ABSTRACT

XFWVZKHWKHUGUXJVFRVPHWLFVRUEXONFKHPLFDOVDUHVXEMHFW DWDUHXVXDOO\SHUIRUPHGRQDQLPDOVWRGHWHUPLQHDQ\SRWHQWLDO QJWKHURXWLQHO\SHUIRUPHGWHVWVLVRQHGHVLJQHGWRHYDOXDWHWKH QWLDOWRFDXVHH\HLUULWDWLRQRULQMXU\WKH'UDL]HWHVW'UDL]HHW L]H WHVW DQG LWV QXPHURXV PRGLÀFDWLRQV KDYH UHPDLQHG WKH HYDOXDWLQJRFXODUWR[LFLW\IRUPRUHWKDQÀYHGHFDGHV6\PSR

,W KDV KRZHYHU EHHQ VXEMHFW WRPXFK FULWLFLVP IRU LWV WR DQLPDOV DV ZHOO DV WKH LUUHSURGXFLELOLW\ RI WKH VXEMHFWLYH KHUHIRUHWKHUHLVDJUHDWQHHGIRUPLQLPL]LQJWKHXVHRIDQLPDOV WHVWV DQG GHYHORSLQJ DPHFKDQLVWLFDOO\ EDVHG LQ YLWUR WHVWLQJ 3URFHHGLQJV

6HYHUDO SURPLVLQ RUDUHFXUUHQW .UXV]HZVNLHWDO IURPFHOOOLQHV*ULIÀ FHOOGHDWK-HVWHUHW RUJDQFXOWXUHV\VWHP FKHPLFDOV ;XHWDO DQGEHQ]DONRQLXPF SHUPHDELOLW\DQG'1 DQG 1)O% FDXVH GLIIHUHQWOHYHOVRIRF WLJKW MXQFWLRQV7- VXUHG E\ VXUIDFH EL GHWHUPLQHGE\WKH' LQGLFDWRUVRIFHOOXOD WUDWLRQVFDXVLQJPLQ ZKLFKLQGLFDWHVHSLW VHYHUH RFXODU LUULWDQ GHVWUXFWLRQRIHSLWKH RI FKHPLFDOLQGXFHG VWUHVVUHODWHGJHQHV DQLPDOWHVWLQJ

7RFRQÀUPWKHX LQDGRXEOHEOLQGP FDXVHG EUHDNGRZQ '1$ELQGLQJ DFWLY HIIHFWVRQFXOWXUHG DOWHUQDWLYHIRUWKH L

Corneal Organ C

%RYLQHH\HVZHU RQ LFH LQ DPRLVWXUH WKH OLPEDO FRQMXQFW EXIIHUHGVDOLQH3%6 VWHULOH ´FXSµ 7KH H HVVHQWLDOPHGLXP0 ODJHQPDLQWDLQHGDW VXSSRUWLQJ JHOZDV FXOWXUHPHGLXPDER XQWLOWKHOLPEDOFRQM )RUHPDQHWDO

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

VHIXOQHVVRIWKLVPRGHOZHHYDOXDWHGWKUHHKDLUFDUHSURGXFWV DQQHU:HIRXQGWKDWVKDPSRRVWKDWDUHPRGHUDWHO\LUULWDWLQJ RI WKH HSLWKHOLDO EDUULHU DQG DOWHUDWLRQ RI$3 DQG 1)O% LWLHV ZKLOH D PLQLPDOO\ LUULWDWLQJ KDLU FDUH SURGXFW KDG QR FRUQHDV7KHVHUHVXOWVLQGLFDWHWKHYDOXHRIWKHV\VWHPDVDQ QYLYR'UDL]HWHVW

METHODS

ulture

HREWDLQHGIURPDORFDODEDWWRLUDQGWUDQVSRUWHGWRWKHODERUDWRU\ FKDPEHU&RUQHDOVFOHUDO ULPVZLWK DSSUR[LPDWHO\PPRI LYD SUHVHQW ZHUH H[FLVHG DQG ULQVHG LQ VWHULOL]HG SKRVSKDWH 7KHH[FLVHGFRUQHDVZHUHSODFHGHSLWKHOLDOVLGHGRZQLQWRD QGRWKHOLDO FRUQHDO FRQFDYLW\ ZDV WKHQ ÀOOHG ZLWK PLQLPXP (0FRQWDLQLQJDJDURVHDQGPJPOUDW WDLO WHQGRQFRO

r&7KLVPL[WXUHZDVDOORZHGWRJHO&RUQHDDORQJZLWKLWV LQYHUWHG DQG WKHQ WUDQVIHUUHG WR D PP GLDPHWHU GLVK7KH XWPOZDVDGGHGGURSZLVHWRWKHVXUIDFHRIFHQWUDOFRUQHD XQFWLYDZDVFRYHUHGOHDYLQJWKHHSLWKHOLXPH[SRVHGWRWKHDLU ;XHWDO

