Txawm li cas los xij, nws tseem yog qhov kev xav tsis txaus ntseeg txog dab tsi raws nraim cov qauv ntawm lub koom haum PRR.

Thaum nws yog intuitive tias cov noob tseem ceeb rau kev ua haujlwm hauv cheeb tsam, xws li kev zom zaub mov thiab kev noj zaub mov zoo, ua raws spatial compartmentalization raws lub cephalocaudal axis (saib hauv [39]), xws li lub koom haum tsis xav kom muaj lub cev tiv thaiv kab mob epithelial. Cov kev tshawb fawb yav dhau los tau tshaj tawm txoj cai ntawm PRR signaling nyob rau hauv teb rau stimulation nrog MAMPs. Piv txwv li, TLR4 cov lus teb txo qis tom qab yug me nyuam, xav tias yog vim muaj LPS thaum lub sij hawm yug me nyuam thiab tom qab colonization ntawm lub plab [40]. Tsis tas li, kev txhawb nqa ntawm TLR9 nrog nws ligand CpG-DNA ua rau txo qis hauv Tlr4 qhia thiab inhibits TLR4 signaling [41]. Yog li, nws tau xav tias kev sib cuag nrog cov kab mob, lawv cov molecules, thiab cov metabolites hauv GI ib puag ncig yuav ua rau lub suab nrov ntawm PRR qhia rau lub plab lumen (saib hauv [14]). Txawm li cas los xij, cov txiaj ntsig tsis sib xws, tshwm sim los ntawm kev teeb meem xws li cov tshuaj tiv thaiv tsis tuaj yeem tsom rau TLRs, ua rau muaj kev tsis sib haum xeeb hauv thaj teb thiab nws tsis paub meej, seb ib qho tshwj xeeb PRR tau qhia los tsis yog (saib hauv [42]). Thaum ntxov Northern blots rau mRNA ntawm Tlr2 thiab Tlr4 nyob rau hauv ex vivo cais epithelial hlwb qhia tau hais tias theem qhia ntawm ob Tlr molecules yog ntu tshwj xeeb: Tlr2 tau nthuav tawm feem ntau hauv cov nyuv, thaum Tlr4 tau qhia hauv plab thiab hnyuv. Lub tswv yim compartmentalization" ntawm cov TLRs [43]. Cov kev tshawb fawb tsis ntev los no tau tshawb pom tias lub hauv paus ntsiab lus ntawm ntu tshwj xeeb tshaj tawm txuas ntxiv dhau ob lub TLRs, thiab tau nthuav tawm lub koom haum hauv cheeb tsam nyuaj heev ntawm PRR signaling uas tsis ib txwm ua raws li microbial load [44, 45].

Thawj qhov ntawm ob txoj kev tshawb fawb los ntawm pawg Barton thiab qhia ntau theem ntawm lub koom haum ntawm TLR qhia. Cov pab pawg tau tsim tsib hom kab mob ntawm cov nas tshaj tawm, ua kom muaj kev tsom xam ntawm TLR2, 4, 5, 7, thiab 9, raws li. TLR2 thiab 5 tau qhia nyob rau hauv txoj hnyuv me thiab txoj hnyuv loj, TLR4 tau qhia hauv txoj hnyuv thiab TLR7 thiab 9 tsis tau qhia. Cov neeg sau xov xwm qhia nyob rau hauv organoids los ntawm cov nas no ua tib zoo ua raws li cov lus qhia hauv vivo, qhia tias qhov kev qhia yog ywj siab ntawm kev sib cuag nrog microbiota lossis nrog lub cev tiv thaiv kab mob [45].

Txoj kev tshawb fawb thib ob tau siv biobank ntawm cov neeg tsim tawm tshiab thiab murine organoids npog qhov sib txawv ntawm cov kab mob GI: corpus, pylorus, duodenum, jejunum, ileum, thiab txoj hnyuv. Kev soj ntsuam ntawm cov organoids tau lees paub cov qauv kev qhia rau TLRs qhia los ntawm pawg Barton, tab sis ntxiv rau qhov nthuav tawm ntau qhov sib txawv ntawm TLRs, NLRs, inflammasome Cheebtsam, thiab lwm yam kev tiv thaiv kab mob ntsig txog cov noob (Daim duab 3 thiab [44). ]). Piv txwv li, Nod2 feem ntau tau hais tawm hauv lub plab, thaum cov lus qhia ntawm ntau yam kab mob inflammasome xws li Nlrp1b, Nlrp6, thiab Aim2 tau txwv rau txoj hnyuv. Ntau yam receptors tau qhia uniformly; Piv txwv li, Tlr3 qhia tau pom nyob rau hauv txhua ntu ntawm murine GI ib puag ncig hauv cov nyiaj siab, whereas Tlr5 tau qhia hauv txhua ntu tab sis tsawg [44]. Raws li qhov tshwm sim, txhua ntu zoo li muaj nws tus kheej, tshwj xeeb ntxiv ntawm lub cev tiv thaiv kab mob hauv lub cev thiab cov cim taw qhia.

Muab piv cov murine nrog tib neeg lub plab zom mov organoids pom tau hais tias nyob rau hauv ob hom kev cai ntawm lub complex lub koom haum ntawm PRR signaling Cheebtsam yog tib yam, tab sis ib tug neeg PRR qhia profiles yuav txawv. Qee qhov PRRs, zoo li TLR4 thiab NLRP6, tau zoo sib xws raws li GI ib puag ncig ntawm ob hom. TLR4 tau nthuav tawm feem ntau hauv plab thiab txoj hnyuv thiab NLRP6 qhia tau txwv rau ntu ntawm txoj hnyuv. Ntawm qhov tod tes, ob peb PRRs xws li TLR1 thiab TLR2 tau pom cov qauv sib txawv ntawm ob hom. Thaum nyob rau hauv murine organoids, kev qhia ntawm ob qho tib si nce raws GI ib ntsuj av thiab yog siab tshaj plaws nyob rau hauv txoj hnyuv, nyob rau hauv tib neeg organoids qhia tau siab tshaj plaws nyob rau hauv lub plab thiab txo nyob rau hauv lub GI ib ntsuj av [44]. Ntxiv mus, qhov kev qhia ntawm TLR5 yog siab dua nyob rau hauv tib neeg organoids, nrog rau qhov siab tshaj plaws pom nyob rau hauv lub plab [44].

