0008-5472.CAN-16-2795.full.pdf

ͳ (QHUJ\EDODQFHPRGXODWLRQLPSDFWVHSLJHQHWLFUHSURJUDPPLQJ(5ĮDQG ͳ (5ȕH[SUHVVLRQDQGPDPPDU\WXPRUGHYHORSPHQWLQ0079QHX ʹ WUDQVJHQLFPLFH ͵ Ͷ

(PLO\/5RVVL6DUDK0'XQODS/DXUD:%RZHUV6XEUHHQ$.KDWLE ͷ

6WHYHQ6'RHUVWOLQJ/DXUD$6PLWK1LNNL$)RUG'DUF\+ROOH\3RZHO+ ͸

%URZQ0DUFRV5(VWHFLR'RQQD).XVHZLWW/LQGD$GH*UDIIHQULHG6FRWW ͹

-%XOWPDQDQG6WHSKHQ'+XUVWLQJ ͺ

ͻ

*Contributed equally to the work ͳͲ ͳͳ _{'HSDUWPHQWRI1XWULWLRQ8QLYHUVLW\RI1RUWK&DUROLQD&KDSHO+LOO1&86$} ͳʹ _{/LQHEHUJHU&RPSUHKHQVLYH&DQFHU&HQWHU8QLYHUVLW\RI1RUWK&DUROLQD&KDSHO} ͳ͵ +LOO1&86$ ͳͶ _{'HSDUWPHQWRI1XWULWLRQDO6FLHQFHV8QLYHUVLW\RI7H[DV$XVWLQ7;86$} ͳͷ _{'HSDUWPHQWRI*HQHWLFV8QLYHUVLW\RI1RUWK&DUROLQD&KDSHO+LOO1&86$} ͳ͸ _{'HSDUWPHQWRI&OLQLFDO&DQFHU3UHYHQWLRQ8QLYHUVLW\RI7H[DV0'$QGHUVRQ} ͳ͹ &DQFHU&HQWHU+RXVWRQ7;86$ ͳͺ _{'HSDUWPHQWRI%UHDVW0HGLFDO2QFRORJ\'LYLVLRQRI&DQFHU0HGLFLQH8QLYHUVLW\} ͳͻ RI7H[DV0'$QGHUVRQ&DQFHU&HQWHU+RXVWRQ7;86$ ʹͲ _{'HSDUWPHQWRI0ROHFXODU&DUFLQRJHQHVLV8QLYHUVLW\RI7H[DV0'$QGHUVRQ} ʹͳ &DQFHU&HQWHU6FLHQFH3DUN6PLWKYLOOH7;86$ ʹʹ ʹ͵ .H\ZRUGV%UHDVWFDQFHU'1$PHWK\ODWLRQHSLJHQHWLFVKRUPRQHUHFHSWRUVDQG ʹͶ GLDJQRVLVSURJQRVLVSUHFOLQLFDOVWXGLHVRIHQGRFULQHUHODWHGFDQFHUVUHFHSWRUV ʹͷ DQGVLJQDOWUDQVGXFWLRQDQLPDOWUDQVJHQLFPRGHOVLQSURPRWLRQDQGSUHYHQWLRQ ʹ͸ ELRORJLFDODQGELRFKHPLFDOPHFKDQLVPVLQSUHYHQWLRQGLHWDQGFDQFHUPROHFXODU ʹ͹ PDUNHUVLQSUHYHQWLRQUHVHDUFK ʹͺ ʹͻ )LQDQFLDO6XSSRUW7KLVVWXG\ZDVVXSSRUWHGE\WKH$PHULFDQ,QVWLWXWHIRU ͵Ͳ &DQFHU5HVHDUFK$%UHDVW&DQFHU5HVHDUFK)RXQGDWLRQDQG5 ͵ͳ &$IURPWKH1DWLRQDO&DQFHU,QVWLWXWH7KHPRXVH51$VHTDQG ͵ʹ PHWK\ODWLRQDQDO\VLVZDVVXSSRUWHGE\WKH870'$QGHUVRQ&DQFHU&HQWHU ͵͵ 6FLHQFH3DUN1*6&RUHVXSSRUWHGE\&35,7&RUH)DFLOLW\6XSSRUW*UDQW ͵Ͷ 53(/5RVVLDQG/:%RZHUVZHUHVXSSRUWHGE\DWUDLQLQJJUDQWIURP ͵ͷ WKH1DWLRQDO&DQFHU,QVWLWXWH5&$60'XQODSZDVVXSSRUWHGE\D ͵͸ 86$050&%UHDVW&DQFHU5HVHDUFK3URJUDP3RVWGRFWRUDO)HOORZVKLS ͵͹ :;:+DQG1$)RUGZDVVXSSRUWHGE\DQ$PHULFDQ,QVWLWXWHIRU ͵ͺ &DQFHU5HVHDUFK3RVWGRFWRUDO)HOORZVKLS ͵ͻ ͶͲ