Culture Treatment

7KUHH KDLU FDUH GLOXWHGZLWK0(0À FRUQHD$IWHUGLVFUHW GLSSLQJLQWR3%6VRO RUHSLWKHOLDOQXFOHDU

Surface Biotinyla

&XOWXUHGFRUQHDV 3LHUFH&KHPLFDOV5 &D&ODQGP00J ODEHOHGFRUQHDVZHUH DQGVHFWLRQHGRQDF FROG DFHWRQH DQG ODE VHUXPDOEXPLQ%6$ (FOLSVH(ÁXRUH

Electrophoretic M

7R SUHSDUHG FHOO FXOWXUHGERYLQHFRUQ ZLWKDVFDOSHOEODGH EXIIHUP0+HSH GLDPLQHWHWUDDFHWLFD WK\OVXOIRQ\OÁXRULGH r&IRUK3URWHLQ UHDJHQWNLW3LHUFH& &**$$IRU$3E &*$*$**IRU1) >K3@$73)RUGHWH ROLJRQXFOHRWLGHVZHU UHDFWLRQPL[WXUHZDV

:HKDYH XVHG D VXUIDFWDQWVDQGRXU DQGPHDVXUHPHQWVR VWUHVVUHVSRQVHWUDQV WREHXVHGDVDPHFK 7RYDOLGDWHWK WR[LFLW\RIWKUHHKDLU

SURGXFWV WHUPHG*$*% DQG*&ZHUH DSSOLHG GLUHFWO\ RU UVW DQG WKHQ DSSOLHG GURSZLVH RQWR WKH FHQWHU VXUIDFH RI WKH HWLPHLQWHUYDOVWKHWUHDWHGFRUQHDVZHUHZDVKHG²WLPHVE\ XWLRQDQGWKHQSURFHVVHGIRUHLWKHUFHOOVXUIDFHELRWLQODEHOLQJ H[WUDFWLRQ

tion-Tight Junction Permeability Assay

ZHUHZHWWHGZLWKIUHVKO\PDGHPJPOVXOIR1+6/&ELRWLQ RFNIRUG,/LQ+DQN·VEDODQFHGVDOWVROXWLRQFRQWDLQLQJP0 &O&KHQHWDO$IWHUDPLQLQFXEDWLRQWKHVXUIDFH ULQVHGZLWK3%6HPEHGGHGLQ2&7IUR]HQLQOLTXLGQLWURJHQ U\RVWDWDWDWKLFNQHVVRIQP7KHVHFWLRQVZHUHÀ[HGZLWKLFH HOHGZLWK UKRGDPLQHDYLGLQ' LQ3%6FRQWDLQLQJERYLQH IRUK7KHVOLGHVZHUHPRXQWHGDQGH[DPLQHGXQGHUD1LNRQ VFHQFHPLFURVFRSHHTXLSSHGZLWKD6SRWGLJLWDOFDPHUD

obility Shift Assay (EMSA)

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

RESULTS AND DISCUSSION

FRUQHDO RUJDQ FXOWXUHPRGHO WR DVVHVV HSLWKHOLDO UHVSRQVHV WR UHVXOWVLQGLFDWHGWKDWWKHFRPELQDWLRQRIFRUQHDORUJDQFXOWXUH

IFRUQHDOHSLWKHOLDOSHUPHDELOLW\DQG'1$ELQGLQJDFWLYLW\RI FULSWLRQIDFWRUVIROORZLQJVXUIDFWDQWH[SRVXUHKDVWKHSRWHQWLDO DQLVWLFDOO\EDVHGDOWHUQDWLYHWRLQYLYRDQLPDOWHVWLQJ;XHWDO HXVHIXOQHVVRIWKHV\VWHPLQWKLVUHSRUWZHHYDOXDWHGRFXODU FDUHSURGXFWVWHUPHG*$*%DQG*&