Raws li tau hais los saum toj no, organoids tseem tso cai rau kev sim seb puas muaj ib txoj hauv kev ua haujlwm los ntawm kev sib ntxiv ntawm ligand thiab kev soj ntsuam tom qab ntawm cov kab hauv qab lub hom phiaj gene qhia. Ua kom tiav cov kev qhia tshwj xeeb hauv cheeb tsam, txoj kev tshawb fawb kuj pom tias tib neeg thiab murine organoids qhia thaj tsam tshwj xeeb muaj nuj nqi: lub plab murine teb rau TLR4 ligand LPS, tab sis tsis yog TLR2 ligand PAM3CSK4 los yog TLR5 ligand flagellin, los ntawm upregulating NF- κB phiaj gene Cxcl2. Murine jejunum upregulated Cxcl2 teb rau PAM3CSK4 thiab flagellin, tab sis tsis teb rau LPS. Tsis tas li ntawd, txoj hnyuv tau qhia Cxcl2 hauv kev teb rau tag nrho cov 3 kuaj ligands [44]. Hauv qhov sib piv, tib neeg organoids los ntawm txhua cheeb tsam tau qhia tib neeg Cxcl2 analog IL-8 teb rau flagellin, tab sis tsis teb rau LPS thiab PAM3CSK [44]. Yog li, tsis tsuas yog cov lus qhia tab sis kuj tseem ua haujlwm ntawm PRRs hauv lub plab yog qhov tsim nyog thiab ntu tshwj xeeb. Cov kev sim no tau ua los ntawm kev ntxiv cov stimulus rau hauv nruab nrab organoid, yog li tsuas yog stimulating sab basal, txij li thaum sab apical ntsib lub kaw-tawm lumen ntawm cov organoids. Lwm qhov kev sim ua kom pom tias muaj ntau lub koom haum ntau dua txij li PRR qhia nyob rau hauv ib ntu tuaj yeem txwv rau cov hom cell tshwj xeeb lossis txawm tias qhov chaw subcellular, xws li tsuas yog cov basal compartment. Cov no tau piav qhia ntau yam hauv qab no.

Txhua cheeb tsam ntawm cov kab mob GI qhia txog nws cov txheej txheem ntawm kev tiv thaiv kab mob hauv lub cev. Txawm hais tias ib qho kev nthuav dav yog qhov nyuaj, cov ntaub ntawv tam sim no qhia tias TLRs tau qhia ntau heev hauv lub plab thiab txoj hnyuv, thaum cov hnyuv me yog cov yam ntxwv los ntawm kev qhia ntawm cov kab mob inflammasome. Tam sim no, lub hauv paus txheej txheem molecular thiab cov txiaj ntsig evolutionary tsis meej.

Peb xav tias lub cev thiab tshuaj tiv thaiv uas npog cov txheej txheem epithelial hauv cov plab hnyuv sib txawv txhua qhov yuav tsum tau saib xyuas cov kev hem thawj tshwj xeeb. Piv txwv li, lub plab thiab txoj hnyuv ob leeg muaj ob txheej txheej, tuab mucus txheej nrog lub puab mucus txheej nrees txuas mus rau lub epithelial txheej, thaum cov hnyuv me yog them los ntawm ib tug nyias, viscous mucus txheej [46]. Cov kev cai sib txawv ntawm cov mucus khaubncaws sab nraud povtseg tej zaum yuav yog vim muaj kev sib txawv ntawm cov plab hnyuv ib ntus (kev zom thiab cov khoom noj khoom haus uptake vs dej resorption). Nws yog conceivable tias cov hnoos qeev tuab cuam tshuam rau hom PRRs tsim nyog ntawm cov chaw tshwj xeeb.

Cov hlwb tshwj xeeb nrog kev tiv thaiv kab mob hauv lub epithelium

Lub tswv yim ntev ntev nyob rau hauv epithelial innate tiv thaiv kab mob yog qhov muaj nyob ntawm cell hom tshwj xeeb hauv kev tiv thaiv kab mob. Prototypes ntawm cov hlwb tshwj xeeb epithelial nrog kev tiv thaiv muaj nuj nqi yog cov microfold (M) hlwb thiab Paneth hlwb - tsis ntev los no tau ua tiav los ntawm ib qho tshwj xeeb goblet cell, sentinel goblet cell. Lub hav zoov ntawm cov hlwb tshwj xeeb no qhia txog cov txheej txheem sib txawv thiab rov ua dua tshiab uas ua kom muaj kev sib npaug ntawm cov khoom noj khoom haus, kev sib koom ua ke nrog microbiota, thiab saib xyuas cov neeg tuaj yeem tuaj yeem. Kev taw qhia sib txawv ntawm organoids tam sim no kuj tso cai rau kev kawm ntawm cov hlwb hauv kab lis kev cai ntawm tes.

M hlwb nyob hauv cov follicle-associated epithelium npog Peyer's thaj ua rau thaj. Lawv muaj ib tug tshwj xeeb morphology nrog tsis tu ncua txhuam ciam teb thiab txo cov qauv microvilli. Lawv lub luag haujlwm yog thauj cov antigens hauv lub plab lumen hla cov txheej txheem epithelial mus rau hauv cov ntaub so ntswg lymphoid rau kev tswj hwm lub cev tiv thaiv kab mob [47]. M cell lawv tus kheej nrog rau cov follicle-associated epithelium tau pom los qhia ntau TLRs [48] (saib hauv [14]). Txawm li cas los xij, M hlwb tau nyuaj rau kev kawm vim tias tsis muaj ntau hauv cov ntaub so ntswg epithelial thiab lawv tsuas pom nyob ze ntawm cov qauv ntawm Peyer's thaj ua rau thaj [47]. Txawm li cas los xij, M hlwb tuaj yeem tsim tawm hauv cov kab lis kev cai organoid siv kev qhia txawv. Rau qhov no, receptor activator ntawm NF-κB ligand (RANKL) yog ntxiv rau qhov nruab nrab, uas txhawb nqa qhov hloov pauv ntawm SpiB, uas yog tus yam ntxwv ntawm M cell sib txawv [49]. Organoids tsim los ntawm nas caj ces tsis muaj peev xwm rau NF-κB subunit RelB tsis tuaj yeem tsim M hlwb tom qab stimulation nrog RANKL, qhia tias NF-κB ua kom yog qhov tseem ceeb rau kev loj hlob ntawm M hlwb [50]. Hauv tib neeg kab lis kev cai, ntxiv rau RANKL, lymphotoxin, thiab retinoic acid yog qhov tseem ceeb rau kev ua kom muaj kev sib txawv ntawm M hlwb. Cov M hlwb no tshwj xeeb coj cov kab mob enteric xws li rotavirus thiab reovirus, qhia qhov tseeb phenocopying ntawm lub ntuj M cell muaj nuj nqi kuj nyob rau hauv organoid kab lis kev cai [51]. Yav tom ntej organoid ua hauj lwm yuav tsum tau txuas ntxiv mus unravel lub interconnection ntawm epithelial NF-κB signaling, M cell txoj kev loj hlob, thiab kev sib txuas lus nrog lub cev tiv thaiv kab mob.

