DTIC ADA423266: Sodium Iodide Symporter Gene Transfer for Imaging and Ablation of Prostate Cancer

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by  Defense Technical Information Center

aD.

avaré Nuaber: DAMO27~02-1-0119

‘ITL3: Sodium Todida symporter Gene Transfer for Imaging and
ablation of Prostate Cancer

PRINCIPAL INVESTIGATOR: Sissy M. Z:tang, Ph.D,

CONTRACTING ORGENIZATION: The Ohio State University Research
Poundazion,
Cozumbus, Ohio 43219-1063

RPORT PAYY: ganvary 2004

WYER Ob HuvORY: Annual

PREPARED FOR: U.S. Avny Medica! Research and Ma
Fort Detrick, Maryland 21702 $012

hel Command

DISTRIEUTION STATEYENT: Approved for Public Release;
Distrizutie: Unlinited

‘he views, oginions and/or findings contained in this zepor:
those of the author(s) and sioyid rot be construed as en off.
Departnent of the Army rorition, policy cr decision anless so
desiguated by other documentation

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Tue oat BHT = REROAY HOPE ANG BATES COVERED

dene sant! Annual (15 Doo 2002 ~ 14 sc 2003)

En aio scare RINNE NERS

Scdlum wedide Symorter Gene Trarefer for Imaging ard DeMDr7-02-1-0113

Sblazion of Frosvate Cancer

Sissy %. dalarg, Pao.

7 FERFSRITS GRGARICATON HARES) AND ADRESSES, “EFERFDRT RCE

She thio State Univereizy Research Foundation REPORT aun

Conshue, One 43230-1063

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F SPREGRIG ONTCRENE 8 BRESORNG MOTOR

‘Reeney nates) ato ADDRES) ‘Keeney Aeron? ALANOER
sf, Army todical Roscarsk ané materie: Common
Fort Detrick, Marylané 21702-5012

7 SORPTEMER TARY WPS
Griginsl contains coler plates: #11 L0:¢ reproductions wil] be tn Black and white,

Apgsaved for Publis Relesge; Dietetbut!an UrLinized

5 AASTARCT eas 200 Waste
The sodicm ioside eyaporter (KIS} meiiates 1oaide uptake in thyrots tel! leylar o@lls and
exovldas @ mecknnti fer offective rdiojodide srentront of residual, securren:, até
Retastetic thyrois cancers, The obsestive of the pucpoaed research 1 Le Lest she
hysothesis that exprece-on of exogenous MIIS ir prastatie kissin wilt, mable radioiogide
to tocelize ané sblate rezicual prostate cnsor Following prostatescony, sich chet
recusre:ce an2 metastasis of lle cigease can be prevented. Tow specifiy aims of this
protect ere tor |1) seufian movastarse: provreneioa to distact Limp nodes and lunge
Soliowize subcutanaous incculation of rata with MMILyin prostatia aéenseazcinoma colla
Seouessioy INE, [21 snvervigate uhether redleledide Lleteyy will prevent. pataeraare
Sod/ex prolong suxv-val in vars Uewting dohastancous “OUiyLi timbre that empress MNES) (3)
Sctcunine the cepresaion lovol of RIE ragaired 20 e::eit selective rediolodide-rediaved
Kilticg of Mamyiu-hi=s prostatic adeoncavcizoma cells in Glyo) ond, 14) vestrice AMIS
cagrossion in prostatic tisaus urter transcriptional regulation of prostabe-sperific

74 SURDECT TERRE TE AER OF AOE
wrostcte canser, sodiun ied-#e symporter, sere transfer, radioiodiae a
TF SEDURTTY CEASSEICATON | 76. SECURITY CEAGEREATON | 18 GECUPY CLASGRATION —| Bi CHVTATON OF RESTRICT
ornaroar ‘or as pace Onausreacr
Tnetai fing Inevaneified Onedaveitied meLimived
TEN ae s9oN a Seog Tan Bie 2
‘8 M charg, 2098 annual report for OAMD7-02-1.0119

