HUE031559T2 - Eljárás angiogenezis, vaszkularizáció vagy érjavítás elõsegítésére vagy tumor-angiogenezis gátlására - Google Patents

Eljárás angiogenezis, vaszkularizáció vagy érjavítás elõsegítésére vagy tumor-angiogenezis gátlására Download PDF

Info

Publication number
HUE031559T2
HUE031559T2 HUE08844273A HUE08844273A HUE031559T2 HU E031559 T2 HUE031559 T2 HU E031559T2 HU E08844273 A HUE08844273 A HU E08844273A HU E08844273 A HUE08844273 A HU E08844273A HU E031559 T2 HUE031559 T2 HU E031559T2
Authority
HU
Hungary
Prior art keywords
kot
angiogenesis
rti
ség ség
ség
Prior art date
Application number
HUE08844273A
Other languages
English (en)
Inventor
Stefanie Dimmeler
Andreas M Zeiher
Angelika Bonauer
Carmen Urbich
Original Assignee
T2Cure Gmbh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by T2Cure Gmbh filed Critical T2Cure Gmbh
Publication of HUE031559T2 publication Critical patent/HUE031559T2/hu

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/11—Antisense
    • C12N2310/113—Antisense targeting other non-coding nucleic acids, e.g. antagomirs
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2320/00—Applications; Uses
    • C12N2320/50—Methods for regulating/modulating their activity