&RUUHVSRQGLQJ$XWKRU 6WHSKHQ'+XUVWLQJ3K'03+7KH8QLYHUVLW\RI

ʹ

:RUGFRXQWH[FOXGLQJUHIHUHQFHVDQGILJXUHOHJHQGVLQFOXGLQJ

Ͷ͸

ILJXUHOHJHQGV Ͷ͹

Ͷͺ

)LJXUHV

Ͷͻ ͷͲ

6XSSOHPHQWDU\)LJXUHV

ͷͳ ͷʹ

7DEOHV

ͷ͵ ͷͶ

6XSSOHPHQWDU\7DEOHV

ͷͷ ͷ͸

5HIHUHQFHV

ͷ͹ ͷͺ

5XQQLQJ7LWOH(SLJHQHWLFVHQHUJ\EDODQFHDQGPDPPDU\FDQFHU

ͷͻ ͸Ͳ

͸ͳ

͸ʹ

͸͵

͸Ͷ

͸ͷ

͸͸

͸͹

͸ͺ

͸ͻ

͹Ͳ

͹ͳ

͹ʹ

͹͵

͹Ͷ

͵

$%675$&7 ͹͸

7KHDVVRFLDWLRQEHWZHHQREHVLW\DQGEUHDVWFDQFHUULVNDQGSURJQRVLVLVZHOO ͹͹

HVWDEOLVKHGLQ(5SRVLWLYHGLVHDVHEXWOHVVFOHDULQ+(5SRVLWLYHGLVHDVH+HUH ͹ͺ

ZHUHSRUWSUHFOLQLFDOHYLGHQFHVXJJHVWLQJZHLJKWPDLQWHQDQFHWKURXJKFDORULH ͹ͻ

UHVWULFWLRQPD\OLPLWULVNRI+(5SRVLWLYHEUHDVWFDQFHU,QIHPDOH0079 ͺͲ

+(5QHXWUDQVJHQLFPLFHZHIRXQGWKDW(5ĮDQG(5βH[SUHVVLRQPDPPDU\ ͺͳ

WXPRULJHQHVLVDQGVXUYLYDODUHHQHUJ\EDODQFHGHSHQGHQWLQDVVRFLDWLRQZLWK ͺʹ

HSLJHQHWLFUHSURJUDPPLQJ0LFHZHUHUDQGRPL]HGWRUHFHLYHDFDORULHUHVWULFWLRQ ͺ͵

&5RYHUZHLJKW2:LQGXFLQJRUGLHWLQGXFHGREHVLW\',2UHJLPHQn

ͺͶ

JURXS6XEVHWVRIPLFHn JURXSWLPHSRLQWZHUHHXWKDQL]HGDIWHU

ͺͷ

DQGPRQWKVWRFKDUDFWHUL]HGLHWGHSHQGHQWPHWDEROLFWUDQVFULSWLRQDODQG ͺ͸

HSLJHQHWLFSHUWXUEDWLRQV5HPDLQLQJPLFHZHUHIROORZHGXSWRPRQWKV ͺ͹

5HODWLYHWRWKH2:DQG',2UHJLPHQV&5GHFUHDVHGERG\ZHLJKWDGLSRVLW\ ͺͺ

DQGVHUXPPHWDEROLFKRUPRQHVDVH[SHFWHGDQGDOVRHOLFLWHGDQLQFUHDVHLQ ͺͻ

PDPPDU\(5ĮDQG(5βH[SUHVVLRQ,QFUHDVHG'1$PHWK\ODWLRQDFFRPSDQLHG ͻͲ

WKLVSDWWHUQSDUWLFXODUO\DW&S*GLQXFOHRWLGHVORFDWHGZLWKLQELQGLQJRUIODQNLQJ ͻͳ

UHJLRQVIRUWKHWUDQVFULSWLRQDOUHJXODWRU&&&7&ELQGLQJIDFWRU&7&)RIESR1

ͻʹ

DQGESR2,FRQVLVWHQWZLWKVXVWDLQHGWUDQVFULSWLRQDODFWLYDWLRQRI(5ĮDQG(5β

ͻ͵

0DPPDU\H[SUHVVLRQRIWKH'1$PHWK\ODWLRQHQ]\PHDNMT1ZDVVWDEOHLQ&5

ͻͶ

PLFHEXWLQFUHDVHGRYHUWLPHLQ2:DQG',2PLFHVXJJHVWLQJ&5REYLDWHV ͻͷ

HSLJHQHWLFDOWHUDWLRQVFRQFXUUHQWZLWKFKURQLFH[FHVVHQHUJ\LQWDNH,QWKH ͻ͸

VXUYLYDOVWXG\&5HOLFLWHGDVLJQLILFDQWVXSSUHVVLRQLQVSRQWDQHRXVPDPPDU\ ͻ͹

Ͷ UHYHUVHH[FHVVERG\ZHLJKWDVDVWUDWHJ\WRUHGXFH+(5SRVLWLYHEUHDVWFDQFHU ͻͻ

ULVN ͳͲͲ

ͷ

,1752'8&7,21

ͳͲʹ

%UHDVWFDQFHULVWKHPRVWIUHTXHQWO\GLDJQRVHGQRQFXWDQHRXVQHRSODVP ͳͲ͵

DPRQJZRPHQLQWKH8QLWHG6WDWHVDQGLVWKHOHDGLQJFDXVHRIFDQFHUGHDWK ͳͲͶ

DPRQJZRPHQZRUOGZLGH$SSUR[LPDWHO\RIDOOEUHDVWFDQFHUVODFN ͳͲͷ

HVWURJHQUHFHSWRUDOSKD(5ĮZKLFKFRQIHUVDZRUVHSURJQRVLVLQFRPSDULVRQ ͳͲ͸

WR(5ĮSRVLWLYHEUHDVWWXPRUV)XUWKHUPRUHWKHH[SUHVVLRQRIHVWURJHQ ͳͲ͹

UHFHSWRUEHWD(5ȕDSXWDWLYHWXPRUVXSSUHVVRULVDOVRORVWLQPRVW(5Į ͳͲͺ

QHJDWLYHEUHDVWWXPRUV(5ȕLVWKHPRUHSUHYDOHQW(5LQQRUPDOPDPPDU\ ͳͲͻ

WLVVXHEXWLWVH[SUHVVLRQLVUHGXFHGGXULQJWXPRUIRUPDWLRQ1XPHURXVVWXGLHV ͳͳͲ

KDYHOLQNHGJUHDWHUEUHDVWWXPRU(5ȕH[SUHVVLRQZLWKDQLPSURYHGSURJQRVLV ͳͳͳ

+RZHYHUWKHGHJUHHWRZKLFK(5ĮDQG(5ȕH[SUHVVLRQLVLPSDFWHGE\ ͳͳʹ

GLHWDU\HQHUJ\EDODQFHDQGRUFRQWUROOHGHSLJHQHWLFDOO\LQEUHDVWWXPRULJHQHVLV ͳͳ͵

UHPDLQVXQFOHDU ͳͳͶ

2EHVLW\DUHVXOWRIFKURQLFSRVLWLYHGLHWDU\HQHUJ\EDODQFHLVDQ ͳͳͷ

HVWDEOLVKHGULVNIDFWRUIRUSRVWPHQRSDXVDOEUHDVWFDQFHUDQGPD\DOVRHQKDQFH ͳͳ͸

ULVNLQSUHPHQRSDXVDOZRPHQZLWKDGGLWLRQDOEUHDVWFDQFHUULVNIDFWRUVLQFOXGLQJ ͳͳ͹

W\SHGLDEHWHV,QDGGLWLRQH[FHVVHQHUJ\LQWDNHDQGLQFUHDVHGDGLSRVLW\ ͳͳͺ

KDYHEHHQDVVRFLDWHGZLWKJUHDWHUEUHDVWWXPRUVL]HLQFUHDVHGSURJUHVVLRQ ͳͳͻ

PDUNHUVDQGWKHUDS\UHVLVWDQFHLQERWKSUHDQGSRVWPHQRSDXVDOZRPHQ ͳʹͲ

,QFRQWUDVWWKHPDLQWHQDQFHRIDQHJDWLYHHQHUJ\EDODQFHYLDFDORULHUHVWULFWLRQ ͳʹͳ

&5SUHYHQWVZHLJKWJDLQDQGLQKLELWVWKHGHYHORSPHQWRIVHYHUDOW\SHVRI ͳʹʹ

FDQFHULQFOXGLQJ(5SRVLWLYHDQG(5QHJDWLYHEUHDVWFDQFHUVLQQXPHURXV ͳʹ͵

͸ VHUXPDQGWLVVXHELRPDUNHUVLQZRPHQFDXVDOO\OLQNHGZLWKUHGXFHGEUHDVW ͳʹͷ

FDQFHUULVN ͳʹ͸

$OWKRXJKOLQNVEHWZHHQHQHUJ\PHWDEROLVPHSLJHQHWLFUHJXODWLRQRIJHQH ͳʹ͹

H[SUHVVLRQDQGVHYHUDOFKURQLFGLVHDVHVKDYHEHHQSUHYLRXVO\HVWDEOLVKHGWKH ͳʹͺ

UHODWLRQVKLSEHWZHHQGLHWDU\HQHUJ\EDODQFHHSLJHQHWLFVDQGEUHDVWFDQFHULV ͳʹͻ

SRRUO\XQGHUVWRRG'1$PHWK\ODWLRQOHYHOVDUHUHJXODWHGLQSDUWE\'1$ ͳ͵Ͳ

F\WRVLQHPHWK\OWUDQVIHUDVHDNMT1ZKLFKSUHGRPLQDQWO\VHUYHVWR

ͳ͵ͳ

PDLQWDLQJHQRPLF'1$PHWK\ODWLRQGXULQJ'1$UHSOLFDWLRQ7KXVGXULQJ ͳ͵ʹ

WLPHVRIKLJKFHOOSUROLIHUDWLRQVXFKDVGXULQJGHYHORSPHQWDNMT1LVKLJKO\

ͳ͵͵

H[SUHVVHG+RZHYHUDNMT1FDQEHFRPHGHUHJXODWHGWKURXJKRXWWKHOLIHFRXUVH

ͳ͵Ͷ

LQUHVSRQVHWRPHWDEROLFLQIODPPDWRU\DQGHQYLURQPHQWDOGLVWXUEDQFHVDQGWKLV ͳ͵ͷ

G\VUHJXODWLRQKDVEHHQOLQNHGZLWKDEHUUDQW'1$PHWK\ODWLRQDQGFDQFHU ͳ͵͸

ͳ͵͹

7KHHIIHFWVRI'1$PHWK\ODWLRQDUHGHSHQGHQWXSRQWKHORFDWLRQRIPHWK\O ͳ͵ͺ

JURXSVLQWKHJHQRPLFODQGVFDSH,QJHQHUDOSURPRWHUPHWK\ODWLRQUHVXOWVLQ ͳ͵ͻ

WUDQVFULSWLRQDOO\VLOHQWJHQHV+RZHYHUPHWK\ODWLRQLQWKHWUDQVFULSWLRQUHJLRQRI ͳͶͲ

JHQHVLVRIWHQSRVLWLYHO\FRUUHODWHGZLWKJHQHH[SUHVVLRQ,QDGGLWLRQ ͳͶͳ

PHWK\ODWLRQWKDWSUHYHQWVDUHSUHVVRUIURPELQGLQJ'1$FDQFRUUHVSRQGWR ͳͶʹ

LQFUHDVHGJHQHH[SUHVVLRQ_{&&&7&ELQGLQJIDFWRU&7&)LVDQ]LQF} ͳͶ͵

ILQJHUSURWHLQDQGKLJKO\FRQVHUYHGWUDQVFULSWLRQIDFWRUZLWKHQKDQFHUEORFNLQJ ͳͶͶ

DFWLYLW\'1$PHWK\ODWLRQDW&S*¶&SKRVSKDWH*¶GLQXFOHRWLGHVLV ͳͶͷ

LQYHUVHO\FRUUHODWHGZLWK&7&)RFFXSDQF\:HSRVLWWKDWWKHGLHW ͳͶ͸

͹ JHQHWUDQVFULSWLRQYLDHQHUJ\EDODQFHDVVRFLDWHGFKDQJHVLQ'1$PHWK\ODWLRQDW ͳͶͺ

RUQHDU&7&)ELQGLQJVLWHV ͳͶͻ

7KHJHQHWLFDOO\HQJLQHHUHG0RXVH0DPPDU\7XPRU9LUXV0079QHX ͳͷͲ

PRXVHPRGHOLVFKDUDFWHUL]HGE\PDPPDU\JODQGRYHUH[SUHVVLRQRIWKH ͳͷͳ

RQFRJHQHKXPDQHSLGHUPDOJURZWKIDFWRUUHFHSWRU+(50079QHXPLFH ͳͷʹ

LQLWLDOO\KDYHKLVWRORJLFDOO\QRUPDO(5ĮSRVLWLYHPDPPDU\JODQGVWKDW ͳͷ͵

VXEVHTXHQWO\GHYHORSUHJLRQVRIGXFWDOFDUFLQRPDin situZLWK(5ĮH[SUHVVLRQ

ͳͷͶ

ORVWLQPRVWFHOOV,IXQWUHDWHGWKHVHPLFHXOWLPDWHO\GHYHORS(5ĮQHJDWLYH ͳͷͷ

+(5SRVLWLYHPDPPDU\DGHQRFDUFLQRPDVEHIRUHPRQWKVRIDJH,QWKH ͳͷ͸

SUHVHQWVWXG\ZHWHVWHGWKHK\SRWKHVLVWKDWGLHWDU\HQHUJ\EDODQFHPRGXODWLRQ ͳͷ͹

DOWHUV(5ĮDQGRU(5βH[SUHVVLRQ'1$PHWK\ODWLRQDQGWXPRULQFLGHQFHLQ ͳͷͺ

IHPDOH0079QHXPLFH ͳͷͻ

ͳ͸Ͳ

0$7(5,$/6$1'0(7+2'6

ͳ͸ͳ

In vivoVWXGLHVLQ0079QHXWUDQVJHQLFPLFH ͳ͸ʹ

$OODQLPDOVWXGLHVDQGSURFHGXUHVZHUHDSSURYHGDQGPRQLWRUHGE\WKH ͳ͸͵

8QLYHUVLW\RI7H[DV,QVWLWXWLRQDO$QLPDO&DUHDQG8VH&RPPLWWHH)HPDOHWR ͳ͸Ͷ

ZHHNROG0079QHXPLFH-$;VWRFNn ZHUHSXUFKDVHGIURP

ͳ͸ͷ

-DFNVRQ/DERUDWRU\%DU+DUERU0(86$DQGIHGDPRGLILHG$,1* ͳ͸͸

VHPLSXULILHGGLHWGHILQHGDVWKHRYHUZHLJKW2:LQGXFLQJGLHWIRUWKLVVWXG\ ͳ͸͹

FDWDORJ'%5HVHDUFK'LHWV,QF1HZ%UXQVZLFN1-86$ ad libitum

ͳ͸ͺ

IRUZHHNRIDFFOLPDWLRQ ͳ͸ͻ

ͺ

%DVHOLQHPLFH)ROORZLQJDFFOLPDWLRQDVXEVHWRIPLFHn ZHUHIDVWHGIRU

ͳ͹ͳ

KRXUVDQGWKHQHXWKDQL]HGE\&2DVSK\[LDWLRQIROORZHGE\FHUYLFDOGLVORFDWLRQ ͳ͹ʹ

%ORRGZDVFROOHFWHGE\FDUGLDFSXQFWXUHDOORZHGWRFRDJXODWHIRUPLQXWHVDW ͳ͹͵

URRPWHPSHUDWXUHDQGFHQWULIXJHGDW[gIRUPLQXWHVVHUXPZDV

ͳ͹Ͷ

UHPRYHGDQGVWRUHGDWR&IRUVXEVHTXHQWDQDO\VHV0DPPDU\WLVVXHVZHUH ͳ͹ͷ

FROOHFWHGIRUIXUWKHUPROHFXODUDQGSDWKRORJLFDODQDO\VLV ͳ͹͸

ͳ͹͹

7LPHSRLQWVWXG\$VXEVHWRIPLFHZHUHVLQJO\KRXVHGDQGUDQGRPL]HG

ͳ͹ͺ

n GLHWJURXSWRRQHRIWKHIROORZLQJWKUHHGLHWWUHDWPHQWJURXSVHDFK

ͳ͹ͻ

PRGLILHGIURPWKH2:GLHWZKLFKLV$,1*VHPLSXULILHGGLHWIRUPXODWLRQIRUD ͳͺͲ

PRQWKWLPHSRLQWVWXG\FDORULHUHVWULFWLRQUHJLPHQ&5DORZIDWORZ ͳͺͳ

FDUERK\GUDWHGLHW'2:GLHWDKLJKFDUERK\GUDWHORZIDWGLHW ͳͺʹ

SURYLGLQJNFDOJ'%RUGLHWLQGXFHGREHVLW\UHJLPHQ',2D ͳͺ͵

KLJKFDUERK\GUDWHKLJKIDWGLHWSURYLGLQJNFDOJ'DOOIURP ͳͺͶ

5HVHDUFK'LHWV,QF&5PLFHZHUHDGPLQLVWHUHGWKHLUGLHWIRUPXODWLRQDVGDLO\ ͳͺͷ

DOLTXRWVRIIRRGWKDWSURYLGHGRIWKHNFDOEXWRIWKHYLWDPLQVPLQHUDOV ͳͺ͸

HVVHQWLDOIDWW\DFLGVDQGDPLQRDFLGVUHODWLYHWRWKH2:JURXS0LFHZHUH ͳͺ͹

ZHLJKHGZHHNO\$IWHUDQGPRQWKVRQGLHWPLFHZHUHDQDO\]HGIRU ͳͺͺ

SHUFHQWERG\IDWXVLQJTXDQWLWDWLYHPDJQHWLFUHVRQDQFHVSHFWURVFRS\(FKR ͳͺͻ

0HGLFDO6\VWHPV+RXVWRQ7;86$$WHDFKRIWKHVHVDPHWLPHSRLQWVPLFH ͳͻͲ

SHUGLHWJURXSZHUHNLOOHGDQGVHUXPDQGQRQWXPRUEHDULQJPDPPDU\WLVVXH ͳͻͳ

ZHUHFROOHFWHGDVGHVFULEHGDERYH1RWXPRUVGHYHORSHGLQDQ\PLFHLQWKHWLPH ͳͻʹ

ͻ

ͳͻͶ

6XUYLYDOVWXG\7KHUHPDLQLQJVXEVHWRIPLFHZHUHVLQJO\KRXVHGDQG

ͳͻͷ

UDQGRPL]HGn GLHWJURXSWRWKH&52:RU',2GLHWUHJLPHQVDQGZHUH

ͳͻ͸

IROORZHGIRUVXUYLYDOIRUXSWRPRQWKV7KHVXUYLYDOFXUYHIRUHDFKGLHWJURXS ͳͻ͹

