Depar t ment of Obs t et r i cs and Gy necol ogy
Tadao Tanaka,Professor Kazunori Ochiai,Professor Kazuhiko Ochiai,Professor Hiroshi Sasaki,Professor Takekazu Onda,Professor Naoki Kamiya,Professor Seiji Isonishi,Associate Professor Shigeki Niimi,Associate Professor Aikou Okamoto,Associate Professor Shigemitu Kobayashi,Assistant Professor Kyousuke Yamada,Assistant Professor Kuniaki Ohura,Assistant Professor Hirokuni Takano,Assistant Professor Satoshi Takakura,Assistant Professor Hideo Shinozaki,Assistant Professor Kentaro Sugiura,Assistant Professor
Gener al Summar y
The ma i n r e s e a r c h t opi c s of our de pa r t me nt a r e t he de v e l opme nt of mol e c ul a r l y t a r g e t e d a g e nt s f or g y ne c ol og i c t umor s , i nc l udi ng ov a r i a n c a nc e r ;c l a r i f i c a t i on of t he me c ha ni s ms of s uc c e s s f ul pr e g na nc y ;a nd t he de v e l opme nt of a s s i s t e d r e pr oduc t i v e t e c hni que s . The s e t opi c s we r e i nv e s t i g a t e d bot h e x pe r i me nt a l l y a nd c l i ni c a l l y .
Res ear ch Act i v i t i es Gy ne c o l o g i c o nc o l o g y
1 . I nt e g r a t e d Copy Numbe r a nd Ex pr e s s i on Ana l y s i s of Che mor e s i s t a nt Ov a r i a n Ca r c i noma s
Wome n wi t h s e r ous ov a r i a n c a nc e r a r e of t e n i nt r i ns i c a l l y r e f r a c t or y t o pl a t i num- t a x ol
―ba s e d t r e a t me nt or be c ome r e s i s t a nt on r e l a ps e . Be c a us e a c c ur a t e l y pr e di c t i ng t he r e s pons e t o c he mot he r a py r e ma i ns pos s i bl e ,we s oug ht t o i de nt i f y s oma t i c DNA c opy numbe r v a r i a t i on( CNV)a s s oc i a t e d wi t h pr i ma r y r e s i s t a nc e i n a dv a nc e d- s t a g e di s e a s e . Ge nome - wi de f r e que nc y a nd t he l e v e l of CNV i n 1 1 8 ov a r i a n t umor s we r e me a s ur e d wi t h s i ng l e nuc l e ot i de pol y mor phi s m mi c r oa r r a y s . A we l l - de f i ne d s ubs e t of 8 5 a dv a nc e d- s t a g e s e r ous t umor s wa s t he n us e d t o r e l a t e CNV t o pr i ma r y r e s i s t a nc e t o t r e a t me nt . The di s c ov e r y - ba s e d a ppr oa c h wa s c ompl e me nt e d by qua nt i t a t i v e pol y me r - a s e c ha i n r e a c t i on a na l y s i s of c opy numbe r of 1 2 c a ndi da t e g e ne s pr e v i ous l y r e por t e d t o be a s s oc i a t e d wi t h c l i ni c a l out c ome i n ov a r i a n c a nc e r . Li ke l y CNV t a r g e t s a nd t umor mol e c ul a r s ubt y pe s we r e f ur t he r c ha r a c t e r i z e d by g e ne e x pr e s s i on pr of i l i ng . Ampl i f i c a - t i on of 1 9 q1 2 ,c ont a i ni ng c y c l i n E( CCNE1 )a nd 2 0 q1 1 . 2 2 - q1 3 . 1 2 , ma ppi ng i mme di a t e l y a dj a c e nt t o t he s t e r oi d r e c e pt or c o- a c t i v a t or NCOA3 ,wa s s i g ni f i c a nt l y a s s oc i a t e d wi t h a poor r e s pons e t o pr i ma r y t r e a t me nt . Fr om pr e v i ous l y r e por t e d a s s oc i a t i ons of c opy numbe r wi t h out c ome ,onl y t he a mpl i f i c a t i on s t a t us of CCNE1 wa s v a l i da t e d a s a ma r ke r f or pr i ma r y c he mor e s i s t a nc e . Che mor e s i s t a nt t umor s wi t h hi g h CCNE1 c opy numbe r a nd pr ot e i n e x pr e s s i on we r e pr e di c t a bl y a s s oc i a t e d wi t h i nc r e a s e d c e l l ul a r pr ol i f e r a t i on,a s we r e a s ubs e t of t r e a t me nt - r e s pons i v e pa t i e nt s ,s ug g e s t i ng a c e l l - c y c l e