Description

ELJÁRÁS ÀNGIOÂièitS, VAS£KDLAliI2.ÁCtí> VA«Y &8JAVÍYÁS EI.6.3EGÍ YÈSÊR R ν&φ
1'üiHÔR-AF*C;îiKÎEN«:Â!S GÁTLÁSÁRA
Ths invention rentes to & method lor infhumefog the mlK-32 expression m a ceil, to pameuisr for promoting the angiogenesis. vascularization or vessel repair, or for inhibiting or blocking tumor angiogenesis, sod the tee of such a me!hod for the therapy of a disease or s condition, furthermore, the mvumion relaies so a pharmaceutical composition for promoting aagiogsnesw, 'Wukrlxstioo or vessel repair, or for inhibiting or blocking tensor magló-geaesis In a edh
The erdothelhsru plays as important role la Ins maMermrice of the Integrity and iimolinnallty of the vessels, in the adod, dre growth of new vessels cecum through arteriogenesis, angiogenesis or vsseologenesls. WMIsl artefiogenesis is defined as the grossit of collateral vessels, angiogenesis is understood as the growth of neve flood vessels front pre-erwhog vessels. During angiogenesis, resting endothelial cells ate activated fey angiogenic factors and start to migrate, proliferate and organise thenrseives into tubular structures (21. The term vascttlogcnesis initially described the rk nova blood vessel formation 1st the embryo from sogioblusts, but now also includes the formation of blood vessels from endothelial preenrsot ceils nr other adult stem ceils .(1), ..Angiogenesis and vaséul agenesis rep' reieni physiological developmental processes winch play ars essential role 1st the reconstitution of the blood flow in ischemic tissyes, anti are a basic step in the growth of turners, The promotion of angiogenesis «ml neo vascularisation was kksilhed as a possible therapeutic siratni«, for example, patients suffering from ischemia, (n tumor an· giogenem, the Itm Hat ton of these processes leads to a ^repression of tumor growth, M IctoBNAs (rnilTNAs) «re small mxo'-soeodmg RDAs that regulate the genu expression on. .(he pest-.transcriptional: -levőit ifehs.ugh a degradation of the target rnRhlA or through translational repression (3). in contrast- to small interfering Rbl&s (xiRDAT which bind'to.emspieinemsty ··ÄA sequences, the binding of the srlRblA to a target docs mi only take plsco at a compk-rnenDry RMA, but ferm» more eomgkx RNA-RhlA-stecttn't's that are thamtodysamlcoUy preferred (4), 'tins 'Tneomplcle" binding allows the bbfeing of one míRDA molecule to different toRMA molecules. The tsgtdatlon of a tier of genes (in emstsashtO: a umnntherapy using a gene er a growth factor! cap represent art advantage If ttyeesby .complex regulatory processes tons be Iniloeneed. So far, more -than 400 nn'RblAs: have been identified I» the human geaorne» but lise relevance of most of these mfRbiAs fór (he eeiimar 'function in physiological sud psOtologk processes Is still unclear. Whereas ft was:. described in the -state of the aft that the dowroreguialkm of She mlRNArgroeessisg enzymes Dicer and Drosfes Impedes angiogenesis (3 - 7), only few specific mlRbsAs have been described whnh infioence endothelial cell junctions and angiogenesis. MÍR--331 and mife-222 block the eudothdlal cell migration, proliferation and angiogenesis -in·· vitra in art indirect manner· through an-interaction with the. stem cell factor receptor c-ki! and regulation of the eNDS expression (b,i}}: 'In.contrast to this, the expression of fetMand miR.-2?h contributes to no in vfe-o angiogenesis (7}..
The Siam of the an attributed a strong tumor angiogenesis-promoting activity to- the mihNA cluster oriR- }? 92, lire miR-17-92 cluster consists of miR-lf-Sp, mlli-bMp, miR-Uk, milT-IRa, ntiR-20s, tniR-19h, and oriR-92-· 1 (3}. This miR-17-92 cluster is up-regulated in Myo-induced tumors, and the individual roiRNAs miR-· IS and mlR-19 .could be identified as molecules winds spseldcidiy hitemet with tbs.expression ofasti-angiogenk proteins. A sgeelffe evaluation of the targets of these miliNAs showed drat rniR-ΙΒ preferably suppresses the expression of the sonnective tissue growth factor (CTöFt, whereas miR-i9 -«iteftós with the .strong sugiogettesk-inhibitor thrórnbosponriin-1 (TSF" I f t 10}.
It Wits surprisingly found in secordatroe. wliii thednvention-thaf miR-92 does not promote she angiogenesis as describee? pfeg sím« of |ss;|r| feyt strongly '** «kr« méviya Tp ifthibid»sr<d'mlÄ3::d?«:sissiSSïs^MscuirM&amp;Sv.Âmsr^^^ 8^^?^^^8WMB%:mdfm^::ggg^|9psb'a<ä$#SyyM'ÄeBiMio fer sö the strife dlls® m~ %?s;*tfea^^ MB IMldbitios 0’%e?fm$w **à wá iRithelk? îbckÂgtkss «}5à»|iÂfc:t)i^k oxide Syni#s 'ö»#ÄW {SÏRTIg wlÄsre hotte- :K£Hi:isi:tfef ;|>tïsÎîîîsîÎil endothelial esdi Räoebbiis fS ï to 13¾. âtk? ikiprlRip wbjcb:oosdsRs th« s^ö^1Wí^1i.í».@i8|íy: anáfhc jMeS'äfcUdk wÖk feíRsfe (í-i). 1:¾ íakíb:Nfebíö|?miBidfe by ikiR-tfe mbfefesís fegrcspttss new íhdrípeukti .sraicgy Oar knptovifig tbc end<sííieh&amp;? «H fonction 3iîd.«s«v8S«Mten^^feÂ^^*^ time, it was found that tte over-expressn»·. «4 sttiR'92 reduces tteovascuitai«uk>&amp;. inscription «f ílio íorcnöon änglbgcnsesctsvltyhss dpssniíeá:;&amp; the state of the an s<> tar.
More specifically. ibe soient ion relates to aiseihod k>r sntlnedesog the ivoR-'y· epresem» sa a cell. sa parue-niitr wîbe-;êôntext ot'anpiogenesij. ensd/or v^«i^#p^f.:A€iör^g.i«ö the invent«)«. she method comprises me lol-k>Wmg sidps; ai provùkng si edi: and hit Reçue.:«. d«î n»R~fe' express"!'' ·» the .c?l -a order ?> p«o*mv .e«;;.«.' :m . x^.covr»^^ fef y^kip^lr by felyôÉiediRàiypRïfesïsse moIs^tÄ Rgbhsaitplfeysbdîofes üä?« «r b3) InntkSssrig thé yiiR^iibestiîessiepyplhs «e!l feROn I safe jfeltlerts o f lit« iomsr ;u?po|boests fey iotsootjemg a corofrucf torn the red. wherein said construct Includes an expressible trdR»$3wOMmmec M is also possible, io tmroy once osiR-Sfe mto the cell. lii sr prefes'fed enfeodhaere, said iordmfefe'an η; v;?i<> ipethed.
The ;e;m reducer;·. or in«;vasii« .a die rmR-92 esptessfeti m the cds Is based on a «omprisitb with the rmR-expres-satm in a «bd-type cell, as ears he pcrlhmnx? using RÏ-RCR or reaS-titne FCR. The respective changent g... 0Sít}vsSíOií e;M5 aiso be determined «.-mg die tiinebosa? effects «I the modified expression level ot miR-v.?. on fee characteristic:* <-« toe sell.
An antsstmse molee«?« ;s Asingie'Stramieil molecule having A spddnce whicîï is esseotisliy reverse comple-niedtarv A?as RKfe Cbsee and which inhibits the biological fiinfelon ot mtR-92 through hybridkarion with; the ooR-92. Frc&amp;rably. the molecule is no antisense RbtA, which opr tot-ally can comprise diemical ivt-xldlcainie::. 1¾¾ constriK! can be « pfe^mid, o. costnid. a ciras or a prc;:«:xaX'SViiRNA, whei-e«t the consumer tfevania·· ecttdslv Ineifefes a mcanv for the oanseripiioe of the •.wpo.niibR; roîR-d.c segnenee as tncURteo. sac» as, tor example.. ?P>xW*· ch c a.o ' ''''•d ' > j p;V iedn . tniR-R..:" as used herein inoittdoa bcoh praeursnr moleocdcs. - och as pre-dda··! , pre-feRni, stfe prer oho .r- .%„!i nrtxessed molectdcs such as ouh-feay and sn;R-:t.ïh tsee also toe sequences according to SkQ h>
Th" ioii'oductien of a sooleeule or a cansiroc· can take place «sing both ptiysicai -mÄops isuCi; a® m;' ctoifiiection or oJectreporatifmV by «Ü #f Φ^Μ'^ΦΦ^Φ »·#^8ΐρίρη.ί phosphate or tipoieatmu. of by means of c ira? ntctinKts petung vtmse»}. in s pcefernfe cndxRiirneth -d !hc -ncetiticio, inRoencing of the oxptessicn <e a ρκ««?.π tn the ce a resuns rraivt mfk-wa« the odR-R2 expression in a-.eti. nt-etein said pt c-htin it; selected Rati the groop <»?: «KO». SI Κι t. oneg· tin alpha 5. imegtm hota h mtegtm atoha v. bpiooty HMt-! 4 T ie .:. ΛΝΟ 3. MKK 4, KU' ?· ?’?. A?·. b.lX'« t and MP IB. A15 these pisili isp tmtxishtd whhsite IâîoI <tf O*? «edothelhti cell the sthsto'geotecUn«, spdOrdss posinstni a«ôS?M^feî?s5aîlo5J»-SïR‘V5' t§Ihttgewnisp»nssocittteá with stgswxdsted dtsepSPs.·:
Ihwtdmiigcoli s* öf« sep io 'm\mó Torst, fcuçhm-% bteptpte, s ceii of s 4#" p| ssii Itesvss smfi Sfifee gspvrpte of tetted as prt of s shssst% sa spaôgtettette kted of etel:.Âfersblÿrslp pelî sa provided is a vaastuter cdGaihÄfpiÄ'sait a ssll aa ÉntMöináöQí^%$e#er stetteteted cap, il)is in each cas« in sistes* aii precss sor ceil* and stem «ails oithé «fe as mdicatesi. I« a pseferred embodisoenî of the i5teii?od. ihe stem edi js w>t a human ssoteyotec stem call, Ettehomtortellte «·Η as provided csrrfee present woteted, feat aba :n <ο·μ>-deiioo with a tissue or an organ. la osa emfeadmisnfe she methoo can also be performed m two.