Paneth hlwb intermingled nrog cov hnyuv qia hlwb ntawm lub hauv paus ntawm lub crypt thiab tau ntev tau raug suav hais tias yog tus saib xyuas ntawm qia cell compartment vim lawv secrete antimicrobial peptides. Piv txwv li, Paneth hlwb secrete alpha-defensins, ib tug txheej txheem qhia tau hais tias yuav tsum tau tswj los ntawm microbial qauv thiab innate tiv thaiv mechanisms [52, 53]. Tsis tas li ntawd, NOD2, uas tau pom thawj zaug hauv thaj av crypt ntawm murine cov hnyuv [54], tswj kev tso tawm ntawm ob peb alpha-defensins los ntawm Paneth hlwb, uas ua rau ua rau kom muaj kev tiv thaiv kab mob. Paneth hlwb kuj qhia TLR5 thiab Paneth cell-enriched organoids qhia tshwj xeeb tshaj yog cov qib siab ntawm TLR5. RNA sequencing tom qab stimulation qhia tau hais tias thaum ib txwm me me plab hnyuv organoids tsuas yog qhia theem nrab ntawm lub TLR5 downstream lub hom phiaj noob nyob rau hauv cov lus teb rau stimulation nrog flagellin, xws li NF-kB-induced cytokines, organoids qhia kom muaj coob tus Paneth hlwb mount ib tug ntau zog. teb, qhia tias Paneth hlwb yog cov ntsiab lus teb rau flagellin hauv cov hnyuv [44, 45]. Nyob rau hauv sib piv rau cov hom phiaj qhia, cov lus teb txaus ntshai tshaj plaws ntawm Paneth hlwb (degranulation, extrusion, thiab cell tuag) tsis tshwm sim los ntawm stimulation nrog TLR ligands tab sis yuav tsum tau stimulation nrog cov kws tshaj lij ntawm lub cev tiv thaiv kab mob-derived cytokine interferon-gamma [55]. Cov txiaj ntsig no los ntawm organoids tau pom zoo nrog cov kev soj ntsuam uas qhia Paneth cell degranulation thiab extrusion tom qab interferon-gamma stimulation hauv vivo [55]. Qhov no elegantly underlines lub kaw lus ntawm kev txheeb xyuas thiab sib npaug ntawm cov txheej txheem epithelial.

Goblet hlwb yog ib qho tseem ceeb rau kev tiv thaiv epithelial vim tias lawv tsim cov glycosylated mucins tseem ceeb heev rau kev tsim cov mucus barrier ntawm lub epithelial txheej (saib hauv [56]). Hauv cov nas, MUC2 tsis muaj peev xwm ua rau muaj qhov mob tshwm sim thiab ua rau muaj kev kis kab mob ntau ntxiv [57, 58]. Tsis ntev los no, ib pawg ntawm goblet hlwb hu ua sentinel goblet hlwb tau piav qhia nyob rau hauv nas nyuv. Siv cov ntaub so ntswg explants, txoj kev tshawb fawb tau txheeb xyuas qhov thickening ntawm cov hnoos qeev hauv cov lus teb rau qhov raug rau TLR1/2, 4, thiab 5 ligands, tab sis tsis yog rau TLR9, NOD1, thiab NOD2 ligands [59], nyob rau hauv congruence rau ib daim ntawv qhia dhau los ntawm Tlr 2. , 4 thiab 5 tau hais nyob rau hauv goblet hlwb [60]. Cov kws sau ntawv tau txiav txim siab tias cov lus teb kuj yog nyob ntawm qhov muaj Nlrp6 inflammasome thiab tsis muaj kev ywj pheej ntawm mucosal lymphocytes siv cov ntaub so ntswg los ntawm ntau yam nas nas [59]. Tsis tas li ntawd, ib tsab ntawv tshaj tawm dhau los tau qhia txog qhov tseem ceeb ntawm NLRP6 inflammasome rau cov hnoos qeev los ntawm goblet hlwb [61]. Kev yees duab qhia tau hais tias cov kab mob goblet tshwj xeeb nyob hauv thaj tsam apical ntawm crypt endocytosed fluorescently tagged LPS [59]. Cov hlwb tshiab no hu ua sentinel goblet hlwb tsis tsuas yog ua kom nrawm nrawm thiab tshem tawm cov epithelial tom qab kev kho mob nrog TLR ligands tab sis kuj xa cov calcium teeb liab mus rau cov neeg nyob sib ze ntawm intercellular cytoplasmic txuas hniav tsim los ntawm qhov sib txawv junctions, yuav txhawb nqa lwm lub goblet hlwb kom nce mucus secretion [59] . Ob leeg goblet hlwb thiab Paneth hlwb muaj nyob rau hauv cov kab mob secretory. Organoids tso cai rau kev sib txawv ntawm ob lub xovtooj ntawm tes, yielding organoids hnyav hnyav nyob rau hauv goblet hlwb lossis Paneth hlwb [62]. Sib piv cov ntawv sau tseg ntawm cov skewed organoids tau pab txheeb xyuas cov neeg tswj hwm ntawm txoj kev sib txawv [63]. Kev tshuaj xyuas ntxiv ntawm cov ntaub ntawv omics nrog rau kev tsom xam ua haujlwm ntawm cov organoids no yuav tso cai rau kev nkag siab zoo ntawm lub luag haujlwm ntawm ob hom cell hauv kev tiv thaiv kab mob hauv lub cev.

Thaum kawg, cov qia hlwb lawv tus kheej kuj tau tshaj tawm los qhia txog PRRs tshwj xeeb xws li TLR4 [64, 65], uas tsis pom nyob rau hauv murine me me hnyuv villi lossis Paneth hlwb [66]. Tsis tas li ntawd, feem ntau ntawm Nod2 qhia nyob rau hauv murine crypt zoo li raug txwv rau qia hlwb [67]. Kev txhawb nqa nrog NOD2 ligand nce kev ciaj sia ntawm cov qia hlwb thiab tsim cov organoids, qhia tias kev txhawb nqa ntawm PRRs kuj tseem tuaj yeem tswj lub plab epithelial regeneration ncaj qha.

Tsis muaj qhov tsis ntseeg, qhov kev siv zog tam sim no los tsim cov atlases ntawm cov noob qhia uas npog txhua hom xovtooj ntawm txhua qhov kev nthuav dav yuav sai sai no muab cov duab tiav ntawm lub koom haum cellular ntawm lub cev tiv thaiv kab mob hauv lub plab thiab hauv tag nrho lub cev [68-71. ].