― i nde pe nde nt r ol e f or CCNE1 i n modul a t i ng c he mor e s pons e . Pa t i e nt s wi t h poor
c l i ni c a l out c ome s a nd wi t hout CCNE1 a mpl i f i c a t i on ov e r - e x pr e s s e d g e ne s i nv ol v e d i n
e x t r a c e l l ul a r ma t r i x de pos i t i on. Our f i ndi ng s i de nt i f y 2 di s t i nc t me c ha ni s ms of pr i ma r y
t r e a t me nt f a i l ur e i n s e r ous ov a r i a n c a nc e r ,i nv ol v i ng CCNE1 a mpl i f i c a t i on a nd e nha n-
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c e d e x t r a c e l l ul a r ma t r i x de pos i t i on.
2 . Me s e nc hy ma l - t o- e pi t he l i a l t r a ns i t i on dur i ng t he i nc l us i on c y s t f or ma t i on f r om huma n ov a r i a n s ur f a c e e pi t he l i um
Mos t s ur f a c e e pi t he l i a l - s t r oma l t umor s of t he ov a r y a r e t houg ht t o a r i s e f r om e pi t he l i a l i nc l us i on c y s t s . Thus ,t he s e c y s t s a r e pr e c ur s or l e s i ons of ov a r i a n c a r c i noma . On t he ba s i s of t hi s hy pot he s i s ,we a i me d t o c ha r a c t e r i z e t he huma n ov a r i a n s ur f a c e e pi t he l i um i n whi c h t he me s e nc hy ma l - t o- e pi t he l i a l t r a ns i t i on oc c ur s i n t he pr oc e s s of i nc l us i on c y s t f or ma t i on. We us e d s pe c i me ns f r om 9 pa t i e nt s wi t h e ndome t r i a l c a nc e r who unde r we nt hy s t e r e c t omy a nd bi l a t e r a l s a l pi ng o- oophor e c t omy . I mmuohi s t oc he mi c a l s t udi e s we r e pe r f or me d of 1 0 nor ma l ov a r i e s c ont a i ni ng 9 2 i nc l us i on c y s t s a nd 4 nor ma l f a l l opi a n t ube s t o e x a mi ne t he e x pr e s s i on of a nt i g e n ma r ke r s ,i nc l udi ng c a l r e t i ni n,podopl a ni n, D2 - 4 0 ,t hr ombomodul i n,huma n bone ma r r ow e ndot he l i a l( HBME) - 1 ,v i me nt i n,e pi t h- e l i a l me mbr a ne a nt i g e n( EMA) ,WT1 ,c a r bohy dr a t e a nt i g e n( CA)1 2 5 ,MOC3 1 ,t umor - a s s oc i a t e d g l y c opr ot e i n( TAG)7 2 ,Be r - EP4 ,a nd E- c a dhe r i n. We f ound t ha t pos i t i v e s t a i ni ng r a t e s f or me s ot he l i a l ma r ke r s i n nor ma l ov a r i a n s ur f a c e e pi t he l i um we r e 1 0 0 % ( 1 0 of 1 0 )wi t h c a l r e t i ni n,8 0 %( 8 of 1 0 )wi t h podopl a ni n,8 0 %( 8 of 1 0 )wi t h D2 - 4 0 ,7 0 % ( 7 of 1 0 )wi t h t hr ombomodul i n, 1 0 0 %( 1 0 of 1 0 )wi t h HBME- 1 , a nd 1 0 0 %( 1 0 of 1 0 )wi t h v i me nt i n;t ha t pos i t i v e s t a i ni ng r a t e s f or e pi t he l i a l ma r ke r s i n t uba l e pi t he l i um we r e 1 0 0 %( 4 of 4 )wi t h HBME- 1 ,1 0 0 %( 4 of 4 )wi t h v i me nt i n,1 0 0 %( 4 of 4 )wi t h EMA,a nd 7 5 %( 3 of 4 )wi t h TAG- 7 2 ,a nd 1 0 0 %( 4 of 4 )wi t h Be r - EP4 ;a nd t ha t pos i t i v e s t a i ni ng r a t e s f or bot h ma r ke r s i n i nc l us i on c y s t s we r e 5 1 . 