Ids ceil as provided preferably stems irons a ssteteaoam m particular ferst a mamma! testeit as, ter exmoplA its Ä, sodr asmmst; or ran. a ssumkew a great ape, a bovine, a pig. a dogssrs car os; patlkmmiy prefered, fenva hwvtast fedueikm of ibe miR-92 expresÁste ;·$ as much as she method leash. to a redoetkm of the sniRte2 espresxba in the cedi eonrpared to the aormai exprewfe of miife2 in a fed-type cdh the reduction of site: exprefeorsdaks* piaca dtrostgh pravutesg a mofefe selected isii iMsgäsmp «osîsisîsisg «famixenxe moRcelec. syteischc steR-92 Inhibitors,. and sramtenpfen factor inhibitors. This restesten: of the ooR-92 expression tends to fe example, ai) bmrease of sise vaxetearfehon and vessel .'stes sS)5î;Cissss is» «ma <. suhsvjwwMi of tin memod .tevordteg to the mx maim. the astíisssse sostfeolels a mofeçute: whieiffe bäMifefeh rn W^Â-memrn mm^Mm P «ne uffe^lDsNGt I tea, Issth osafcr sirmg^t:as well ssMfeifas stringent conditions. Ibis includes the ps>ssib;hiy ittat 4 fec'feyM4k«ál«4fe.iá*8vstósea^ molecule «fehlte ^mk- felfe neveAhei&amp;te tb mR dhomate the Ifedioö m sfeesute. tfeefebfedssp« tim base ifebfefe** dsgradafefeui mtlfe2 tekén:ptepe. Ih partieotesi; ffee sotb csosStî msjlecuis can tetve n length of up to hi mííiCPf4ite> pstierstfeijf;sup ts) si) ßccieötliies. pmlteisiat iÿ pn^lchiM ds IS: to W teteteolisics, feariebterlv 8sf so iahiditltet of tis«: isoosatete :)oiR42 pteessrsssp fete: adfesse fttoleedle mn have up to §0 opofeteteas, te psrttetdar «p todp ïtoofeolisles. Tlssaolssitoal tepglh of sttælt m atetestese trtoleotessts silvasstugesHisiy !2 ssuciste-islev, prsifessRsly i 5 nueicoiiiiev.
So aspoterted etshediättem. thetestfisensesototeatdesteciodosss ssspäeoeosfhätt Issrovorssi «omfPemsotery *© a ^sitsetice arcotping; to one of SEQiD HO 1 ts> 5. I heteinf íío have o)lamsteh(es> am formed, whioit Seadk.t©:«s eife cletti sohlbfttes of the soiR-Rit Is a pretesi ,j»iWW A ioveitfieo, Ihe mlhmm tmúrnüh h s mstesstle with:: s: vsgiteuco sccerdisi ië BEQ m m%M if Uipmém ihm Ùmrnm&amp;me mtimtte issotesles m tessf one d);*i)tjc4i OHNhíicaiiotí, sswh .=n. fo) exantple Λ ]ea&amp;n>n« :2iM0|.m«thÿh spho^r»^ ph<^lK,tfemi«N..a»ds^ r€te rnteKíxyethvi-CMteorst-uroup. Th.«? «siiSvtiVe ;suiecu?e oa« .ako ,pr »ddllugtall^ menoie ioeked ostete in suis-· ({.,ΗΛ.ί Ctespoîtetas fer evsmde. a very much pm'cmá etnlutdinteot of the a?)tkesvs:e nsotedè iMhofedlo^thg hteiosote, eled) Inc indes ;t Sicsiuse'ce ses esdise.' x? Sf s,t i|) HO *' )'äto s-.uenttcai itteOitudsO.·)», QARíGCCOOOACAAGÜGCA^AMArChssi (ShQ 0> NÖfe| uhereto }Ικ esptfai .ette«s îsid-ente Z '-O-tttetA;-! »o*.<dth<d estv!CO,,de.,, teesltiteittehl t ÇT) tedtestessa hotid Imttveeo atpaceitt tmteedideA ® ehstievK'fd gi'osjp. A teCAGmmh píolepedtehiteilisdenf: sAlfesi ssliseos« moteesiäesinchtdistiiO sequerteesaocordisgioEfO IOHO k with chaussesi mockitesOote. is däC teiiï>A i;ïç uK>ies:tee: .A.CA^OCC^OACÁAOPÖCA^á^A.-íiö'! Í0$ *D NO Ks) ::Ä '^»^^kèmÊ *μφ*μ* w« ^öt im»m m w m««** ehsVmstsotd gsfossp. FW*™« v^i » « «b«» «ο*** !«'“toä * ’“i>rÄS » m ® m ' ' énig^2vC>-;m^^lm£sál|ká imssfosfodes.
Methods 1er âs^fesis of spfob söfefisemútevtílsís tattern»® to te fo^se« sm skill, i«fal Ä MMmàzmiteé ^ ím wmrnmm md.^rnA *"**' wtevMl.t*etapyof»ah««w«ta.«»«XitSon, !»*=άΙ«»«*or^-^oiífifenany pMhötftgfcaO«^^ M boss *s< for *μΦ, mjwdiaí fct (foronk „cbemk hmûÆmm, ipsrlphssod·' :or comsssssy setassy ocsiissfoss.. Iscbcssse in&amp;i$% stroke), palaaiegfo»* ·4«>&amp;ΐ%βί*νδ»ΐ» vSvSv»*,s>.*; λ vxaa^lv», lessor angles», sseSafosses form*!««. dissisesse «tirmpafoy, foi»tcsry Ä^· sotfooscSas-ofo, s«‘nodaxy diseases to artismscfotosss. («case ixssussary syssfofosnc\ sexsxasfofo sísssKo, 5,0»^«. v.a}sxsxs^up>;fo>,s >>» <K*matu«0 agin* or Ägag-assoclakd fosses. « *di 3* fleurodegwwuiiw Awa**, such as Moins» Afcfofoypr,, ssssi Morbus Pas'fosssos.
Further»«»«, foomedxsäas i«ikie:k®ä &amp;rb««s <>? <^k <* ******* cdh ä'i;ds a<" *hr ex»pfe, p^aisgsogcsse cobs, org.:cs :=;xod< p«*»* dfo% towrrwv drived odd 0? «foliating P*«w»t «dis). mil m$mà tm-m $m Ä fo Wài sfotfssf ffoofo Ä‘?fo *»h ÖSs for s****?\*, for the prrtductsoa or snsssel foifosemsssfo materül. iß yattkuter for tissue ripsssuriibo therapy. orte secsforch, ^KmwfdimßsR-α 2 o gxvasksî lomsmssfo as is the mdkrd iöe ftsipwssioß oIobRAO P t«enî»8«d «omp^«4. wifo she «ansutj express^' of sssR-92 ids a yrçiOÿp Ä folg fos ««ample. can take placc throaab an fomasfô oi foc express«»: by sseassx ,of |stîdÉiCm|;of g corsxínscí Imrksdmg as; »xpornsîMst mrË-ki sagaudso íísio ta« ool<< fo i! fÎV fosre« esssfoxforsess of the sseshod assroKSsssg to dis sswersîfosx six: ssúK-foí comafocS stsefodes ;m exfsrsisxssxe «χρκηηχ avcofofog so 80s or SHQ M> MO 1 Is $, or cas fixons sack a sequence, rsspscbvdy. Advaatageoos cotixfoisseats ut sueli a coostrac; have beu-s descrikai above, AlerastfoeO, adfo-92 «so sisoik tolnfooces olreesy nstöfoa edi A maisad ssOfos: Oder dassfobei kind for isilsibitsog the tomsr aagiogssxsis csa de uses for a tber&amp;py ûî a -^^^es«r:Â:-jfpa^»î<>^Si|::ea«KdliïbjE«:=sciiâ-©iâfci^l tfo«? tbs grs>ap esnaisfoig of ayersfoiObqg sopttl^r^fo; undesoed »ago i<^es>es!%^tiors::M4ch^sfo Sfiil&amp;ssuoaifoîss, ;f hfiö^S(íí;tt|{fe«| «^«posiÔïit
Tbc fovaifoojy foifoerfoaxi foiáfoa íö A pkapdMfisfoeai ^ dfouang: fos sfiiltfol 3sdv% m exixessíosvfo ácsi bytheforsfi of áss abdsdosestfolseufo ssgafosf mslOdA ao, pipíorfy svíí-ilsetlCs brfofofor asstlfos' is ss-ausyriptfoss Ifocfor Isslssbsforj qr-a» ssgcisl fer socs-cssfog fos nssR-foá sxpf«ssfo« so a ces es dbe fos·:;·! í>f s çofistrisrS: sfor espsessirig: m«:R'Ä>· llserabyj tlse sästsstsfid eaa a»jîfr.fe«· m.· ïmluà&amp;m .«.xprassfofe sé-qssessca ascsxdfog tousse IXIiMP ; I tu 1 Its osss amfetxitBsriÇt'h« sfidisslesl irs Iha pbarssiaccfitfoal sesoposfoon: Sscfesdes ss ssguefsee accfsfosag so SEQ iD MO fo I or s .s t ssr sdoslsossai pseSes ted xssibsxlibfotiO of sAe eossqacts m- of sbe sssdiscsss« tssofoetsfop referesssc lis made fo foo.d»«enp$<m.as-;·söwt? »s Àt-essfoedlseresi.
Idc fovesîfots fosifecsmsac sAfelgs foia ssÎfilIbdifof pïxxIisaifigïiso.sboYefoetCifodS fotstïu^tâïsaq cosfiposdifps», fhs pbfii'ssjsseesjSfoid fiósssposítlíát tosoMiáig sosfoç bseefoioss: csa b® |«*yçsA sa foe S'orsst (>t folssesg sbssgceg IIPisi, gmodfotssj: íséroaofes, ssfossfoss sölfstíbsss, síísssIsíosss, sys.pcfosfo.ns, sokstlusss,. or fo <a on tbc conssssg ssusscs sai Of m implantable medical device, for example a stetst, respectively.
The i!s.e ncsorsfhag to the Inventur of the agent sat the. pharmaceutical composition. respectRoly, ysk Mkf place using ss es .table' teoWR : h. u.k ^cC'''î-'xîÎt to5he txxuatkstx κ ' h,' 'aarvaVtaa'-ed -a a h koo' > son;" ou» «ht ns.uk' À-mn· UîïïoîS'î- such as, ibr exemple, rabiots., dm goes.. pilis, granulates, aerolites. emulsions, suspensions and sottrnons. us-:s!s·. uses — ssosa-toarâ:, phasnsesieunenlk acceptable cmners. tx sols eats- Hevens, the Thetape'nnsaih etteetsve ssct.ni «.oascesatrutsoia. vs áé teferenne k- da thempënncalh elYetaivs c tarn pounds, shah ke présén; each nt a eoeeemnninn of onvk ft,I sxt .. K· 'is vt -A r'etevapK s- ,sbora p ' 0¾ · ·." ·ό yo ·“··. of ' 'wt et ; Si'sutn ! .' in <oa,n nos tin; use sülhetem at order to .Khn.se the biog ctayyntnRk'Sk n> tegraAd in the tarne; tasssse.
The Kitlpilstsötts spnlhtriphptpld,^' :^t^&amp;:lSing;-I^Î};:'Üit^· ^êïrrs eiím^S^s^tjöiií^· u ,s n'enn.-.-0.-.-Se ο .Άνι,ΐ' 's 'ν ^ .0' ο „ ο sis. ο. η. .ne s sv s κ < > u.i A»s'r > <. r gavik. solvents; v.;ir. he osHlss pcshcux upbtsk F-oriher mentioned its auxiliary sgeats shad he n.en ata-er, non-mate »mivertss, sited tes. for exusaaple, pant dir, V u O'odtíKtr' V t u W„ μ\Ί. '1 " V '' <M\Sh ^e>Ni\u'”i -k *’> ^ .n s .a sus .susnnk. ratnta; stone ou.-ss ,. ·, {.noons, .shrunk;. ta'enm, etaak* uxnthxsao Onra dmts5c s. Hahy .hstxnve shleic »chi, eitle«), stipes (eg. xsKtuxe, hacks* e stsnl gl access), esaulgaîtars. (eg. paaiyosyoihyleue-ibtiy acid-esters, poi>osYe%Pas?-hsttY alcohol-ether*}, dispersing agesats (e.g. iigsat». suilate spent iispsosk triethyf cellulose, scads, so ; "·η s s-î'xh'''>'hdo-v·, .· sd np.natsn i; j ·ο,χα;,"— m . k.e tk ' a-earas, skxrs. a.-o, .no. sont; t · uhuk > ) ne íd'nhsosmríxís takes pace tn .; used muunor, pnierndk sactllx tat purenkidh >.n pumesdot porhnpudx - , i h Is'i ' ..>> s .a . . !î\.' X a " Ip t\0 lv <. tV »V. 't s' ; ' kJUSt a !' ?0 >' . en Os indseated. tablets can of nannte aUo contain additives, sated as, tot exttmptt?, sodi-ataa dtrsle, cakinm carlxm«- and d;en at p;-".pt;Stn kme.nef asuh ; agak ;'t .«tdimev 'itet! <'· at'«. 'î .aotkí-íp:. ροΐ.-kS :t.;k h. ox'i.ap,·, and h e t|s.' a , so . f ^ n ' ' >.n. ' > 0 ·. i a k . s a ! v. > ' \ > t V >. u d '\ ; xihedüg thlliíty. Id ids# sÄ^tadpp^'^p^n^tshVfit-^^^^^^^s^htitötksnisi kitxiiktvg agytitS; tiigidnsp oathhs ;S MV J , d lb e ! as e >a 0 a . 0 ' e 0 hew v >. - f> a. ; se ok > 0 tit 0 <„ assno soitatskc tiqstid corrisr taaatonah; etsta he used:
Meth-sds for iiaeis-py tax prophylaxis |t'. WO V » ü e te · t>; et Ok' S' k' i v \''' o\ ' '.'Ud. 5; . i Ui'.i ! ruiikta nr νηί|ΐθί2 or an tnsk-kl; atsktgutaisa (snua à', fs··; e': a;npT:, an· .amijettaf tovieoak' or.; trottyorapftoti iactíar in-•visitors or a•phtmaammoal nn}npxokk>n ;n aeoordance sauia k:o desetiption as thove os cosutitted hnn'ita, kt the edhtext of such at method, hi d^i^iVt^h dddfl mg Ki ekö mg pet \g nods seeiyht pet d.p oak heρκ!- niitiiaKirdsl;