Table of Contents

cover Pago 1
SF 298. Poge 2
TABLE OF GONTENTS.... Poge 3
INTRODUCTION. Page 4
BODY. Page 48
KEY RESEARCH ACCOMPLISHMENTS Page 6
REPORTABLE OUTCOMES. Pago 6
ONCLUSIONS. Page 6-7
REFERENCES. Page 7
APPENDIX 4 Page 8-15
APPENDIX 2s once Page 16-24
APPENDIX. Page 25-21

Pages
r ‘Sef Jhiang, 2008 anual wp for DAWMDIT-02-1-0119

INTRODUCTION

The sodium iod'de symporter (NIS} mediates iodids uptake in thyrold follicular calls and
provides @ mechanism for effective radioiodide treatment of rosidual, recurrent, and
Tetastatle thyrold cancers. The objective of the proposed research is to test the
hypothesis lhl expression of exogenous hNIS in prastatic Hssue will enable radioiodide
to localize and ablate residual prostate cancer following prostatectomy, such that
Teourrence and metastasis of the disease can be prevented. The specific aims of this
project are to: (1} confirm metastatle progression to distant lymph nadas and lungs
following subcutaneous inooulation of rats with MATLyLU prostatic. adenocarcinama
cols exprossing hNIS; (2) investigate whether radlofodide therapy will prevent
metastases andior prolong survival in rats bearing subcutaneous MATLyLu tumors that
express HNIS; (3) determine the expression level of hNIS required 10 elicit velectve
radlo‘odide-medisted kiling of MATLyLU-ANIS. prostatic adenocarcinoma cells in vivo:
and, (4) restrict HNIS expression in prostatic tissue Under transcriptional regulation of
prostate-specific promoter.

BODY

Task 4. Confirm metastatic progression to distant iymph nodes and lungs following
subcutanaous inoculation of rats wih MATLyLu prostatic adenocarcinoma cells
expressing hNIS: Completed in Year 1 with one published paper

KMD La Perle, D Shen, TLF Buckwaltar, 8 Wiliams, A Haynam, G Hinkle, R Pozderao,
C Capen, and SM Jiang (2002) "i Vivo Expression and Funct on of the Sodium
logide Symporter Following Gene Transfer in the MATLyLu Rat Model of Netastatle
Prostata Cancar’, The Prostate, 80: 170-178.

Task 2 Investigate whether radioiodide thorapy will prevent metastazes andiar prolong
survival in rats beating subcutaneous MATLyLu tumors that express ANIS (Months 12-
24, completed)

Appendix 1: KMD La Perle, EAG Blorime, CC Capen, and SM Jhang (2003) “Ettect of
Exegenous Human Sodiurvlouide Symporter Express'on an Growth of MatLyLu Cells’, Thyroid
3433440

Appendix 2: DHY Shen, DK Marsee, J Schaap, W Yang, J-Y Cho, G Hinkle, HN Nagaraja, RT
loos, RF Barth, end Sh Jhiang (2004) “Effec’s of dose, intervention time, and radior weld on
sodium iodida symportar (NIS} targeted radionuclide Frerapy", Gene Therapy 112 161-169,

‘As reported in the previous annual report, we faund that NIS oxprosston inhibits
MATLyLu cell growth (La Perle et al, 2063}, We also found (Rat the time course of tumor
progression and spread of metastases vary significantly among animals even in the
Same experimental group (La Perte et al, 2002, and La Per'e ot al, 2003}, Thus, we have
chosan an alternate aniinal model fo investigate the theraneutc affectivaness of NIS-

Page 4
‘SM -hisng, 2008 anntal part for DARD17-C2-1-0119,

maciated radioiodide therapy (Shon ot al, 2004). The effectiveness of NIG os a gene
therapy agent ies in ifs ablity fo madiata the uptake of a variaty af radionuclides, such
as 1, "|, and “Re, We have conducted a study comparing the therapeutic
effectiveness of these diferent radioncclides In NIS-expressing gllomas, which have
telaiively uniform survival tima among untreated animals. We show that: (1) the
therapeutic effectivenass af "Vin prolanging the survival tima of rats hearing FS@/MNIS
gliomas is dose- and treatment-ime-dapandent, (2) the number of remaining NIS
expressing tumor cells decreased greatly in RGZ/NIS gllomas post "I treatment and
‘was inversely relaled o survivat tine; (2) 8 ani each of "lis as effective as 16 mCi
"1 alone, despite @ smaller tumor absorbed dose; (4) "*ReO., a polent i erritir, is
more efficlent than “| to enhance the survival of rats bearing F9B/hNIS gllomas. These
studies demonstate the importance of radiopharmaceutlca” selection, dose, and tinlng
of treatment (0 optimize the therapeutic effeciveness of NiS-largeted radionuclide
thorapy.