1 %( 4 7 of 9 2 )wi t h HBME- 1 ,4 4 . 6 %( 4 1 of 9 2 )wi t h v i me nt i n,6 5 . 2 %( 6 0 of 9 2 )wi t h TAG- 7 2 ,a nd 8 8 . 0 %( 8 1 of 9 2 )wi t h Be r - EP4 . Ov a r i a n s ur f a c e e pi t he l i um ha s bot h me s e nc y hma l a nd e pi t he l i a l c ha r a c t e r i s t i c s . I n c ont r a s t ,i nc l us i on c y s t g a i ns mor e e pi t he l i a l c ha r a c t e r i s t i c s wi t h t he l os s of me s e nc y hma l c ha r a c t e r i s t i c s . The s e f i ndi ng s s uppor t a me s e nc hy ma l - t o- e pi t he l i a l t r a ns i t i on dur i ng i nc l us i on c y s t f or ma t i on f r om ov a r i a n s ur f a c e e pi t he l i um.
3 . Mi c r oRNA ( mi RNA)e x pr e s s i on pr of i l e s f or c a nc e r s ,i nc l udi ng t hos e of t he l ung , br e a s t ,s t oma c h,pr os t a t e ,a nd c ol on,we r e e x a mi ne d t o i nv e s t i g a t e t he mi RNA i nv ol v e - me nt i n c a r c i nog e ne s i s . We a r e now i nv e s t i g a t i ng t he r ol e s of mi RNA i n t he r e s i s t a nc e of huma n ov a r i a n c a nc e r c e l l s t o pa c l i t a x e l . Our f i ndi ng s ma y ha v e s i g ni f i c a nt i mpl i c a - t i ons f or t he r a pe ut i c s t r a t e g i e s a i mi ng t o ov e r c ome c a nc e r c e l l c he mor e s i s t a nc e . 4 . A r a ndomi z e d t r i a l of r e t r ope r i t one a l c l os ur e v e r s us ope ni ng ha s be e n r e g i s t e r e d t o pr e v e nt l y mphe de ma a f t e r l y mphe de ne c t omy f or pa t i e nt s wi t h ut e r i ne c e r v i c a l c a nc e r or e ndome t r i a l c a nc e r . A t ot a l of 1 5 0 c a s e s wi l l be r e g i s t e r e d;6 4 c a s e s ha d be e n r e g i s t e r e d by Ma r c h 2 0 0 9 .
5 . Thi s s ma l l pr oof - of - pr i nc i pl e s t udy ha s de mons t r a t e d t ha t t he r e a r e pr of i l e s i n t he s e r um of J a pa ne s e ov a r i a n c a nc e r pa t i e nt s t ha t c a n be us e d t o c l a s s i f y t he pr e s e nc e of c a nc e r . The i nf or ma t i on i s s i mi l a r t o t ha t i n ov a r i a n c a nc e r s e r a f r om t he Uni t e d St a t e s be c a us e N- di me ns i ona l c l us t e r s bui l t on Uni t e d St a t e s s e r a s pe c t r a c r e a t e d c l us t e r ma ps pr e di c t i v e of t he J a pa ne s e s a mpl e s . The mode l pe r f or me d be t t e r t ha n a ny of t he e x i s t i ng s i ng l e bi oma r ke r a s s a y s ,a l t houg h t r ul y us e f ul mode l s a wa i t a muc h l a r g e r s a mpl e s i z e a nd t he us e of i nde pe nde nt v a l i da t i on s a mpl e s e t s t o de mons t r a t e t he i r r obus t ne s s . The s e r e s ul t s e nc our a g e us t o s t a r t a l a r g e - s c a l e , mul t i s i t e c ol l e c t i on of s e r a f r om J a pa ne s e pa t i e nt s wi t h ov a r i a n c a nc e r t o de v e l op a J a pa ne s e ov a r i a n c a nc e r s e r um pr of i l e a s s a y .