:HÀUVW DVVHVVH FDUHSURGXFWV LQFXO VHYHUDO OD\HUV RI FH PHPEUDQHOLHVLPPH WKHXQGHUO\LQJVWURP ÀEUREODVWV O\LQJ EH FRYDOHQWO\FURVVOLQN FRUQHD)LJXUH% ZDVFRQWLQXDOO\ODEH OD\HURIWKHFRUQHDH FRQGLWLRQ LV LPSHUP FXOWXUHG FRUQHDV SR PROHFXOHVDQGPLFUR

&KHPLFDOVFDXVL DQG FDXVH EUHDNGRZ REVHUYHGWKDWWKHVX DQG*%'UDL]HVFRU ZKHQWKH\ZH WKHFHQWHURIWKHFXO PLQ WR PLPLF LQ Y UHPRYHGTXLFNO\E\ *$IRUPLQUHVXOWH UHYHDOHG WKH ORVV R EUHDNGRZQRIHSLWKH DQG SHQHWUDWLRQ RI VXUIDFH ODEHOLQJ DQG ,QFXEDWLRQRIXQGLOX RIHSLWKHOLXPSULPD VWDLQLQJVXJJHVWHGQ

G GLVUXSWLRQ RI HSLWKHOLDO WLJKW MXQFWLRQ LQGXFHG E\ WKHVH KDLU WXUHGERYLQHFRUQHDV0DPPDOLDQFRUQHDOHSLWKHOLDFRQVLVWRI OOV )LJXUH $ *LSVRQ DQG 6XJUXH $ EDVHPHQW GLDWHO\EHQHDWKWKHEDVDOODPLQDRIWKHFRUQHDOHSLWKHOLXPDQG DLVFRPSRVHGRIPDQ\ODPHOODHRIFROODJHQÀEHUVZLWKÁDWWHQHG WZHHQ DGMDFHQW ODPHOODH 8VLQJ VXOIR1+6/&ELRWLQ WKDW VWRSURWHLQVZHGHPRQVWUDWHGWKHLPSHUPHDEOHQDWXUHRIWKH ,QQRUPDOFRUQHDRQO\WKHRXWHUPRVWVXUIDFHRIHSLWKHOLXP OHGZLWKELRWLQDVLQGLFDWHGE\DÁXRUHVFHQWOLQHRQWKHDSLFDO SLWKHOLXPZKLFKVXJJHVWVWKDWWKHFRUQHDXQGHUQRUPDOFXOWXUH HDEOH WR N' VXOIR1+6/&ELRWLQ 7KXV DV LQ YLYR WKH VVHVV D IXOO\ IXQFWLRQDO EDUULHU WKDW SUHYHQWV IUHH SDVVDJH RI RUJDQLVPVLQWRFHOOOD\HUV*RRGHQRXJK QJH\HLUULWDWLRQDUHOLNHO\WRDIIHFWFRUQHDOHSLWKHOLDO LQWHJULW\ Q RI EDUULHU IXQFWLRQ &RQVLVWHQW ZLWK WKLV K\SRWKHVLV ZH UIDFHODEHOLQJSDWWHUQLVDOWHUHGE\WZRKDLUFDUHSURGXFWV*$ HDQGUHVSHFWLYHO\PRGHUDWHO\LUULWDWLQJ*UDQWHWDO UHHLWKHUDSSOLHGGLUHFWO\RUGLOXWHGÀUVWDQGWKHQDSSOLHGRQWR WXUHGFRUQHDV)LJXUH:HFKRVHDVKRUWH[SRVXUHWLPH LYR DFFLGHQWDO H[SRVXUH WR RFXODU LUULWDQWV ZKLFK DUH XVXDOO\ WHDUVDQGEOLQNLQJ7UHDWPHQWRIFXOWXUHGFRUQHDZLWKQRQGLOXWHG GLQKHDY\ODEHOLQJRIWKHHQWLUHHSLWKHOLXPZKLOH(3VWDLQLQJ I HSLWKHOLDO FHOOV $W ORZHU FRQFHQWUDWLRQ *$ FDXVHG OLDOWLJKWMXQFWLRQHYLGHQFHGE\ODEHOLQJRIHSLWKHOLDOFHOOOD\HUV