Antisense mnkeyle stgaitsst -akR-');; ) ne sosenilor at.sta jeimes to .-n .joii.aen'e tnoieuale ansnas; oalRaf2, oYtapfish.g à sequence ashsch dxpradtees tö a aaeslecoie of n sequence aeeotdista to SEQ IDshiÖ I lu S? Arets täte moleddlsife car consists safdpÎdJhinuek-•atäties. in partktalar \S to 22 «»deotjdes Such att aaxtiautnse molecnle aelvatatagenusty contprisea at least Oasxsefees Sees, pfeicrahly »t !en;a à5 n-sdeotides The hyhrkitalota enn éfet^ewfcrf^entoHm'Âpgyhtc.eÉâ&amp;m: hfeth ods fda* ikhoisdihihg tit e ; .hyhrkiipät iota ρρ kfiown to the pet sun of sh ill; xuxh a molecule can opuouaih, in-lndx evtaaka· rneditkaiions; tWttenlas'ly prdfeed is a anolscide äesorde hag Kt St.ts id kiQ g or i { '.vinet'. use hües at Last 0.¾ ot th<. aboxe-tnentimted chemical mödltlcá'k>nskít«^míxi Pásatissnse iiaddslsdititons; sie the muidenks aeeordbg;îtà È'^: ÍT>; hilï S;utad: Id, which Ä smdÄÄ» tee meiecte« aseotesng te SHQ ïü HO *fiî' *>
According te; ter isivemlote xrate a mteeia.sk. cas fee rded vpp^ísgatícte Ante vssste repair.
Accotera;;· s<> tee tevesuon. raSRte:; of slöfci**Ä a. te#es>e? svîorteng So «k of SEQ 10 HO I te S esu; te; usas for iRhteAteg:; pW sisgteiesrpA Site is particular ter Producing s pharmaceutical ompositkm as tie-psi Rite sfevi, |ó pátert stesc not rskäfe te :^c«sses:fey cteoing Waste ivísps oí process Sor mödiiyißg tee germ Hoc aßneik ld«te% «jiffeamœ betep as weit m aw &amp;Î huma« aWryoa terlnHoateii W WraiteteialwlAACS or pswcexxp Iprteoii^ing tea geoeSteidsnfite of araraW atóá-are HMy te Ç&amp;OS0 tefitn Wterteg v4faw4^-^m0&amp;m$k4 bziiipät-tä-Mm or mimtkmë atteiWteäissosoistegferarasskh AO "A.'· '-
Iks lavWWik fcxpiweà is ira»* d«teiÄs.Mrtei*$Μ»ρΙ«.ρ wüte»' feé^ Hoteed tóim m&amp;mplu< ti?«: cessdte of tee experimeras as d*«s ribed sa tin C\.ra;pSs'; ·;ο dopkied Whe.iogur&amp;Si Figuras
The j- sgeras »hew. FWce R lieds of tes over-expression oíonR-te: ou te W® IteWttes of eodoSheW celts <»s provided cs?R#. ( A) Civeraexgressfeo oWIRW is ff oAnilteSitessisfeotedasdoteolte!: Wig. p/C): Hítetóoös of spjrssH terossiiö« I» a spHsroidgB} arai a s i WF) tirées' oi' pre-Îsoilf-92 oîî ite p| ao# adhesWfF};. figure 2: Fiidei oiiSse 0 ver-e vprrateos·! off»ÍRc-93 <5ΐϊ angtepfiafis te vteo; (A) Ovev-eJcpRjisi©» ntmiK^Mn-pr©“mí ti^íísi&amp;cteáeii'ésailsőiráteíáís'-: p/CïD) tehsitekra of tee assgtegeWis is tern BöVit were tensteeted wltha contest:or wste pre-: mi. I x Ff ceils teste iafriateod &amp; or ante Wteigef pta|jsR Itee angiogenesis was dëter-.mined os ii&amp;F·sutiéra {B;i%«sí
F iparé 3: itfífecl of «-nlWS:: f tetetette m. teo te vife». arte te? ws® angioge WÍE fteiSSisAuteisra pfise /rfÄ^pmsißgraÄg áffö^dthjditeiígö^iéotidPabisKí^aífifntR^ :pVte/D/E| Bfteef sif íira Wetek Hsateri of ssroo-itegsrasif 00 Síi® te tevootesvvsscsiarteaiirssc P) Bxprr-srsteotel desipc Áte Expaaasiosr oísoiMteS in dlifesttecpös Çtertj· arate, Splew fes^'F ^.•feWec»· iïü the |terföXíön aoeoPlao:w tee rexoHs of me&amp;söríog·. Hi* costeCiPrsfiost ®i iteroogissRis); |o) repfo^ sopiailve pseisireg ÉPateiîsgJ ter Hi; iesîra! teggite Ra-sriaiiop of ves$aSs:. iigusx; 4:: ifetei iseat iosr ofj» 1¾¾ tsrgoi ; mofecai««·: (A) Goac ete>^askm pmiie MKI€ telis>wisg tesiistes-sftef:®vp^milte92. (ß;C) €«:Ι&amp;Β}&amp;Μί!^ίο|%^-:^Γ^βί8ί»:Λ.&amp; ptetesr mpîméfcm tsateg Ämtern Bterimö-ü'^Ft- .#· ter ;raiu;K:eai<··; of pre.miR-92 '{D'E'j'Doïecifcn ;>i'ihe effeeia of pre-rasR-92 rss> Hse «sprersiori of mtera-is; osmg HACS,
Figuï* à: iEitevS of sniRo>,>;ä e-sr sise {jssgiogessa-sir· ο; ν,-'ο-ο fa-o) vsni rte '· A·:;; td-B i/s vi/ro: SAO'rssiR-yo vvss ovgs'texp'esaed te MUVFC asoliise stegiogestesfe. wsv determteotMo-mm ;«: lise spiteroM m^déí({#«d.makidéi sway {b%'Æ3, %<ÖJA versos satekssi fCol Farteje atsows rap roseraatfvi; síx&amp;snples, lo vivo; Ri;teiPs;B:-|2s 5¾ epäRpl piieroRMA im&amp;fèiüâ ^PFlß?we:;Ä«d wlte sisiaspis asif: Arasspiaoied ifik! orke. Iltevvcassi femias-íoö raas dteooÄcd ossog SN coïte «s irasósgtei«d (dg; teermhte b vivo io ieetki-psHused Y*s»b fel, ste by steîenninksg the hemoglobin cornets R}· Hto, VOtef OOíVí'ói (Ciî'j.:
Pigxo-e δ g|H®5 of?tdk-'P2a kshsfeksoe ors te· engtevecte /ο v/ko. a/i>! sosR-PPa was blocked by oveMK}>rv»SKm of 2'U-s«s%| gadapp* ftte te «ngteetteH* was detected is-te 8ph^y:m>dis{.:m;:v-lte gó;y. ^ssíCíiB. c»o«l ob|oo«·· cîe&amp;iàJes (C. o> sfVOMeCsf'P). ¢5 ««R-^2a was inhibited through te tettette; with smPtgoíair~Í2sg esni te so||o|goesis was tes tes mised is vko> ist the spheroid sx-xlel. N:;$. *p<O.Ob vssrïse, PBS stosksol.
FsxWs ?: Bftet <?f »htsgöroit-FSa or* te-tetete' of new vessek isiisg «xtgigei îoosiel pwcy ttefensytesb n* ohoxrsäioxxiei:|ä'0· Äödtgmste&amp;t S dsîgrenfeothTo! «Biagomirs or live scivest F0S wer«: mjtód bv. <sn 4a|~ g % S#; mgdvgsiMh aad te msmgsifpbigsïwes-e $sptetcôko&amp; day 1, íte mteersmf im*steatstf«slis fBádg al, parliäeii lestiö-pi^itiva vessels floatete hswlfetecsMitet fie) wvscteerassstoBNto, pspiis ve:rsas:FB$fer iattegemirxioatrolS fCb:J> Aalagskskstote ssrosteek: #erg &amp;j«tet$:Í¥. ö» dify (0 0,,4, g 9 gte hfcái kspÉ Ischeste, «n$!te psrtefcsy-^Mtesiiùftsiiusi^ ibte floppier:$&amp; gsiample El iFigörei F sbsswsitbe spaeii&amp;liy of aoUigooslävgä&amp; iOa ihs exotessiísíxöíoo^^sdFsiattáiílifibíeot öiseí íOX:' eroEBAs. 90.05 versos enbtrte $30&amp;:
Figrsrei: Eigora 4: kitbet of aiUagomiMMa ob te heart testen after saysvstrtel «a fater às:logoxOr'OSa¥ eoakel steifte'#íte aoiveót f*BS svgr®; iiOfeciést Oy. attes«$Pr tes of si-ο cartes infer# os; day 0, 2. 5, '0 9 Ri stsg'f g bv.·}. snd she heartittteton »maetertäte «s Bay: 14 ussssg a MSbrxate6ari.:.af8;tew'ätg5te^ttet^te'öf'h^Ä:im«tte pamtstete fix coitectth?· sysia pressim «otesssstoK crrß^a»f)5Äagtek“§is4tete ardtek Bif Ä»w beiRo beak tectsns parstraetcss.. svhen compared wish Ose asksigos-nh'-Cs? ássd PBS group. 0} shows te ssgiss fktsssti sacres® of the capillary iss diftkesk regimts s>f te heart alte sntagsteb-dte kaMmeat, E) shows She reducisoo of the ;0:se of ta fasts aas! bbrosb irs the aîïisgon5sr>9®a tsxofed: group : Rsi experisnsnts A>5. ''ρ-0 0> versos ο os!trs.d (Co).
BigureE; Eiffel öftefefad raetee-rs of the miiR~ 1ÍA92 eteisr ssmte sogkigmeskk? fern. ai;ISifett::oites;sxf sho tsver-'gxpsvssiasj oPOse hKlMisr&amp;i stdcroBMAsoss te.-wglő$cwesk to te spot-· rs>id ars;sy (N Ό Ro ó.OSr »5 P.arass'issa ssOOcS ot Uoísoí'tssih; svhicfs svero sras;sibcses; svdi; sxiR--fl <a task· i 0 on dît asigKsgtsxvBS of «Rdsstelsal colis. As depstied, taasor colls wssso sraasfocs-esii. axsd dx-ss Ose supemasasií svar askissd ts> oÄteiäi ceiis piOVEO so tster to tasí teseÄi;or^aeElfi*::tetteffte«¥er^pröste»f«tef 17 Se&amp;sfe ks ass sapsxasssiOf ?h® äS5:|lssgsßesB. P.siisxspk's
SfRvksrlikls ássdMethods Cell exsHtios? i’Hssnitsv ssisibiiscssi vsiSfï ehttetebe* tels.·: (MUVP.Ç) ocre purchased fttitn'CanthfOt, and cuitttred until the ütird pttäaags to eodpteidf hasat C8n#r«s|:l«pjp|ôîne^teAv|teÿ#^Âte, böVte hraloextifags, ορό gKíRíis tsstof aná U) w<, fetal calf ien»« <f CS; Qibco). After te sfetehtweat with j^slrtyte ssetís: Wsté sah-totst; m § <rip esíihst e dislies tor at least 24 ίο 48 hems.
Trssîssfeefk,« ftëftvlriiibitio* otâàmx fflimi M« sÄrvd up m a cofiOuenoe of *0 % to ?0 Ά «*«»* ma tr:>n^K-e-m w-lfh the «fmffic ««fcS^AWii® oligonlxmudeotides miamid mïR42 fï'CAOGCCOO-ÙACAAi.HjGCAAliA-T, NFQ ÏD NO H) «r fi F A *5 AAACA.TCAAm*- 1·ΟΑ4.λ. Un. ;\AAi.'l= -o . S'-.Q Si.) Usí i mm syhihrôd ^vmsmm^mâ 50 omul/! were ftv»focied y8ri5lî,tR1 ,Γ CMoBilec) according » the prttoi of the sM&amp;tfer.airov^xmesckui ofimR-92, HfiVBO were «ok^d fó a contee of 50 %. I0 oamgf:i>^-miR-§2 or eosgmF|no-ftOR (AmbUses wa* traiusf'ecscd *un »pofec^mtne. -RlfAlMAX. ^Mtrogcpjl ««.-eordihg loihs passai dtsh^m^^
Apia go ro i r-strategf·· Π» smgh- Mramkd RNA as used herein, rnrmied of 2 î to 23 ime?«#s and was synthesis by VÖC Bto-iech as described. i λ, AU mslmai nmddx svon: held =n a CJ7BL1Í? background. bight xveek old mice «Ä» n«ous!> injected ,»kmg d··: abdomme! middle litis on das 0 svilh *'·*<> ,d«#iiipi.b?w^s«t nMm ρ«Μ§> > aso is'echui mil s-du ijdectf··!» of a sj»us viiuo.«» oi an .mtogoma 97 on stay k X and > Anttgpmrr A·. wavadmuiLteieti >n. dosages of 8 mu per kg hndy o eight in O.i nil phosphate hafiered valsne (BBS; per injection. I hsat- >md .piattukd plugs" s1· ere Imrs ested on Jay sh. The tissue wax iro?en in 18¾¾¾ ndfögsn, stop, stored #.-:¾ ^ !<A kNA amuysk,: For hentwpk-hht analysis. .» ,.m:itnv.e} pîuo' was rémesed aflat savsp day*, add hopksgepwod: st? .·~η) μ.» aekuuzed wmer Λ i·.·.-' rent! · inga: lot-, tik- niperrífu.un was used io n t.nabkm assay vSignni-'Akäik.hi hx mcasanug die honiin gtobin eonreîniadoîi. Sunk solutions of the h?magjktfcpt wesst used ior generadog a tstasioaus sauve. Ïhe lesohs were expressed reins: sc to ihe tots! protein in -he supernaiant. The second „rmungei pmg wax osca for die tpaajtiincatton dC|fygd|ifg cells hsiRgd-j^B-'fSairllfig.
Western idol aassty# für Wesse; ; : Biot analysis. HUVC.Cs were lysed id «?% hsis bulur (Sigmsd Tor 20 minutes no ice Alter cemri legal ion 1er là minhes a? 20.001? x g {* H.) die ptoiain cooiens of the samples teas deiemnoad aecotduig to the laeihod ol Bradford, identical alnot-nis di'pipiaid svesa loaded on is SIfh-yKtlyacrynsioioe gel, and htdited Opto à PvDF- ο; niitovvdikhxie-ifietnhratiis. Westens #St^i:pesfeirtnpd «Stpg.*btt|jodl^ against tOibgrltt &amp;§ tfáb%t poly-,.nu invgr ' ' a- f twr.: } 2^K f'lu'mfcöft), MK.K.4 (labhu poivek-maf intsekK4 5 « edi stgn#mg); dvöS (rnokse möitócfooal atiti-èSipS, BfJRIÍ ff»bbif;pöI^lo»al.A#í‘SmTik, it>sfd«yef:fo·' hallhifhlölfse nk>nöeidoai:3íífi-'fUhal;i®f Itf.SÖÖ, pnindfeB.;