LOOXVWUDWHVWLPHWRHYHQWGDWDZLWKWKHHYHQWFRQVLVWLQJRIHLWKHUGHDWKRUWKH ͳͻͺ

SUHVHQFHRIDPDPPDU\WXPRU!FPLQDQ\GLUHFWLRQWKH,$&8&DSSURYHG ͳͻͻ

PD[LPDOWXPRUVL]H1RQPDPPDU\WXPRUUHODWHGGHDWKVZHUHFHQVRUHG ʹͲͲ

0LFHZHUHSDOSDWHGIRUPDPPDU\WXPRUVZHHNO\2QFHGHWHFWHGWXPRU ʹͲͳ

GLDPHWHUVZHUHPHDVXUHGLQWZRGLPHQVLRQVWZLFHZHHNO\ZLWKHOHFWURQLF ʹͲʹ

FDOLSHUV:KHQWXPRUGLDPHWHUUHDFKHGFPLQHLWKHUGLPHQVLRQRUDIWHU ʹͲ͵

PRQWKVRIVWXG\LQWKHDEVHQFHRIWXPRUPLFHZHUHNLOOHG7XPRUDQGRUGLVWDO ʹͲͶ

PDPPDU\WLVVXHZHUHFROOHFWHGDQGSURFHVVHG2QHKDOIRIHDFKFROOHFWHGWLVVXH ʹͲͷ

VDPSOHZDVIL[HGLQQHXWUDOEXIIHUHGIRUPDOLQIRUKRXUVWUDQVIHUUHGWR ʹͲ͸

HWKDQROIRUDWOHDVWKRXUVHPEHGGHGLQSDUDIILQDQGFXWLQWR—PWKLFN ʹͲ͹

VHFWLRQVIRUKHPDWR[\OLQDQGHRVLQ+ (VWDLQLQJRULPPXQRKLVWRFKHPLFDO ʹͲͺ

DQDO\VLV7KHRWKHUKDOIZDVSODFHGLQDFU\RWXEHIODVKIUR]HQLQOLTXLGQLWURJHQ ʹͲͻ

DQGVWRUHGDWR&IRUVXEVHTXHQWPROHFXODUDQDO\VHV%ORRGZDVFROOHFWHGE\ ʹͳͲ

FDUGLDFSXQFWXUHDQGVHUXPZDVLVRODWHGIRUDQDO\VLV ʹͳͳ

ʹͳʹ

$QDO\VHVRIFLUFXODWLQJHQHUJ\EDODQFHUHODWHGKRUPRQHVDQGȕHVWUDGLRO

ʹͳ͵

6HUXPVDPSOHVIURPDOOPLFHDWEDVHOLQHDQGDOOPLFHSHUGLHWJURXS ʹͳͶ

DWWKHDQGPRQWKWLPHSRLQWVZHUHFROOHFWHGDIWHUPLFHZHUHIDVWHGWR ʹͳͷ

ͳͲ 6HUXPVDPSOHVZHUHDQDO\]HGIRUOHSWLQLQVXOLQLQVXOLQOLNHJURZWKIDFWRU,*) ʹͳ͹

DQGDGLSRQHFWLQE\/XPLQH[EDVHGEHDGDUUD\DVVD\0LOOLSRUH%LOOHULFD0$ ʹͳͺ

86$UHDGRQ0DJ3L[PXOWLDQDO\WHGHWHFWLRQV\VWHP%LR5DG+HUFXOHV&$ ʹͳͻ

86$7KHPHDQLQWHUDVVD\FRHIILFLHQWRIYDULDWLRQ&9RIPXOWLSOH[HGEHDG ʹʹͲ

EDVHGDVVD\VIRUPHWDEROLFKRUPRQHGHWHFWLRQKDVEHHQVKRZQWREHLQ ʹʹͳ

SXEOLVKHGVWXGLHV6HUXPȕHVWUDGLROZDVPHDVXUHGE\(/,6$$OSKD ʹʹʹ

'LDJQRVWLFV6DQ$QWRQLR7;86$ ʹʹ͵

ʹʹͶ

5HDOWLPHTXDQWLWDWLYHUHYHUVHWUDQVFULSWLRQT573&5DQDO\VHVRI(5Į

ʹʹͷ

DQG(5ș

ʹʹ͸

7RWDO51$ZDVH[WUDFWHGXVLQJ75,5HDJHQW6LJPD$OGULFK6W/RXLV ʹʹ͹

0286$DFFRUGLQJWRPDQXIDFWXUHU¶VLQVWUXFWLRQVIURPWKHIODVKIUR]HQ ʹʹͺ

PDPPDU\WLVVXHVDPSOHVFROOHFWHGDWEDVHOLQHn PLFHDQGHDFKRIWKH

ʹʹͻ

WLPHSRLQWVn PLFHGLHWJURXSWLPHSRLQW51$ZDVDOVRH[WUDFWHGIURP

ʹ͵Ͳ

QRQWXPRUEHDULQJIODVKIUR]HQPDPPDU\WLVVXHFROOHFWHGIURPPLFHLQWKH ʹ͵ͳ

VXUYLYDOVWXG\XSRQWKHLUWHUPLQDWLRQEHWZHHQDQGPRQWKV51$ ʹ͵ʹ

FRQFHQWUDWLRQZDVVSHFWURSKRWRPHWULFDOO\GHWHUPLQHGXVLQJDQDQRGURS7KHUPR ʹ͵͵

6FLHQWLILF/RJDQ8786$DQGTXDOLW\ZDVFRQILUPHGXVLQJDQ$JLOHQW ʹ͵Ͷ

%LRDQDO\]HU6DQWD&ODUD&$86$51$ZDVUHYHUVHWUDQVFULEHGZLWK ʹ͵ͷ

0XOWLVFULEH57$SSOLHG%LRV\VWHPV&DUOVEDG&$86$DQGUHVXOWLQJF'1$ ʹ͵͸

ZHUHDVVD\HGLQWULSOLFDWHXVLQJ7DTPDQŠ*HQH([SUHVVLRQ$VVD\VIRU(5Į ʹ͵͹

(5ȕDQGDNMT1$SSOLHG%LRV\VWHPV3&5UHDFWLRQVZHUHPRQLWRUHGE\D

ʹ͵ͺ

ͳͳ ZHUHQRUPDOL]HGWRWKHKRXVHNHHSLQJJHQHȕDFWLQDQGDQDO\]HGXVLQJWKHGHOWD ʹͶͲ

GHOWDF\FOHWKUHVKROGPHWKRG ʹͶͳ

ʹͶʹ

+LVWRSDWKRORJLFDQGLPPXQRKLVWRFKHPLFDODQDO\VHV

ʹͶ͵

7XPRUVZHUHH[DPLQHGIRUKLVWRSDWKRORJLFDOPDUNHUVRIWXPRUSURJUHVVLRQ ʹͶͶ

LQFOXGLQJYDVFXODULW\SUHVHQFHRIEORRGYHVVHOVDQGSUROLIHUDWLRQQXPEHURI ʹͶͷ

PLWRWLFILJXUHVSHUILHOGLQ+ (VHFWLRQVE\DERDUGFHUWLILHGYHWHULQDU\ ʹͶ͸

SDWKRORJLVW9DVFXODULW\ZDVJUDGHGLQDEOLQGHGIDVKLRQRQDFDWHJRULFDOVFRUH ʹͶ͹

IRUWKHHQWLUHVOLGH QRLQWUDWXPRUDOEORRGYHVVHOVSUHVHQW ORZQXPEHURI ʹͶͺ

YHVVHOVSUHVHQW PHGLXPQXPEHURIYHVVHOVSUHVHQW KLJKQXPEHURI ʹͶͻ

YHVVHOVSUHVHQW0LWRWLFILJXUHVZHUHFRXQWHGLQQRQRYHUODSSLQJILHOGVRIYLHZ ʹͷͲ

DQGDPHDQQXPEHURIPLWRWLFILJXUHVZDVGHWHUPLQHGIRUHDFKPRXVH9DOXHV ʹͷͳ

IURPHDFKPRXVHZHUHXVHGWRFDOFXODWHPHDQYDVFXODULW\VFRUHDQGPLWRWLF ʹͷʹ

ILJXUHVIRUHDFKGLHWJURXS ʹͷ͵

,PPXQRKLVWRFKHPLFDOVWDLQLQJRIPDPPDU\WLVVXHZDVSHUIRUPHGn

ʹͷͶ

PLFHGLHWJURXSXVLQJDSULPDU\DQWLERG\IRU(5Į&DWDORJVF6DQWD&UX] ʹͷͷ

%LRWHFKQRORJ\6DQWD&UX]&$86$DWDQG(5ȕ$EFDP ʹͷ͸

&DPEULGJH0$86$DW7KHVHFRQGDU\DQWLERG\ZDVKRUVHUDGLVK ʹͷ͹

SHUR[LGDVHODEHOHGDQWLUDEELWDQWLERG\'$.2&\WRPDWLRQ&DUSLQWHULD&$ ʹͷͺ

86$ ʹͷͻ

ͳʹ

'1$PHWK\ODWLRQDQDO\VLV

ʹ͸ͳ

'1$ZDVH[WUDFWHGIURPDUDQGRPVDPSOHn JURXSRIPDPPDU\

ʹ͸ʹ

WLVVXHVIURPEDVHOLQHPLFHDQG&5DQG2:PLFHLQWKHPRQWKWLPHSRLQWDQG ʹ͸͵

VXUYLYDOVWXG\XVLQJ8OWUD3XUH70SKHQROFKORURIRUPLVRDP\ODOFRKROSHU ʹ͸Ͷ

PDQXIDFWXUHU¶VLQVWUXFWLRQV/LIH7HFKQRORJLHV/LEUDU\SUHSDUDWLRQDQG ʹ͸ͷ

VHTXHQFLQJIRUEDVHOLQH&5DQG2:LQWKHPRQWKWLPHSRLQWDQGVXUYLYDO ʹ͸͸

VWXG\ZHUHSHUIRUPHGDW870'$QGHUVRQ&DQFHU&HQWHU¶V'1$0HWK\ODWLRQ ʹ͸͹

$QDO\VLV&RUHDQG6FLHQFH3DUN1H[W*HQHUDWLRQ6HTXHQFLQJ)DFLOLW\DFFRUGLQJ ʹ͸ͺ

WRSXEOLVKHGSURWRFROVDVSUHYLRXVO\GHVFULEHG6XSSOHPHQWDU\7DEOH6 ʹ͸ͻ

6DPSOHVIURP',2PLFHZHUHQRWDQDO\]HGJLYHQWKHFRVWRI55%6DQGWKH ʹ͹Ͳ

VLPLODULWLHVEHWZHHQ2:DQG',2PLFHLQ(5ĮDQG(5ȕP51$DQGSURWHLQ ʹ͹ͳ

H[SUHVVLRQ*HQHSURPRWHUUHJLRQVZHUHFDOFXODWHGEDVHGRQ5HI6HTJHQH ʹ͹ʹ

DQQRWDWLRQVZLWKUHJLRQVVWDUWLQJNEXSVWUHDPRIWKHDQQRWDWHGWUDQVFULSWLRQ ʹ͹͵

VWDUWVLWH766DQGH[WHQGLQJEDVHSDLUVGRZQVWUHDPRI766'LIIHUHQWLDO ʹ͹Ͷ

PHWK\ODWLRQZDVFDOFXODWHGE\ILOWHULQJVDPSOHVEDVHGRQUHDGFRYHUDJH• ʹ͹ͷ

WKHQSHUIRUPHGDWWKHVLQJOHEDVHOHYHO0HWK\ONLW5SDFNDJHZDVXVHGWRDSSO\ ʹ͹͸

ORJLVWLFUHJUHVVLRQDQGWKHOLNHOLKRRGUDWLRWHVW2EVHUYHGSYDOXHVZHUHDGMXVWHG ʹ͹͹

ZLWKWKHVXFFHVVOLNHOLKRRGLQGH[PHWKRG6/,0&S*GLQXFOHRWLGHVWKDW ʹ͹ͺ

H[KLELWHGGLIIHUHQWLDOPHWK\ODWLRQSDWWHUQVEHWZHHQ&5DQG2:JURXSVZHUH ʹ͹ͻ

FURVVUHIHUHQFHGZLWKDQQRWDWHGJHQHUHJXODWRU\UHJLRQVZLWKLQDQGVXUURXQGLQJ ʹͺͲ

ESR1DQGESR2LQMus musculusRXWOLQHGE\(QVHPEO7RJHQHUDWHDKHDW

ʹͺͳ

PDSZHLGHQWLILHG&S*GLQXFOHRWLGHVZLWKVLJQLILFDQWO\KLJKHUPHWK\ODWLRQLQ&5 ʹͺʹ

ͳ͵ DYDLODEOHIRUEDVHOLQHDQG&5DQG2:PLFHDWWKHPRQWKWLPHSRLQW

ʹͺͶ

'LIIHUHQWLDOO\PHWK\ODWHG&S*GLQXFOHRWLGHVZHUHFOXVWHUHGXVLQJKLHUDUFKLFDO ʹͺͷ

FOXVWHULQJZLWKFRPSOHWHOLQNDJHDQGD(XFOLGLDQGLVWDQFHPHDVXUH ʹͺ͸

&RUUHVSRQGLQJGHQGURJUDPDQGKHDWPDSVIRUSURPRWHULQWURQH[RQDQGRWKHU ʹͺ͹

ZHUHSURGXFHGXVLQJWKHheatmap.2IXQFWLRQIURPWKHJSORWVSDFNDJHLQ5

ʹͺͺ

YHUVLRQ ʹͺͻ

ʹͻͲ

'LIIHUHQWLDOH[SUHVVLRQDQDO\VLVXVLQJ51$VHTDQG,QJHQXLW\3DWKZD\

ʹͻͳ

$QDO\VLV,3$

ʹͻʹ

51$ZDVH[WUDFWHGDVGHVFULEHGDERYH51$OLEUDULHVZHUHSUHSDUHG ʹͻ͵

XVLQJWKH,OOXPLQD7UX6HT6WUDQGHG7RWDO51$6DPSOH3UHSDUDWLRQNLWDFFRUGLQJ ʹͻͶ

WRPDQXIDFWXUHVLQVWUXFWLRQV7KHOLEUDULHVZHUHVHTXHQFHGXVLQJD[EDVHV ʹͻͷ

SDLUHGHQGSURWRFRORQWKH,OOXPLQD+L6HTLQVWUXPHQW7KHUHDGVZHUH ʹͻ͸

PDSSHGWRPRXVHJHQRPHPPE\7RS+DWYHUVLRQ7KHQXPEHU ʹͻ͹

RIIUDJPHQWVLQHDFKNQRZQJHQHIURP5HI6HTGDWDEDVH8&6&*HQRPH ʹͻͺ

%URZVHUZDVHQXPHUDWHGXVLQJ+76HTFRXQWIURP+76HTSDFNDJH ʹͻͻ

YHUVLRQS+76HT7KHGLIIHUHQWLDOH[SUHVVLRQEHWZHHQFRQGLWLRQVZDV ͵ͲͲ

VWDWLVWLFDOO\DVVHVVHGE\5%LRFRQGXFWRUSDFNDJH(GJH5YHUVLRQ*HQHV ͵Ͳͳ

ZLWKIDOVHGLVFRYHU\UDWH”ZHUHFDOOHGVLJQLILFDQW)RUSDWKZD\DQDO\VLV ͵Ͳʹ

JHQHVZLWKGLIIHUHQWLDOH[SUHVVLRQLQWLVVXHIURP&5YV2:PLFHDIWHUPRQWKV ͵Ͳ͵

RQGLHWDQGRULQVXUYLYDOVWXG\PLFHZHUHLQWHJUDWHGLQWR,3$SDWKZD\GHVLJQHU ͵ͲͶ

4LDJHQ9HQOR1HWKHUODQGVWRGUDZFRQQHFWLRQVRIUHJXODWRU\UHODWLRQVKLSV ͵Ͳͷ

XVLQJYDOLGDWHGVFLHQWLILFILQGLQJV ͵Ͳ͸

ͳͶ

6WDWLVWLFDODQDO\VHV

͵Ͳͺ

9DOXHVDUHSUHVHQWHGDVPHDQ“VWDQGDUGGHYLDWLRQVG)RUDOOWHVWV ͵Ͳͻ

*UDSK3DG3ULVPVRIWZDUHZDVXVHG*UDSK3DG6RIWZDUH,QF/D-ROOD&$DQG ͵ͳͲ

PZDVFRQVLGHUHGVWDWLVWLFDOO\VLJQLILFDQW'LIIHUHQFHVEHWZHHQGLHWJURXSV

͵ͳͳ

LQERG\ZHLJKWZHUHDQDO\]HGE\UHSHDWHGPHDVXUHVDQDO\VLVRIYDULDQFH ͵ͳʹ

$129$IROORZHGE\7XNH\¶VSRVWKRFWHVW'LIIHUHQFHVEHWZHHQGLHWJURXSVLQ ͵ͳ͵

LQVXOLQOHSWLQDGLSRQHFWLQ,*)ȕHVWUDGLRODQGPDPPDU\(5ĮDQG(5ȕ ͵ͳͶ

P51$DQGSURWHLQH[SUHVVLRQDWHDFKWLPHSRLQWZHUHDQDO\]HGE\RQHZD\ ͵ͳͷ

$129$IROORZHGE\7XNH\¶VSRVWKRFWHVW.DSODQ0HLHUVXUYLYDOFXUYHVZHUH ͵ͳ͸

SORWWHGDQGWKHGLIIHUHQFHLQRYHUDOOVXUYLYDOEHWZHHQWKHJURXSVZDVDQDO\]HG ͵ͳ͹

E\WKHORJUDQNWHVW ͵ͳͺ

͵ͳͻ

5(68/76

͵ʹͲ

'LHWDU\HQHUJ\EDODQFHPRGXODWLRQLPSDFWVERG\ZHLJKWERG\FRPSRVLWLRQ

͵ʹͳ

DQGVHUXPPHWDEROLFKRUPRQHV

͵ʹʹ

)HPDOH0079QHXPLFHZHUHUDQGRPL]HGWRUHFHLYHGLHWDU\HQHUJ\ ͵ʹ͵

EDODQFHPRGXODWLRQYLD&5RYHUZHLJKWLQGXFLQJ2:RUGLHWLQGXFHGREHVLW\ ͵ʹͶ

',2GLHWUHJLPHQVDQGZHUHPRQLWRUHGDVSDUWRIDDQGPRQWKWLPHSRLQW ͵ʹͷ

VWXG\n PLFHGLHWIRUHDFKWLPHSRLQWRUIRUXSWRPRQWKVLQDVXUYLYDO

͵ʹ͸

VWXG\n PLFHGLHWKHUHDIWHUUHIHUUHGWRDV³VXUYLYDOVWXG\PLFH´&52:

͵ʹ͹

DQG',2GLHWUHJLPHQVUHVXOWHGLQOHDQRYHUZHLJKWDQGREHVHSKHQRW\SHV ͵ʹͺ

UHVSHFWLYHO\$IWHUPRQWKRIGLHWWUHDWPHQWGLIIHUHQFHVLQDYHUDJHFDORULFLQWDNH ͵ʹͻ

ͳͷ $ZLWK&52:',27KLVFRQWLQXHGIRUXSWRPRQWKVLQWKHVXUYLYDO ͵͵ͳ

VWXG\PLFHDIWHUZKLFKWXPRUGHYHORSPHQWLQ2:DQG',2PLFHPDGHERG\ ͵͵ʹ

ZHLJKWPHDVXUHPHQWVXQVWDEOH&5PLFHKDGVLJQLILFDQWO\ORZHUSHUFHQWERG\IDW ͵͵͵

)LJXUH%FRPSDUHGZLWK',2PLFHDIWHUDQGPRQWKVRQGLHW2:PLFH ͵͵Ͷ

KDGLQWHUPHGLDWHSHUFHQWERG\IDWEHWZHHQ&5DQG',2 ͵͵ͷ

&RPSDUHGZLWK&5PLFH2:DQG',2PLFHZHUHPHWDEROLFDOO\ ͵͵͸

G\VUHJXODWHGDVDVVHVVHGE\HQHUJ\EDODQFHUHODWHGKRUPRQHV7DEOH',2 ͵͵͹

PLFHFRPSDUHGZLWK&5PLFHKDGKLJKHUVHUXPOHYHOVRILQVXOLQOHSWLQLQVXOLQ ͵͵ͺ

OLNHJURZWKIDFWRU,*)DQGȕHVWUDGLRODQGORZHUVHUXPOHYHOVRI ͵͵ͻ

DGLSRQHFWLQFRQVLVWHQWZLWKREHVLW\DVVRFLDWHGPHWDEROLFSHUWXUEDWLRQV2: ͵ͶͲ

PLFHJHQHUDOO\GLVSOD\HGLQWHUPHGLDWHOHYHOVRIWKHVHPHWDEROLFIDFWRUVDIWHU ͵Ͷͳ

DQGPRQWKVRQGLHW ͵Ͷʹ

͵Ͷ͵

'LHWDU\HQHUJ\EDODQFHPRGXODWLRQLPSDFWVPDPPDU\(5ĮDQG(5ȕ

͵ͶͶ

H[SUHVVLRQ ͵Ͷͷ

0DPPDU\WLVVXHZDVFROOHFWHGIURPWLPHSRLQWVWXG\PLFHDIWHUDQG ͵Ͷ͸

PRQWKVDQGIURPVXUYLYDOVWXG\PLFHEHWZHHQDQGPRQWKVXSRQWKHLU ͵Ͷ͹

WHUPLQDWLRQ7LVVXHVZHUHDQDO\]HGIRU(5ĮDQG(5ȕH[SUHVVLRQDQGYDOXHVDUH ͵Ͷͺ

UHSRUWHGDVUHODWLYHWRH[SUHVVLRQLQPDPPDU\WLVVXHVFROOHFWHGIURPDEDVHOLQH ͵Ͷͻ

VDPSOHRIPLFHNLOOHGEHIRUHLQLWLDWLRQRIGLHWDU\PRGXODWLRQ&5PLFH ͵ͷͲ

FRPSDUHGZLWK2:DQG',2PLFHKDGVLJQLILFDQWO\KLJKHUPDPPDU\P51$ ͵ͷͳ

OHYHOVRI(5ĮDIWHUDQGPRQWKVRQGLHWDQGLQVXUYLYDOVWXG\PLFH)LJXUH$ ͵ͷʹ

ͳ͸ FRPSDUHGWR&5PLFHE\PRQWKDQGFRQWLQXHGWKURXJKRXWVWXG\)LJXUHV% ͵ͷͶ

DQG) ͵ͷͷ

5HODWLYHWREDVHOLQHPDPPDU\(5ȕP51$OHYHOVZHUHLQFUHDVHGDWOHDVW

͵ͷ͸

IROGRQDYHUDJHLQWLVVXHVIURP&5PLFHEXWGHFUHDVHGLQWLVVXHVIURP2: ͵ͷ͹

DQG',2PLFH)LJXUH&$WWKHPRQWKWLPHSRLQWDQGHYHU\WLPHSRLQW ͵ͷͺ

WKHUHDIWHU&5PLFHKDGVLJQLILFDQWO\JUHDWHUOHYHOVRIERWK(5ȕP51$DQG(5ȕ ͵ͷͻ

SURWHLQFRPSDUHGZLWK2:DQG',2PLFH)LJXUHV&'DQG)1RGLIIHUHQFHV ͵͸Ͳ

EHWZHHQ2:DQG',2PLFHLQ(5ĮH[SUHVVLRQ(5ȕH[SUHVVLRQRUWKHUDWLRRI ͵͸ͳ

(5ĮWR(5ȕZHUHIRXQG6LJQLILFDQWGLHWGHSHQGHQWGLIIHUHQFHVLQWKH(5ĮWR ͵͸ʹ

(5ȕUDWLRZHUHGHWHFWHGDWHDFKWLPHSRLQWLQWLVVXHVFROOHFWHGIURP&5YV2: ͵͸͵

PLFHDVZHOO&5YV',2PLFH)LJXUH( ͵͸Ͷ

͵͸ͷ

'LHWDU\HQHUJ\EDODQFHPRGXODWLRQLPSDFWV'1$PHWK\ODWLRQZLWKLQESR1

͵͸͸

DQGQHDUESR2

͵͸͹

8VLQJUHGXFHGUHSUHVHQWDWLRQELVXOILWHVHTXHQFLQJ55%6ZHDQDO\]HG ͵͸ͺ

GLHWHIIHFWVRQWKHPHWK\ODWLRQVWDWXVRI'1$LVRODWHGIURPWKHPDPPDU\WLVVXH ͵͸ͻ

RIEDVHOLQH&5DQG2:PLFHDWWKHPRQWKWLPHSRLQWDQGLQVXUYLYDOVWXG\ ͵͹Ͳ

PLFH'XHWRWKHVLPLODULWLHVEHWZHHQ2:DQG',2PLFHLQ(5ĮDQG(5ȕP51$ ͵͹ͳ

DQGSURWHLQH[SUHVVLRQVDPSOHVIURP',2PLFHZHUHQRWDQDO\]HG ͵͹ʹ

0DPPDU\'1$PHWK\ODWLRQZDVJHQHUDOO\KLJKHULQ&5PLFHWKDQ2: ͵͹͵

PLFHSDUWLFXODUO\LQ&S*GLQXFOHRWLGHVDW&7&)ELQGLQJVLWHVRUIODQNLQJUHJLRQV ͵͹Ͷ

ZLWKLQESR1DQGQHDUESR26XSSOHPHQWDU\7DEOH6,QPDPPDU\WLVVXH

͵͹ͷ

ͳ͹ SHUFHQWDJHRIPHWK\ODWHG'1$ZDVVLJQLILFDQWO\GLIIHUHQW&5!2:DWGLVWLQFW ͵͹͹

LQWURQLF&S*GLQXFOHRWLGHV&KU&KU&KUDW ͵͹ͺ

DQDQQRWDWHG&7&)ELQGLQJVLWHZLWKLQESR1ZKLFKHQFRGHV(5Į'LHW

͵͹ͻ

GHSHQGHQWHIIHFWVRQ(5Į'1$PHWK\ODWLRQZHUHQRWREVHUYHGLQVDPSOHVIURP ͵ͺͲ

WKHPRQWKWLPHSRLQW)LJXUH$DQG6XSSOHPHQWDU\7DEOH6 ͵ͺͳ

,QPDPPDU\WLVVXHIURP&5PLFHFRPSDUHGZLWK2:PLFHFROOHFWHGDW ͵ͺʹ

HLWKHUWKHPRQWKWLPHSRLQWRUWKHVXUYLYDOVWXG\&S*GLQXFOHRWLGHVQHDU ͵ͺ͵

ESR2WKHJHQHHQFRGLQJ(5ȕKDGVLJQLILFDQWO\KLJKHUPHWK\ODWLRQZLWKLQRU

͵ͺͶ

QHDUD&7&)ELQGLQJVLWH)LJXUH%6SHFLILFDOO\VLJQLILFDQWGLHWGHSHQGHQW ͵ͺͷ

HIIHFWVRQ'1$PHWK\ODWLRQZHUHGHWHFWHGLQVDPSOHVIURPWKHVXUYLYDOVWXG\DW ͵ͺ͸

&S*GLQXFOHRWLGHVQHDUESR2&KUGRZQVWUHDPRIESR2DQG&KU

͵ͺ͹

DQG&KUERWKXSVWUHDPWKDWDOOIHOO“NERID&7&) ͵ͺͺ

ELQGLQJVLWHZKLFKZHGHILQHDVD&7&)IODQNLQJUHJLRQ:HDOVRREVHUYHG ͵ͺͻ

VLJQLILFDQWO\KLJKHUPHWK\ODWLRQLQ&5PLFHDWWKHPRQWKWLPHSRLQWDWRWKHU ͵ͻͲ

VLWHV&KUDQG&KUERWKGRZQVWUHDPWKDWIHOOZLWKLQ ͵ͻͳ

DQDQQRWDWHG&7&)ELQGLQJVLWH)LJXUH%DQG6XSSOHPHQWDU\7DEOH6 ͵ͻʹ

͵ͻ͵

'LHWDU\HQHUJ\EDODQFHPRGXODWLRQLPSDFWV(5ĮDQG(5șH[SUHVVLRQLQ

͵ͻͶ

SDUWWKURXJKHSLJHQHWLFPHFKDQLVPV

͵ͻͷ

:HLQYHVWLJDWHGJOREDOGLHWGHSHQGHQWGLIIHUHQFHVLQPDPPDU\'1$ ͵ͻ͸

PHWK\ODWLRQLQ&5YHUVXV2:PLFH$IWHUPRQWKVRQGLHW2:PLFHKDG ͵ͻ͹

&S*GLQXFOHRWLGHVZLWKVLJQLILFDQWO\KLJKHUPHWK\ODWLRQDQG&S* ͵ͻͺ

ͳͺ )XUWKHUPRUHVXUYLYDOVWXG\PLFHSRVVHVVHGPRUHSURQRXQFHGGLHWGHSHQGHQW ͶͲͲ

GLIIHUHQFHVLQPDPPDU\'1$PHWK\ODWLRQ2:VXUYLYDOVWXG\PLFHKDG&S* ͶͲͳ

GLQXFOHRWLGHVZLWKVLJQLILFDQWO\KLJKHUPHWK\ODWLRQDQGZLWKVLJQLILFDQWO\ ͶͲʹ

ORZHUPHWK\ODWLRQFRPSDUHGZLWK&52IWHQWKHVHJHQHVZLWKKLJKHUPHWK\ODWLRQ ͶͲ͵

LQ&5VXUYLYDOFRPSDUHGWR2:VXUYLYDOKDGKLJKHUPHWK\ODWLRQOHYHOVLQEDVHOLQH ͶͲͶ

DQGLQ&5DQG2:PLFHDWWKHPRQWKWLPHSRLQWVXSSRUWLQJDGHYLDQFHLQ2: ͶͲͷ

VXUYLYDOPHWK\ODWLRQ)LJXUHV$DQG% ͶͲ͸

0DPPDU\JODQG51$VHTDQDO\VLVGHPRQVWUDWHGWKDW'1$F\WRVLQH ͶͲ͹

PHWK\OWUDQVIHUDVHDNMT1H[SUHVVLRQZDVVLJQLILFDQWO\KLJKHULQ2:

ͶͲͺ

FRPSDUHGZLWK&5VXUYLYDOVWXG\PLFH6XSSOHPHQWDU\7DEOH67RYDOLGDWH ͶͲͻ

51$VHTUHVXOWVZHDQDO\]HGDNMT1P51$OHYHOVE\T573&5LQWKH

ͶͳͲ

PDPPDU\WLVVXHRI&52:DQG',2PLFHDWDOOWLPHSRLQWV)LJXUH&DNMT1

Ͷͳͳ

P51$OHYHOVZHUHQRWVLJQLILFDQWO\GLIIHUHQWDPRQJ&52:DQG',2PLFHDIWHU Ͷͳʹ

DQGPRQWKVRQGLHW+RZHYHULQVXUYLYDOVWXG\PLFHPDPPDU\WLVVXHV Ͷͳ͵

IURP2:DQG',2PLFHKDGVLJQLILFDQWO\KLJKHUDNMT1P51$OHYHOVFRPSDUHG

ͶͳͶ

ZLWKPDPPDU\WLVVXHVIURP&5PLFH Ͷͳͷ

51$VHTDQDO\VLVLQQRQWXPRUPDPPDU\WLVVXHIURPWKHVXUYLYDOVWXG\ Ͷͳ͸

DOVRLGHQWLILHGVHYHUDOFRPSRQHQWVRIWKHVLJQDOWUDQVGXFHUDQGDFWLYDWRURI Ͷͳ͹

WUDQVFULSWLRQ67$7QXFOHDUIDFWRUNDSSDOLJKWFKDLQHQKDQFHURIDFWLYDWHG% Ͷͳͺ

FHOOV1)ț%()WUDQVFULSWLRQIDFWRU()DQGLQVXOLQVLJQDOLQJSDWKZD\VWKDW Ͷͳͻ

FDQLPSDFWH[SUHVVLRQRIDNMT1DQGZHUHVLJQLILFDQWO\GLIIHUHQWLQ2:WKDQ&5

ͶʹͲ

PLFH6XSSOHPHQWDU\7DEOH6(DFKRIWKHVHSDWKZD\VFDQLPSDFWH[SUHVVLRQ Ͷʹͳ

RIDNMT17KXVWKHPHWDEROLFDQGLQIODPPDWLRQUHODWHGSHUWXUEDWLRQV

ͳͻ PHDVXUHGLQ2:DQG',2PLFHUHODWLYHWR&5PLFHPD\XQGHUOLHWKHREVHUYHG Ͷʹ͵

GLHWGHSHQGHQWFKDQJHVLQH[SUHVVLRQRIDNMT1.