Cell polarity thiab sab-specific innate immune teb

Thaum kawg, nws tseem ceeb heev uas yuav tsum xav txog tias cov kab mob plab hnyuv epithelial yog cov polarized heev, nrog rau sab tshwj xeeb apical sab ntsib lub lumen ntawm lub plab nrog nws cov microbiota, thiab sab basolateral ntsib cov ntaub so ntswg. Raws li homeostasis, MAMPs ncav cuag sab apical nkaus xwb. Txawm li cas los xij, thaum cov kab mob epithelial raug ua txhaum cai, cov kab mob tuaj yeem tuaj yeem cuam tshuam rau sab basolateral. Nws tau yog li no tau pom zoo tias cov hlwb epithelial tuaj yeem xaiv cov lus teb rau cov tshuaj tiv thaiv tsuas yog thaum txhawb los ntawm sab basolateral, kom phim qhov kev hem thawj los ntawm lub teeb liab. Piv txwv li, TLR9 tau tshwm sim los ua kom muaj kev sib txawv ntawm kev taw qhia thaum txhawb nqa los ntawm apical lossis basolateral sab hauv cov kab mob qog noj ntshav [72] thiab TLR5 tsuas yog ntxias cov NF-κB cov lus teb noob IL-8 thaum txhawb los ntawm sab basal [73 ].
Thaum cov kev tshawb fawb yav dhau los siv cov tshuaj tiv thaiv kab mob tiv thaiv TLRs tau tshaj tawm cov lus qhia tshwj xeeb ntawm ib sab nkaus xwb (saib xyuas hauv [14, 16]), kev tshuaj xyuas ntawm TLR cov neeg sau xov xwm nas siv staining ntawm HA tag tsis tau lees paub qhov no tab sis es tsis txhob ua TLR2, 4, thiab 5. receptors ntawm ob qho tib si apical thiab basal sab ntawm cov nyuv proximal nrog rau qee qhov intracellular TLR4 [45]. Cov kev sib txawv pom tseeb no yuav yog vim muaj cov txheej txheem sib txawv.

Organoids tam sim no tso cai rau kev sim ua haujlwm ncaj qha ntawm kev tiv thaiv kab mob sab nraud, vim tias cov cellular polarization tau khaws cia hauv organoids. Raws li cov qauv txheej txheem thaum organoids loj hlob nyob rau hauv ib qho extracellular matrix, sab apical ntsib lub lumen ntawm lub organoid thiab sab basal ntsib lub extracellular matrix [74–76]. Thaum loj hlob sab nraum lub matrix extracellular, lub polarity tuaj yeem thim rov qab [77, 78]. Thaum cov hlwb los ntawm cov organoids tau cog rau hauv cov txheej txheem kab lis kev cai ntawm tes, xws li kab lis kev cai tais diav lossis transwells, sab apical ntsib lub lumen ntawm qhov dej [44, 79–82].

Ntau qhov kev tshawb fawb tau siv organoids los ntsuas qhov kev ua haujlwm ntawm PRRs tshwj xeeb yam tsis tau hais txog qhov sib txawv ntawm apical thiab basal stimulation. Cov kev tshawb fawb no suav nrog cov stimulus nyob rau hauv nruab nrab ntawm cov organoids, uas, nyob rau hauv tus qauv tej yam kev mob, stimulates basal sab ntawm lub cell. Siv cov txheej txheem no, upregulation ntawm NF-κB downstream lub hom phiaj noob tau txheeb xyuas tom qab basal stimulation nrog ligands ntawm TLR4 nyob rau hauv lub plab, TLR2 thiab 3 nyob rau hauv cov hnyuv me, thiab TLR2, 3, 4, thiab 5 nyob rau hauv cov hnyuv ntawm nas [44, 45, 83] thiab rau ligands ntawm TLR2 thiab 5 nyob rau hauv lub plab thiab TLR5 nyob rau hauv cov hnyuv me thiab cov hnyuv ntawm tib neeg [44]. Tsis tas li ntawd, basal stimulation ntawm murine nyuv organoids nrog TLR4 agonists induced cellular sib txawv, tshwj xeeb tshaj yog rau cov secretory lineage [64], thaum NOD2 agonists induced nce ciaj sia taus ntawm qia hlwb [67, 84], thiab taurine stimulated NLRP6- nyob ntawm kev tswj hwm. ntawm inflammasome downstream hom phiaj IL-18 [85]. Hauv tag nrho cov kev tshawb fawb no, apical stimulation tsis tau sim.

Tsuas yog qee qhov kev tshawb fawb tau hais txog cov haujlwm tshwj xeeb ntawm PRRs. Nws yog ib qho tseem ceeb uas tsis yog cov kev tshawb fawb siv cov kab mob qog noj ntshav polarized, txog tam sim no, tsis muaj ib qho ntawm cov kev tshawb fawb siv organoids tau txheeb xyuas qhov tshwj xeeb ntawm kev ua haujlwm ntawm ib qho NF-κB-dependent proinflammatory gene. Transwell monolayers muab tau los ntawm tib neeg txoj hnyuv organoids qhia cov qib zoo sib xws ntawm NF-κB hom phiaj IL-6 thaum txhawb los ntawm apical lossis basal sab nrog ligands ntawm TLR1/2, 3, 4, 5, 7/8, thiab 9 [82]. Murine gastric organoids kuj tau teb rau ib qho apical stimulation nrog TLR4 ligand LPS nyob rau hauv ntau qhov kev ntsuam xyuas, nrog rau hauv transwells thiab microinjection ntawm LPS rau hauv lub lumen ntawm organoids [44]. Murine lub plab hnyuv me me epithelial hlwb tsis tau qhia txog NF-κB lub hom phiaj noob icam1 hauv cov lus teb rau ntau yam ligands, txawm tias lawv tau ntxiv rau cov kab mob tsis zoo, yog li txhawb rau sab basal, lossis ntxiv rau ib leeg, cov hlwb sib txawv, yog li txhawb txhua tus. sab [55].