HT-PCR la oxster K- deieimi-k the dlfterendsl miKNA espfeissfen in pt|¥i|iÇs;^Àf#b-:tf^:feïe4 #iiA2'Æhiethyf; ar-Usensc óSIgei'íboskícIeeiíidea agaiást pïîR-iSa of píss-todi -V2, whole· RM A sens isoiatővt 24 h'-íöfs pttef die líenskic· non usina TR ko! llmikuynn ikcosusag to die gfotocoS pf fite mamlSiefiaer. Asi RT'-Uf H was putfotnwd assop the ní<í-Vs«a: ^ qRT-F'v'R ad UN A oeiecdoa fii {Atshid«).. and primer aetr. spécifié: for ihe aptplifleafiosi of i'xa-miRdfg f Alohionj fisrie eyds; 3 pfipaies s; 95 BC,'S5 fA seeoniis as "£f 3fi;:secohäSAt.fis) wL>
Migraik»«· .assay
In order to ueiennlne the nugiasion ot a-kk-ihesml .e- s tU'\ Í i won dot,a ho 1 uah l-spO», Sun eu, d u;·-fhg hdsRlfügaiipfk and reAsspendel::|s FÖß yii EBM wlh hd BSA;::::epsfhlsd anti ρ%ό«ί| shlddhé Ιΐ|3ρ# #h#h«f Pf a: modified Baydtm eiuunher f5 x |0S cells per ehamber, pore sin: 8 pm, 813 Birs»eienees}. which was maied with 3.3 •ksd übrosectiH. T he chamber was placed uds> a otdtui® ;dsii with 34 wohs eorummag F8M witn U. ; '-* B.SA and human sfSSCUThr endothelial grosvth factor < Vng'ml}. s tmowing au tr.cub'Ukm ks; 3 nour» m a lue non-: srarsafino veils os< She iípper íAle os sd,' eha''': " wen >.mon·, m\b· ο·α tsj\ ,« 4 à wum-'i c\lls "U ue lossee Sídé Wipf« íÍA«d selth 4 % :lbnna!dehyï;e:. ;Fpv gasfitifiesifiöp,: file: óéH pph|sf w'#^ ^|ib phaiyKixM* (DA PO- liSMT pf'M «<^*.1 in fiv« r«a&amp;M»b ghpsgb syil«iOSc«>pss ilglds.
Vessel formalioe atfD* ΗΟ¥:Β€χ|1 s {J^wçMfedltw«tfln a 13 mfí*0kúm$vM&amp;Mm smm has^mőftí:::m$mbís«e:s^íl^iíPi>'.Bímcfea^The, esrdoifeslPisete «Arä* h%mm îïi 0¾¾ «iSï<k«% öhö;®íí í5si<;í^scöflc Adds usiisg a cviæppss^coninifed nkarosapw «sàsg tasptogsmi RB» J:0 sZdss),
Spiserolsi basst stsigiogssssJs assay
Endö^Äftei ^ßmMsatt* ctel&amp;sl oá! iaoxber we*s gndw»*»» 42$3$ the: ftrv&amp;m sngkv genesis \ms WBïe:gro®5ï ih?m e;î<;5> spheroid siting a digital imaging soíhvan: (ACopiofh Ze;s*), whereby lit sphecords: were awdysed per experiments« group sad experiment
Md:T¥bïbdiîv assay
Fix measiírmg the vsabbiiy of the eeils, ihp (3^4ib-sliiagb:iyllbsax<>5"2-yi;F .isS“0ipbßfgia-ZH -iein5xöiiafi5bitSi5:;sa) yh0Tl»p yy:y ilsesh |J: ·χ»1 Μ ΓΤ 'Vií-> added into each WeiH&amp; hours after trrenikAon, toki the edd weie iocu-!\í;B'I fix a -vass..« v’w' ih.w J!' \m-u ^ks ·\; with Ph\ » iw-d η» emuik "X wxnxxxwe xbh kW. wjà
X ,»0 USXW i Hi. ) Sx is, in , V 'A > Í \ W,·' W ,·Χ -Wir HV'OiXCti iC-'JH XX Smied ai OHU
Cell mairie sdheslo»
Del! eglMe piaies Wli»® :wèlk mJ?é évier iigfet at 4: W;..m$i 1; $g&amp;il solable rsc^biosid hmm® uXkgen ! (Rouie. Mannheim. üei«:euy} or 2,S μ»1 borna« ííhroneetm (Roche, Mannheim Genrusnyt sa PBS; sind wüï I |g|w/v):heaídösudreafe4 p boars, Sb igibomtR (HSAI :ÂSVEC$ :wfee mïmé with: Hnoraseoln: aeétoæy meihyteswr pCECF-AM) orCelTssÄr Greso |Motecsfe Iréhes, -Eugene* Dre§·» arai wei» resnspsndsd fe j· 8M svii BdlS % HSA aber lise detssehmssst Wäh It^fsto. Tks% 5&amp;Ö0R Ä pgp well wm-msétâ m the eoahM xíiílls: io 1 ί.Η) μ! EBM with öJS % HSA, msd iacubakai tor £0 missies Si 37 ®C, Aida: wasising óul öl tbe sksv iRibereal veid rising; warm £BM, tbe ,^ere.«í: cefc!Wg: gaíxitilted jbrsie: tiroes with,·.«. 8uôr*s@e»ee plate reader irloostab BMO î.ab 'rcchiK>k>gies, Ofieohiisg, Geonany).
Row <:v|o«ierry aoaGsd fsw a peroisabdlaabois, |®WCs. drsti were !:ra8s.Í5ciéé:w8lí p.r«*«ÄSl· or eosBrel :were deiaGied: wbl· %p« ax, fised ίο I: % Ixsiakiebyde ibr ü mőnte,· aad îr«»ü«à wHls o;l *y iriiospi-iOtl, The: yells yperoreabllieail aitd ifcdipÄÄl&amp;gd) Sv«®: lloebdl usiog ;f%. 8llA:,: ásol sÄid. will mièg &amp;$: (abriHlDdRe-FilC l’M, ifsioninoieeh) or enegri;; bl ϊ:χΡΙ-€023·λ?€ 1:2b. BD) autibodwr. Ί He ooih wereboalyseil wiib:s I ACS Caoio 11 •K-vieet'BDb
Iff rdsw -%iatr|gel plog7 assay
This, asssy wasperloroied as ikssrrbed (24g: iroi: With the :Mb>wmg foodlrkaiions: llG'lECs wefs tixssfefed witP:pre-:obRW*, orIbt is eoairolas deserlled afe»v«i :11)hoars stkr tlie.bOirsDe'· >iob tiw eelîs -Wt^lteiNíod with ‘Well fraeler iCbCDil' |bry:iwoggfry::::deiirebedi woshed aril: cooked, 1: x ÎQ' eeiis were iPospsIdbil Ιχ 3:0 pi RRS aed ssixeé With DKi: pi b.ma&amp;^eCfeerosM sneraferane osGrlW' pD Blos&amp;i«Bee*| c-;iXaiixag i5 anils hepgrin Pigma-ÀWrM»), The efcil~3î?sis?ig«ynbe .was iolpeied sobeotioïeoosiy mse sD ío Oîghi Wpk idd feioaie sihymie -más··· siiee iHtirdri). aioxg She »bskarsma! middle; iise; Heáieglóbió aasisysisi aird ÄI-::;ainir:g were eeifonrsed us described alxsve.
AtlYnssntds {»HSA prstbhag
HtlVBi's w tfan*d«c$€d *&amp;t piV-ükR'92. or for « control. Whole RHA was ísoiatc-J sdte" '48' hoo,í%áttá: jiisí g«'w e\p;«:s:s!n prtdsk a.ssaseexared us:ng an Ally ««2 wgww whipves««sión axs&amp;y
Refaits :Fíá->8ÍR-92 biot'lo t-pdeíhí'íisí «91 funeltons ki vkm m4 m «>η |y. <ΐπ^;· ^ «.<,{ ihtf çfieu öfrolR-92 an eodotM««) cells, HUVFCx wem· im&amp;sbækd A ibe iÄWAps'emsaor Ι»μλ5Κ··92, and the «&amp;; of tins imsieeti»«. vnad&amp;emmted In dlrfetont ií^-sííMjas^p/llsíeifeiimí ovapAApessides öfttÄÄ was itt® beírnád .«slag $£lRCfc ({%so IÁ}·. l'bo shiRWS ovenekpatsstón sigmík&amp;ody Mocked IMsifór.* «^tloa Rí vesse! mmsm Itt·.#'(Fijgom 1¾ aad inhíMtdí the îbrrnÉio». efa vasoä aeiwark k vtàfrfaek flog«« !£$, whkb lnäJfes^.tfe^:;mlI.-9l!% a^gaim «eí^ílís-Ébr.d€Öí«r=ass^4s^«n«sl»::^«:to onferds d« ÎSCÎ8 possible eiii;vsÄiky .*» msssarod. Tbesbbf aller sIg- :4IÄe««es; cowk! Fe deseaed, compared vAsts aon'tatt&amp;ffettâd íl% eell mb-
-gpdib» Is o£;|»ge·Impssrwe® |brfeas^of^^::selv%‘^f:ÄS itt vim, is stMlios lise saigsÄni oniiWEC aod® Essai cobIIÉöss and as :s tysponse so YEGP was böensbaskl,IbréosbRWd naisReá lba:migiadön ífíguse íB).;8»d ífei iXs\ K Ntjji . si>> > ’ «η' Í. JK El m1 M, >3 v -ÎWV On\G«K fh\ ! ' ν\Ί,κ ν' ' whJ Ά\ geil rgspöäise:, MsRís;:ls PiRShsRihot« îtïs a^*i9í:Of:tEea'8iB-Í3: «íd ihp ssagiogPhsssih m VA« was. determined Fer thi·.,. ttUVEC» transfected wish pi«n;iR-9;i wem impi&amp;oted ln a knattigei plag" iflfo nudei s>< kvo. The stííkkas.'y of dis' kihibi-ioa «;« s« h eoskroilsd in a .sob--isat:bt>i; of the hapknUeb c-ei H (Figure 2Λ). Aa shown in dsa tsp^sdinad ve: piksRes üöf#t^i''2B add bk 2C, pmtMlBÄ blacks äs; growth: οΠηο ivessbls bî vbs> -l« »sj: alïiclaal maetssf. bi addilioxsiba säg»lfesÄ:mns:sd as caöblbe: sliawK bymmmmm tbs !w!^glÄi.-ß®n«e»&amp;«öbÄ d>® tttbfebia« áÉmíUSMmvesms/» \>&amp;m--mê4itt$kr· M was fsrtbecsaaïs esattïbïsdi wbeibaf Ibe Isbibltioa af äslR-92 causes íba sbo^alaíioa af vessel pïîwtb. MIE-> $2 was bbbbsk-d % 2'··«'.Inneihyi antiseas* abgortbanacicotkle il> ittethyl-iniR-b2). and the fbrenason <d vessel siiyctîJîés b? Vkm was dalsa«feed asißg a sphei«id assay l i-niethyl-aviR-dl increases the sprc-iie fomakie,) m vim* (Figsare 3A). wÄh l»da:.aies ib;k. she snldbítíon <.if nisR-bi: eoidd represcsR a new therapen; 1;.- slssfegy b» ivapmensg Sde angl<.'genesis. In erika ta kési ?bá bypoiliesls, adR-92 was systai-deally iabdmed swing so-eslled "antaga-nirs'', siagie-skwvted RHA oligoandeobdes. w:bidy campsevd xvdh sgx;dlk miRNAs., bsvc a eamplssnesiary sepaeace, Rbendesl stödbkallísns lesd to sst ibästeased sisbllisy. and ehofaiead'-eoipagabosdii as iusproved uptake mus lbs eed:S:flA>::.Aatagamira dlreeted sgslbsl m)R-f2 wese:;admialstes:eä op ibree siitp, m B ï'fee fÿst:·
Rdnk ádátinsiíraíidb íöfisikákáíibrs liopypysd: ÖRí vssselkgfdwtb add die petbAioa: dl lbs. Abadàgef fllag as «vo i'biglii'di 3:|2/D). As aiiAssH, dbe :ibbifetl|«#s0f'jilBk92dPo^&amp;^|· dbdsahells! cell fass ebb ax m vitm add intfsoavas Ibe vessel gfsiwdä àso'ivss.
Rleal lftc a b « it o f as 1^921 As rge kpeses lAiiRbsAa eoníroi tasígfí: pipes throp^,.a :#ge#i|i|lötii. dlltst'íltiibt'stAR^ eb tbcdágb tmbslatsoísat rő]Rdsásöas,: la dbldf Wf ifeieRaiae die ÉSgdi a:sEA;As tbat ase degiiMcdkin iesta>ase m iatRAP sveAbspresssoa, a ctafi aaatysss with ass Afîymétrtx sssRMA gene i-wpsesskin arnsy with 59 ,s>R l gesses ^HO - U13 3 Idas 2 i was peiidä -sied. 1 he a-saly-sh of ibe regid-sted -aHNAs idcabried dlffctcat key eisvysnsis that coastal she ea-dskbebsd tuaesstas, isscladsssy -eHGS SiRTK iïik-gssav. «ad gmwsh lacsarc: soebasattgeopksfedS {Figui^4A). A ftadian afsbs dovaacgulased gcovs is easapkiiks with aa ssaslysis of poteasisi ssiRAiAlsigess pesfoseaed in skico ( fadic I ) bi order ro casasix; iU~ - esabs of the sCiveaS: the passcla «kpfsssiis of she mpecbve ge;s:,:-s sva:.,· deiecied saasg \Y<«;«T: blot ta t A t s «iaiysas la with the. results SU. predicted. {Hs SIET] Süd: pssssM |5T ;jpsNwM>2 ( FifBt« A B tó B). h^v! <>,1 ΛΗ > *. F'»’ ! iuu\ ! vv'-^KtlW’tVIli'NkuV > *<K <* tóiK-92 - Target« integric s5
kttógi'SO UV mrj MRld ku\: PCAt 1:.0(.: ;
íEEEííE
