Txoj Kev Kom Txhim Kho Kev Kawm Ua Ntej Ntawm Cov Hnub Nyoog Txog Cov Kab Mob Neurodegenerative: Kev Pom Zoo Ntawm Cov Tsiaj Thiab HiPSC-Drived Models Part 4

hiPSC-los ntawm cov qauv vitro ntawm PD kuj tau qhia ntau yam hauv kev nthuav qhia cov kab mob pathophysiological txuas nrog kev hloov pauv hauv tsev neeg PD hauv cov neeg mob-los ntawm hiPSC DA neuronalmodels.

PD yog kab mob neurodegenerative uas feem ntau muaj qee yam cuam tshuam rau tus neeg mob lub cim xeeb thiab kev txawj ntse. Txawm li cas los xij, nrog kev txhim kho txuas ntxiv ntawm cov tshuaj niaj hnub no, peb tuaj yeem siv ntau txoj hauv kev los pab cov neeg mob kom muaj kev nco zoo.

Ua ntej, kev tawm dag zog ib txwm tuaj yeem txhim kho kev txawj ntse thiab kev nco ntawm PD cov neeg mob. Kev tawm dag zog tuaj yeem txhawb nqa cov ntshav ncig thiab lub hlwb noj zaub mov ntawm cov paj hlwb hauv lub hlwb, uas pab tswj kev sib txuas thiab kev sib koom tes ntawm ntau qhov chaw ntawm lub hlwb, yog li txhim kho tag nrho cov kev txawj ntse.

Qhov thib ob, kev noj haus kuj yog tus yuam sij rau kev txhim kho kev nco ntawm PD cov neeg mob. Kev noj zaub mov muaj fiber ntau, rog tsawg thiab cov roj cholesterol tsawg yog tus yuam sij rau kev tswj lub hlwb. Cov zaub mov no tuaj yeem txhawb cov ntshav ncig hauv lub hlwb thiab muab cov khoom noj muaj txiaj ntsig zoo, uas tuaj yeem tiv thaiv cov neurons thiab ua rau qeeb ntawm PD.

Thaum kawg, tus cwj pwm zoo thiab so kom raug kuj yog tus yuam sij rau kev txhim kho kev nco ntawm PD cov neeg mob. Txawm hais tias PD yuav ua rau muaj kev cuam tshuam rau lub cev thiab lub siab, yog tias koj tuaj yeem tswj hwm tus cwj pwm zoo thiab muab koj tus kheej so, koj tuaj yeem txo qhov kev ntxhov siab ntawm lub hlwb thiab lub hlwb thiab tswj koj lub cim xeeb zoo dua.

Hauv luv luv, txawm hais tias PD yuav cuam tshuam rau kev nco, nws tsis tau txhais hais tias peb hloov tsis tau. Los ntawm kev tawm dag zog kom zoo, noj zaub mov thiab tswj kev puas siab puas ntsws, PD cov neeg mob tuaj yeem tswj tau qee theem ntawm kev txawj ntse thiab kev nco. Nws tuaj yeem pom tias peb yuav tsum txhim kho kev nco, thiab Cistanche tuaj yeem txhim kho kev nco, vim Cistanche yog cov khoom siv tshuaj suav tshuaj suav nrog ntau yam tshwj xeeb, ib qho ntawm kev txhim kho kev nco. Cov nyhuv ntawm Cistanche los ntawm ntau yam khoom xyaw uas nws muaj, suav nrog tannic acid, polysaccharides, flavonoid glycosides, thiab lwm yam. Cov khoom xyaw no tuaj yeem txhawb lub hlwb kev noj qab haus huv ntau txoj hauv kev.