Txawm li cas los xij, saib dhau ntawm NF-κB cov lus teb, kev tshawb fawb tsis ntev los no tau txheeb xyuas qhov ua haujlwm tshwj xeeb ntawm TLR3 thiab nws qhov tseem ceeb hauv kev kis kab mob [82]. Cov txiaj ntsig ntawm kev sim nrog tib neeg txoj hnyuv organoid-derived monolayers tau pom tias qhov kev qhia ntawm tus kab mob tseem ceeb-tiv thaiv cov noob hom I thiab hom III interferon tau raug kho tom qab basal stimulation nrog TLR3 agonist, tab sis tsis yog tom qab stimulation nrog lwm TLR agonists. Thaum kis tus kab mob, zoo ib yam, cov lus teb ntawm interferon ntawm cov organoids muaj zog ntau dua thaum kis los ntawm sab basal piv rau kev kis kab mob los ntawm sab apical. Qhov no tau pom nyob rau hauv organoid-derived monolayers nrog rau hauv organoids microinjected nrog tus kab mob. Txoj kev tshawb fawb ntxiv tau txheeb xyuas lub clathrin-sorting adapter AP-1B raws li lub molecule lub luag hauj lwm rau lub polarized qhia ntawm TLR3. Raws li txoj cai, cov nas tsis muaj peev xwm hauv Ap{11}}b tau pom tias muaj kev tiv thaiv kab mob hnyav dua tom qab kis kab mob [82]. Qhov no tau lees paub qhov kev ua haujlwm polarized ntawm PRRs thiab hais txog qhov tseem ceeb ntawm qib kev cai no hauv kev hloov kho kab mob ntxiv thiab kev tiv thaiv.

Txog tam sim no, tsis muaj "ib qho me me haum rau tag nrho" cov lus teb rau lo lus nug polarity, thiab kev daws qhov chaw tshwj xeeb ntawm PRR signaling tseem yog ib txoj haujlwm nyuaj rau yav tom ntej.

Tolerance raws li ib tug teb rau colonization thiab raws li ib tug default developmental program

Cov txheej txheem pab txhawb rau lub koom haum ntawm epithelial innate tiv thaiv yog tseem tsis meej. Lub ntsiab lus tseem ceeb hauv qhov no yog qhov ua rau muaj kev zam txim tom qab kev ua kom muaj menyuam tsis muaj menyuam thaum yug menyuam, lub sijhawm hu ua qhov rais ntawm lub sijhawm (saib hauv [86–88]). Lub tswv yim no postulates lub sij hawm priming ntawm lub innate thiab adaptive lub cev tiv thaiv kab mob tom qab yug me nyuam, uas teem rau theem rau kev tiv thaiv homeostasis thiab tom ntej host-microbial kev sib cuam tshuam.

Lub cev tiv thaiv kab mob neonatal thiab epithelial innate immunity yog tshwj xeeb tsim los ua qhov kev hloov pauv ntawm kev tsis muaj menyuam mus rau kev ua neej nyob nrog microbiota. Thaum yug los, tib neeg lub plab hnyuv loj hlob tuaj nrog plab hnyuv villi thiab crypts uas muaj Paneth hlwb. Lub murine neonatal plab hnyuv epithelial txheej yog qhov tsis paub tab thiab dhau qhov kev hloov pauv loj uas ntsuas tau ntawm qib transcriptome piv cov theem sib txawv ntawm kev loj hlob [89] thiab pom nyob rau hauv cov ntaub so ntswg architecture thiab cell sib txawv: Lub crypt-villus axis tseem tsis tau tsim thiab cell proliferation yog. qis dua, tsis muaj cell migration lossis exfoliation. Nws tsis muaj Paneth hlwb paub tab; Txawm li cas los xij, enterocytes tsim cov cathelicidin zoo li antimicrobial peptide (CRAMP) [90]. Paneth hlwb tshwm sim thaum crypts tsim 2 lub lis piam tom qab yug me nyuam [91]. Thaum lub sij hawm weaning, lub epithelium yog tag nrho tsim nrog crypts thiab villi, enterocytes, goblet hlwb, thiab enteroendocrine hlwb; Paneth hlwb tau noj cov tshuaj tua kab mob peptide ntau lawm; thiab goblet hlwb tau nce cov mucins los tsim cov mucus txheej (saib hauv [92]).

Qhov kev hloov pauv ntawm lub hnub nyoog ntawm lub epithelium mus koom tes nrog kev txo qis ntawm TLR5 qhia nyob rau hauv cov hnyuv me me epithelium. Nyob rau tib lub sijhawm, kev qhia ntawm TLR3 nce ntxiv thaum lub sijhawm xeeb tub. Lwm cov PRRs, xws li TLR2, 4, thiab 9, tau qhia nyob rau theem zoo sib xws hauv cov nas me me thiab cov laus [45, 93].

Cov txheej txheem ua rau kev tswj hwm ntawm PRR qhia thiab kev ua haujlwm tom qab yug me nyuam tseem tsis meej. Raws li tau hais los saum toj no, ntau qhov kev tshawb fawb tau tawm tswv yim kev koom tes ntawm ib puag ncig, tshwj xeeb tshaj yog microbial colonization, rau kev tswj hwm ntawm PRR qhia tom qab yug me nyuam [40, 41] (saib hauv [14]). Txawm li cas los xij, cov kab mob tsis muaj kab mob vs cov nas tsis muaj kab mob tsis pom qhov sib txawv hauv TLR qhia hauv cov hnyuv me los yog cov hnyuv, qhia tias tsis muaj kev tswj hwm ntawm TLR3 los yog downregulation ntawm TLR5 nyob rau lub sij hawm thaum ntxov no nyob ntawm microbiota [45, 93]. Tsis tas li ntawd, nyob rau hauv organoids, qhov kev qhia ntawm ntau, tab sis tsis yog tag nrho, PRR signaling Cheebtsam twb tau txhais nyob rau hauv organoids los ntawm cov ntaub so ntswg uas tsis tau muaj nyob rau hauv kev sib cuag nrog microbial khoom [44]. Qhov no qhia tau hais tias ib feem loj ntawm lub koom haum ntawm lub cev tiv thaiv kab mob txoj hauv kev tau txhais ntawm nws tus kheej ntawm kev sib cuag nrog microbiota thiab zoo li tau txiav txim siab los ntawm cov txheej txheem kev loj hlob, xws li cov uas tau teev tseg saum toj no, uas ua rau cov ntaub so ntswg dav dav raws li GI ib puag ncig. . Qhov no tsis suav nrog kev kho kom zoo ntxiv ntawm PRR qhia los ntawm ib puag ncig ib puag ncig thaum muaj hnub nyoog laus.