XrafXïOk OTiR-O? OVUt'W ! '.ifv'PV ÏVI MKJofiJ·! \ Ci R\ ! Ss'd Olg -O'J' 'xilUirvO' ''udotóldíUK'sdl -’U\ fias« tó v*f*'* ük! .a ï a p p, ,".,wfn* ut v- * h S ·*’' -S u*· mg '9' neR R "· '5 "*n ' mt'exs '" of..n .omyouns ieao * tó ;;:.a improved vcssci growth >;·; i#ssf1$s S u< civnxpfÿ tn dsc pro-aogkuRmk acnvdy <d Uk· mtR-d T-92 elustór '*S:4f^crifeiïd (iO,ï. S V U*.«' i Iti * i , ^ ‘ tip v! ' pv- ! Ut OiSlX , i\ <K tll I Sl'OW)» '!>»- 5 malőr rak': m the esmothdisl veil bkdm;· Binon·;« de genes s-· identified usütc a udeoxuray. $o paOkuEr äks duno-roguiaReu of ski >S, SIR í k and uxtóurm a> on U*e protein kivi eouki be confirmed Mtce thsd őre deficient f r ttv« ptok'uxc eh ou a-* * npatn\i xa'xuhxr R. nettót' a' o m a> *m out red nhthty Sν' portóul ·χ I «eovaseaiartttóiiú«. '' J. ÍU'· * <s V 0 t v. p t MU ' V * \ Vu. ·. t 00 k aiA I1 P IV't'·' \ ik UVk'1 <i u ív , " ä ' n\ cd -.k' «'V. ΜΗ] ') tXOU' W X Vtó ' X v 1 ! 'l!V'l. \ lu'lït.'S'ViHK Ili v tía'uUtóv
vil Pi P Si ,;'k" U \ \OU\rde J X j "'. }'( ' ;V,u VUUl tó' O'' U\p I *v uP 5 'Sm' ..: i egitóe OXgX* 't i**s the mH Ux'tk'O vvjth Sin- ceü matrix and thus Impair lUttf-apopKMic stgnabng »ad cell migration MT !&amp;). The gwvth factor Oftâ -uvula'".. <k vex pvr heJk and 5 r-'Utów vtsksotx wt'd as ΜΜΓ4, vopItpI xe«oi οωοκάο^ί í kp ÜOj, U-cords:?'pJ>< Tnt K'9* Ιϊρρ.ί ρρ s with s paru s cf p-înex íkat con ttot thi loda'ks'llai coll tonettóks ot >idk·-: osp laxíh ks akiSRv cd oxíR'-kd to sofsaence dttfvreo: oftiX'tóít poo'íd'-'x ftä1 xds'Vüívpx· of *nx- nriHNA-hascd tippapinipv strategy i>v } tó < V-' sill <VMs. ÍMI4\I« Λ < Si.Ui i,uv 0 ' 'u t í s Ov*k <*' f Λ*ιν ο* a slnyk' ιοον \;:ρο tóc \.ρη sox p*a,s ms. et I > roots ' o«·. ' iio srxssop -i \5 tl^ instof ao s' - *i *; ' oasiso at\0'sx s ass ki!'Piv to'i.\|î5s V fti!i„'iit> 'd νΊ \s. 1 i s ol '
As a svssilt. fhi· Infhpp'i'kis «f vîIR -0^.- roptisoots 3 rst"o iin'rapeuisc atfists·^ tor í* looko] at ctKästiheHai <ólI tóiKíiotík fik' viistótók osv of ttfsntgwnors 3s skiop herein ;·:- pastica].;nl\ sait-xble tot sakyoiKittg tó ihe aoRk l k tos isk X ‘'s*slv>* ' i'T'Mi'f." '. SK V S'O x.ssvi k>tóí X Ofxí WÏX ' V 10' I .0 'k O-O X tó ti, avsxv&amp;saikiäisXikixo and vevőd repair. With ivko'eace to the gooes ivhsch are known tor their atheropwitective effect; vxa'h as. for esatapk. ckOS, the bkeksik of miR-92 ts of use to ti><: anti-atherojatSerotic therapv Since a ise-nuoioteeii'·'' role i;* rseaioilegeattótflvc tiistixset; le.g. Motlxos Psukiitsuo) is also tittrihnted tó SiH t I (115). oxilt-9;; :mt;igioxxi!S eao he sitccesshilly sav·::: :ski* ia this ihsease scenario, to contrast to this, the over-espressioft of mlE-91! Is useful- forossuipk· for hiacking tumor crtist gone.'ts. sins·' d .tssOgli reduces sessel grcootk.
Ckcd Liitissture 1 í'\í-;ííJ>Ό 7 PB>0) Meehsss-Bix esket ge AB''V>.- <';B Ä -*n '-’-i«-"·' ' ? -A\. o í'íX '§ ' 2 Adams, 8.H. má Álltaié, K, {2W1} Mofcctsk? η$αΙ*ϋβ of angiogesoais and 5>-s«j>h*ngH>g*tjnss '<>? &amp;·ν Λί'.ο sV-'6v/8 slg 4M4>' V Bartók Γ* Γ s, '00- ! Ms-6 kk! V goswou. -, HBg-ov.'v »-oPcrn^m ioro líon. Í.V/Í I 16BUíliB9? 4 'Btw- s. OBT'Usa >' <. m-'WP, 4 a - . "í '' l> 5 fmi&amp; WJ Síi ssl cm$) Oiís;?· is wsx|atr«d lm whryaxK aegloggneáss tkrhg mease lesePp-s^pJ JS# í Vív«? 2df)íid|s9im^:'.> £ S«afvx. V si (200?) Dket ík^ede»! *«k«<iHN.\n 'eesuske gsassg seKpsssskiosM sheävi-.elk -,V- -, Sív Ok 033, S 5«4 - i S73 W KueiftnriKr, A, staí. (2007) Rak of Okét sms! Ptmfssi :fo ssktoihelMi métsMM &amp;ήφρκφ%$ιψ· À £fe Aev 10) 0), 50·6a g Poisserns. L, ci ai. pool·} MictoRkÄH «odotsste Site asSgissgmiis; ρπ>ρ«ίϋο·> oHlUVhCv Ä6*’ 108 (9),:3:068--307 i 9 Ve-darmi, i... ei ssl. <2007> 8 xp-xmskm ·.>( thv st-sR-i7"62 SRtiydsarsss in -Ihrom«' lijsgfeliiifessgsmsa pMkj CD34* ediv OsW 10# (M}: 4j96-44í)> 5sí [Sews, M Síi ssl. pisil) Asigínssiisssssís sí tumso ssssgsssges'siísk !>y ss Mye-ssRPasetl smemRNA í-fsssser. Ask öewt 31 P), 1060-0)61 JI Amham, I „ es ai. (illts Hktisí: Mt *>^âass;:mÂ1ste: aagssspsiigsis-feíms^Msíio'síissiUSiOksihemk^. J-mxmvisi! sOS síi).2567-2878 M -Ak-ber. A. sí? ai pOoè) Botsai mis: stsPsxkssi-eibl alak stxkie sysuhase for mobiíiaaims·» of«*«* ássd p<> gsmiun· cclh. Aw AfeJ9{\ i.í, S37(1-S 376
B Pmetsse. M. ¢5 al.p0071 SÍRT} controls s; mis uhstüsslsi$|fepaie iméíiMrs:4sék§m%mtez·^%MbíWmmrÚ£V· 2i si'O; P44B6.SR 14 \< sí ^ i \ s o",R * *' > í ν'* ΐΛ l'hus < ' \iu\ <oumI s 1 s...g.vs.s<>sí «nd s.t.^svgv?'— si:; by íssSovjís; sUphalbesai ísíx: prossiíí 1:A. ?&amp;."«'! hst.-i 375 (431, 75030- í.ei.A. il K;'.sí.·Pisi·', S. as sí. (200:7 Sskfic-ing ofaiieroRNA-t as v-λο wish 'sässlogoxsiss-''· Ahíííív 43f< (,·0s>gy 687-689 46 PlSíííTselex ái. sssd Zeshes, A.Mi:i( SOOp Ols íc oxisie - ss-s S6sisssheiisl cell sisi'vivsí Isssíps·. (,«'0 Danik üsf-er 67 664-668 ït feîAÎ (I. el ai, (3o60): Tíí^&amp;enpssíísai äfesesssg)ami Jefifspflj,' gppib UP: u-m.MA&amp; Isipetslsisiiiiss»!'!» iicseesviase. Nalus-e 403 (67? i i, 795-800. ;|g mrn^Zrn ssi. Ç3Ü5) :¾ alpha 5 %m 1 b«6|rsa ssws^ wvjyal sfisaÜs ass ibmssasiss: sssslags^guiates BsO-é v:>;f!‘ií\sÍ6íí, pt&amp;: 3v<x/8;-; Ir .8 ,-i ?2 ( i 3 s, fs 161-6165
Il Asahsta. T. es al. Π 698) Tse2 rec«^or Ssgassdse Aaskspsleiiss^ 1 god Aagiosssis^iaP, *‘*'^»9-9« V s ui:· aOsicssI ppsisstkss! s>6s>vseeaisiiBosses. Qrc Avs 88, 2)8-290 (26 Sang. Q.X, ( I'^S) Complex tok of maifix m<;la}i»>prot«ioa8^ is sogH'se««?^ A - ? ' ς Ί 21 S'Bte, Λ. et si, (2007} MkroRNA-18? t-osnf^ls cardiac l|}H7Sssmpy> Aüí Akvf 1?· s> ·- s· '3-658 :22: tB-sST Γ >5od A.e-U^oo H.o. s jOs)H) Js^uoo« at'essdßOteibl eells is ma}íieell«l&amp;rs{>htó:ás^ey««É bpe®*· tövis ssnsi iadtices slsSiVt'sssstiaS'sa·-,,,.? CPs .66-/ 143 s>), 5341-1352:
:28 Dss'hl, R ,χ aj poor? ihe hiva-x tssv-hyissssssils^ts« MtUi.s-w-^|^am::t^I»iÂFOÎ-':«îkte3a'tl»l ccU sp^ist S'osiHtr ei' t'l 1' ," '4 ', 1S- " Sl^S 24 IPsæiîîgi Aii. yt a!:.(|l|t}|p»:y<dymiiot:.^fif»A0 tfäßseipfPss4s*sîïs Is stagfeias'sasss stso psatssosslssfesvsaeos: isnfeigg, j Çlîk liïvgsi ? 5 5(9), 23S3--2392, 25 i5iii :·:>. A, mú Kelly, l.W. (220?) Tfe fe-i'ggg ôf Amoas, AifeosgAl.T 461-402
SFQgEFF R 1,1 St 1RS 91! 09 .JolEosE: 1192 AggSg Ckyij25>e-- lro 2ae:îï\1 tSfc Frag kf Art a&amp;: FAllo <1203 AoRhoRl fer promstg.gg ímg::.-3g«ma:;90«:, ^ssojy Irrigst Aon %Γ ο'οοοοί ropäl:,?“ or loo Ifelhitisg toffoor Arogiogogrooros <lo.2> FS22AS3 <4 9F> DF. IF 2ÁF3 Oil 114 , 3 2151.9 2 0 Or: -!>-3® oiggg 9.1
r'9?v:>' I::FΟΑγΟΟ'Ια vs rg i on 3 > 5 <2.100- I ..-- ;A\„ '1' 2
<212 9 9 FA '<20. 9> Hogso gagssag <3002 1
í:limc:-.i9car .sggoougggs.o: ag.:F.g2;aaao; .fggguggqfe· :0.qgaoKfg;asu; agarscogaFia 6 A cfejAcfeal gogiRgigg 18 223-3 9 '2.: 9212:8 9:5 ,0 0 .0 ·:' ;0 : · .· -.
Ailla IFFaa ïolplaaos 9 -0 :: 3 9 8 . íAalílcxjaAg gosigogaaolv ΟιοοοΟΑοιον? fegfeaaruä feaAfegiife ivAAlaaofeK:: £9 ooaqgovragog:; qasga 15: <2109: 3 <2119 .2:6
9212.9 AMA >21 3 > F g ;na ο; gp i.:.g so g. <4- 0:0.2- .3
a gagor ago: g g. go: g-ggoog gps g g gag g g g®:0 g < g g a g < 2g a g < v g 0 lag ma. cgS goaagaa a AO sg :goi gór s o: a a' g ‘a a: sog; g g a gars a : - g g g o: g çrç'g 'g. g g gogo: 96
:0....:.:.:0.3.--. S o:2'l. I> 22
<2.12> FF A <212> Fosra .ssp'feng. <4:02:9 4 sogososgggaaa goä.go:o.g-go:.o::a .ejο 22 <21U > S oAIll 22 .922:23 RFA. <213:> Boss? <«<<<<< <33ö> 3 uanagoannc y n< ca ηηκηη: <:< 3< < 11 3 ' » <2?I> 21 <2(2> 2m <213> r: :: liliaia 1. <22ü> <223a ;<12-32 elinni «ntininnits ölignrlnonnklnni'.ld <100> 3 nngæ::c<niga naatanjcnaa a <1 <:22δ> ? <211 > 2 2' <23 21 3231 <2131 árt itliaiai <22Ü> <22 2a 21? <1 lön 1 .· 2.* 1 -ant.3..sannan, ail.«cl ?««naki.cot<d <·2:222 <22 i> alac lantnía <22.2> (11 , , 1221 <22 3> 2 ' li-snat ll-KÍídca iinát naklcctixi <«0u> ? η<232«<:9;<2 gácgpág&amp;dg ηη << <21.0 a 2 '· ·: - ' ·. -' ’' <212 a 292 <213 a tart:?, f. ici«? <22ö> <223> <12“SÍ elinni -in tinte««« allí|nx'i.?><nnk.lentid
<4öS< S acagntnrgg« acatgatn·«« na 22 <21 St S <21 X< 21. <212> sia. <2131 2:rt?ficlal <2201 <222? s.·:;3.2- 02 a i .te n i, 2 ‘ 0 - seá t..;. i - nőd«® 1t «t t a n t: l a a a n s a ο 11 g?r i fe a n « <1 an t i n <22 01 < 221> ::. (<< fatitnt®: <2221 1.11,.(21 <2231 Faxte Inti aá t - 23t.2® <220» <22 i> ?sisK;_;£«st«rs <222> (2):.,(3) < 2 3 2> rsss 3 íXit Ï »ss t-»SS: èxs »220» »2» i:5' » ·.·;·. f»&amp;t».r» <222» (IS), ,112) <2 2 3» "s>S:K f »tixiéts - Ills és <2 2 3.» »2 21 > »; i s»_ f « » tp.r S: <222»·: i lS) , . C2»i < 2.2 3 » Fb s » t sss i ;s s<: - x » t és ^2· >» v: »* »221» si»» i:e;»txs<Sí »322» <2il) , (.2:1 ) »22 3» Ρ»»:1 sol: Léé»:. -- <:»<»» <22(1> »221» pdssï fxxéésxre - s. SS » S .-: .:. : . . s -S. S. s < 2 2 3: » :3 ’ -- 2» 1. s s x : s r L xi sx» »p si s : s» ’,·:: ·:: S S S- <221:» ;<i»c fs»t»x·« .< X -.s ·. : s -: S .. -s- s s ·χ·λ.·,·:, s. .· s .» c .· ·, v . .·.· · » 22 3 » Fé» 2 -s » s:: :L si é é <<»»;» 3 s»:».;»;»»»:»» sis;äps;gi:s;»>sx5 a: 21
<222» ID » < X- X .» -s » »2:22:» ïéva »213» ért: i f isiéi: »2 2 il» <22 3»' :<<.<,-·- 22 »2 Is::si , 2! »"«»: L 1--c»d·:··:; x totst srï'stié s-sx»; s »iLlésjribénsxklsislSiisî <223» <221 > xísíxíC »»κϊ;:»:;:»? <222» il),.;») <2 23.» s »; s a i»>t 1 sé t --»: (M: é s <220 > <221 > m i s;», ft: ¢51 » r » _.·' '1 O -»- s . .· ·',· S ,· ·'» \
'· ·:·. <1x2 ·· s , X <2.£3> Fos?Zf ot: le-éit~ köté« 3. ·.< .< ·'< .- íi:j.ö'C f<;a ·:.<· -;e