ͶʹͶ

Ͷʹͷ

&5LQKLELWV0079QHXPDPPDU\WXPRUGHYHORSPHQW

Ͷʹ͸

&RPSDUHGZLWK2:DQG',2GLHWUHJLPHQV&5ZDVDVVRFLDWHGZLWK Ͷʹ͹

VLJQLILFDQWO\LQFUHDVHGVXUYLYDO)LJXUH$6XUYLYDOZDVFRPSDUDEOHEHWZHHQ Ͷʹͺ

2:DQG',2PLFH$IWHUPRQWKVRIVWXG\RQO\2:PRXVHDQG',2PLFH Ͷʹͻ

FRPSDUHGZLWK&5PLFHUHPDLQHGDOLYHDQGWXPRUIUHH$PRQJWKHWXPRU Ͷ͵Ͳ

EHDULQJPLFHWKHPDPPDU\WXPRUVIURP2:n DQG',2n PLFH

Ͷ͵ͳ

FRPSDUHGZLWK&5PLFHn ZHUHJHQHUDOO\PRUHYDVFXODUDQGFRQVLVWHGRI

Ͷ͵ʹ

PRUHSUROLIHUDWLQJFHOOV)LJXUH% Ͷ͵͵

Ͷ͵Ͷ

',6&866,21

Ͷ͵ͷ

7KLVVWXG\DVVHVVHGZKHWKHUGLHWDU\HQHUJ\EDODQFHPRGXODWLRQUDQJLQJ Ͷ͵͸

IURPOHDQ&5WRRYHUZHLJKW2:WRREHVLW\',2DOWHUVPDPPDU\(5 Ͷ͵͹

H[SUHVVLRQHSLJHQHWLFUHSURJUDPPLQJDQGRUPDPPDU\WXPRUGHYHORSPHQWLQ Ͷ͵ͺ

IHPDOH0079QHXWUDQVJHQLFPLFH:HILUVWFKDUDFWHUL]HGGLHWGHSHQGHQW Ͷ͵ͻ

PHWDEROLFSHUWXUEDWLRQVLQVXEVHWVRIGLHWWUHDWHGPLFHHXWKDQL]HGDWEDVHOLQH ͶͶͲ

DQGPRQWKVRQVWXG\&5PLFHUHODWLYHWR2:DQG',2PLFHKDGGHFUHDVHG ͶͶͳ

ERG\ZHLJKWDGLSRVLW\DQGREHVLW\DVVRFLDWHGVHUXPPHWDEROLFKRUPRQHV ͶͶʹ

LQFOXGLQJLQVXOLQOHSWLQ,*)DQGȕHVWUDGLRODVZHOODVLQFUHDVHG ͶͶ͵

DGLSRQHFWLQDIWHUPRQWKRIGLHWWUHDWPHQWFRQVLVWHQWZLWKSUHYLRXVVWXGLHV ͶͶͶ

ʹͲ :HVKRZIRUWKHILUVWWLPHWKDW&5EXWQRW2:DQG',2UHJLPHQV

ͶͶ͸

SUHVHUYHVPDPPDU\(5ĮDQG(5ȕH[SUHVVLRQLQ0079QHXPLFH/RVVRI(5Į ͶͶ͹

DQG(5ȕDQGLQFUHDVHG(5ĮWR(5ȕDVREVHUYHGLQ2:DQG',2PLFHKDYH ͶͶͺ

HDFKEHHQOLQNHGZLWKSRRUEUHDVWFDQFHUSURJQRVLVLQFOLQLFDOVWXGLHV ͶͶͻ

2XUILQGLQJWKDWWKH(5ĮWR(5ȕUDWLRDSURJQRVWLFLQGLFDWRULQEUHDVWFDQFHUFDQ ͶͷͲ

EHPDQLSXODWHGE\HQHUJ\EDODQFHPRGXODWLRQKDVQRWWRRXUNQRZOHGJHEHHQ Ͷͷͳ

SUHYLRXVO\UHSRUWHG Ͷͷʹ

:HDQGRWKHUVKDYHVKRZQWKDWREHVLW\FDQLQGXFHDEHUUDQW'1$ Ͷͷ͵

PHWK\ODWLRQRIJHQHVLQYROYHGLQJURZWKIDFWRUDQGLQIODPPDWRU\VLJQDOLQJ ͶͷͶ

7RDVVHVVZKHWKHUWKHHQHUJ\EDODQFHGHSHQGHQWHIIHFWVRQ(5ĮDQG(5ȕ Ͷͷͷ

H[SUHVVLRQDUHFRQWUROOHGHSLJHQHWLFDOO\ZHFKDUDFWHUL]HGHSLJHQHWLFDOWHUDWLRQV Ͷͷ͸

LQPDPPDU\WLVVXH'1$IURPEDVHOLQHPLFHIURP&5DQG2:PLFHDIWHU Ͷͷ͹

PRQWKVRQGLHWDQGIURPVXUYLYDOVWXG\PLFHDIWHUWRPRQWKVRIGLHW,Q Ͷͷͺ

VXUYLYDOVWXG\PLFHGLIIHUHQWLDOO\PHWK\ODWHG&S*GLQXFOHRWLGHVZHUHREVHUYHGLQ Ͷͷͻ

D&7&)ELQGLQJVLWHZLWKLQWKHESR1JHQHDQGLQ&7&)ELQGLQJVLWHVRUIODQNLQJ

Ͷ͸Ͳ

UHJLRQV“NERID&7&)ELQGLQJVLWHXSVWUHDPDQGGRZQVWUHDPRIESR2,

Ͷ͸ͳ

FRQVLVWHQWZLWKVXVWDLQHGWUDQVFULSWLRQDODFWLYDWLRQRI(5ĮDQG(5β0DPPDU\ Ͷ͸ʹ

H[SUHVVLRQRIDNMT1ZDVVWDEOHLQ&5PLFHEXWLQFUHDVHGRYHUWLPHLQ2:DQG

Ͷ͸͵

',2PLFHVXJJHVWLQJ&5SUHYHQWHGHSLJHQHWLFUHSURJUDPPLQJRIDNMT1 WKDW

Ͷ͸Ͷ

RFFXUVZLWKH[FHVVHQHUJ\LQWDNHDQGZHLJKWJDLQ7KHHIIHFWRIDOWHUHGDNMT1

Ͷ͸ͷ

H[SUHVVLRQOLNHO\LPSDFWVWKHH[SUHVVLRQRIPDQ\JHQHVLQFOXGLQJESR1DQG

Ͷ͸͸

ESR2+RZHYHUDSODXVLEOHPHFKDQLVPIRUWKHVXVWDLQHGH[SUHVVLRQRIWKH

Ͷ͸͹

ESR1DQGESR2JHQHVLQUHVSRQVHWR&5LVWKHPDLQWHQDQFHRIDNMT1

ʹͳ H[SUHVVLRQDQG'1$PHWK\ODWLRQ)LJXUH7KLVFDQSURKLELW&7&)ELQGLQJ Ͷ͸ͻ

WKHUHE\SUHYHQWLQJDOORVWHULFUHSUHVVLRQDQGGHFUHDVHGLQWHUDFWLRQVEHWZHHQ Ͷ͹Ͳ

HQKDQFHUVDQGSURPRWHUV3UHYLRXVVWXGLHVKDYHVKRZQWKDW'1$ Ͷ͹ͳ

PHWK\ODWLRQFDQLPSHGH&7&)ELQGLQJSRVLWLYHO\LQIOXHQFLQJWUDQVFULSWLRQYLDD Ͷ͹ʹ

ORVVRIUHSUHVVLRQRIVSHFLILFJHQHVVXFKDVGAD1DQGXAF1DQG

Ͷ͹͵

LQFUHDVHGHQKDQFHUSURPRWHULQWHUDFWLRQVLQIGF2DQGc-MYC

Ͷ͹Ͷ

:HDOVRDVVHVVHGWKHHIIHFWVRIGLHWDU\HQHUJ\EDODQFHPRGXODWLRQRQ Ͷ͹ͷ

PDPPDU\WXPRUGHYHORSPHQWLQDFRKRUWRIPLFHUDQGRPL]HGWRWKHWKUHHGLHWV Ͷ͹͸

n GLHW&5UHODWLYHWRWKH2:DQG',2UHJLPHQVUHVXOWHGLQVLJQLILFDQWO\

Ͷ͹͹

LQFUHDVHGVXUYLYDOLQ0079QHXPLFHLQDVVRFLDWLRQZLWKLQFUHDVHGPDPPDU\ Ͷ͹ͺ

(5ĮDQG(5ȕH[SUHVVLRQDQG'1$PHWK\ODWLRQDWRUQHDU&7&)ELQGLQJVLWHV Ͷ͹ͻ

6SHFLILFDOO\RIWKH0079QHXPLFHIHGWKH&5GLHWIRUXSWRPRQWKVRQO\ ͶͺͲ

GHYHORSHGVSRQWDQHRXVPDPPDU\WXPRUVZKLOHWKHPHGLDQVXUYLYDORIWKH Ͷͺͳ

2:DQG',2JURXSVZDVOHVVWKDQPRQWKV)LJXUH$7RRXUNQRZOHGJH Ͷͺʹ

WKLVVWXG\LVWKHILUVWWRGHPRQVWUDWHWKHDQWLFDQFHUHIIHFWVRIDFKURQLF&5 Ͷͺ͵

UHJLPHQFRPSDUHGZLWK2:RU',2LQ0079QHXPLFH+RZHYHU0L]XQRet al.