Qhov tseem ceeb ntawm txoj cai raws sij hawm ntawm kev sib cuam tshuam ntawm epithelial microbial ua kom pom tseeb thaum lub epithelium tsis paub qab hau yog ntxov ntxov ntsib nrog microbial colonization: cov me nyuam mos ua ntej muaj feem cuam tshuam rau kev loj hlob ntawm necrotizing enterocolitis (NEC), uas muaj cov plab hnyuv necrosis, kab mob sepsis, thiab ntau yam kab mob. ua tsis tiav. Txawm hais tias lub pathogenesis yog suav tias yog multifactorial, ntau cov kev tshawb fawb tau qhia tias nws txhim kho nyob rau hauv cov lus teb rau qhov tsis sib xws ntawm proinflammatory signaling thiab kho mechanisms nyob rau hauv lub plab ntxov ntxov (saib hauv [94]) thiab ib qho kev koom tes ntawm PRRs tau pom zoo [41, 65, 95. — 97]. Ntau qhov kev tshawb fawb tau siv tib neeg fetal organoids [24, 98, 99], ib txwm murine organoids raug cov kab mob thiab hypoxia rau tus qauv NEC [100], los yog organoids los ntawm murine NEC qauv zoo li los ntawm NEC cov neeg mob [101]. Cov kev tshawb fawb yav tom ntej yuav siv cov qauv tsim thiab tshiab organoid los piav qhia ntxiv txog kev koom tes ntawm epithelium hauv NEC.

Ua qauv epithelium-dependent yam ntawm IBD nrog organoids

Kev poob ntawm txoj hnyuv epithelial teeb meem kev ncaj ncees yog ib qho kev txhais ntawm IBD, piv txwv li CD thiab UC, thiab zoo li tshwm sim los ntawm kev sib cuam tshuam ntawm cov caj ces, ib puag ncig, kev hloov ntawm plab microbiota, thiab kev hloov pauv ntawm lub zos thiab lub cev tiv thaiv kab mob (saib xyuas hauv [27]). Cov kws kho mob tam sim no feem ntau tsom rau cov tshuaj tiv thaiv kab mob (aberrant) hauv IBD, uas cuam tshuam nrog cov txiaj ntsig siab ntawm cov tsis teb thiab cov kev mob tshwm sim (saib hauv [27]). Kev txhim kho kev nkag siab ntawm epithelium tshwj xeeb kev koom tes rau pathophysiology ntawm IBD yog yuav tsum tau txheeb xyuas cov hom phiaj kho tshiab uas tuaj yeem tuaj yeem cuam tshuam ncaj qha rau hauv plab hnyuv epithelial barrier restoration thiab yog li mucosal kho.

Txhawm rau kom nkag siab txog cov kab mob epithelial, ntau pab pawg tau tsim cov chaw nyob biobanks uas tsim los ntawm cov neeg mob uas muaj UC lossis CD. Txawm hais tias txoj hauv kev no yog qhov pom tseeb, txog tam sim no, tsuas yog qee qhov kev tshawb fawb tau tshaj tawm cov txiaj ntsig ntawm organoids tau los ntawm pawg neeg mob no [102-104]. Qhov no tuaj yeem piav qhia los ntawm kev soj ntsuam tias organoids los ntawm cov neeg mob nrog IBD nyuaj rau tsim. Peb tus kheej qhov kev paub yog tias cov organoids tsim los ntawm cov neeg mob CD loj hlob qeeb dua thaum thawj cov kab lus thiab qee cov qauv tau ploj - uas tau cuam tshuam nrog cov kab mob ntau dua hauv kab lis kev cai [105].

Cov yam ntxwv ntawm cov organoids muab tau los ntawm cov neeg mob nrog IBD tau nthuav tawm ib qho phenotype nrog txo qhov loj me thiab budding muaj peev xwm, nce tus nqi ntawm cell tuag, luminal khib nyiab, thiab ib nrab inverted polarization ntawm epithelial hlwb [106]. Kev sib piv thoob ntiaj teb ntawm organoids los ntawm UC lossis CD cov neeg mob thiab kev tswj xyuas kev noj qab haus huv tau pom tias qhov sib txawv ntawm kev hloov pauv thiab methylation pom nyob rau hauv txoj hnyuv epithelium tau khaws cia hauv vitro [102, 103, 107]. Tsis tas li ntawd, ib qho kev tshawb fawb ua ntej los ntawm organoids tsim los ntawm cov neeg mob CD qhia txog kev hloov pauv ntawm cov kab mob hauv plab hnyuv hauv IBD. Qhov no yog raws li kev soj ntsuam uas cov organoids tsim los ntawm cov kab mob CD ua haujlwm tau khaws cov kev qhia siab ntawm txoj hnyuv epithelial qia cell markers [108]. Qhov no yog rau qee qhov kev lees paub hauv kev tshawb fawb tsis ntev los no uas cov kab mob hauv plab tau muab los ntawm cov neeg mob IBD cov neeg mob tau pom qhov ntev-kev nthuav qhia tus qauv ntawm cov tshuaj tiv thaiv kab mob tam sim no [109].

Kev tsom mus rau cov kev hloov hauv plab hnyuv epithelial barrier muaj nuj nqi, nrog rau kev poob ntawm nruj junctions thiab desmosomes, uas feem ntau pom nyob rau hauv IBD [110], nws tau pom tias organoids los ntawm CD cov neeg mob tuav no phenotype ntawm junctional alterations nyob rau hauv kab lis kev cai tej yam kev mob [105]. Qhov no yog qhov tshwj xeeb tshaj yog thaum cov organoids tau tsim los ntawm qhov chaw mob hnyav [105]. Kev txo qis hauv cov proteins sib txuas kuj tseem tuaj yeem ua rau cov organoids los ntawm kev noj qab haus huv pub dawb los ntawm kev siv cov proinflammatory cytokine TNF- thiab / lossis IFN- [106, 111]. Txawm li cas los xij, cov qauv ruaj khov ntawm kev hloov pauv ntawm cov protein sib txuas hauv organoids los ntawm cov neeg mob IBD tau pom tsuas yog nyob rau qib protein tab sis tsis nyob rau qib mRNA [105].

Cov kev soj ntsuam no qhia tias ntau qhov kev hloov pauv tau kho hauv organoids los ntawm cov neeg mob IBD. Kev soj ntsuam tias qee qhov, tab sis tsis yog tag nrho, kev hloov pauv mus tas li tsuas yog pom ntawm cov protein, tab sis tsis yog RNA-theem, qhia tias muaj kev hloov pauv mus tas li hauv kev hloov pauv hloov pauv lossis cov protein degradation hauv organoids los ntawm cov neeg mob IBD. Txawm li cas los xij, qhov no tseem yuav tau tshawb xyuas kom meej.