Claims (2)

  1. <222> (3)>- (¾) <223 > í'ósxfotioát^ketéís :22ο > <221 > í«is<__2<2t<ré :<.SÇ<< : /: 'S : .· . :. .·:'! 1 ; <223* Foss ;· ilósos-kölo <22ii <221 > ?! sas J:aa K« <222 s 220}, , (21 i <2 2 2 > Fo<:;l;;ot:i,oàt - lotis <22ô> <22X> :»:iiso lixal: saos <2221 (211..(22} <22 3i Fe< s £ o; 21 oá s: - 2 öt: és; <222> <2211 oies: ^((sslar®: <22 2< (22 } , . ( 2 2; <22 31 3 < k ο I s: s; a t « Si sc s: sps> x t <2201 <2211 ci s< . 2« i << « <; V s. ·> Ί 1 s: '1 ·> 2 ' y.y ,i, s", s sV .S' ί > } X <V ; <223> ils àfsCilât:. <i()0> Xi s.ossxo xosq ossoxopoos xo 22 <21 3 2 S. 1 <2xx> ai <31 2 > gm ' .. : S - '' S' : ’. ..V ^ 1 .· .·. S <223> <2231 mii-'-22: ollsxü., 2-í:0iise;úiX~mo<3íisltO'.i:J:. scs :(as: :oo :ol.i:gli.j:\x:^>:ÿ3SxsS5..U;ss.ti<i "X si· si- Vη·^· <22 II ïSiic... issir. airs; <22 21 {1} . . (21 ; <22<> < 'i-osthi’I SKssii lise Iss oses; PukX&amp;cstid <10 Q> II so o;o.'o:o< ·'<: so.oca as: >:s 21 Sieafcadatei: îgényi>o»t»fe I. ils ykfàoptès laxstelMsâdé vsgy éxjiviás slössgltésárs, sasdÿ iariimsiss s kÖvKkesi (epéxeksi • cp blatosbssa, és - siü92 .ellesi maiaKSsa 'moleküls bajaíteíáss mxp|b><. 2 Az ?. igénypont rminti eljárás »M a biztosítod sejt »îieteo|^|-^Îë«lfe Mtlbiioo «isiMOk &amp; %;? elé#, :íí!tó|ii||#stet ssâ|rt«^àÂ^ hpm^tetikm m$, mmrnm* :^)¾ gygfeaásösssl^ llfi|sfe|?s Äteji k««àa:emirimÂ!&amp;-'te^î01 A Az 1-è. Iprsylyssiöl: ^|rr»tdy|;k§ a&amp;bl aznaÄtmss! ;m0».ök ^ÿa» ««Äk, ÄSp·®: •SSO H> NO. l-$ «.«rinn RNS-r?mbkuk*k bármelyhez hsl^dî^t $iÀz îH |gé^V|>e^#;:bi^|fík«va35^Í«^ ölját'ás, «hei m aÄasesm mbiofebia ©ly&amp;t m»l«jksrf^ -ss#«%as% szekvencbb a SEQ tö NO. 6., g. vagy i t. s?^sâ.mteaeià;·· $,:Az idkigbvypööÄ bám'eiyfe sgsmklspMs!;:as>ol:ag a SlÎQ IP MO- M, ^ejfôsft síV-kvcncíák 6àmj»i^kè^l"kOâ».ptemmlçr szskvessbr btstesz. %. A:? i ·<·>. sgOrypobok bànndybe szerinti eljbóv. ahoi az ssmszeosz moMóM fessas legfeljebb 3;δ naklaeiid, MibaÁsaa i 5-22 mskkotid. íi. Az I-?. Igénypontok bármelyike szenok cpob alkalmadba èfefeyenesùô anyag előállítására &amp;-Ws»«..itíifelá |«-iÉl^atóa^^sy^2k^kk'^.:és éryavKás stömoi»iltására, ahol ab Söifeaeosz oxdstefe óságában foglal agy szekvenciái. amely J»íbnd«zákklik SEQ ii> NO t.< ovnndse^ ^rmti νλ-Uvsx vau nfeleiu Iával. aW az snuszeusz molekulának fegieMsbb 30 ansfeo-<dj&amp; vád,
  2. 10. Astíszensz axbekda sikdouudsz Isakê-ost- patotögius afigiogvrvZis, itisroKaKlerdzts. u-ero.tzXKuvrzÍsnak kószfemctn aiiböíOsgtss Htegfeg, feäfesfesfel Öfeze:fe||ö Hiegfegelh VsMtiitA ##bö4^8-Ä^'' 1$>· rodyikéoek kezefeaéíe szolgád c-yügv&amp;znti készítmény efedittäsät», .ah«)]· az untts<v:tsz mofekula megeoan togfe. íg> szekvenciát, amely SEÇ is.) NO s-5. bármelyike szerinti szekvenoá|á rofeel-okih'V bibriöszáf. «Hol az arm-sacRsz medekolänak legfeljebb 30 nukleotkfja van. H A líklsfezívpenfesaefesíl éiksIsbíáA bkot uz iszkomfe s Hydkgzpfc közöl vfeasziod: ooHazofeks ódat κ · sus. krónikus iszkdnsás- safeMspáfe pmféfelfe artéria dzánkksa vagy szsykoszons&amp;tterrs-e-záródás. bzxdosás in-fetáfus vagy extrák (agyi várkeringést zavar?: vagy ahol sferoszkkozhríak köszönitek roásodlagos Htvgság akövek ke-xöfcfkÖÄ:váíasztóí skfe sâdvkos«snèèr*sÂOrêtn«> nnokanferos snSarköts, uztrók ás v.utdnxnlftjxuta; vagy ífeoí g sïsyros;fe|eBeïiaî.ivbetegség aMvetkszAk kiás01 vafeszHO: MlÄfefi^kdr U WteWÄ iái A iO- vagy .1L igásygoat- sagrbti AikaWazH sk*)l ág: árdsszénsz;-RKdekida öiyasoisóOagp, asne;yfíék szskvenbab s SEO ID MO. ó, 8. vag> H. s^bái szekvencia. Isaioíökbs angloggóéábs SísOHklarúkíSi.abroHkbHsisiíákikö&amp;AbbsO fagsödkgos kstggyeg, sdikiOgsscl ásszgkíg-· ad bV'tességskv v»l$a«M nearo^genetanv H-egségek, -ai«i m- sniiszarssz nudekaia «mtaitMa «g.V »z^venesus, smdy SHQ ID NO, I O. äOmxHAe vzsrngl srekvsncbs« m»!H«lához hibrkhHi. ahol az ansivzcnsz nxdektdának tegfcljidsb 30 í-ükk-odAa vsa. |4, A 3 .A !gss5;VP<jrsi sz-áiíítii antkizíglsz Stdekkls Sií att íbegltskkögÁli aMák idfeáárái : 3θθί áz ssc,kcbhg 4 : kOvs-.'-kezbk közűi vái:»zlo!t: mU>kaHÍíáiív inikik-n:;. kréníkfs AzkOnbs gMvbciegyeg, pcriteraits arténa efásodba szÂoszmtanêt is-efeátfó-Ms. iazkánnás úríárktas. vagy sztsOk; vagy aböl akö'OS.KkSso?.!s«a.k köszotrhsítd nbaósgagös oe-tecszg a kuvdHs^k közút v.>i&amp;s?{olt: akut $z»vk<5U?otvct".s?mdK>»i5a,'ö««ksnäbbs t»{afbíp>.^:^^á#áiöíÖ.t<>ji^ÍH vagy·' adói a KögriK!egive..x'ativ betegség a következők köztti választott: Aitátetmef-kor és ParktmöR-kőr; IS Á 13s VáH 1:4. :8gógyiOOö|: báiíOOillá Szárioli sSkisgsöázAttidkkUib^ söd ga: antiszSBSg: Söbbkiga szeg? \e:K.ia]8aSkQii>MO. 6., 8. vagy i L igcöytxínt vzaribi szekvenda.
HUE08844273A 2007-10-30 2008-10-30 Eljárás angiogenezis, vaszkularizáció vagy érjavítás elõsegítésére vagy tumor-angiogenezis gátlására HUE031559T2 (hu)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
DE102007052114A DE102007052114B4 (de) 2007-10-30 2007-10-30 Verfahren zur Modulation der Funktion, des Wachstums oder der Differenzierung einer Zelle