Task 3. Determine the expression levet of hNIS required to elicit sslective radiciodida-
medlated killing of MatLyLU-hNIS prostatic adenocarcinoma cal's in vivo {months 18-36,
In progress)

We hypcthesize that radioiode uptake in vivo correlates with NIS expression levels
within a certain range. We have been trying to establish MatLyLu clonal cells with
doxyoycline inducible NIS expression so that we could determine the range of NIS
expression levels required to aliit radioiodide-madiated killing effects. Unfortunately,
wwe have failed ta acquire doxycyctine-induciblo MatLyLU-nNIS clones despite several
attempts. However, se have been abla to establish doxyeyctine-inducib‘e hnNIS clones
using other cell lings. We are nav trying to switch to other human prostate cancer cell
lines to accomplish this task,

Task 4. Restrict hNIS expression in prostatic tissue under transcriptioral regulation of
prostate-spectf c promoters (Months 1-36, in pragress)

Appendix 3: DK Marsee, DHY Shen, LR MacDonald, DD Vadysitisack, X Lin, G Hinkle, RT
Koos, S14 Jhdang (2004) *Fnaging of Pulmnana-y Tumors Following RIS Gene Transfer Using
‘Single Photon Emission Computed Tomography’, Concer Gene Therapy in press,

Since the most challenging issues of gene therapy remains vector delivery to the target
tissue, we have beon trying fo use NIS as imaging renorter gene to optimize vector
delivery efficiency, in addition to engineer adenovirus camying NIS under the contro! of
prostate-spectfc pramoter. Due fo the high mortality assoctated with pulmonary tumor
metastases, we have focused our MatLyLu studies on the use af NIS in gene therapy
for lung tumor nodules (Marsee et al, 2004), In this study, we established NIS-
expressing pulmonary tumois In nude mice by intravenous injection of tumor calls and
investigated the minimal tumor sizo roquired for in vivo detection of pulmonary tumors
by single photon emission computed tomography (SPECT) with pinhole collimation. to
ander to define the axatumic location of NiS-expressing tumor nadules detactable by
SPECT, we performed simultaneous, duslisotope imaging. We injected 1 mCi sonTo+

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StH Jhlang, 2008 annual sepor for DAMD17-02-1-0119

MAA via teil veln to Image pulmenary perfusion and injected 1 mCI Na"
intrapertoneally to Image NIS-expressing tumors. Fused images showed that "Tc
MAA perfusion defects correlated with NIS-mediated "uptake. Post-mortem analysis
revealed that tumors 3 mm in diameter could ba dotected by SPECT with pintole
collimation, These studies demonstrate the feasibilty of SPECT to datact pulmonary
‘umors expressing exogenous NIS in rice.

We have also been exploring different methods to efficiently dellver adenoviaises to
MalLyLu puknonery turnor nodules. fn vitro studies demonstrated that MatLyLu cells are
highly susceptible to adenoviral infection. Inilally, we attempted ta deliver adenoviruses
via intravenous delivery. Viral gere expression was found Ih the liver, but not in elther
the lung parenchyma or pulmonary tumor nodules. We algo performed Intratracheal
injection to celiver virusés to pulmonary wmors. Whereas the lung parenchyma
showed strong viral gene exoression, the wmor nedules were not Infected. These
results suggest that MatLyLu pulmonary tumors do not communicate with the alveotar
alrspace, likely du to the localizatio of Maly. tumors on the surface of the lung, We
are currently in the process of performing intrapleutal adenoviral delivery as a means to
infect pulmonary metastases.

KEY RESEARCH ACCOMPLISHMENTS.

