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DTIC ADA423340: Antigen-Specific Immunotherapy Using Lentivirus-Transduced Hematopoietic Progenitor Cell: A Novel Approach for the Treatment of Metastatis Prostate Cancer PDF

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Preview DTIC ADA423340: Antigen-Specific Immunotherapy Using Lentivirus-Transduced Hematopoietic Progenitor Cell: A Novel Approach for the Treatment of Metastatis Prostate Cancer

axaré umber: GRRDL?-02-1.-99) Till: antigen-specific Iammoterspy Using Leuvivirus-Traas Tenatepoietic Progen:tor Cell: & Yovel Apprasch “0 Treatnan: 0! Neascat=s Proptate Cancer “the PROSCUEAL VES OR: binzhao cheng, PL. COMTRACTIN Jobna Hopsins University Baltingee, 02 21205 ariel fomrend y Medical Research and Je, Mavylazé 21702-3012 varr POON STAORMENT: Approved 2 Dit ribuio: Public Release; dalinived cha views, cpinions aad/oz (ind! repers ave Uhowe of Uke walker fi anG whwuld aol be consLeuvd ue ae official Baparinent of Lhe Apmy oosilion, policy woes gignated ky other documentasion 20040602 062 Form Aparaved . REPORT Ont He. O7e- 088 DOCUMENTATION PAGE Linznea Cheng, 2h-s ohne opting vaivorsity Re-tingre, ND 2Z0E, LB, Aum Modicn’, Hosor~ob 974 Nobevig! Coxmand Feel, Satrien, maryland 21752-5003 ginal coszaiua coley 3lator: Aid, PLUG ropreductinns wi"! he fs black ane uti Snn cved Tor ub: Recent: Ole2-bulion Dal iatbad hs gays eee i ot eI Las Lanai the wrens eee tome tae ively gece in DCs diteexiated rno tom naostued bem topieic progoier cals They bave: 1} ate en ial veces oepressing 3 ‘del ns: angen Intucas hora zhain (HA): dovelag » meched oes pansies bens boos ma-TOR PEEL (al shat conti nme at toys il lepine ies? err hl he as aed ad ‘moosicse coll can peste preszut tk 14 adel minor aso and sisal Tc responses; 4) devised apart snags to Pint pvr fland spe ata on 1 expestsg opr nel Laut! and 3 dices the cea a eadcee ab appeive e-csublicied neuoroxpeesie be HHA angro, This adv powes«proockpriplea unmene compe ia mals th Eantglaavion of aniger goveaznduced hominy lageor elle aa #0 component in acces poke ane Rte igor irmimerhespy of agree rumors Ky eacoses by i gry sr Iho Hepes elle ere paint Teschng Macon vesctch joel Vator Meise 0. RED. Ginn vareinen, antigen preeer-ing calle; lentiviral vearoms sirstozoletic prgimite: celle) gens tacrary evened Tachaest sted Unelazeitied ating RET TSEC ‘Bata go han. FS Table of Contents Cover... SF 298. able of Contents... Introductio Body. Key Research Accomplishments. Reportable Outcomes. Conclusions. References... ANpENEICES a esse Introduction ‘This ie the Binal report fara New: Investigator Avwatd (DAMDI 5) wih the proposal exited “Aarigen specific immmnotacrapy using Ientivinis transd-ced hemarapoietic priyenitnevel’s navel sayproach forthe ieatment of metastatic prostate cautot” Rody Backeround: Over forty thousand Americans dic annually Fram metstatic prontalecansemm even after ‘sxtonsive teattnents, and reevot dats suggested thot he deuth rate wos underestimated. Standard ‘hems sal rahul therapies have been inellevtive at prolonging tke survival ofthese patits aud are ‘lien associated with severe vide ellis. New larms oF unerapies therefore aro wipaatly needed for these ying patients, Biological therapucs, pa-icularly immunotherapy which boosts the body's self defense against tumor cells ate inrcesingly used as adjunct or alternative treatments for prostate an alher faimcers, Muny fherapewtie tamer vuccination appracehes ave focused on engineesing antigen resenting calls (APC), partinlarly dendsitic cells (DCs). However, recent stdics using ey ¥iv0 Antigen-foaded DCs as cancer vaccines have shoven limited suocess. Objective! Hypothesis; We hypothesize thal humalopuieli stemépreyeniter eels (TISPCs) wansduced wilh anpecific susnarantigen gene fallowed by wansplaaration can be differentiated in vivo into functional DCS that express a tuuior antigo: ac high Ievels, Large