Nyem paub 10 txoj hauv kev los txhim kho kev nco

Piv txwv li, hiPSC-derived neurons los ntawm tus neeg mob nrog LRRK2 GS2019S kev hloov pauv tau nthuav tawm cov kab mob ntsig txog cellular phenotypes, suav nrog kev ua kom muaj zog ntxiv rau oxidative stress170 thiab txo qis neurite ntev thiab tus lej.171

Lwm cov piv txwv suav nrog hiPSC-derived DA neurons tau los ntawm cov neeg mob uas muaj PARK2 kev hloov pauv uas nthuav tawm mitochondrial dysfunction thiab oxidative stress morphology172 thiab hiPSC-derived neurons nrog A53T kev hloov pauv thiab SNCA triplication uas nthuav tawm qhov pom kev nce siab thiab nce ntxiv ntawm 7-3.

Ntxiv mus, ntau qhov kev tshawb fawb tau kho cov kab mob phenotypes los ntawm kev cuam tshuam los kho cov noob hloov pauv, suav nrog LRRK2,174PINK1,175 SNCA, 140,176 thiab GBA1.177 Cov qauv zoo li no qhia txog kev siv hluav taws xob ntawm hiPSC thev naus laus zis hauv kev txheeb xyuas thiab kev lees paub ntawm cov tshuaj tshiab lub hom phiaj. rau gene therapyapproaches.

hiPSC-los ntawm cov qauv rau cov NDDs sib txawv txij li tau nthuav dav mus rau ntau qhov sib txawv ntawm kev coj noj coj ua 2D thiab 3D kev teeb tsa, nthuav dav cov qauv thiab muaj peev xwm tshawb fawb thiab kev siv ua ntej.

Tam sim no Configurations ntawm hiPSC-Drived NDD Models2D Ib leeg thiab Co-Culture Cell Models

Ib qho tseem ceeb ntawm kev ua haujlwm siv 2D kab lis kev cai suav nrog hiPSC-los ntawm ib hom xovtooj ntawm tes tau tshaj tawm nyob rau xyoo kaum xyoo dhau los. Cov qauv ntawm tes ib leeg no tau qhia txog kev nthuav tawm lub luag haujlwm ntawm cov tib neeg ntawm tes hauv NDDs, xws li cov leeg nqaij pob txha atrophy, 178,179 AD, PD, thiab amyolateral sclerosis (ALS) 180 (Daim duab 1), thiab ntxiv mus, tau ua kom muaj ntau dua. kev nkag siab txog yuav ua li cas cov kab mob cuam tshuam nrog cov noob ua haujlwm ntawm qib lom neeg los cuam tshuam cov cellular perturbations.

hiPSC cellular qauv tau raug xaiv nyob rau hauv AD kev tshawb fawb kom nkag siab zoo dua lub luag hauj lwm ntawm noob caj noob ces polymorphisms thiab kev hloov nyob rau hauv ib tug neeg cell cell thiab yuav ua li cas pab tau los yog cuam tshuam cov kab mob. Hauv ib qho hiPSC-based ADmodel, Kondo thiab al.181

pom tau tias nce Ab oligomer accumulation txuas rau nce endoplasmic reticulum dysfunction thiab oxidativestress nyob rau hauv hiPSC-derived neurons thiab astrocytes tau los ntawm sADand fAD cov neeg mob uas muaj APP E693D kev hloov. hiPSCderived 2D tus neeg ua qauv kuj tau pom muaj txiaj ntsig zoo hauv kev kawm cov caj ces ntawm APOEε4, qhov loj tshaj plaws thiab feem ntau cov kev pheej hmoo ntawm AD, hauv ntau hom neural cell qauv.182

Lin thiab cov npoj yaig183 tau siv 2D kab lis kev cai los nthuav tawm tias isogenic hiPSCderived APOEε4 astrocytes nthuav tawm cov kev hloov pauv loj hauv cov geneexpression hais txog cov roj cholesterol ntau ntxiv thiab sab hauv ntawm Ab42, ob qho tib si yog qhov tseem ceeb hauv homeostatic zog ntawm astrocytes, txheeb ze rau isogenic APOEε3 hlwb. Tsis tas li ntawd, isogenic ε4 astrocytes tau pom qhov txo qis hauv kev nthuav qhia thiab zais APOE, ua rau txo qis lipidation.