ELRWLQ LQWR VWURPD 7KXV *$ SULPDULO\ FDXVHG GLVUXSWLRQ RI WKH LQWHQVLW\ RI GLVUXSWLRQ ZDV UHODWHG WR LWV FRQFHQWUDWLRQ WHG*%OHGWRWKHSHQHWUDWLRQRIELRWLQLQWRWKHXQGHUO\LQJOD\HUV ULO\WKHEDVDOOD\HUDQGIDLQWODEHOLQJRIWKHVWURPDZKLOH(3 R ORVVRIHSLWKHOLXP ,QFXEDWLRQRI*% IRUPLQ OHG WR

YLVLEOHGLVUXSWLRQRI WKHVWURPD8QOLNH* LUULWDWLQJH[KLELWHGP ZDVXVHGDQGQRDSS WRJHWKHU RXU UHVXOWV ODEHOLQJZDVFRQVLVWH PLQHG E\ WKH LQ YLY FRUQHDLVDQHIIHFWLY

VXUIDFHODEHOLQJDQGODEHOLQJRIPXOWLSOHHSLWKHOLDOOD\HUVDQG $DQG*%FRUQHDVH[SRVHGWR*&'UDL]HVFRUHPLQLPDOO\ LQLPDOGLVUXSWLRQRIVXUIDFHELRWLQ\ODWLRQZKHQXQGLOXWHG*&

DUHQWHIIHFWVZKHQGLOXWLRQZDVXVHG)LJXUH7DNHQ VXJJHVW WKDW KDLU FDUH SURGXFW²LQGXFHG DOWHUDWLRQ RI VXUIDFH QWZLWKLUULWDQF\SRWHQWLDORIWKHVHFRQVXPHUSURGXFWVDVGHWHU R'UDL]H WHVW )XUWKHUPRUH VXUIDFH ELRWLQ\ODWLRQ RI FXOWXUHG HZD\WRHYDOXDWH7-SHUPHDELOLW\DQGDIWHUDODUJHQXPEHURI

FKHPLFDOV ZLWK NQR VXUIDFWDQWEDVHGFRQ

:HDOVRDVVHVVWK FDUHSURGXFWVLQWKHF WRGLIIXVHLQWRGHHSH WLRQµDQGHSLWKHOLDO ORJLFDOO\ UHOHYDQWRX GLIIHUHQWLDOH[SUHVVLR WLRQIDFWRUVFRRUGLQD '1$VHTXHQFHVORFD 1)O% -DQVVHQ HW UHVSRQVH WUDQVFULSWL UHVSRQVH JHQHV DQG H[SUHVVLRQ ,Q WKLV FKDOOHQJH RI FXOWXUH DVVD\(06$ZLWK 1)O% '1$ELQGLQ FRUQHDV7KHUHZDV FHOOV,QFUHDVHLQ*$

ZQ RFXODU WR[LFLW\ DUH WHVWHG WR JUDGH WKH RFXODU WR[LFLW\ RI VXPHUSURGXFWVDQGRWKHUFKHPLFDOV HDOWHUDWLRQRI1)O%'1$ELQGLQJDFWLYLW\FDXVHGE\WKHKDLU RUQHDOHSLWKHOLDOFHOOV'LVUXSWLRQRI7-VZRXOGDOORZFKHPLFDOV UHSLWKHOLDOOD\HUVZKHUHVHQVHQHXURQVUHVLGHFDXVLQJ´LUULWD FHOOVWUHVVUHVSRQVHV,W LVJHQHUDOO\EHOLHYHGWKDWPRVWWR[LFR WFRPHVUHTXLUHQRWRQO\GLIIHUHQWLDOJHQH H[SUHVVLRQEXWDOVR QRIPXOWLSOHJHQHV)DUUDQG'XQQ$FWLYDWHGWUDQVFULS WHO\PRGXODWHWDUJHWJHQHH[SUHVVLRQYLDELQGLQJWRWKHVSHFLÀF WHGLQWKHSURPRWHURUHQKDQFHU$3.DULQHWDODQG DO 3LQNXV HW DO DUH ZHOOFKDUDFWHUL]HG VWUHVV RQ IDFWRUV NQRZQ WR WUDQVDFWLYDWH D ODUJH QXPEHU RI VWUHVV WKHUHIRUH ÀW WKH SURÀOH RI LQGLFDWRUV RI VWUHVVLQGXFHG JHQH VWXG\ 1)O% '1$ELQGLQJ DFWLYLW\ LQ UHVSRQVH WR FKHPLFDO G FRUQHDV ZDV PHDVXUHG XVLQJ HOHFWURSKRUHWLF PRELOLW\ VKLIW H[WUDFWVIURPHSLWKHOLDOFHOOV)LJXUHVKRZHGDOWHUDWLRQRI J DFWLYLW\ IROORZLQJ KDLU FDUH SURGXFW FKDOOHQJHV WR FXOWXUHG RQHPDMRUVKLIWHGEDQGREVHUYHGLQXQWUHDWHGFRUQHDOHSLWKHOLDO FRQFHQWUDWLRQOHGWRDGHFUHDVHLQ$3'1$ELQGLQJDFWLYLW\