ͶͺͶ

IRXQGWKDWLQWHUPLWWHQWFDORULHUHVWULFWLRQGHFUHDVHGPDPPDU\WXPRULQFLGHQFHLQ Ͷͺͷ

0079QHXPLFH2XUILQGLQJVRIWKHDQWLFDQFHUHIIHFWVRI&5FRPSDUHG Ͷͺ͸

ZLWK2:RU',2UHJLPHQVDUHFRQVLVWHQWZLWKUHSRUWVRIDOLQNEHWZHHQGLHWDU\ Ͷͺ͹

HQHUJ\EDODQFHPRGXODWLRQDQGPDPPDU\WXPRULJHQHVLVLQRWKHUSUHFOLQLFDO Ͷͺͺ

PRGHOVRIPDPPDU\FDQFHU Ͷͺͻ

7ZRSUHYLRXVSXEOLFDWLRQVFRPSDUHG',2YHUVXVFKRZGLHWVVLPLODUWRRXU ͶͻͲ

ʹʹ WXPRULJHQHVLVLQ0079QHXPLFH&OHDU\et al.UHSRUWHGWKDW',2DQGFKRZIHG

Ͷͻʹ

PLFHKDGVLPLODUPDPPDU\WXPRUGHYHORSPHQWDQGVXUYLYDOFRQVLVWHQWZLWKRXU Ͷͻ͵

REVHUYDWLRQRIQRVLJQLILFDQWGLIIHUHQFHLQWXPRUGHYHORSPHQWRUVXUYLYDOLQ2: ͶͻͶ

YHUVXV',2PLFH,QFRQWUDVW&KHQet al.UHSRUWHGDVLJQLILFDQWGLHW

Ͷͻͷ

GHSHQGHQWGLIIHUHQFHFKRZ!',2LQVXUYLYDOUDWHV2XUILQGLQJVZLWK&5 Ͷͻ͸

PD\KHOSUHFRQFLOHWKHVHDSSDUHQWO\FRQIOLFWLQJUHVXOWVVLQFHZHIRXQGKLJKO\ Ͷͻ͹

VLJQLILFDQWGLIIHUHQFHVLQVXUYLYDOLQ&5PLFHUHODWLYHWR2:DQG',2PLFH&KHQ Ͷͻͺ

et al.OLQNHGWKHSURFDQFHUHIIHFWVRI',2UHODWLYHWRFKRZLQWKHLUVWXG\WR

Ͷͻͻ

LQFUHDVHGVLJQDOLQJWKURXJKWKH,..ȕP725DQG9(*)SDWKZD\VZKLFK ͷͲͲ

VWLPXODWHSUROLIHUDWLRQDQGVXUYLYDO:HKDYHSUHYLRXVO\HVWDEOLVKHGWKDW',2 ͷͲͳ

LQFUHDVHVDQG&5GHFUHDVHVFLUFXODWLQJ,*)LQVXOLQDQGWKHLUGRZQVWUHDP ͷͲʹ

VLJQDOVWKURXJKWKH,*)DQGLQVXOLQUHFHSWRUW\URVLQHNLQDVHV,*)LV ͷͲ͵

DSRWHQWPLWRJHQZKLFKSURPRWHVVLJQDOLQJWKURXJKWKH,..ȕP725DQG9(*) ͷͲͶ

SDWKZD\VXOWLPDWHO\SURPRWLQJJURZWKDQGDOVRLQKLELWLQJDSRSWRVLV,QZRPHQ ͷͲͷ

FLUFXODWLQJ,*)LVSRVLWLYHO\DVVRFLDWHGZLWKWHUPLQDOGXFWOREXODUXQLWLQYROXWLRQ ͷͲ͸

PDPPRJUDSKLFGHQVLW\DQGEUHDVWFDQFHUULVN ͷͲ͹

,QWKHSUHVHQWVWXG\ZHIRXQGWKDWDIWHUPRQWKRIGLHWWUHDWPHQW&5 ͷͲͺ

PLFHUHODWLYHWR2:DQG',2PLFHKDGGHFUHDVHGVHUXP,*)DQGLQVXOLQ ͷͲͻ

7DEOH+RZHYHUWKH',2DQGFKRZIHGPLFHLQWKH&OHDU\VWXG\GLGQRWGLIIHU ͷͳͲ

LQ,*)7KXVRQHSRVVLEOHH[SODQDWLRQIRUWKHREVHUYHGGLIIHUHQWLDOWXPRU ͷͳͳ

UHVSRQVHVPD\LQYROYHWKHGLHWGHSHQGHQWHIIHFWRUODFNWKHUHRIRQV\VWHPLF ͷͳʹ

PHWDEROLVPSDUWLFXODUO\JURZWKIDFWRUVDQGWKHLUGRZQVWUHDPVLJQDOV'LIIHUHQWLDO ͷͳ͵

ʹ͵ VSHFLILFGLIIHUHQFHVLQWXPRUUHVSRQVHVWR',2DVWKHPHGLDQVXUYLYDOIRUWKH ͷͳͷ

&OHDU\VWXG\FKRZIHGPLFHDQGRXU2:PLFHZHUHFRPSDUDEOHaPRQWKV ͷͳ͸

DQGaPRQWKVUHVSHFWLYHO\LQFRQWUDVWWRWKH&KHQVWXG\FKRZIHGPLFHa ͷͳ͹

PRQWKV ͷͳͺ

,QWHUDFWLRQVEHWZHHQGLHWDU\HQHUJ\EDODQFHPRGXODWLRQDQG'1$ ͷͳͻ

PHWK\ODWLRQPD\LQIOXHQFHWKHH[SUHVVLRQRINH\EUHDVWFDQFHUUHODWHGJHQHV ͷʹͲ

VXFKDV(5Į(5ȕYLDGLHWLQGXFHGFKDQJHVLQDNMT1H[SUHVVLRQDQG'1$

ͷʹͳ

PHWK\ODWLRQDWRUQHDU&7&)ELQGLQJVLWHV)LJXUHLQWHJUDWHVILQGLQJVIURP51$ ͷʹʹ

VHTDQDO\VLV6XSSOHPHQWDU\7DEOH6XVLQJ,3$WRLOOXVWUDWHDSURSRVHGPRGHO ͷʹ͵

RIDGLHWUHVSRQVLYHQHWZRUNFRQWULEXWLQJWRDOWHUHGJHQHH[SUHVVLRQUHODWHGWR ͷʹͶ

VHYHUDOWUDQVFULSWLRQIDFWRUSDWKZD\VHJ()67$7DQG1)ț%WKDWVHUYHDV ͷʹͷ

UHJXODWRUVRIDNMT1,QIODPPDWLRQSURPRWHVDNMT1H[SUHVVLRQZKLFKKDV

ͷʹ͸

EHHQVKRZQWRSRVLWLYHO\FRUUHODWHZLWK,/H[SUHVVLRQLQWXPRUVDQGEORRG ͷʹ͹

_{2YHUH[SUHVVLRQRI}

DNMT1PD\EHDPHFKDQLVPXWLOL]HGE\FDQFHUFHOOVWR

ͷʹͺ

HYDGHUHJXODWLRQE\UHQGHULQJWXPRUVXSSUHVVRUVWUDQVFULSWLRQDOO\LQDFWLYH ͷʹͻ

7KXVPHGLDWRUVRIREHVLW\DVVRFLDWHGLQIODPPDWLRQPD\SURPRWHLQFUHDVHG ͷ͵Ͳ

H[SUHVVLRQRIDNMT1,WKHUHE\ FRQWULEXWLQJWRDEHUUDQW'1$PHWK\ODWLRQDQG

ͷ͵ͳ

WUDQVFULSWLRQRIEUHDVWFDQFHUUHODWHGJHQHVVXFKDV(5ĮDQG(5ȕ ͷ͵ʹ

7RRXUNQRZOHGJHWKHUHDUHQRSUHYLRXVUHSRUWVUHJDUGLQJWKH ͷ͵͵

PDLQWHQDQFHRIPDPPDU\(5ĮDQG(5ȕSRVLWLYLW\ZLWKD&5UHJLPHQ$V ͷ͵Ͷ

PHQWLRQHGSUHYLRXVO\RXU&5PLFHKDGORZHUOHYHOVRIHQHUJ\EDODQFHUHODWHG ͷ͵ͷ

PHWDEROLFIDFWRUVLQFOXGLQJ,*)LQVXOLQDQGOHSWLQFRQVLVWHQWZLWKQXPHURXV ͷ͵͸

ʹͶ WKHLUGRZQVWUHDPVLJQDOLQJSDWKZD\VFDQDOVRLPSDFWDNMT1EXWRXUVWXG\LV

ͷ͵ͺ

OLPLWHGLQWKHDELOLW\WRHVWDEOLVKWKHLUSUHFLVHUROH)XWXUHVWXGLHVDUHSODQQHGWR ͷ͵ͻ

DVVHVVWKHSRVVLEOHOLQNVEHWZHHQV\VWHPLFPHWDEROLFIDFWRUVVXFKDVLQVXOLQ ͷͶͲ

,*)OHSWLQDQGȕHVWUDGLRODQGWKHLQFUHDVHGPDPPDU\(5ĮDQG(5ȕ ͷͶͳ

DVVRFLDWHG'1$PHWK\ODWLRQDWRUQHDU&7&)ELQGLQJVLWHVLQ&5PLFH ͷͶʹ

$OWHUQDWLYHO\WKHGHFUHDVHGOHYHOVRIȕHVWUDGLROREVHUYHGLQ&5PLFHFRXOG ͷͶ͵

FRQWULEXWHWRSURORQJHGVXUYLYDOLQGHSHQGHQWRIHSLJHQHWLFHYHQWVDVKLJKOHYHOV ͷͶͶ

RIȕHVWUDGLROFDQSURPRWHFHOOSUROLIHUDWLRQDQGWXPRUSURJUHVVLRQ ͷͶͷ

$GGLWLRQDOVWXG\OLPLWDWLRQVLQFOXGHDUHVWULFWLQJ55%6DQG51$VHTDQDO\VLVWR ͷͶ͸

&5DQG2:PLFHDOWKRXJK2:DQG',2PLFHZHUHVLPLODUUHJDUGLQJ(5ĮDQG ͷͶ͹

(5șP51$DQGSURWHLQH[SUHVVLRQDQGVXUYLYDODQGEQRWPHDVXULQJSK\VLFDO ͷͶͺ

DFWLYLW\DOWKRXJKZHKDYHSUHYLRXVO\VKRZQWKDW&5UHODWLYHWRFRQWURORU',2 ͷͶͻ

GLHWVGRHVQRWLQFUHDVHORFRPRWRUDFWLYLW\LQPLFH ͷͷͲ

,QFRQFOXVLRQZHIRXQGLQ0079QHXWUDQVJHQLFPLFHWKDWPDPPDU\ ͷͷͳ

WXPRUGHYHORSPHQWDQG(5ĮDQG(5ȕH[SUHVVLRQDUHGLHWDU\HQHUJ\EDODQFH ͷͷʹ

GHSHQGHQWLQDVVRFLDWLRQZLWKHSLJHQHWLFUHSURJUDPPLQJ6SHFLILFDOO\DGLHW ͷͷ͵

DNMT1PHWK\ODWLRQD[LVPD\VHUYHDVDSULPDU\UHJXODWRURIJHQHWUDQVFULSWLRQ

ͷͷͶ

YLDGLHWLQGXFHGFKDQJHVLQ'1$PHWK\ODWLRQDWRUQHDU&7&)ELQGLQJVLWHV ͷͷͷ

7KHVHSUHFOLQLFDOILQGLQJVVXJJHVWWKDWLQWHUYHQWLRQVUHGXFLQJWKHLPSDFWRI ͷͷ͸

H[FHVVZHLJKWRQHSLJHQHWLFG\VUHJXODWLRQRI(5PD\UHSUHVHQWDQHZVWUDWHJ\ ͷͷ͹

IRUWKHSUHYHQWLRQRUFRQWURORI+(5SRVLWLYHEUHDVWFDQFHULQRYHUZHLJKWDQG ͷͷͺ

REHVHZRPHQ ͷͷͻ

ʹͷ $FNQRZOHGJHPHQWV ͷ͸ͳ :HZDQWWRZLVKDVSHFLDOWKDQNVWR/XFLD-&KLDR-HQQLIHU/LPDQG6WHSKDQLH ͷ͸ʹ &KHQIRUWKHDQLPDOFDUHIHHGLQJDQGPRQLWRULQJ$XGUH\-5DVPXVVHQDQG ͷ͸͵ /DXUHQ+DUULVRQSURYLGHGRUJDQL]DWLRQDOKHOSDQGDQLPDOFDUHVXSHUYLVLRQ:H ͷ͸Ͷ WKDQN'U-LDQMXQ6KHQWKH'LUHFWRURIWKH0ROHFXODU%LRORJ\)DFLOLW\DW0' ͷ͸ͷ $QGHUVRQ&DQFHU&HQWHU6FLHQFH3DUNIRUDVVLVWDQFHZLWKWKHPRXVH51$VHT ͷ͸͸ DQGPHWK\ODWLRQDQDO\VLV ͷ͸͹ ͷ͸ͺ 5HIHUHQFHV ͷ͸ͻ

6LHJHO5/0LOOHU.'-HPDO$&DQFHU6WDWLVWLFVCA Cancer J Clin

ͷ͹Ͳ ͷ͹ͳ )HUOD\-6RHUMRPDWDUDP,(UYLN0'LNVKLW5(VHU60DWKHUV&et ͷ͹ʹ al.*/2%2&$1Y&DQFHU,QFLGHQFHDQG0RUWDOLW\:RUOGZLGH ͷ͹͵

,$5&&DQFHU%DVH1RInternational Agency for Research on Cancer

ͷ͹Ͷ ͷ͹ͷ -HPDO$%UD\)&HQWHU00*LDFLQWL/&ODXGLR33/RSH]0*LRUGDQR$ ͷ͹͸ (SLJHQHWLFLQIRUPDWLRQDQGHVWURJHQUHFHSWRUDOSKDH[SUHVVLRQLQEUHDVW ͷ͹͹

FDQFHUOncologist±

ͷ͹ͺ

6NOLULV*30XQRW.%HOO60&DUGHU3-/DQH6+RUJDQ.et al.

ͷ͹ͻ

5HGXFHGH[SUHVVLRQRIRHVWURJHQUHFHSWRUEHWDLQLQYDVLYHEUHDVWFDQFHU ͷͺͲ

DQGLWVUHH[SUHVVLRQXVLQJ'1$PHWK\OWUDQVIHUDVHLQKLELWRUVLQDFHOOOLQH ͷͺͳ

PRGHOJ Pathol±

ͷͺʹ

+RQPD1+RULL5,ZDVH76DML6<RXQHV07DNXER.et al.&OLQLFDO

ͷͺ͵

LPSRUWDQFHRIHVWURJHQUHFHSWRUEHWDHYDOXDWLRQLQEUHDVWFDQFHUSDWLHQWV ͷͺͶ

WUHDWHGZLWKDGMXYDQWWDPR[LIHQWKHUDS\J Clin Oncol±

ͷͺͷ

,ZDVH+=KDQJ=2PRWR<6XJLXUD+<DPDVKLWD+7R\DPD7et al.

ͷͺ͸

&OLQLFDOVLJQLILFDQFHRIWKHH[SUHVVLRQRIHVWURJHQUHFHSWRUVDOSKDDQG ͷͺ͹

EHWDIRUHQGRFULQHWKHUDS\RIEUHDVWFDQFHUCancer Chemother

ͷͺͺ

Pharmacol6XSSO6±

ͷͺͻ

0RULPRWR/0:KLWH(&KHQ=&KOHERZVNL57+D\V-.XOOHU/et al.

ͷͻͲ

2EHVLW\ERG\VL]HDQGULVNRISRVWPHQRSDXVDOEUHDVWFDQFHUWKH ͷͻͳ

:RPHQ¶V+HDOWK,QLWLDWLYH8QLWHG6WDWHVCancer Causes Control

ͷͻʹ ± ͷͻ͵ 9RQD'DYLV/5RVH'37\SHGLDEHWHVDQGREHVLW\PHWDEROLF ͷͻͶ LQWHUDFWLRQVFRPPRQIDFWRUVIRUEUHDVWFDQFHUULVNDQGQRYHODSSURDFKHV ͷͻͷ

WRSUHYHQWLRQDQGWKHUDS\Curr Diabetes Rev±

ͷͻ͸

ʹ͸ REHVLW\DQGW\SHGLDEHWHVFRQWULEXWHWRLQFUHDVHGEUHDVWFDQFHUULVNLQ ͷͻͺ

SUHPHQRSDXVDOZRPHQCardiovasc Diabetol

ͷͻͻ

+HDO\/$5\DQ$0&DUUROO3(QQLV'&URZOH\9%R\OH7et al.

͸ͲͲ

0HWDEROLFV\QGURPHFHQWUDOREHVLW\DQGLQVXOLQUHVLVWDQFHDUHDVVRFLDWHG ͸Ͳͳ

ZLWKDGYHUVHSDWKRORJLFDOIHDWXUHVLQSRVWPHQRSDXVDOEUHDVWFDQFHUClin

͸Ͳʹ

Oncol±

͸Ͳ͵

/LWWRQ-.*RQ]DOH]$QJXOR$0:DUQHNH&/%X]GDU$8.DX6: ͸ͲͶ

%RQG\0et al.5HODWLRQVKLSEHWZHHQREHVLW\DQGSDWKRORJLFUHVSRQVHWR

͸Ͳͷ

QHRDGMXYDQWFKHPRWKHUDS\DPRQJZRPHQZLWKRSHUDEOHEUHDVWFDQFHUJ

͸Ͳ͸

Clin Oncol±

͸Ͳ͹

+XUVWLQJ6'6PLWK60/DVKLQJHU/0+DUYH\$(3HUNLQV61&DORULHV ͸Ͳͺ

DQGFDUFLQRJHQHVLV/HVVRQVOHDUQHGIURP\HDUVRIFDORULHUHVWULFWLRQ ͸Ͳͻ

UHVHDUFKCarcinogenesis±

͸ͳͲ

&ROPDQ5-$QGHUVRQ50-RKQVRQ6&.DVWPDQ(..RVPDWND.-͸ͳͳ

%HDVOH\70et al.&DORULFUHVWULFWLRQGHOD\VGLVHDVHRQVHWDQGPRUWDOLW\LQ

͸ͳʹ

UKHVXVPRQNH\VScience±

͸ͳ͵ )DELDQ&-.LPOHU%)'RQQHOO\-(6XOOLYDQ'..OHPS-53HWURII%.et ͸ͳͶ al)DYRUDEOHPRGXODWLRQRIEHQLJQEUHDVWWLVVXHDQGVHUXPULVN ͸ͳͷ ELRPDUNHUVLVDVVRFLDWHGZLWK!ZHLJKWORVVLQSRVWPHQRSDXVDO ͸ͳ͸

ZRPHQBreast Cancer Res Treat

͸ͳ͹

+DUYLH0:ULJKW&3HJLQJWRQ00F0XOODQ'0LWFKHOO(0DUWLQ%et al.

͸ͳͺ

7KHHIIHFWRILQWHUPLWWHQWHQHUJ\DQGFDUERK\GUDWHUHVWULFWLRQYGDLO\ ͸ͳͻ

HQHUJ\UHVWULFWLRQRQZHLJKWORVVDQGPHWDEROLFGLVHDVHULVNPDUNHUVLQ ͸ʹͲ

RYHUZHLJKWZRPHQBr J Nutr±

͸ʹͳ

'RQRKRH'5%XOWPDQ6-0HWDERORHSLJHQHWLFV,QWHUUHODWLRQVKLSV ͸ʹʹ

EHWZHHQHQHUJ\PHWDEROLVPDQGHSLJHQHWLFFRQWURORIJHQHH[SUHVVLRQ J

͸ʹ͵

Cell Physiol ±

͸ʹͶ

3RUWHOD$(VWHOOHU0(SLJHQHWLFPRGLILFDWLRQVDQGKXPDQGLVHDVHNat

͸ʹͷ

Biotechnol±

͸ʹ͸

:DWHUODQG5$0LFKHOV.%(SLJHQHWLFHSLGHPLRORJ\RIWKHGHYHORSPHQWDO ͸ʹ͹

RULJLQVK\SRWKHVLVAnnu Rev Nutr±

͸ʹͺ

6KDUPD1.9DUPD90D/+DVVWHGW6-'DV6.2EHVLW\DVVRFLDWHG ͸ʹͻ

PRGXODWLRQRIPL51$DQGFRUHJXODWHGWDUJHWWUDQVFULSWVLQKXPDQ ͸͵Ͳ

DGLSRVHWLVVXHRIQRQGLDEHWLFVXEMHFWVMicroRNA±

͸͵ͳ

0XW]H./DQJHU56FKXPDFKHU)%HFNHU.2WW.1RYRWQ\$et al.

͸͵ʹ

'1$PHWK\OWUDQVIHUDVHDVDSUHGLFWLYHELRPDUNHUDQGSRWHQWLDO ͸͵͵

WKHUDSHXWLFWDUJHWIRUFKHPRWKHUDS\LQJDVWULFFDQFHUEur J Cancer

͸͵Ͷ

±

͸͵ͷ

<X'+:DUH&:DWHUODQG5$=KDQJ-&KHQ0+*DGNDUL0et al.

͸͵͸

'HYHORSPHQWDOO\SURJUDPPHG¶&S*LVODQGPHWK\ODWLRQFRQIHUVWLVVXH ͸͵͹

DQGFHOOW\SHVSHFLILFWUDQVFULSWLRQDODFWLYDWLRQMol Cell Biol

͸͵ͺ

±

͸͵ͻ

-RQHV3$7KH'1$PHWK\ODWLRQSDUDGR[Trends Genet±

͸ͶͲ

/DL$<)DWHPL0'KDVDUDWK\$0DORQH&6RERO6(*HLJHUPDQ&et al.

͸Ͷͳ

'1$PHWK\ODWLRQSUHYHQWV&7&)PHGLDWHGVLOHQFLQJRIWKHRQFRJHQH ͸Ͷʹ

%&/LQ%FHOOO\PSKRPDVJ Exp Med±

ʹ͹ 2QJ&7&RUFHV9*&7&)DQDUFKLWHFWXUDOSURWHLQEULGJLQJJHQRPH ͸ͶͶ

WRSRORJ\DQGIXQFWLRQNat Rev Genet±

͸Ͷͷ

:DQJ+0DXUDQR074X+9DUOH\.(*HUW]-3DXOL)et al.