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Fe t o mat e r nal me di c i ne
1 . Ant i phos phol i pi d s y ndr ome( APS)i s a c l i ni c a l e nt i t y ma ni f e s t e d by a r t e r i a l a nd v e nous t hr ombos e s a nd r e c ur r e nt mi s c a r r i a g e s a nd i s c a us e d by a nt i phos phol i pi d a nt i bodi e s . Re c e nt l y ,APS ha s a l s o be e n obs e r v e d wi t h s ome c ompl i c a t i ons of pr e g - na nc y ,e . g . ,pr e g na nc y - i nduc e d hy pe r t e ns i on,i nt r a ut e r i ne g r owt h r e s t r i c t i on,a nd l a t e f e t a l de a t h. Howe v e r ,l i t t l e i s known a bout how APS i s i nv ol v e d i n t he s e c ompl i c a - t i ons . The Fc r e c e pt or f or I g G ( Fc γ r e c e pt or )i s i mpl i c a t e d i n s ome a ut oi mmune di s e a s e s . To i nv e s t i g a t e t he pa t hol og i c a l s i g ni f i c a nc e of t he Fc γ r e c e pt or i n APS a nd c ompl i c a t i ons of pr e g na nc y ,we ha v e a t t e mpt e d t o e s t a bl i s h a n e x pe r i me nt a l mode l f or APS us i ng Fc γ r e c e pt or knoc k- out mi c e .
We e x a mi ne d t he pr e s e nc e of a nt i phos phol i pi d a nt i bodi e s i n pa t i e nt s who ha d obs t e t r i - c a l c ompl i c a t i ons ,a nd i nv e s t i g a t e d pl a c e nt a l pa t hol og y .
2 . Ma ny pa t i e nt s wi t h r e c ur r e nt pr e g na nc y l os s be c ome i nf e r t i l e or ha v e r e pe a t e d s pont a ne ous a bor t i ons a f t e r i nf e r t i l i t y t he r a py . The s e t r a ns i t i ona l c ondi t i ons ha v e not be e n r e s e a r c he d s o f a r . We ha v e i nv e s t i g a t e d t he di f f e r e nt pos s i bl e c a us e s a nd c l i ni c a l ma ni f e s t a t i ons of t he s e c ondi t i ons f r om t he pe r s pe c t i v e of r e pr oduc t i v e f a i l ur e . 3 . Es t a bl i s hme nt of a n i mmor t a l i z e d huma n e x t r a v i l l ous t r ophobl a s t c e l l l i ne by r e t r ov i r a l i nf e c t i on of E6 / E7 / huma n t e l ome r a s e r e v e r s e t r a ns c r i pt a s e
I nv e s t i g a t i on i nt o t he f unc t i on of huma n t r ophobl a s t s ha s be e n r e s t r i c t e d by a l a c k of s ui t a bl e c e l l mode l s . We a i me d t o obt a i n l ong - l i v e d huma n nor ma l t r ophobl a s t c e l l l i ne s t ha t woul d s e r v e a s i de a l i n v i t r o c e l l mode l s . Pr i ma r y huma n t r ophobl a s t c e l l s we r e de r i v e d f r om t he pl a c e nt a of a woma n who ha d unde r g one e l e c t i v e a bor t i on dur i ng t he 7 t h we e k of g e s t a t i on. The c e l l s we r e i mmor t a l i z e d by i nf e c t i on wi t h r e t r ov i r a l e x pr e s s i on v e c t or s c ont a i ni ng t y pe 1 6 huma n pa pi l l oma v i r us E6 a nd E7 i n c ombi na t i on wi t h huma n t e l ome r a s e r e v e r s e t r a ns c r i pt a s e( hTERT) . Cha r a c t e r i z a t i