Publications (1)

Publication Number Publication Date
HUE031559T2 true HUE031559T2 (hu) 2017-07-28

Family

ID=40375407

Family Applications (2)

Application Number Title Priority Date Filing Date
HUE13179144A HUE037494T2 (hu) 2007-10-30 2008-10-30 Eljárás angiogenezis, vaszkularizáció vagy érjavítás elõsegítésére, vagy tumor-angiogenezis gátlására
HUE08844273A HUE031559T2 (hu) 2007-10-30 2008-10-30 Eljárás angiogenezis, vaszkularizáció vagy érjavítás elõsegítésére vagy tumor-angiogenezis gátlására

Family Applications Before (1)

Application Number Title Priority Date Filing Date
HUE13179144A HUE037494T2 (hu) 2007-10-30 2008-10-30 Eljárás angiogenezis, vaszkularizáció vagy érjavítás elõsegítésére, vagy tumor-angiogenezis gátlására

Country Status (16)

Country Link
US (4) US8258113B2 (hu)
EP (3) EP2684955B1 (hu)
JP (4) JP5898843B2 (hu)
CA (1) CA2704290C (hu)
CY (2) CY1120360T1 (hu)
DE (1) DE102007052114B4 (hu)
DK (2) DK2217704T3 (hu)
ES (2) ES2618203T3 (hu)
HR (2) HRP20170346T1 (hu)
HU (2) HUE037494T2 (hu)
LT (2) LT2217704T (hu)
NO (1) NO2684955T3 (hu)
PL (2) PL2217704T3 (hu)
PT (2) PT2684955T (hu)
SI (2) SI2684955T1 (hu)
WO (1) WO2009056116A1 (hu)

Families Citing this family (27)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ITRM20030376A1 (it) 2003-07-31 2005-02-01 Univ Roma Procedimento per l'isolamento e l'espansione di cellule staminali cardiache da biopsia.
US11660317B2 (en) 2004-11-08 2023-05-30 The Johns Hopkins University Compositions comprising cardiosphere-derived cells for use in cell therapy
SI2056845T1 (en) 2006-08-08 2018-02-28 Rheinische Friedrich-Wilhelms-Universitaet Bonn STRUCTURE AND USE 5 'PHOSPHATE OF OLIGONUCLEOTES
DE102007052114B4 (de) 2007-10-30 2011-01-05 T2Cure Gmbh Verfahren zur Modulation der Funktion, des Wachstums oder der Differenzierung einer Zelle
US9738680B2 (en) 2008-05-21 2017-08-22 Rheinische Friedrich-Wilhelms-Universität Bonn 5′ triphosphate oligonucleotide with blunt end and uses thereof
US9249392B2 (en) 2010-04-30 2016-02-02 Cedars-Sinai Medical Center Methods and compositions for maintaining genomic stability in cultured stem cells
US9845457B2 (en) 2010-04-30 2017-12-19 Cedars-Sinai Medical Center Maintenance of genomic stability in cultured stem cells
WO2012065024A1 (en) * 2010-11-11 2012-05-18 University Of Miami Compositions, cells, kits and methods for autologous stem cell therapy
EP2508530A1 (en) 2011-03-28 2012-10-10 Rheinische Friedrich-Wilhelms-Universität Bonn Purification of triphosphorylated oligonucleotides using capture tags
EP2861238A4 (en) 2012-06-05 2016-03-16 Capricor Inc OPTIMIZED METHODS FOR GENERATING CARDIAC STEM CELLS FROM CARDIAC TISSUE AND THEIR USE IN CARDIAC THERAPY
WO2013192576A2 (en) 2012-06-21 2013-12-27 Miragen Therapeutics Oligonucleotide-based inhibitors comprising locked nucleic acid motif
EP2882445B1 (en) 2012-08-13 2019-04-24 Cedars-Sinai Medical Center Exosomes and micro-ribonucleic acids for tissue regeneration
EP2712870A1 (en) 2012-09-27 2014-04-02 Rheinische Friedrich-Wilhelms-Universität Bonn Novel RIG-I ligands and methods for producing them
EP2759595B1 (en) * 2013-01-24 2016-09-14 Pierre Fabre Médicament S.A.S. Composition comprising an encapsulated antagomir
JP6446026B2 (ja) 2013-03-15 2018-12-26 ミラゲン セラピューティクス， インコーポレイテッド 架橋二環式ヌクレオシド
US11253549B2 (en) 2014-05-23 2022-02-22 JangoBio, LLC Methods to rebalance the hypothalamic-pituitary-gonadal axis
US11439668B2 (en) 2014-05-23 2022-09-13 JangoBio, LLC Methods to differentiate stem cells into hormone-producing cells
EP3200808A4 (en) 2014-10-03 2018-05-16 Cedars-Sinai Medical Center Cardiosphere-derived cells and exosomes secreted by such cells in the treatment of muscular dystrophy
US20160122762A1 (en) * 2014-10-27 2016-05-05 University Of Iowa Research Foundation Methods of treating atherosclerosis
SG10201906716QA (en) * 2015-01-20 2019-08-27 Miragen Therapeutics Inc Mir-92 inhibitors and uses thereof
EP3402543B1 (en) 2016-01-11 2021-09-08 Cedars-Sinai Medical Center Cardiosphere-derived cells and exosomes secreted by such cells in the treatment of heart failure with preserved ejection fraction
US11351200B2 (en) 2016-06-03 2022-06-07 Cedars-Sinai Medical Center CDC-derived exosomes for treatment of ventricular tachyarrythmias
CN107586784B (zh) * 2016-07-08 2020-08-25 田小利 一种分离的rnasek基因及其在制备抗血管衰老药物中的应用
EP3515459A4 (en) 2016-09-20 2020-08-05 Cedars-Sinai Medical Center CELLS DERIVED FROM CARDIOSPHERES AND THEIR EXTRACELLULAR VESICLES TO DELAY OR REVERSE AGING AND AGE-RELATED DISORDERS
WO2018195210A1 (en) 2017-04-19 2018-10-25 Cedars-Sinai Medical Center Methods and compositions for treating skeletal muscular dystrophy
US11660355B2 (en) 2017-12-20 2023-05-30 Cedars-Sinai Medical Center Engineered extracellular vesicles for enhanced tissue delivery
WO2019167995A1 (ja) * 2018-02-28 2019-09-06 国立大学法人東京医科歯科大学 虚血病変部位特異的な遺伝子治療法

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2533701A1 (en) * 2003-07-31 2005-02-17 Isis Pharmaceuticals, Inc. Oligomeric compounds and compositions for use in modulation of small non-coding rnas
EP2290071B1 (en) * 2004-05-28 2014-12-31 Asuragen, Inc. Methods and compositions involving microRNA
WO2006133022A2 (en) * 2005-06-03 2006-12-14 The Johns Hopkins University Compositions and methods for decreasing microrna expression for the treatment of neoplasia
EP2455493B1 (en) * 2006-07-13 2014-01-08 The Ohio State University Research Foundation Micro-RNA-based methods and compositions for the diagnosis and treatment of colon related diseases
US8466117B2 (en) * 2006-07-28 2013-06-18 The Johns Hopkins University Compositions and methods for modulating angiogenesis
WO2008070082A2 (en) * 2006-12-04 2008-06-12 The Johns Hopkins University Stem-progenitor cell specific micro-ribonucleic acids and uses thereof
WO2009004632A2 (en) * 2007-07-05 2009-01-08 Yeda Research And Development Co. Ltd. Methods of identifying components of a biological pathway and use of said components in regulating diseases associated with altered cell proliferation
BRPI0813527A2 (pt) * 2007-07-18 2014-12-30 Univ Colorado Expressão diferencial de micro-rnas em corações humanos que não falham versus que falham
DE102007052114B4 (de) 2007-10-30 2011-01-05 T2Cure Gmbh Verfahren zur Modulation der Funktion, des Wachstums oder der Differenzierung einer Zelle

Also Published As

Publication number Publication date
ES2651287T3 (es) 2018-01-25
US20150018407A1 (en) 2015-01-15
CA2704290C (en) 2018-05-15
PT2684955T (pt) 2017-12-13
JP2018083817A (ja) 2018-05-31
US8912158B2 (en) 2014-12-16
US9862949B2 (en) 2018-01-09
NO2684955T3 (hu) 2018-02-03
DE102007052114A1 (de) 2009-05-07
DE102007052114B4 (de) 2011-01-05
EP2217704A1 (de) 2010-08-18
WO2009056116A1 (de) 2009-05-07
US8258113B2 (en) 2012-09-04
DK2217704T3 (en) 2017-03-13
PL2217704T3 (pl) 2017-06-30
JP2011500858A (ja) 2011-01-06
HRP20170346T1 (hr) 2017-04-21
ES2618203T3 (es) 2017-06-21
EP2684956B1 (de) 2017-08-23
EP2217704B1 (de) 2016-12-07
SI2684955T1 (en) 2018-01-31
JP2014196317A (ja) 2014-10-16
US20100324118A1 (en) 2010-12-23
EP2684955B1 (de) 2017-09-06
SI2217704T1 (sl) 2017-05-31
CA2704290A1 (en) 2009-05-07
PT2217704T (pt) 2017-03-15
EP2684956A1 (de) 2014-01-15
JP2016199565A (ja) 2016-12-01
CY1120226T1 (el) 2018-12-12
CY1120360T1 (el) 2019-07-10
HRP20171874T1 (hr) 2018-01-12
HUE037494T2 (hu) 2018-08-28
US20120322856A1 (en) 2012-12-20
US20160237433A1 (en) 2016-08-18
LT2684955T (lt) 2017-12-27
PL2684955T3 (pl) 2018-05-30
US9279123B2 (en) 2016-03-08
LT2217704T (lt) 2017-04-10
JP5898843B2 (ja) 2016-04-06
EP2684955A1 (de) 2014-01-15
DK2684955T3 (en) 2017-12-18

Similar Documents

Publication Publication Date Title
HUE031559T2 (hu) Eljárás angiogenezis, vaszkularizáció vagy érjavítás elõsegítésére vagy tumor-angiogenezis gátlására
AU2016244333A1 (en) DLL3 modulators and methods of use
Goldmann et al. A comparative evaluation of NB30, NB54 and PTC124 in translational read‐through efficacy for treatment of an USH1C nonsense mutation
Mirotsou et al. Secreted frizzled related protein 2 (Sfrp2) is the key Akt-mesenchymal stem cell-released paracrine factor mediating myocardial survival and repair
EP2673001B1 (en) Nts-polyplex nanoparticles system for gene therapy of cancer
UA126549C2 (uk) Композиція і спосіб для з&#39;єднання позаклітинних доменів типу і та типу іі як гетерологічного химерного білка
HUE025013T2 (hu) Szulfonamid-származékok
Fukui et al. Gene therapy for the inner ear
Tanaka et al. Modeling alveolar soft part sarcoma unveils novel mechanisms of metastasis
Zhang et al. The pathogenic mechanism of diabetes varies with the degree of overexpression and oligomerization of human amylin in the pancreatic islet β cells
BR112020018364A2 (pt) Sistemas e métodos para o tratamento de hemoglobinopatias
KR20080036015A (ko) 글루코오스 유도성 인슐린 발현 및 당뇨병 치료 방법
Katovich et al. Angiotensin‐converting enzyme 2 as a novel target for gene therapy for hypertension
CN110381982B (zh) 与myomixer促进的肌细胞融合相关的组合物和方法
BR112020012921A2 (pt) Oligonucleotídeos antissenso de alfa-sinucleína e usos dos mesmos
Kolosov et al. A role for tricellulin in the regulation of gill epithelium permeability
Ceinos et al. Mutations in blind cavefish target the light-regulated circadian clock gene, period 2
WO2014123614A2 (en) Antiviral rift valley fever virus virus peptides and methods of use
PT876491E (pt) Gene do síndroma qt longo que codifica para kvlqt1 e a sua associação com mink
ES2708624T3 (es) Composiciones y métodos para tratar la enfermedad arterial periférica
Liu et al. POLR2A blocks osteoclastic bone resorption and protects against osteoporosis by interacting with CREB1
US9757402B2 (en) Method for measuring cell-to-cell transmission of α-synuclein aggregates using bimolecular fluorescence complementation system and method for screening a substance for preventing or treating neurodegenerative disease using the same
Rethnam et al. Differential expression of neuropeptide Y Y1 receptors during pulpal inflammation
Recino et al. Immunosuppression overcomes insulin-and vector-specific immune responses that limit efficacy of AAV2/8-mediated insulin gene therapy in NOD mice
Jovanovic et al. Sustained chemogenetic activation of locus coeruleus norepinephrine neurons promotes dopaminergic neuron survival in synucleinopathy