͸Ͷ͸

:LGHVSUHDGSODVWLFLW\LQ&7&)RFFXSDQF\OLQNHGWR'1$PHWK\ODWLRQ ͸Ͷ͹

Genome Res±

͸Ͷͺ

5RZVH*-7HPSHUR509DQ/LWK0/+ROOLQJVZRUWK0$*HQGOHU6-͸Ͷͻ

7ROHUDQFHDQGLPPXQLW\WR08&LQDKXPDQ08&WUDQVJHQLFPXULQH ͸ͷͲ

PRGHOCancer Res±

͸ͷͳ

7KRPDV'*%UHVORZ1*DUW--7UHQGDQGKRPRJHQHLW\DQDO\VLVRI ͸ͷʹ

SURSRUWLRQVDQGOLIHWDEOHGDWDComput and Biomed Res

͸ͷ͵

%URZQH5:.DQWDUFL$/D0RQWH0-$QGUHZV&$+RYH\.0)DONQHU

͸ͷͶ

./et al3HUIRUPDQFHRIPXOWLSOH[F\WRNLQHDVVD\VLQVHUXPDQGVDOLYD ͸ͷͷ

DPRQJFRPPXQLW\GZHOOLQJSRVWPHQRSDXVDOZRPHQPLoS ONE

͸ͷ͸

H

͸ͷ͹

5RVVL(/GH$QJHO5(%RZHUV/:.KDWLE6$6PLWK/$9DQ%XUUHQ( ͸ͷͺ

et al2EHVLW\DVVRFLDWHGDOWHUDWLRQVLQLQIODPPDWLRQHSLJHQHWLFVDQG

͸ͷͻ

PDPPDU\WXPRUJURZWKSHUVLVWLQIRUPHUO\REHVHPLFHCancer Prev Res

͸͸Ͳ

͸͸ͳ

<DWHV$$NDQQL:$PRGH05%DUUHOO'%LOOLV.&DUYDOKR6LOYD'et al

͸͸ʹ

(QVHPEONucleic Acids Res'

͸͸͵

.LP'3HUWHD*7UDSQHOO&3LPHQWHO+.HOOH\56DO]EHUJ6/7RS+DW ͸͸Ͷ

DFFXUDWHDOLJQPHQWRIWUDQVFULSWRPHVLQWKHSUHVHQFHRILQVHUWLRQV ͸͸ͷ

GHOHWLRQVDQGJHQHIXVLRQVGenome Biol

͸͸͸

3UXLWW.'7DWXVRYD7%URZQ*50DJORWW'51&%,5HIHUHQFH ͸͸͹

6HTXHQFHV5HI6HTFXUUHQWVWDWXVQHZIHDWXUHVDQGJHQRPHDQQRWDWLRQ ͸͸ͺ

SROLF\Nucleic Acids Res

͸͸ͻ

=KDQJ4:DQJ+<:RHWPDQQ$5DJKXQDWK312GXP1:DVLN0$ ͸͹Ͳ

67$7LQGXFHVWUDQVFULSWLRQRIWKH'1$PHWK\OWUDQVIHUDVHJHQH ͸͹ͳ

'107LQPDOLJQDQW7O\PSKRF\WHVBlood±

͸͹ʹ

=KDQJ%*+X/=DQJ0':DQJ+;=KDR:/L-)et al +HOLFREDFWHU

͸͹͵ S\ORUL&DJ$LQGXFHVWXPRUVXSSUHVVRUJHQHK\SHUPHWK\ODWLRQE\ ͸͹Ͷ XSUHJXODWHG'107YLD$.71)ț%SDWKZD\LQJDVWULFFDQFHU ͸͹ͷ GHYHORSPHQWOncotarget ͸͹͸ .LPXUD+1DNDPXUD72JDZD77DQDND66KLRWD.7UDQVFULSWLRQRI ͸͹͹ PRXVH'1$PHWK\OWUDQVIHUDVH'QPWLVUHJXODWHGE\ERWK()5E ͸͹ͺ

+'$&GHSHQGHQWDQGLQGHSHQGHQWSDWKZD\VNucleic Acids Res

͸͹ͻ ± ͸ͺͲ 1RJXHLUD/0'XQODS60)RUG1$+XUVWLQJ6'&DORULHUHVWULFWLRQDQG ͸ͺͳ UDSDP\FLQLQKLELW0079:QWPDPPDU\WXPRUJURZWKLQDPRXVHPRGHO ͸ͺʹ

RISRVWPHQRSDXVDOREHVLW\Endocr Relat Cancer±

͸ͺ͵

2OLYR0DUVWRQ6(+XUVWLQJ6'3HUNLQV616FKHWWHU$.KDQ0&URFH& ͸ͺͶ

et al.(IIHFWVRIFDORULHUHVWULFWLRQDQGGLHWLQGXFHGREHVLW\RQPXULQHFRORQ

͸ͺͷ

FDUFLQRJHQHVLVJURZWKDQGLQIODPPDWRU\IDFWRUVDQG0LFUR51$ ͸ͺ͸

H[SUHVVLRQPLoS OneGRLMRXUQDOSRQH

͸ͺ͹

0DGHLUD00DWWDU$/RJXOOR$)6RDUHV)$*HEULP/+(VWURJHQ ͸ͺͺ

ʹͺ HQGRFULQHWKHUDS\UHVSRQVLYHQHVV²DUDQGRPL]HGQHRDGMXYDQWWULDO

͸ͻͲ

FRPSDULVRQEHWZHHQDQDVWUR]ROHDQGWDPR[LIHQIRUWKHWUHDWPHQWRI ͸ͻͳ

SRVWPHQRSDXVDOEUHDVWFDQFHUBMC CancerGRL

͸ͻʹ ͸ͻ͵ 1DGDO6HUUDQR06DVWUH6HUUD-3RQV'*0LUy$02OLYHU-5RFD3 ͸ͻͶ 7KH(5DOSKD(5EHWDUDWLRGHWHUPLQHVR[LGDWLYHVWUHVVLQEUHDVWFDQFHU ͸ͻͷ

FHOOOLQHVLQUHVSRQVHWR%HWDHVWUDGLROJ Cell Biochem

͸ͻ͸

͸ͻ͹

%DUUHV5.LUFKQHU+5DVPXVVHQ0<DQ-.DQWRU).URRN$et al.

͸ͻͺ

:HLJKW/RVVDIWHU*DVWULF%\SDVV6XUJHU\LQ+XPDQ2EHVLW\5HPRGHOV ͸ͻͻ

3URPRWHU0HWK\ODWLRQCell Rep±

͹ͲͲ

<DQ+7DQJ*:DQJ++DR/+H76XQ;et al.'1$PHWK\ODWLRQ

͹Ͳͳ

UHDFWLYDWHV*$'H[SUHVVLRQLQFDQFHUE\SUHYHQWLQJ&7&)PHGLDWHG ͹Ͳʹ

SRO\FRPEUHSUHVVLYHFRPSOH[UHFUXLWPHQWOncogene±

͹Ͳ͵

9LFWRULD$FRVWD*9D]TXH]6DQWLOODQ.-LPHQH]+HUQDQGH]/0XxR] ͹ͲͶ

*DOLQGR/0DOGRQDGR90DUWLQH]5XL]*8et al.(SLJHQHWLFVLOHQFLQJRI

͹Ͳͷ

WKH;$)JHQHLVPHGLDWHGE\WKHORVVRI&7&)ELQGLQJSci Rep

͹Ͳ͸ ͹Ͳ͹ +DUN$76FKRHQKHUU&-.DW]'-,QJUDP56/HYRUVH-07LOJKPDQ60 ͹Ͳͺ &7&)PHGLDWHVPHWK\ODWLRQVHQVLWLYHHQKDQFHUEORFNLQJDFWLYLW\DWWKH ͹Ͳͻ +,JIORFXVNature ͹ͳͲ )LOLSSRYD*1)DJHUOLH6.OHQRYD(00\HUV&'HKQHU<*RRGZLQ*et ͹ͳͳ al.$QH[FHSWLRQDOO\FRQVHUYHGWUDQVFULSWLRQDOUHSUHVVRU&7&)HPSOR\V ͹ͳʹ GLIIHUHQWFRPELQDWLRQVRI]LQFILQJHUVWRELQGGLYHUJHGSURPRWHU ͹ͳ͵

VHTXHQFHVRIDYLDQDQGPDPPDOLDQFP\FRQFRJHQHVMol Cell Biol

͹ͳͶ ± ͹ͳͷ 0L]XQR1.5RJR]LQD236HSSDQHQ&0/LDR'-&OHDU\03 ͹ͳ͸ *URVVPDQQ0(&RPELQDWLRQRI,QWHUPLWWHQWFDORULHUHVWULFWLRQDQG ͹ͳ͹

HLFRVDSHQWDHQRLFDFLGIRULQKLELWLRQRIPDPPDU\WXPRUVCancer Prev Res

͹ͳͺ ± ͹ͳͻ (QJHOPDQ5:'D\1.*RRG5$&DORULHLQWDNHGXULQJPDPPDU\ ͹ʹͲ GHYHORSPHQWLQIOXHQFHVFDQFHUULVN/DVWLQJLQKLELWLRQRI&++H2X ͹ʹͳ

PDPPDU\WXPRULJHQHVLVE\SHULSXEHUWDOFDORULHUHVWULFWLRQCancer Res

͹ʹʹ ± ͹ʹ͵ 1RJXHLUD/0/DYLJQH-$&KDQGUDPRXOL*9/XL+%DUUHWW-&+XUVWLQJ ͹ʹͶ 6''RVHGHSHQGHQWHIIHFWVRIFDORULHUHVWULFWLRQRQJHQHH[SUHVVLRQ ͹ʹͷ PHWDEROLVPDQGWXPRUSURJUHVVLRQDUHSDUWLDOO\PHGLDWHGE\LQVXOLQOLNH ͹ʹ͸

JURZWKIDFWRUCancer Med

͹ʹ͹

&OHDU\03*UDQGH-3-XQHMD6&0DLKOH1-'LHWLQGXFHGREHVLW\DQG ͹ʹͺ

PDPPDU\WXPRUGHYHORSPHQWLQ0079QHXIHPDOHPLFHNutr Cancer

͹ʹͻ

±

͹͵Ͳ

&KHQ&7'X<<DPDJXFKL++VX-0.XR+3+RUWREDJ\L*1et al.

͹͵ͳ

7DUJHWLQJWKH,..EHWDP7259(*)VLJQDOLQJSDWKZD\DVDSRWHQWLDO ͹͵ʹ

WKHUDSHXWLFVWUDWHJ\IRUREHVLW\UHODWHGEUHDVWFDQFHUMol Cancer Ther

͹͵͵

±

ʹͻ 7KH(QGRJHQRXV+RUPRQHVDQG%UHDVW&DQFHU&ROODERUDWLYH*URXS ͹͵ͷ ,QVXOLQOLNHJURZWKIDFWRU,*),*)ELQGLQJSURWHLQ,*)%3DQG ͹͵͸ EUHDVWFDQFHUULVNSRROHGLQGLYLGXDOGDWDDQDO\VLVRISURVSHFWLYH ͹͵͹

VWXGLHVLancet Oncol± ͹͵ͺ +RUQH+16KHUPDQ0(3IHLIIHU50)LJXHURD-'.KRGU=*)DON57et ͹͵ͻ al.&LUFXODWLQJLQVXOLQOLNHJURZWKIDFWRULQVXOLQOLNHJURZWKIDFWRUELQGLQJ ͹ͶͲ SURWHLQDQGWHUPLQDOGXFWOREXODUXQLWLQYROXWLRQRIWKHEUHDVWDFURVV ͹Ͷͳ

VHFWLRQDOVWXG\RIZRPHQZLWKEHQLJQEUHDVWGLVHDVHBreast Cancer Res

͹Ͷʹ

GRLV

͹Ͷ͵

&KHQ0)/LQ3<:X&)&KHQ:&:X&7,/H[SUHVVLRQUHJXODWHV

͹ͶͶ

WXPRULJHQLFLW\DQGFRUUHODWHVZLWKSURJQRVLVLQEODGGHUFDQFHUPLoS ONE

͹Ͷͷ H ͹Ͷ͸ 0XUSK\702¶'RQRYDQ$0XOOLQV12¶)DUUHOO\&0F&DQQ$0DORQH. ͹Ͷ͹ $Q[LHW\LVDVVRFLDWHGZLWKKLJKHUOHYHOVRIJOREDO'1$PHWK\ODWLRQDQG ͹Ͷͺ DOWHUHGH[SUHVVLRQRIHSLJHQHWLFLQWHUOHXNLQJHQHVPsychiatry ͹Ͷͻ Genet ͹ͷͲ 5R\'0:DOVK/$&KDQ7$'ULYHUPXWDWLRQVRIFDQFHUHSLJHQRPHV ͹ͷͳ Protein Cell ͹ͷʹ 3DGRYDQL0/DYLJQH-$&KDQGUDPRXOL*93HUNLQV61%DUUHWW-& ͹ͷ͵

+XUVWLQJ6'et al'LVWLQFWHIIHFWVRIFDORULHUHVWULFWLRQDQGH[HUFLVHRQ

͹ͷͶ

PDPPDU\JODQGJHQHH[SUHVVLRQLQ&%/PLFHCancer Prev Res

͹ͷͷ ± ͹ͷ͸ ͹ͷ͹ )LJXUH/HJHQGV ͹ͷͺ )LJXUH'LHWDU\HQHUJ\EDODQFHPRGXODWLRQDIIHFWVERG\ZHLJKWDQGSHUFHQW ͹ͷͻ

ERG\IDWLQ0079QHXPLFH$%RG\ZHLJKWPHDQ“VGLQPLFHUHFHLYLQJ ͹͸Ͳ

FDORULHUHVWULFWLRQ&5RYHUZHLJKWLQGXFLQJ2:RUGLHWLQGXFHGREHVLW\',2 ͹͸ͳ

GLHWUHJLPHQVRYHUPRQWKVn PLFHGLHW+HWHURJHQHLW\LQERG\ZHLJKWV

͹͸ʹ DIWHUPRQWKVLQFUHDVHGDVPLFHEHFDPHPRULEXQGWKHUHIRUHZHLJKWGDWD ͹͸͵ EH\RQGWKLVSRLQWDUHQRWVKRZQ6WDWLVWLFDOGLIIHUHQFHVLQERG\ZHLJKWVEHWZHHQ ͹͸Ͷ JURXSVZHUHGHWHUPLQHGE\UHSHDWHGPHDVXUHVDQDO\VLVRIYDULDQFH$129$ ͹͸ͷ %3HUFHQWERG\IDWDVVHVVHGE\TXDQWLWDWLYHPDJQHWLFUHVRQDQFHVSHFWURVFRS\ ͹͸͸