LQDGRVHGHSHQGHQWP RIWKHFRQWURODQG WKHUPRUH D GH FRUQHDZDVH[SRVHG ELQGLQJ DFWLYLW\ EHW WRJHWKHUDOWKRXJK WK FRPSOH[ZHREVHUYH WRFRQVXPHUSURGXFW WKHOLDOFHOOVWRWKHVH 'UDL]HWHVW;XHWDO FXOWXUHGFRUQHDLQUHV YLDELOLW\ WKDW DIIHFWV VHFRQGDU\WRGHFUHDV D FDXVDO UHODWLRQVKLS ZKLFKVXJJHVWVWKDW1 WR[LFRORJLFDOHQGSR

%\FRPSDULVRQR DELOLW\ DQG$3 DQ VFRUHVDQ WKDWWKHGHJUHHVRIWK ZLWKWKHUHVXOWVRILQ

,Q VXPPDU\ RXU LUULWDQF\ SRWHQWLDO RI 8VLQJ WKUHHKDLUFDU VKDPSRRLQGXFHG7VXUHGE\VXUIDFHELRW PLQHG E\ WKH 'UDL]H FDXVLQJRFXODULUULWDQ GHVWUXFWLRQRIHSLWKH RI FKHPLFDOLQGXFHG VWUHVVUHODWHGJHQHV YLYR'UDL]HWHVWIRUH

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ÁXHQFH GLUHFWO\ RU HGQXPEHURIYLDEOHFHOOV1HYHUWKHOHVVRXUUHVXOWVGHPRQVWUDWH EHWZHHQ DOWHUDWLRQ RI 1)O% DFWLYLWLHV DQG RFXODU WR[LFLW\ )O%ELQGLQJDFWLYLW\PD\VHUYHDVDVSHFLÀFDQGTXDQWLÀDEOH LQWIRURFXODULUULWDWLRQSRWHQWLDORIWHVWFKHPLFDOV I*$*%DQG*&LQGXFHGDOWHUDWLRQV LQHSLWKHOLDOSHUPH G 1)O%'1$ ELQGLQJ DFWLYLW\ ZLWK WKH DYHUDJH 'UDL]H WHVW GIRU*$*%DQG*&UHVSHFWLYHO\WKHUHVXOWVLQGLFDWHG HDOWHUDWLRQVLQGXFHGE\WKHVHSURGXFWVDUHLQJRRGDJUHHPHQW YLYRWHVWLQJ DLP KDV EHHQ WR GHYHORS DQ H[ YLYRPRGHO WR DVVHVV RFXODU WHVW FKHPLFDOV XVLQJ D VLPSOH FRUQHDO RUJDQ FXOWXUH V\VWHP HSURGXFWV*$*%DQG*&ZHREVHUYHG WKDW WKHGHJUHHRI GLVUXSWLRQDQGWKHH[WHQWRIVPDOOPROHFXODUSHQHWUDWLRQPHD LQ\ODWLRQFRUUHODWHGWRLQYLYRLUULWDQF\PHDVXUHPHQWVDVGHWHU WHVW DQG ZH GHPRQVWUDWHG WKDW FKHPLFDOV DW FRQFHQWUDWLRQV F\LQGXFHGDGHFUHDVHLQ'1$ELQGLQJDFWLYLW\ZKLFKLQGLFDWHV OLXP7KXVFRUQHDORUJDQFXOWXUHFRXSOHGZLWKPHDVXUHPHQWV FRUQHDO HSLWKHOLDO EDUULHU GLVUXSWLRQ DQG WUDQVDFWLYDWLRQ RI UHSUHVHQWVDVXLWDEOHH[YLYRPRGHODVDQDOWHUQDWLYHWRWKH LQ YDOXDWLQJRFXODULUULWDWLRQSRWHQWLDOIRUFRQVXPHUSURGXFWV

REFERENCES