unmburs of antigen-expressing DCs that mature fn vi cur effectively autvate lurcoe-speciticT eels, eesolting ia more potect and loup Tasting antétamar imanunity. This novel approach to generate large amber of antigen-specific DCs fe vivo may overcome limitations observed in carly 1X-based clinival trish. Specitis sims: Wo witltost whether chese in viv differentiated, antygr-exprossing DCs muinlsin theic functional capaci in priming unlitomar immune responses und ulimalely reverse igninue Loleiance £0 lamar antigens. Specifically, we will: [} examine Uae inmoume rexponses “a specilic mode! eanuar sotigen in mics with reconstituted immune system by HSPCs eransceced with the angers 2) examine if the tansduced DCs ean generate imnunity to provsat and eradicate the amtigen-expreswing tumar cells in movse wnodels; 3) examine if his wppronch ean preverand eradicate the lamor cells expressing prostate pecifiy membrane gnligen (PSMA) in sinilar mouse randel Study designs: Ta addition to PSMA, we will aso use a wellcharsoterized modst mumor autigen, influenza Hemagelutinn (ILA), in order to track directly antigen-specific imaruae responses and quickly asscss ancnnnnor efficacy usiug tpvisgerie mice expressing T call raeplars weenwmniving the TTA angen, Tic system eTlhnes i ha preisely delerrsine “imeliaal ales af TA-expressiny DCs in iaducing T cet] activarion verses tolerance, and dofine optimal vacctaation suatezics, HSPCs from mouse bone marrow ‘will be tansduced e* vivo witk lentivital vootors expressing PSMA. or HA ueder the control of a DC- solective promoter, and thon transplanted into synyencie repent rive, We will evaluate ibe efficaty wud spevifcity of DCx differenlialed frone transduced YISPCs with either TA o¢ PSMA gene in priming imatmne responses to eliminate planted TRAMP-C2 mouse prostace tumor cells engineered 40 cxpressing cither HA or PSMA). ‘These santies wil serve tn enlifythe hes! wacxination suteyy tat will subsequently be tested in cliniga tia asa treatmen! for metaslatie prostate caneet, targeting PSMA or other emerging Drie turwir ankigens, Since PSMA is alsa over-espresed inthe neovascalamucc of @ vation of ‘malignant neoplasan, this immune therapy’ approach may also apply to other cancers sue. as unctastatic brsast canects, ‘Tesi-d: To examine the immune responses toa mo¢el rumor sntigen select:vely expressed! in DC's Tifferentsued in viva fom iranecuced HSPCs, A well-churucierized axilel ume untigen,inlluen.a Ihenaagghutin (TA), uleng with transyenie mice (lines expressing T cell receptors -ecognizing the LA antigen aud lines expressing HA ir prostate cpithelisl cells), will be used. This system allows us to 3 “precisely derermune fictional roles of HA. expressing DCs (differentiated fv vivo from transduced HSPCs) in indueing ‘coll activation versus tolerrnce. Sinue PORP noon endef ond only Farle the Task 4, we have been toeusing sm Task I i: the past Key Research Accomplishments 1. We have successfolly construct Lentiveal vectors exntessicg HA (the mode! ruracr antigen usin tis stil) aml several cami vec expressing GFP or NGPR (incline nerve grveih ctor ‘eceplor). High tle vias sacks van be made. All the jasent gent funetioned a designed. Th addition to nae hematopoietic seemipeogenitor eels (ISPCS), we als foued thatthe same approach can be applied to mouse TISPCs (Re. 1-2}, "Ths is portant Decause mouse nodes ae used as pee-linieal ules inthis proposal (Rel. 1-2). We devised the method to etticiently ‘uansdice igolted HSPC from monse bone marrow cells that ae found to morc difficult ta be ‘ranstood by lentiviral vetors, tn fact. the established outisirus-modivtd gone transduction technology allowed us to investigate a related approach to improve bone inee=ow transplantation ol. 34), 3. We uilved a lentivewsw tansdave mouse HSPCS, ten wansplunted the mide cli and ana! yred expression the teuigent in the dendrite ells (DCS) in relevant Iriarte to determine