Txhim khu kev loj hlob ntawm neurite nyob rau hauv ob qho tib si lengthand tooj nyob rau hauv neurons, ntxiv rau txhim kho Ab secretion, inisogenic APOEε4 neurons kuj pom tau tias yog ib tug pathogenic pab nyob rau hauv cov qauv no.183

Hauv kev tsim cov kab lis kev cai astrocyte tau los ntawm cov tib neeg neurotypical isogenic rau APOEε3 thiab APOEε4alleles, Zhao et al.161 pom tias APOEε4 astrocytes nthuav tawm APOE lipidation qis dua piv nrog kev noj qab haus huv APOEε3 tswj, ntxiv qhov tseem ceeb ntawm kev pom tau los ntawm cov qauv zoo li no.

Kev siv 2D co-cultures kuj tau ua rau txoj kev tshawb fawb ntawm APOEε4 nyob rau hauv cov ntsiab lus ntawm cov tib neeg ntawm tes thiab kev sib cuam tshuam ntawm tes thiab yuav ua li cas cov no tuaj yeem ua rau AD pathogenesis.

Txuas ntxiv ntawm qhov kev sim saum toj no, raws li kev sib koom ua ke ntawm hiPSC-los ntawm APOEε4 astrocytes nrog cov neurons, morphological thiab kev ua haujlwm zoo ntawm cov neurons raug cuam tshuam thiab pom tias inhibition ntawm maturation piv rau lawv cov isogenic ε3 tswj.161

Kev sib cuam tshuam ntawm cov hlwb sib txawv neuronal thiab glial ua lub luag haujlwm tseem ceeb hauv txoj kev loj hlob thiab kev loj hlob ntawm AD thiab PD, yog li leveraging ntawm hiPSCtechnology los tsim cov qauv hauv vitro uas ua rau kev sib cuam tshuam ntawm tes-cell, thiab cellular microenvironment tau ua pov thawj tias cov ntaub ntawv muaj txiaj ntsig zoo.

Cov txiaj ntsig ntawm 2D tus qauv thiab cov qauv kev coj noj coj ua tau zoo sib xws hauv kev kawm txog kev koom tes ntawm ntau PD-ua rau kev hloov pauv mus rau kab mob ntsig txog cellular phenotypes, xws li txo qis cell viability thiab mitochondrial dysfunction.

Qhov no tau tshwm sim thawj zaug hauv kev tshawb fawb los ntawm Nguyen et al.,170 uas tau pom tias PD tus neeg mob tau txais cov qauv hiPSCneuronal nrog LRRK2 G2019S kev hloov pauv tau pom tias nce qib-syn, txhim kho cov noob koom nrog hauv oxidative stressresponse, nrog rau kev nce siab rau H2O{{ 6}}induced oxidative kev nyuaj siab thiab cell tuag.

Ib yam li ntawd, Sánchez-Danés et al.171 tau soj ntsuam kev hloov pauv hauv autophagy hauv DA neurons tau los ntawm PD tus neeg mob hiPSCslacking LRRK2 G2019S kev hloov pauv thiab pom qhov nce a-synand autophagic dysfunction.

Kev nkag siab ntau dua ntawm kev sib cuam tshuam ntawm cellular li cas underpin pathophysiology tau ua tiav nroghiPSC-derived co-culture qauv systems. Piv txwv li, hiPSCderived astrocytes tau pom tias muaj txiaj ntsig kev tiv thaiv hauv cocultured neural progenitor cells (NPCs) tau los ntawm PD cov neeg mob tom qab raug rau mitochondrial co toxins, rotenone thiab potassiumcyanide, pom los ntawm kev cawm ntawm kev sib txawv ntawm qhov tsis txaus thiab mitochondrial dysfunction.