Dab tsi yog qhov ua rau muaj kev hloov pauv mus tas li hauv cov qauv qhia ntawm cov kab mob epithelium yog tsis meej. Ib qho txiaj ntsig ntawm microbiota tau xav tias ntev ntev; Txawm li cas los xij, kev tshawb fawb tsis ntev los no tau pom tias cov txiaj ntsig ntawm microbiota ntawm lub epithelium tau ploj mus dhau sijhawm [112]. Txoj kev tshawb no tau muab piv rau ntau qhov chaw nas vim tias qhov sib txawv ntawm cov microbiome muaj nyob ntawm nas cov cheeb tsam tau raug txheeb xyuas raws li qhov tsis txaus ntseeg hauv kev tshawb fawb vivo. Txhawm rau kom paub meej qhov txiaj ntsig no, txoj kev tshawb fawb tau muab piv rau cov kab mob epithelial cais tawm thiab cov kab lis kev cai organoid los ntawm cov nas tsis muaj kab mob thiab ob qhov tshwj xeeb cov kab mob tsis muaj nas nyob nrog cov microbiota sib txawv. Thaum lub epithelium uas tau raug cais tawm tshiab tau pom tias muaj kev cuam tshuam ntawm microbiota ntawm RNA thiab cov protein ntau, cov nyhuv no tau ploj tom qab ob peb lub lis piam ntawm kab lis kev cai ntawm cov hnyuv me me [112]. Tsis tas li ntawd, kev sib piv thoob ntiaj teb ntawm cov noob qhia ntawm cov organoids tsim los ntawm thaj chaw mob lossis tsis yog mob ntawm tib cov neeg mob IBD tau pom tias IBD organoids ntawm thaj chaw mob tau ploj mus rau cov kab mob inflammatory uas twb muaj lawm tom qab ob peb lub lis piam hauv kab lis kev cai. Cov ntawv sau tseg ntawm cov organoids tau muab faib ua ib tus neeg mob; Yog li, qhov sib txawv mus tas li ntawm IBD thiab kev tswj hwm kev noj qab haus huv tseem nyob. Tus mob phenotype hauv IBD organoids tuaj yeem rov ua dua los ntawm kev ntxiv ntawm cytokine cocktail [107]. Zuag qhia tag nrho, qhov no qhia tau tias kev hloov pauv mus tas li pom hauv organoids los ntawm cov neeg mob IBD tsis muaj kev sib cuag nrog microbiota lossis cytokines.

Yog li, txawm hais tias nws yog qhov tsim nyog los xav tias qee qhov kev hloov pauv mus tas li hauv plab hnyuv epithelial hlwb los ntawm cov neeg mob IBD tuaj yeem tshwm sim los ntawm kev hloov pauv hauv lub cev tiv thaiv kab mob, cov pov thawj rau qhov no tam sim no tsis tshua muaj. Tsis tas li ntawd, tsis yog tag nrho cov kev hloov pauv ntawm epithelial pom nyob rau hauv cov cheeb tsam o nyob rau hauv lub plab yog ruaj ruaj nyob rau hauv lub purified epithelium, qhia tau hais tias tseem ceeb pab los ntawm lub zos ib puag ncig. Nyob rau hauv lub neej yav tom ntej, cov kev tshawb fawb tshiab nrog ntau cov qauv organoid, kuj suav nrog lub cev tiv thaiv kab mob, inflammatory cytokine stimulation, thiab co-cultures nrog microorganisms, yuav tsum pab daws qhov kev xav no kom ntxaws.

Cov lus xaus, kev pom yav tom ntej, thiab Outlook

Hauv cov ntsiab lus, txawm tias muaj ntau yam kab mob hauv GI ib puag ncig tseem nkag siab tsis tiav, ua pov thawj ntxiv rau lub luag haujlwm tseem ceeb ntawm lub plab hnyuv epithelium hauv cov kab mob ntawm ntau ntawm lawv - txawm hais tias nws lub luag haujlwm tshwj xeeb tseem tsis meej.

Qhov kev paub tsawg ntawm lub cev tsis muaj zog ntawm lub plab hnyuv epithelium tau raug ntaus nqi rau qhov tsis muaj cov qauv kev sim tsim nyog. Nrog rau kev siv tshuab organoid, ib kauj ruam tseem ceeb tau ua los daws qhov teeb meem no. Organoids tsim los ntawm txhua cheeb tsam ntawm lub plab zom mov yuav ntxiv qhov tseem ceeb rau cov kev paub uas twb muaj lawm. Raws li qhia los ntawm cov kev tshawb fawb yav dhau los, tiam ntawm organoids muab cov txiaj ntsig tseem ceeb ntawm kev muaj peev xwm soj ntsuam cov lus teb ntawm cov kab mob plab hnyuv loj uas tsis yog cov kab hloov pauv ntawm tes, uas feem ntau yog tsim los ntawm cov qog nqaij hlav hauv plab. Ib qho tshwj xeeb tshaj plaws ntawm cov organoids yog tias lawv tuav cov yam ntxwv tshwj xeeb ntawm ntu ntawm cov kab mob plab uas lawv tau tsim los ntawm ib feem ntawm lawv tus kheej ntawm tes thaum muaj hnub nyoog laus. Raws li kev tshawb fawb tam sim no, lub regional tus kheej yog kho nyob rau hauv lub plab hnyuv qia hlwb. Hauv cov ntsiab lus no, ib qho teeb meem ntxiv yuav yog los txiav txim siab seb yuav ua li cas thiab thaum twg thaum lub sij hawm kev loj hlob no intrinsic programming tshwm sim.

Thaum saib cov ntsiab lus tshwj xeeb ntawm yuav ua li cas lub epithelium yuav teb los yog cuam tshuam nrog ib puag ncig, organoid thev naus laus zis tam sim no tau ua kom pom kev sib txawv thiab ntu tshwj xeeb qhia thiab kev ua haujlwm ntawm PRRs hauv plab hnyuv epithelium. Tag nrho cov txiaj ntsig ua haujlwm rau cov txheej txheem kev tswj hwm nyuaj hauv tag nrho cov plab hnyuv siab raum tseem tsis meej thiab yuav tsum tau hais txog yav tom ntej. Rau qhov no, nws tseem yuav yog qhov tseem ceeb rau coculture organoids nrog lub cev tiv thaiv kab mob (saib hauv [113]), cov hlwb ntawm lub paj hlwb enteric [114], thiab luminal yam xws li kab mob co-cultures (saib hauv [115]) (Daim duab . 4).