the etficacy ofthis approach. UISPC were transduced with GP asa reporter gene under the consol af a rang cansiucive promoter Ref. 1-2 5). Traesduction was folewed by transplantation inc Ithly inant reipicess, After engrefecont, DCs were isolatoc from spleen and lymph xis, anal FACS arulysis wes sorfomed ta deteine the seroeatage of DCs {CDIfe MMC TP cele that expressed GTP. ALS wacks pst inumplant an uveraye of 34% ofthe DC expressed the gene inthe spleen and 254 in the LN (Fg. | in Ref, S/Appendiv 1). Thus ceaascuction of precursors, diersndation of wansduved precursors into DC, and tafficking of DCs to secondary lymptoi organs all sere sfficiont. Simliar potcensiges of wansduced cell wax bir inthe CDT 1e_ rnetion (Fig. in Appel 1) as well asthe lytaphaey's faction ater collagenase digestion, indicating hal transduction wan abt selective for DC progenitors. Similar numbers of splenocytes wese obcained from nop-taasplanted aad tansplanted mice, with equivalent percentages of B and T Ipmiphocyres, indice tha recoestintioa is well underway’ at this ae (not all deta show). ‘Tho proportion of CD 1s DCs expressing the wansgene dd not change after sdainisiraton of agonist ant-CT40 ano, indicating tha system io DC wetvation did not selesively affect wanspene® vs transgene” DC progeny 4. Using de HUA a ¢ model ture antigen, we have exaived specie antigen presenta and Te activacin in mice postax vivo gene rancduction (with HA gave) and trasplanaton a mause HPCs, Dis, Yan Cui, Drow Padall and | (co-investigatrs ofthis application) togetber wich my cofkengues a Jebus Hopiins Oncology Contr, psfermed lage scales of tamor vaccinations auc ‘swiention experiments with HA-cxpressing experiment: tumors, We fomnd tot 1) eh ransplanied ‘nl HA. gene ~ansduced vlln cin persistently prcat the HA mall eumor antigen an salt cel responses 2a tiparite strategy is requited ro nant powertsl and specific ateck an TTA- cxprestng experimental tures ané 3) tis combined approach can eradicate an agpressive pre- ‘tablished tor expressing the HA antigen, Details of immune analyses have bee just published (Get, 5) and inched as Anaendix Reportable outeares Manascsipi: Bel. 5 ‘Yan Cei, Bru Kellcher, Erin Staley, Ephraim Fuchs, Kevin Gorski, Hyam Levitsky. Ivan Borrello, Cur T.Civin, Stephen Sehoenberger, Linzhan Cheng, Drow M. Pardoll, Katharine A, Whareenby, 4 Tumor immunciterspy with at 10:882. n taansduced hemstapoietic stem culls. Nulure Medicine, 2003, 1. Lemtvirnl vostors expressing dhe HA artigen aad otter reporter geucs (GEP or NGFR) 2, Lentivinl vectors expressing the PSMA gene, used by Drs, Samuel Denmeade, John Ie, ‘Robert Pili (currently PCRP awardees) at Johns Hopsins Oncology, and Dr. Party Pomper a: Jolns Hoping University Radielogy ‘Methods 1. Eificioar nd stuble yer transler into eapuse HSPCs capable of reconstituting the blood and samme systems in irate recipient mice 2, Efficient und sustained transgene eapressian in engsafeed ovlls corived from tamnsduced HSPCS, ‘including dendrite cells that play a cereral rote in modnlaring imazonc responses 3. A tripanite steategy {BMT with antigen transduced HSPCs, expansion and activation of derived [DCs in in viva by FIG and CD4p activators end donor lymphocyte infusion) to eradicate aggressive tamer. “Eployment und regsarch epporcunitins Dr, Yin Cul, posidhcteral fellow esrinentored by Dis. Linzhao Checg snd Drew Paedoll, tad co lr of This appication, is de test author ofthe papcr cited in ef. 5, She has aevepted ea independent research postion as Aseistant Professor in Louisiana State University Health Science Center, Dept. of Medicine, Gene Tuerapy Progeam. She is pluning to conlinae asiag gene cherapy approach fo develop new mnnvothorspics for agursssive cancers Conetu ‘Thi in the Fins anal repoet for a New Investigator Aswaud, We have mad sigan progress for ‘LaskeLtha: wins fimcled by PCRP ps listed ahve. The funding (575.000, cect) bas been