Lwm txoj kev tshawb fawb siv neuron-astrocyte co-cultures los tshawb xyuas qhov cuam tshuam ntawm PD-ua rau LRRK2G2019S kev hloov pauv ntawm neuron-astrocyte crosstalk. Cov kws sau ntawv tau tshaj tawm tias kev sib koom ua ke ntawm kev tswj hwm hiPSC-derived DA neurons nrog cov neeg mob tau txais astrocytes nqa G2019S kev hloov pauv induced neurodegeneration thiab tsub zuj zuj ntawm a-syn hauv DA neurons.185

Txoj kev tshawb no tau ua piv txwv txog kev siv hluav taws xob ntawm kev sib koom ua ke hauv kev tshuaj xyuas qhov sib txawv ntawm cov noob sib txawv / kev hloov pauv los ntawm cov xovtooj ntawm tes los nthuav tawm cov xovtooj ntawm tes thiab cov xovtooj ntawm tes uas ua rau muaj kev cuam tshuam ntawm cov kab mob sib txawv.

Txawm li cas los xij, 2D co-cultures tsim microenvironment rau kev sib cuam tshuam ntawm cov hlwb sib txawv thiab tseem muaj kev txwv hauv lawv lub peev xwm los ua qauv kev nyuaj ntawm lub hlwb raws li cov ntaub so ntswg peb sab thiab xws li, muaj qhov cuam tshuam hauv kev txhais lus.

3D HiPSC-Drived Models

Cov thev naus laus zis tshiab tau ua rau cov kws tshawb fawb pib tsim thiab tshuaj xyuas 3D hiPSC-los ntawm cov qauv ntawm NDDs, suav nrog cov kab lis kev cai thiab organoids, uas yog ob qho tib si biochemically thiab physiologically robust.Organoid qauv. 3D tib neeg lub hlwb organoid qauv yog tsom mus ntes lub complex spatial lub koom haum ntawm tib neeg lub hlwb kom ze li sai tau nyob rau hauv ib tug nyob rau hauv vitro qauv.

Tau txais los ntawm hiPSCs, cov hlwb organoids yog tus kheej tsim thiab tus kheej-patterning 3D. Ntxiv rau lawv tag nrho cov epigenomic thiab transcriptional kos npe, 186,187 lub physiological zog ntawm 3D organoid qauv, xws li neuronalresponsiveness rau glutamate, instinctive Ca2+ signaling, thiab kev loj hlob ntawm cellular maturation phenotypes, kuj tau pom los ze ze iav tib neeg lub hlwb. .188–190

Cov kev tshawb fawb tsis ntev los no tau tsim thiab ua tus cwj pwm AD thiab PD 3D hiPSC-ua qauv los ntawm cov qauv, uas tau pom tias muaj cov yam ntxwv ntawm cov kab mob hauv lub cev zoo.

Piv txwv li, Gonzalez li al.191 tau tsim ib qho ADbrain organoid qauv uas nthuav tawm kev nce qib ntawm Ab peptide nrog cov plaque-zoo li cov qauv thiab amyloidogenic zog, nrog rau cov tsos ntawm phosphorylated tau thiab NFTs.

Linet al.183 nthuav dav ntawm cov kev tshawb pom no los ntawm kev qhia isogenicAPOEε3- lossis APOEε4- nqa cov hlwb zoo li microglia mus rau yav dhau los tsim cerebral organoids cultivated los ntawm hiPSCs uas muaj AD-associated APP duplication mutation thiab soj ntsuam cov txheej txheem ntev heev. nyob rau hauv APOEε4 microglia-zoo li cov hlwb piv rau APOEε3 tswj, qhia txog qhov muaj peev xwm tsawg hauv kev hnov ​​​​lus thiab kev muaj peev xwm teb rau extracellular Ab.

