FACULTY OF HEALTH SCIENCES DEPARTMENT OF MEDICAL BIOLOGY

Progressive lupus nephritis: Biological consequences of renal DNaseI gene silencing

Dhivya Borra Thiyagarajan A dissertation for the degree of Philosophiae Doctor August 2013

3URJUHVVLYHOXSXVQHSKULWLV %LRORJLFDOFRQVHTXHQFHVRIUHQDODNaseIJHQHVLOHQFLQJ   %\ 'KLY\D%RUUD7KL\DJDUDMDQ 





A dissertation for the degree of Philosophiae Doctor

8QLYHUVLW\RI7URPV¡ 







)DFXOW\RI+HDOWK6FLHQFHV









'HSDUWPHQWRI0HGLFDO%LRORJ\











$XJXVW 1

                

2

7DEOHRI&RQWHQWV 7DEOHRI&RQWHQWV .......................................................................................................................................... 3 $FNQRZOHGJHPHQW ......................................................................................................................................... 5 $EEUHYLDWLRQV ................................................................................................................................................ 7 /LVWRI3DSHUV ................................................................................................................................................. 9 6/(±DQRYHUYLHZ ...................................................................................................................................... 11 &ODVVLILFDWLRQRI6/(................................................................................................................................... 12 (SLGHPLRORJ\ .............................................................................................................................................. 12 (WLRORJLFDOIDFWRUV........................................................................................................................................ 13 3DWKRJHQHVLVRI6/(.................................................................................................................................... 14 ,PSDLUHGFOHDUDQFHRIDSRSWRWLFFHOOVin vivo .......................................................................................... 15 /XSXV1HSKULWLV ........................................................................................................................................... 17 (SLGHPLRORJ\ .............................................................................................................................................. 17 (WLRORJ\ ....................................................................................................................................................... 17 &ODVVLILFDWLRQRIOXSXVQHSKULWLV .................................................................................................................. 18 3DWKRSK\VLRORJ\RIOXSXVQHSKULWLV ............................................................................................................. 19 $SRSWRVLVDXWRLPPXQLW\DQGOXSXVQHSKULWLV ............................................................................................. 21 1XFOHDVHV ................................................................................................................................................. 22 '1DVH,DQGLWVUROHLQDXWRLPPXQLW\....................................................................................................... 25 %LRORJLFDODQGSDWKRSK\VLRORJLFDOFRQVHTXHQFHVRIVLOHQFLQJRIWKHUHQDO'1DVH,JHQH ................... 25 +RZLV'1DVH,UHJXODWHGLQOXSXVQHSKULWLV" ...................................................................................... 26 5ROHRIF\WRNLQHVLQOXSXVQHSKULWLV .................................................................................................... 28 $LPVRIWKHVWXG\ ........................................................................................................................................ 33 6XPPDU\RIWKHSDSHUV ................................................................................................................................ 34 3DSHU, ........................................................................................................................................................ 34 3DSHU,,....................................................................................................................................................... 35

3

3DSHU,,, ..................................................................................................................................................... 36 *HQHUDO'LVFXVVLRQ ...................................................................................................................................... 37 &RQFOXGLQJUHPDUN ...................................................................................................................................... 44 )XWXUHSHUVSHFWLYHVLQDQDO\]LQJOXSXVQHSKULWLV......................................................................................... 45





4

$FNQRZOHGJHPHQW 7KH SUHVHQWHG ZRUN ZDV FDUULHG RXW LQ WKH IRUPHU 0ROHFXODU 3DWKRORJ\ 5HVHDUFK *URXS QRZ 51$DQG0ROHFXODU3DWKRORJ\5HVHDUFK*URXS ,QVWLWXWHRI0HGLFDO%LRORJ\DW8QLYHUVLW\RI 7URPV¡ 1RUZD\ IURP 6HSWHPEHU  WR $XJXVW  , WKDQN WKH 8QLYHUVLW\ RI 7URPV¡ IRU ILQDQFLDOVXSSRUWDQGIRUWKHRSSRUWXQLW\WRXVHWKHPRGHUQHTXLSPHQW¶VLQWKHODERUDWRULHV $FKLHYLQJD3K'GHJUHHLVDZRQGHUIXOIHHOLQJDQGDKDSSLHVWPRPHQWLQWKHOLIH%XWZLWKRXWWKH KHOSRIVXSHUYLVRUVIDPLO\IULHQGVDQGFROOHDJXHVLWLVQRWSRVVLEOHVR,WDNHWKLVRSSRUWXQLW\WR WKDQNWKHIROORZLQJV )LUVWDQGIRUHPRVW,OLNHWRH[SUHVVP\JUDWLWXGHWRP\VXSHUYLVRU2OH3HWWHU5HNYLJ2OH3HWWHU WKDQN \RX IRU RIIHULQJ PH WKH 3K' SRVLWLRQ LQ WKLV FRPSOH[ ILHOG RI OXSXV DQG PROHFXODU LPPXQRORJ\