on of t he c e l l l i ne wa s pe r f or me d. I mmunoc y t oc he mi c a l s t a i ni ng f or huma n c hor i oni c g ona dot r ophi n c ha i nβ ,c y t oke r a t i n 7 ,huma n l e ukoc y t e a nt i g e n G,a nd CD9 i ndi c a t e d a n e x t r a v i l l ous t r ophobl a s t i c phe not y pe . Tr a ns we l l i ns e r t i nv a s i on a s s a y s howe d t he i nv a s i v e ne s s of t hi s c e l l l i ne ,a nd g e l a t i n z y mog r a phy s howe d s e c r e t i on of ma t r i x me t a l l opr ot e i na s e s 2 a nd 9 . Ka r y ot y pe a na l y s i s s howe d a l mos t nor ma l c hr omos oma l numbe r wi t h s ma l l de v i a t i ons r a ng i ng f r om 4 6 t o 4 8 ,a nd a nude mous e a s s a y s howe d no t umor i g e ne c i t y . Thi s ne wl y i mmor t a l i z e d c e l l l i ne , HChEpC1 b, wi l l pr ov i de a us e f ul mode l f or t he s t udy of e x t r a v i l l ous t r ophobl a s t f unc t i on.
4 . Dy na mi c s of bi opy r r i ns i n pr e g na nt wome n a nd ne wbor ns
Bi opy r r i ns a r e bi l i r ubi n ox i da t i v e me t a bol i t e s t ha t c a n be me a s ur e d i n pr e g na nt wome n t o moni t or ps y c hol og i c a l s t r e s s . Ur i na r y bi opy r r i n l e v e l s we r e s i g ni f i c a nt l y e l e v a t e d by de l i v e r y f r om 4 . 2 2 ±0 . 4 7 t o 7 . 3 3 ±0 . 6 8( U / g ・ Cr e ) . I nduc t i on of de l i v e r y i nc r e a s e d pos t pa r t um bi opy r r i n l e v e l s by 8 9 %( 1 0 . 7 4 ±1 . 3 7 v s 5 . 6 7 ±0 . 8 0( Ug ・ Cr e ) . The s e da t a s ug g e s t t ha t pr e g na nc y a nd de l i v e r y i nc r e a s e ps y c hol og i c a l s t r e s s a nd t ha t de l i v e r y i nduc t i on i nc r e a s e s s t r e s s e v e n mor e .
Re p r o duc t i v e e ndo c r i no l o g y
1 . Mos t of t he me c ha ni s ms f or a c hi e v i ng pr e g na nc y ha v e be e n c l a r i f i e d owi ng t o
a dv a nc e s i n a s s i s t e d r e pr oduc t i v e t e c hnol og y . Ne v e r t he l e s s , t he me c ha ni s m of i mpl a nt a -
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t i on r e ma i ns unc l e a r .
CD1 4 7 i s e x pr e s s e d a t hi g h l e v e l s on c e l l s ur f a c e s of v a r i ous t umor s a nd s t i mul a t e s ma t r i x me t a l l opr ot e i na s e s . We hy pot he s i z e d t ha t CD1 4 7 ma y pl a y a n i mpor t a nt r ol e i n i mpl a nt a t i on. The a i m of t hi s s t udy wa s t o de t e r mi ne t he e x pr e s s i on a nd hor mona l r e g ul a t i on of t he CD1 4 7 g e ne dur i ng t he huma n i mpl a nt a t i on pe r i od i n c ont r ol l e d ov a r i a n hy pe r s t i mul a t i on c y c l e s . We f ound t ha t l e v e l s of CD1 4 7 a nd ma t r i x me t a l l o- pr ot e i na s e 2 mRNA i n huma n e ndome t r i um we r e s i g ni f i c a nt l y de c r e a s e d dur i ng t he s e c r e t or y pha s e i n c ont r ol l e d ov a r i a n hy pe r s t i mul a t i on c y c l e s .