Tsis ntev los no, Zhao et al.192 tau tsim cerebral organoid qauv siv hiPSCs muab los ntawm cov neeg mob AD thiab kev noj qab haus huv tswj nqa cov genotypes APOE sib txawv. Tus qauv APOEε4 organoid tau nthuav tawm cov kab mob ntsig txog phenotypes piv rau APOEε3 organoid muab los ntawm ib tus neeg pub dawb thiab isogenic APOEε3 organoid qauv tsim los ntawm genome kho ntawm APOEε4 kab.

Qhov cuam tshuam tsis zoo ntawm APOEε4 genotype hauv cov qauv organoid recapitulated keyAD pathological nta, suav nrog kev poob hauv synaptic kev ncaj ncees, apoptotic cell tuag, Ab thiab phosphorylated tau tsub zuj zuj, thiab nce qib APOE soluble, yog li ua kom pom qhov tsim nyog ntawm lub kaw lus rau kev ua qauv tsis yog-fAD.

Tsis tas li ntawd, cov txiaj ntsig no muab kev txhawb nqa ntxiv rau lub luag haujlwm pathogenic ntawm theAPOEε4 hauv AD thiab nws lub peev xwm ua lub hom phiaj siv tshuaj rau AD.192 Ib yam li ntawd, Smitset al.193 tau tsim ib qho kev ua haujlwm midbrain DA organoid qauv ntawm PD tau los ntawm PD cov neeg mob cov ntaub so ntswg uas muaj LRRK GS2019 kev hloov.

Cov qauv organoid no tau tsim thiab zais DA thiab ua tiav tau nthuav tawm cov cim qhia txog pathologies hauv organoids, xws li qhov ntau thiab qhov nyuaj ntawm midbrain DA neuronss193Txawm li cas los xij, GT txoj hauv kev kho LRRK kev hloov pauv tsis txaus rau amelioration ntawm cov kab mob phenotypes hauv cov qauv no.

Bologninet al.194 kuj tau tsim ib tug 3D DA neuronal qauv muab los ntawm ib tug PDpatient nrog LRRK2 G2019S kev hloov, uas nthuav tawm cov cim ntawm mitochondrial tsis ua haujlwm thiab hloov morphology, txo DAdifferentiation thiab branching complexity, thiab elevated cellular kev tswj cov kab mob ua ntej. Qee qhov kev cuam tshuam no tau cawm los ntawm kev tswj hwm ntawm LRRK2 inhibitor 2 (Inh2).

Txawm li cas los xij, nws yog ib qho tseem ceeb uas yuav tsum nco ntsoov txog ntau yam kev txwv uas tau tshaj tawm hauv kev siv 3D organoid qauv, tshwj xeeb tshaj yog hais txog tus kheej thiab tsis tswj hwm ntawm cov cell loj hlob thiab kev sib txawv uas ua rau muaj kev sib txawv.

Qhov kev cia siab ntawm tus kheej ntawm kev sib sau ua ke hauv organoid qauv tuaj yeem ua rau tus qauv-rau- qauv hloov pauv, suav nrog cov qauv ntawm tes heterogenous, spatial orientation, thiab kab mob phenotype, ua rau kev rov tsim dua ib qho kev txhawj xeeb tseem ceeb hauv 3Dco-kev coj noj coj ua thiab organoid qauv.195

Kev nce qib tsis ntev los no yog, txawm li cas los xij, qhia txog kev txhim kho hauv kev rov ua dua tshiab hauv 3D organoid qauv, 196 qhov sib txawv hauv cov cim ntawm lub xov tooj ntawm tes tseem nyob, 195 thiab kev vam meej txuas ntxiv yog xav tau.