+ Demonstrate the importance ef radiopharmaceutical selection, dase, and timing of
treatment fo optimize the therapeullc effectiveness of NiS-targeted radionuclide
therapy

+ Demonstrate the feasiiity of single photon emission computed tomography
(SPECT) fo non-invasively detect pulmonary metastases of MatLyLU-NNIS tumors
wit a dlameter of 3 mm in size in mie.

REPORTABLE OUTCOMES
Manuscript 1: KMD La Perle, EAG Blonme, CC Caper, and SM Jhiang (2003} "Effect of

Exogenous Human Sodiumlodide Symporter Expression on Growth of MatLytu Cells", Thyroid
13133-4140

Manuscript 2: DHY Shen, OK Marsee, J Schaap, W Yang. J-Y Cho, G Hinkle, HN Nagaraja,
RT Kloos, RF Barth, ark! SM Jhiang (2002) Efecis of dove, intervention line, and radionuciide
on sodium iodide symportar (NIS) targeted radionuctide terony’, Gone Tharapy 14; 161-169.

Manuscript 2: DK Marsee, DHY Shen, LR MacDonald, DD Vadysiisack, X Lin, G Hinkle, ST
Klos, SM Jhiang (2004} Imaging of Pulmonary Tumors Following NIS Gere Transfer Using
Single Photon Emission Computed Tomograptiy’, Cancor Gene THerany lh orass.

CONCLUSIONS

© Selecting the optimal raciopharmacauticals, as welt as the optimal dose and timing
of treatment, could further optimize NIS-targeted radionuclide therapy for prostate
cancer.

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‘SM Jnlang, 2003 annual reper for DAMIDI7-02-1.0119

+ NIS can serve as an imaging reporter gane to optimize vector delivery for prostate
cancar gene therapy.

REFERENGES

KMD Le Perle, D Shen, TLF Buckwaltor, B Willams, A Haynam, G Hinkle, R Pozderac,
CC Capen, and 8M Jhiang (2002) “In’ Vivo Expression and Function of the Sodium
lotiide Symporter Following Gere Transfer in the MATLyLu Rat Madel of Metastatic
Prostate Cancer", The Prostate, 50: 170-178.

KIND La Perle, EAG Blomme, CC Capen, and SM Jhiang (2093) "Effect of Exogenous Harman
Sodiumifocide Symportse Exprescion on Grownh of MatLy.il Calle’, fhyrole 43134-4140

HY Shon, DK Matsoe, J Schaap, W Yang, J-Y Cha, G Hinkle, HN Nagaraja, RT Kloos, RF
Barth, and SM Jhlang (2004) "Elfects of dose, intervention tims, and radionucide on sodium
jodie symporter(NIS} targeted radionuclide therapy". Gene Therapy 11: 164-168.

DK Marsee, DHY Shen, LR MacDonald, DD Vadysirisack, X Lin, G Hinkle, RT kloas, St uhiang
(2004) "imaging of Pulinonery Tumo’s Folloulng NIS Gene Transfer Using Single Phaten
Emission Computed Tomograpty", Cancer Gane Tharapy in press.

APPENDICES

‘Three manusoripts are appended.

Pane?
Appendix 1

Effect of Exogenous Human Sodium lodide Symporter
Expression on Growth of MATLyLu Cells

Risa WD. Le Pte.) Die AG Rammras Crees @ Caren, awl Sis Wang!

“Tha sodium iodide symporter (Nis) medias iodis wptake in Hye calls ena enable He fftve radii
Gide tetmnent uf dngoid cancer, Ler ic mich intrest in fait edoinlide thenops i other cancers by
IS ger tami, This study showed Tit exogennce DIS cxprseon decreased MATT yu al prosaic ae
‘octsaneme cell growth, Lumor gro ans uutnetatie progression Were sight any delayed in smugencic
fete ingtel oni mized or lon papilations of SAIL YLAGNIS rolls eneapared ty rots with control famurs
NAT TaN tumors in muck mite had a lose, albeit latistcaly sisfcant, ote rate tho antl
Irmons The KET lebeling inn at Nlepoutve oss a3 Laver than io NiSumyalive ateas of tat mers le
freed feom amined popwiilon af MALLyLu BIS cally Cacwtb of cong) populations of MAILyT w-NTS celts

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