Ib qho tseem ceeb ntxiv yog tias cov organoids tsim los ntawm cov neeg mob cov ntaub so ntswg cuam tshuam los ntawm cov kab mob GI xws li IBD tuav qee cov yam ntxwv uas pom hauv cov ntaub so ntswg sib xws lawv tau muab los ntawm. Qhov no muaj qhov tshwj xeeb muaj peev xwm ntxiv rau kev tshem tawm cov kab mob epithelium- thiab kab mob tshwj xeeb rau kev ua haujlwm ntawm cov kab mob GI-tsis yog cov uas cuam tshuam nrog kev mob tshwm sim los ntawm kev hloov pauv, tab sis kuj muaj cov kev hloov pauv hauv cov kab mob malignant. Ob leeg ntawm lawv tuaj yeem ua rau muaj kev koom tes tshwj xeeb ntawm epithelium-derived innate tiv thaiv kab mob. Rau qhov no, kev tsim kho ntawm "nyob biobanks" yuav yog ib kauj ruam tseem ceeb. Raws li lub zeem muag rau yav tom ntej, xws li biobanks muaj sia nyob tuaj yeem txuas nrog cov biobanks uas twb muaj lawm, uas tam sim no tsuas yog "tuag" biomaterials. Qhov no yuav sawv cev rau lwm kauj ruam tseem ceeb tsis yog rau kev tshawb fawb nkaus xwb tab sis kuj tseem pab txhawb kev kuaj mob tus kheej thiab kev kho mob rau cov neeg mob.

Lub thawv 1. Innate immunity thiab epithelial hlwb nyob rau hauv cov kab mob GI tsis paub meej tias qhov kev cuam tshuam yog vim lub cev tsis muaj zog tiv thaiv kab mob hauv cov kab mob epithelial los yog cov tshuaj tiv thaiv kab mob.

Txhawm rau tshem tawm qhov kev sib cuam tshuam ntawm cov kab mob kev tiv thaiv kab mob thiab cov kab mob epithelial, ntau cov kev tshawb fawb tau siv cov epithelium tshwj xeeb knockouts, los yog tsis ntev los no, epithelial organoids. Hauv cov qauv nas, tsis muaj kev tshem tawm ntawm cov epithelial tshwj xeeb ntawm PRRs ua rau muaj kev mob tshwm sim. Txawm li cas los xij, cov nas uas muaj cov kab mob tshwj xeeb tshwj xeeb ntawm MyD88 yog qhov muaj feem cuam tshuam rau kev sim colitis thiab ua rau muaj kev cuam tshuam loj heev, tsis muaj peev xwm goblet thiab Paneth cell teb [121], thiab txo qis cov mucin thiab antimicrobial peptides [121, 122]. Cov kab mob hauv plab me me tsis tuaj yeem ua rau cov kab mob inflammatory rau ntau cov purified PRR ligands [55], txawm hais tias qhov no tsis tuaj yeem ua tau dav dav thiab nyob ntawm hom, qhov chaw, thiab hnub nyoog ntawm cov ntaub so ntswg cov organoids yog tsim los ntawm [44, 45].

Qhov tsis muaj qhov tshwm sim ntawm qhov tshwm sim tshwm sim hauv epithelial cell-specific PRR knockout qauv txhawb nqa qhov kev xav tias lwm yam tsis yog cov lus teb dav dav ntawm cov epithelium muaj kev cuam tshuam rau epithelial homeostasis. Piv txwv li, kev txhawb nqa nrog NOD2 agonist muramyl dipeptide (MDP) nce tus naj npawb ntawm cov organoids loj hlob tawm ntawm cov qia hlwb, qhia tias lub cev tsis muaj zog tiv thaiv kab mob txhawb kev ciaj sia ntawm cov qia hlwb [67, 84]. Tsis tas li ntawd, cov ntaub ntawv los ntawm nas qhia qhov tseem ceeb ntawm kev tiv thaiv apoptotic los ntawm NF-κB teeb liab hauv kev teb rau lwm yam stimuli, xws li TNF- [123]. Interestingly, nyob rau hauv tib neeg, polymorphisms nyob rau hauv innate immune genes nrog rau NOD2 thiab TLR4 yog txuam nrog ib tug muaj kev pheej hmoo los tsim IBD [124] thiab blockage ntawm TNF- yog tam sim no qhov zoo tshaj plaws kev kho mob rau IBD nyob rau hauv ib co neeg mob (saib xyuas hauv [125]).

Ib daim duab tshwm sim, uas qhov qis qis ntawm lub cev tiv thaiv kab mob hauv lub cev yog qhov tseem ceeb rau kev tso tawm cov hnoos qeev, thaiv kev ncaj ncees, thiab kev muaj sia nyob ntawm epithelial cell. Nws qhov kev puas tsuaj tuaj yeem tso cai rau kev hloov cov kab mob hauv plab hnyuv los ntawm lumen mus rau hauv cov ntaub so ntswg subepithelial, ua rau mob.

Kev lees paub

Peb ua tsaug Ömer Kaya rau kev pab nrog cov duab kos thiab Rike Zietlow rau kev kho.

Sau ntawv pab

Lub tswv yim: SB Initial concept: SB thiab Ö.K.; Cov ntaub ntawv tshawb fawb: Ö.K., NS, thiab SB; Initial Draft: Ö.K. Revising, rewriting, added sections: NS, SB Txhua tus kws sau ntawv tau hloov kho cov ntawv sau kawg.

Nyiaj txiag

Qhib Kev Siv Nyiaj tau qhib thiab tsim los ntawm Projekt DEAL. Txoj haujlwm no tau txais kev txhawb nqa rau Deutsche Forschungsgemeinschaft (DFG GRK 2157; 3D Tissue Models for Studying Microbial Infections by Human Pathogens, Project 10, to SB), Interdisciplinary Center for Clinical Research (IZKF; www.med.uni-wuerzburg.de/izkf /startseite) hauv Würzburg (Grant AD-427 rau NS thiab SB), thiab DFG SPP1982 SCHL1962/5-2 rau NS.

Cov lus tshaj tawm

Kev tsis sib haum xeeb

Cov kws sau ntawv tshaj tawm tsis muaj kev sib tw nyiam.

Qhib Access

Kab lus no tau tso cai raws li Creative Commons Attribution 4.0 Daim ntawv tso cai thoob ntiaj teb, uas tso cai siv, sib koom, hloov kho, faib, thiab luam tawm hauv ib qho nruab nrab lossis hom ntawv, tsuav yog koj muab credit rau tus thawj sau ) thiab qhov chaw, muab qhov txuas mus rau Creative Commons daim ntawv tso cai, thiab qhia seb puas tau hloov pauv. Cov duab lossis lwm yam khoom siv thib peb hauv tsab xov xwm no suav nrog hauv tsab xov xwm Creative Commons daim ntawv tso cai tshwj tsis yog tau qhia lwm yam hauv kab qiv nyiaj rau cov khoom siv. Yog tias cov ntaub ntawv tsis suav nrog hauv tsab xov xwm Creative Commons daim ntawv tso cai thiab koj qhov kev npaj siv tsis raug tso cai los ntawm txoj cai lij choj lossis tshaj qhov kev tso cai siv, koj yuav tsum tau txais kev tso cai ncaj qha los ntawm tus tuav ntaub ntawv.

Cov ntaub ntawv

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