Co-Culture Systems. Kev nce qib biotechnological tsis ntev los no tau ua rau lub cim ntawm kev sib tw peb-dimensional scaffolding uas tso cai rau hiPSC-los ntawm cov hlwb kom loj hlob hauv cov qauv kev coj noj coj ua, uas ua rau muaj kev tswj xyuas ntau dua piv rau cov organoids tau tham saum toj no, tshwj xeeb yog hais txog celltype, maturation, muaj pes tsawg leeg, thiab homogeneous raug ntawm lub hlwb.197

Cov qauv kev coj noj coj ua uas muaj xws li cov qauv ntawm motorneurons thiab cov neuromuscular junction tau los ntawm cov neeg mob ALS; 180 DA neurons los ntawm PD cov neeg mob uas paub txog kev hloov pauv (piv txwv li, LRRK2194); tri-cellular qauv ntawm neurons, astrocytes, thiab microglia ua qauv rau cov neuroinflammatory AD ib puag ncig; 198 thiab cov leeg nqaij pob txha.199

Cov qauv no muab cov kws tshawb fawb nrog cov cuab yeej txhim kho los tshawb xyuas lub luag haujlwm ntawm cov ntaub so ntswg spatial-cell-cell thiab cell-matrix kev sib txuas, uas ua lub luag haujlwm tseem ceeb hauv NDD pathogenesis. Ib txoj kev tshawb fawb tsis ntev los no los ntawm Blanchard et al.200 tsim ib lub hiPSC-los ntawm 3D co-culture qauv ntawm BBB uas muaj cov hlwb endothelial hlwb, astrocytes, thiab pericytes tomodel AD vascular-txog pathologies nrog siab fidelity.

Qhov 3Dmodel no tau nthuav tawm lub luag haujlwm tseem ceeb ntawm pericytes hauv APOEε4- cov hlwb tau los, uas tau nthuav tawm kom nce BBB txuam nrog Ab peptides thiabfibril amyloid vim yog nce NFAT-calcineurin pericytesignaling.

200 Park et al.198 kuj tau tsim ib tug hiPSC 3D tri-cultureAD qauv uas muaj cov neurons, astrocytes, thiab microglia nyob rau hauv ib tug microfluidic platform, uas ntse recapitulates hallmark AD yam ntxwv, nrog rau tsub zuj zuj ntawm phosphorylated tau ntawmAb aggregates, raws li zoo raws li xws li neuroinflammatory nta. microglial recruitment, nitric oxide tso tawm, thiab tom qab oxidativestress.

Txawm hais tias 3D co-culture systems aims kom kov yeej cov kev txwv uas tau hais tseg hauv cov qauv organoid, lawv kuj muaj kev cuam tshuam. Tam sim no hiPSC-los ntawm cov qauv tsim cov kab mob neuronal uas tsis muaj cov yam ntxwv tseem ceeb ntawm lub hlwb cov ntaub so ntswg uas lawv lub hom phiaj los sawv cev, suav nrog qhov tsis muaj vascularization thiab xav tau kev ncig ntawm cov as-ham thiab tshem tawm cov khoom pov tseg hauv metabolic, 201 thiab qhov sib txawv ntawm cov neeg sawv cev ntawm cov neeg sawv cev ntawm cov neuronal thiab glial cell. pej xeem, nta uas yog arguable zog nyob rau hauv kev siv 3D organoidcultures.

Tsis tas li ntawd, 3D co-culture qauv xav tau cov khoom siv dag zog thiab kev sib txuas ntawm cov hlwb, thaum organoid qauv tsim ntau "naturally." Txawm li cas los xij, kev siv hiPSC-derivedco-cultures tsis tsuas yog tso cai rau cov kws tshawb fawb los tshawb xyuas lub luag haujlwm tseem ceeb ntawm txhua hom cell hauv tiam AD thiab PD tus yam ntxwv kab mob tab sis kuj muaj kev cuam tshuam ntawm cov hlwb tau los ntawm cov genotypes sib txawv, qhia txog cov cuab yeej hloov pauv uas hiPSCs sawv cev rau hauv. Themolecular kev tshawb fawb ntawm NDDs.

Qhov zoo thiab cov kev txwv ntawmhiPSC-raws li hauv vitro qauv yog tshawb nrhiav kom ntxaws ntxiv hauv ntu hauv qab no.

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