PHDQ“VGDWDQGPRQWKVRQGLHWn PLFHGLHWWLPHSRLQW6WDWLVWLFDO

͹͸͹

͵Ͳ

2:RU',2YV&5PPP2:YV',2♦P

͹͸ͻ

♦♦P♦♦♦P

͹͹Ͳ

͹͹ͳ

)LJXUH'LHWDU\HQHUJ\EDODQFHPRGXODWLRQDIIHFWVPDPPDU\(5ĮDQG(5ȕ

͹͹ʹ

JHQHDQGSURWHLQH[SUHVVLRQ5HODWLYHPDPPDU\(5Į$P51$DQG%SURWHLQ ͹͹͵

OHYHOVPHDQ“VGLQ&52:RU',2PLFHDWWKHDQGPRQWKWLPHSRLQW ͹͹Ͷ

DQGDWVXUYLYDOn PLFHGLHWWLPHSRLQW'DWDDUHSUHVHQWHGDVUHODWLYHWR

͹͹ͷ

EDVHOLQHOHYHOVn PLFH5HODWLYHPDPPDU\(5ȕ&P51$DQG'SURWHLQ

͹͹͸

OHYHOVPHDQ“VGLQ&52:DQG',2PLFHDWDQGPRQWKVRQGLHWDQG ͹͹͹

DWVXUYLYDOn PLFHGLHWWLPHSRLQW3URWHLQOHYHOVZHUHDQDO\]HGE\,+&ZLWK

͹͹ͺ

SHUFHQW(5ĮDQG(5ȕSRVLWLYHQXFOHLGHWHUPLQHGDQGUHVXOWVSUHVHQWHGDV ͹͹ͻ

UHODWLYHWREDVHOLQHOHYHOVn PLFH(7KH(5ĮWR(5ȕUDWLRZDVFDOFXODWHG

͹ͺͲ

E\GLYLGLQJWKHSHUFHQWSRVLWLYHQXFOHLIRU(5ĮE\SHUFHQWSRVLWLYHQXFOHLIRU(5ȕ ͹ͺͳ

)5HSUHVHQWDWLYHLPDJHVRI(5ĮDQG(5ȕ,+&DWEDVHOLQHDQGIRUHDFKGLHW ͹ͺʹ

JURXSDWPRQWKVRQGLHW6WDWLVWLFDOGLIIHUHQFHVLQ(5ĮRU(5ȕP51$DQG ͹ͺ͵

SURWHLQEHWZHHQGLHWJURXSVZHUHGHWHUPLQHGE\RQHZD\$129$2:RU',2 ͹ͺͶ

YV&5PPP

͹ͺͷ

͹ͺ͸

)LJXUH'LHWDU\HQHUJ\EDODQFHPRGXODWLRQDIIHFWV'1$PHWK\ODWLRQRIWKH ͹ͺ͹

LQWURQUHJLRQVRIESR1 DQGXSVWUHDPDQGGRZQVWUHDPRIESR2$ESR1LQWURQ

͹ͺͺ

PHWK\ODWLRQPHDQSHUFHQW'1$PHWK\ODWLRQ“VGLQWKHPDPPDU\WLVVXHRI&5 ͹ͺͻ

DQG2:PLFHDWWKHPRQWKWLPHSRLQWDQGVXUYLYDOVWXG\PLFH'1$ ͹ͻͲ

͵ͳ DQG&KUZKLFKDOOIDOOZLWKLQD&7&)ELQGLQJVLWHDUH ͹ͻʹ

VKRZQ%0HDQSHUFHQW'1$PHWK\ODWLRQ“VGDW&S*GLQXFOHRWLGHVXSVWUHDP ͹ͻ͵

&KU&KUDQGGRZQVWUHDP&KU&KU ͹ͻͶ

&KURIESR2LQWKHPDPPDU\WLVVXHRIPLFH

͹ͻͷ

PDLQWDLQHGRQ&5RU2:GLHWUHJLPHQVIRUPRQWKVRUWKURXJKVXUYLYDODUH ͹ͻ͸

VKRZQ7KHVHDOOIDOOZLWKLQD&7&)ELQGLQJVLWHRU&7&)IODQNLQJUHJLRQDV ͹ͻ͹

LQGLFDWHG6WDWLVWLFDOGLIIHUHQFHVGHWHUPLQHGE\ORJLVWLFUHJUHVVLRQDQGWKH ͹ͻͺ

OLNHOLKRRGUDWLRWHVWSYDOXHVZHUHDGMXVWHGZLWKWKHVXFFHVVOLNHOLKRRGLQGH[ ͹ͻͻ

PHWKRGPPP

ͺͲͲ

ͺͲͳ

)LJXUH'LIIHUHQFHVLQGLHWDU\HQHUJ\EDODQFHLPSDFWJHQRPHZLGH

ͺͲʹ

PHWK\ODWLRQSDWWHUQVLQWKHPDPPDU\WLVVXH$+HDWPDSDQGFOXVWHULQJ ͺͲ͵

GHQGURJUDPRIPDPPDU\WLVVXH'1$PHWK\ODWLRQ&S*GLQXFOHRWLGHVZLWK ͺͲͶ

VLJQLILFDQWGLIIHUHQFHVLQPHWK\ODWLRQ&5VXUYLYDOYV2:VXUYLYDODUHVKRZQ ͺͲͷ

DQGFOXVWHUHGZLWKLQJHQRPLFORFDWLRQLHSURPRWHUH[RQLQWURQRWKHU%DVHOLQH ͺͲ͸

&5DQG2:DIWHUPRQWKVRQGLHWDQG&5DQG2:VXUYLYDOVWXG\PLFHDUHHDFK ͺͲ͹

UHSUHVHQWHGn JURXSZLWKRQHFROXPQSHUnVKRZQ7KHPHWK\ODWLRQ

ͺͲͺ

IUHTXHQF\SHUFHQWDJHUDQJHVIURPWR$YDOXHRIµ¶LVFRPSOHWHO\ ͺͲͻ

XQPHWK\ODWHGDQGµ¶LVIXOO\PHWK\ODWHG6WDWLVWLFDOGLIIHUHQFHV&5YV2: ͺͳͲ

VXUYLYDOGHWHUPLQHGE\ORJLVWLFUHJUHVVLRQDQGWKHOLNHOLKRRGUDWLRWHVWSYDOXHV ͺͳͳ

ZHUHDGMXVWHGZLWKWKHVXFFHVVOLNHOLKRRGLQGH[PHWKRG%5HSUHVHQWDWLRQRI ͺͳʹ

JHQRPLFORFDWLRQVRIGLIIHUHQWLDOO\PHWK\ODWHG&S*GLQXFOHRWLGHVPDSSHG ͺͳ͵

͵ʹ UHJLRQDQGGLGQRWPDSWRDSURPRWHUH[RQRULQWURQDQGDUHFODVVLILHGDV ͺͳͷ

µRWKHU¶&0DPPDU\DNMT1P51$OHYHOVPHDVXUHGE\TXDOLWDWLYHUHDOWLPH

ͺͳ͸

3&5LQPLFHPDLQWDLQHGRQ&52:DQG',2GLHWUHJLPHQVIRUDQG ͺͳ͹

PRQWKVRUWKURXJKVXUYLYDO'DWDDUHSUHVHQWHGDVUHODWLYHWREDVHOLQHOHYHOVn

ͺͳͺ

PLFH6WDWLVWLFDOGLIIHUHQFHVLQDNMT1H[SUHVVLRQGHWHUPLQHGE\RQHZD\

ͺͳͻ

$129$PPP

ͺʹͲ

ͺʹͳ

)LJXUH'LHWDU\HQHUJ\EDODQFHPRGXODWLRQDIIHFWVVXUYLYDOWXPRUYDVFXODULW\ ͺʹʹ

DQGSUROLIHUDWLRQ,Q0079QHXPLFH$.DSODQ0HLHUVXUYLYDOFXUYHVIRU&5 ͺʹ͵

2:DQG',2PLFHn PLFHGLHWRYHUDPRQWKSHULRG0LFHZLWK

ͺʹͶ

QRQWXPRUUHODWHGGHDWKV&52:DQG',2PLFHZHUHFHQVRUHG ͺʹͷ

6WDWLVWLFDOGLIIHUHQFHVLQVXUYLYDOUDWHGHWHUPLQHGE\ORJUDQNWHVW% ͺʹ͸

3KRWRPLFURJUDSKVRIPDPPDU\WXPRUVHFWLRQVVWDLQHGZLWKKHPDWR[\OLQDQG ͺʹ͹

HRVLQDQGGLVSOD\HGDW[RU[PDJQLILFDWLRQ$PRQJWKHWXPRUEHDULQJPLFH ͺʹͺ

WKHPDPPDU\WXPRUVIURP2:n DQG',2n PLFHFRPSDUHGZLWK

ͺʹͻ

&5PLFHn ZHUHPRUHYDVFXODUJURXSPHDQ“VGYDVFXODULW\VFRUHV“

ͺ͵Ͳ

“DQG“UHVSHFWLYHO\DQGFRQVLVWHGRIPRUHSUROLIHUDWLQJ ͺ͵ͳ

FHOOVJURXSPHDQ“VGQXPEHURIPLWRWLFILJXUHVLQGLFDWHGE\EODFNDUURZKHDG ͺ͵ʹ

““DQG“UHVSHFWLYHO\6WDWLVWLFDOGLIIHUHQFHVLQ ͺ͵͵

YDVFXODULW\DQGPLWRWLFILJXUHVGHWHUPLQHGE\RQHZD\$129$PP

ͺ͵Ͷ

P

ͺ͵ͷ

͵͵

)LJXUH3URSRVHGPRGHORIDQHQHUJ\EDODQFHUHVSRQVLYHQHWZRUNDVVRFLDWHG

ͺ͵͹

ZLWKDNMT1UHJXODWLRQ'1$PHWK\ODWLRQDQGWUDQVFULSWLRQDOUHJXODWLRQRI(5Į

ͺ͵ͺ

DQG(5ȕ2EHVLW\DQGDVVRFLDWHGHQHUJ\H[FHVVUHVXOWVLQVHYHUDOPHWDEROLF ͺ͵ͻ

SHUWXUEDWLRQVWKDWDUHDPHOLRUDWHGE\FDORULHUHVWULFWLRQ7RLGHQWLI\UHJXODWRU\ ͺͶͲ

UHODWLRQVKLSVEHWZHHQDNMT1DQGRWKHU JHQHVWKDWZHUHGLIIHUHQWLDOO\H[SUHVVHG

ͺͶͳ

LQRXU51$VHTDQDO\VLVZHXVHGWKH3DWK'HVLJQHUIXQFWLRQRI,QJHQXLW\ ͺͶʹ

3DWKZD\$QDO\VLV,3$:HIRXQGWUDQVFULSWLRQIDFWRUV67$71)N%DQG() ͺͶ͵

WRKDYHGLUHFWUHODWLRQVKLSVZLWKDNMT1DFWLYDWLRQ8SVWUHDPRIWKHVH

ͺͶͶ

UHODWLRQVKLSV,3$DOVROLQNHGJURZWKIDFWRUVVXFKDVLQVXOLQDQG(*)DVZHOODV ͺͶͷ

F\WRNLQHVLQFOXGLQJ71),/ȕDQG,/WRWKH()67$7DQG1)N%VLJQDOLQJ ͺͶ͸

SDWKZD\V:HSURSRVHWKDWRQHFRQVHTXHQFHRIREHVLW\DVVRFLDWHGJURZWK ͺͶ͹

IDFWRUDQGF\WRNLQHVLJQDOLQJLVLQFUHDVHG'107DFWLYDWLRQZKLFKLQWXUQFDQ ͺͶͺ

PRGXODWH'1$PHWK\ODWLRQDQGLPSDFWWUDQVFULSWLRQRILPSRUWDQWJHQHVLQEUHDVW ͺͶͻ

FDQFHUVXFKDV(5ĮDQG(5ȕ0HWK\ODWHG&S*GLQXFOHRWLGHVDUHLQGLFDWHGE\ ͺͷͲ

UHGFLUFOHVXQPHWK\ODWHG&S*GLQXFOHRWLGHVDUHLQGLFDWHGE\ZKLWHFLUFOHV ͺͷͳ

ͺͷʹ

ͺͷ͵

ͺͷͶ

ͺͷͷ

ͺͷ͸

ͺͷ͹

ͺͷͺ

ͺͷͻ

͵Ͷ 'LIIHUHQFHVEHWZHHQGLHWJURXSVLQLQVXOLQOHSWLQDGLSRQHFWLQ,*)DQGȕ ͺ͸ͳ

HVWUDGLRODWHDFKWLPHSRLQWZHUHDQDO\]HGE\RQHZD\$129$IROORZHGE\ ͺ͸ʹ

7XNH\¶VSRVWKRFWHVW2:RU',2YV&5PPP

ͺ͸͵

2:YV',2♦P♦♦P♦♦♦P

ͺ͸Ͷ

0RQWKV

RQ'LHW &5 2: ',2

,QVXOLQ QJP/

%DVHOLQH “

“ “ “

“ “ “

“ “ _“♦♦

/HSWLQ QJP/

%DVHOLQH “

“ “ “

“ _{“ “}

“ “ _“

$GLSRQHFWLQ QJP/

%DVHOLQH “

“ “ _“

“ _{“ “}

“ _{“ “}

,*) QJP/

%DVHOLQH “

“ _{“ “}

“ _{“ “}♦

“ _{“ “}♦

ȕ

HVWUDGLRO SJP/

%DVHOLQH

“ “ “

“ _{“ “}

͵ͷ

ͺ͸ͷ

Published OnlineFirst April 3, 2017. Cancer Res

Emily L. Rossi, Sarah M Dunlap, Laura W. Bowers, et al.

MMTV-neu transgenic mice

expression and mammary tumor development in

β

and ER

α

ER

Energy balance modulation impacts epigenetic reprogramming,

Updated version

10.1158/0008-5472.CAN-16-2795

doi:

Access the most recent version of this article at:

Material Supplementary

http://cancerres.aacrjournals.org/content/suppl/2017/03/30/0008-5472.CAN-16-2795.DC1

Access the most recent supplemental material at:

Manuscript

Author

edited.

Author manuscripts have been peer reviewed and accepted for publication but have not yet been

E-mail alerts Sign up to receive free email-alerts related to this article or journal.

Subscriptions

Reprints and

.

[email protected]

Department at

To order reprints of this article or to subscribe to the journal, contact the AACR Publications

Permissions

Rightslink site.

Click on "Request Permissions" which will take you to the Copyright Clearance Center's (CCC) .

http://cancerres.aacrjournals.org/content/early/2017/04/01/0008-5472.CAN-16-2795