Publications
Itoh H,Iwasask M,Nakajima Y,Endo Y,Hanaoka T,Sasaki H,Tanaka T,Yang B,Tsugane S.A case-control study of t he association between urinary cadmium concentration and en- dometriosis in infertile Japanese women.Sci Total Environ 2008;402:171‑5.
Omi H,Okamoto A,Nikaido T,Urashima M, Kawaguch R,Umehara N,Sugiura K,Saito M, Kiyono T,Tanaka T.Establishment of an im- mortalized human extrav illous trophoblast cell line by retroviral infection of E6/E7/hTERT and its transcriptionalprofile during hypoxia and reox- ygenation.Int J Mol Med 2009;23:229‑36.
Saeki T,Sano M,Komoike Y,Sonoo H,Honjyo H, Ochiai K,Kobayashi T,Aogi K,Sato N,Sawai S, Miyoshi Y,Takeuchi M,Takas hima S.No increase of breast cancer incidence in Japanese women who received hormone replacement therapy:overview of a case-control study of breast cancer risk in Japan.Int J Clin Oncol 2008;13:8‑11.
Katsumata N,Fujiwara Y,Kamura T,Nakanishi T, Hatae M,Aoki D,Tanaka K,Tsuda H,Kamiura S, Takehara K,Sugiyama T,Kigawa J,Fujiwara K, Ochiai K,Ishida R,Inagaki M,Noda K.Phase II clinical trial of pegylated liposomal doxorubicin (JNS002)in Japanese pat ients with mullerian carcinoma(epithelial ovarian carcinoma,primary carcinoma of fallopian tube,peritoneal car- cinoma)having a therapeutic history of platinum- based chemotherapy:a Phas e II Study of the Japanese Gynecologic Oncology Group.Jpn J Clin Oncol 2008;38:777‑85.
Tada H,Teramukai S,Fukushima M,Sasaki M.
Risk factors lower limb lymphedema after lymph
node dissection in patients with ovarian and uterine carcinoma.BMC Cancer 2009;9:47.
Etemadmoghadam D,deFazio A,Beroukhim R, Mermel C,George J,Getz G,Tothill R,Okamoto A,Raeder MB,Harnett P,Lade S,Akslen LA, Tinker AV,Locandro B,Alsop KChiew YE, Traficante N,Fereday S,Johnson D,Fox S, Sellers W,Urashima M,Salvesen HB,Meyerson M,Bowtell D;AOCS Study Group.Integrated copy number and ex pression analysis of chemoresistant. ovarian carcinomas.Clin Cancer Res 2009;15:1417‑27.
Okamoto S,Okamoto A,Nikaido T,Saito M, Takao M,Yanaihara N,Takakura S,Ochiai K, Tanaka T.Mesenchymal to epithelial transition in the human ovarian surface epithelium focusing on inclusion cysts.Oncol Rep 2009;21:1209‑ 14.
Isonoshi S,Ogura A,Kiyokawa K,Suzuki M, Kunito K,Hirama M,Tachibana T,Ochiai K, Tanaka T.Alpha-fetoprotein(AFP)-producing ovarian tumor in an elderly woman.Int J Clin Oncol 2009;14:70‑3.
Schetter AJ,Leung SY,Sohn JJ,Zanetti KA, Bowman ED,Yanaihara N,Yuen ST,Chan TL, Kwong DL,Au GK,Liu CG,Calin GA,Croce CM, Harris CC.MicroRNA expression profiles as- sociated with prognosis and therapeutic out- come in colon adenocarcinoma.JAMA 2008;
299:425‑36.
Kawaguchi R,Tanaka T. Priming of peripheral monocytes with prolactin sensitizesIFN-gamma- mediated Indolamine 2,3-dioxygenase expres- sion without affecting IFN- gamma signaling.J Reprod Immunol 2008;77:117‑25.
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