crucial Support me ts participate the team effort at okins Jopkins Oncology Center and to develop novcl ‘euncer immumotherapies. Together with other aur Hopkins colleagaos, Dx, Yan Cui, Drow Purdoll ‘nd 1 (oo-investigatos inthis app'ication). Ws have: 1) mails ‘onfiviral vectors expressing 6 model tumor antigen HA; 2) developeilu method 10 efficiently teansdiee mouse bons marrow progeticor ‘ells that esenstituteimamine and blaod systems ‘n iradiared and wareplimtll ices demonstrated thst she wangplanted a transduced calls cam persistently aresent the TTA model turner antigen and stimulate T coll responses, 4 devised w triparilesrategien un anouct powerful and specific altack on TIA expressing enperimental turaorst and 5) domanstrated the cfficy to orients tu aegcessive poe-established tumor expressing the HA antigen, ‘Chis study provides a proof-ol- principle in immune competent animals models that transpkitation of anligen gene transduced Ihematopoistic progenitor cells ads s new corsponent to ackieve potent and sustained aatigen- snectfic immunotherapy of aguressive turaoes. Key discoveries by tis group and thefc Hopkins collegues are published in a leading welicine jovanal Nuture Medicine ANate My Jab has sited resenrch lnewses aller we joined the Johns apkius Jasdtute for Cell Lngincoring, jn Fall 2093. TTosvever, D:. Deowe Pandoll (a co-nvestiaator of this proposal) and ny ceher Former calleagucs at Jobs Hopkias Oncology Center re eomtinuing the effoet oF immuno-therapies to teat _motastafie anoers including prostule vancers My current address The Johns Jlepkins lnsinue for Cell Engineering Broadway Research Building, Room 747 733 N, Broadway Baltimoce, MID 21085 References 1. Cul ¥. Golob 1, Kelicher Is. Yo‘, Bundall DM snd Cheng T (2001). Targeting wansgene expression ta amtigon prssenting cell lTereniiaed in vivo [own lea-virus-trnsduced, engrafting man and sniouse hematopoietic stemprogenitar ccs. Blood, Vol, 98. 214s, 2. Cul Y, Kelleber E, Golab J. Partoll BM ard Cherg 1 (2001), Spesifie antigen presentation and T cell activation iu mice post ex vivo gene transduction anc cansplantatian of bone mattow cols, Blood. Vol. 8, 235i. 3. Whartenby KA, Seeacy HN, Kim H, Rucke F, Gorski KS, Cheng L., Paudall “tramgluction af donnr hemauophisic sem-pragenlor cells with Fas allogeneic hone maseoss transplant, Blond, 100: 3137-3154, 4. YoZ and Cheng L (2002), Restctec transgene oxprossion in human CD341 cells mediated by novel Jemvira vectors, Expr. Hemacelogy, ol. 30, 177s, 5. Yan.Cui, Brin Kellcher, Frin Sizley, Ephraim Fuchs, Kevin Goeski, Hyam Levitsky, Ivan Borclio, Curl I. Civin, Stephen Schoenberses: Linzhao Cheng, Drew M. Pardoll, Katharine A, Wharton. ‘Temor imawnothcrapy with antigen tunsdeved heraaiapoietic stem wells, Nature Medicine, 2003, 1982, Mand Civia C (2002, wneud engraftment in Appendix: Ra 3 ARTICLES name, medicine Immunotherapy of established tumors using bone marrow transplantation with antigen gene-modified hematopoietic stem cells ve CGUET Chit, Steen PSchoenbegerHinhno Cheng! re M Purdl? &aharine A Whartenby! 2°, Erin Kale, ein Ste! Eprsin Fuchs, Koen Seay Lavish, van Hartel i 1 sa eas cancer mmuotnagy it else ince ea expe tea pean fot { SigtN Val an 000 et nn ten nr sy cet a ogee eon. 4 Eioganened tne tery net si win tater eicton few bts aie wena tmp Sete int isto gan patti Teste ice an rr oat een Be ne 3 ters toon empresa sen se na oie at en neared hee {edcaltinimdsicdrcpew mc win exes memser oso eet aimee rotor E SUSSGRUo.repeaae ml te amitenor tones ompision nny tec Woes $ i fT Hy aketerapat Teal Tate see ort ptt nigenpesi immu fe a8 re Dee ey Heed. screed aro ae” aa sory tatannae Ennesion fangs 8 08 In ARTICLES ARTICLES QR eater teseePuttehing croup mipsinmnstmcomtetoniiin

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Most books are stored in the elastic cloud where traffic is expensive. For this reason, we have a limit on daily download.