Campbell, A.I.M.

Kuliszewski, M.A.

Latter, D.A.

Monge, J.C.

Stewart, D.J.

Elsevier - Publisher Connector 

Direct DNA injection of vascular endothelial growth factor induces angiogenesis in non-ischemic myocardium 

JACC February 1998 ABSTRACTS - P,~tcr 25A 
by ELISA. Ab¢ixlmab had a dose-dependent inhibitory effect on both ~ arid 
dense PeR that paralleled the Inhibition of aggregation, Decreased PGR may 
enhance both the long and short term therapeutic benefits o! abciximab by 
minimizing mural thrembo~ formation and decreasing the amount of mitogen 
mlasse at It~ IIte Of vascular damage, 
At~xm~lb % Inl1~Mloft Of~t lOn t % Inh i~ ot ~eteaee 
G'g/mL) ATIP" P.TG t PAl.1 t 
0:5 5~3 14¢13.5 174-5.1 6~4.5  
1.5 t5 i  ?.? 24 *2~4 10 ,110  ~5.  ]'6 
2,S 54¢6§  64 t 1~,~ I~ "t" 5 3 76 * 0.9 
50 99,  t 4 1'~ ~: t:).8 07 ~ ~.5 82,  t 2 
tO0 100 ~06 ~Bt ±~0 67*33  fit ~r t7  
'TRAP(4tOa.M): t~0~(5 ~M) Data~s~ave ~: ~ ~tandard~ewatmn 
O~ne Transfer 
Sunday. March 29, 1998, 5:00 p,m.-?:00 p,m, 
Georgia Wodd Congmu Center, West Exhibit Hall Level 
Presentation Hour: 5:00 p,m.-7;00 p,m, 
~ G e t ~  Tlrlmsh~' to  the  l~ l tmonew VS~UIMuI~ Us ing 
~ l l y  I ~ n g ~  SmOOth Mul¢ le  Cells 
/~I.M. ~ ,  J.C Mo~ge, D.,~ Lalt~, K Teict~1-Kulmzewska, 
DJ. Stewae, Tenence Doemeay Heart Centre S~ M/chaers Hoep~at, 
Unemej~ o~ Toe~ Toe~ Canada 
Ba~f :  To Olcumve~ the prob~¢rm olm vtto Vam~ectmn and avOad Ihe 
use ol viral vedo~ o~ pmfems, we sought o astatd~ ~ smooth muscte 
ceas (SMC) transleofed ex vevo could be de~t~red wa the system~: venous 
c i ~  into the ptdmo{~a~ bed. acbm~mg lOCal t[ansgene exl~es~ron m 
me lung 
Pttma~ cultures Of p~J~morraty afle w SMC's tram Fislna[ 344 
rats were labeled wilh a guomscent, m e m ~  dye (CMTMR), 
er tmnsfected w~ I~e bem-garacP.s~;e mpo~er gene ueder the comro! 
of 1he CMV md~no~/pmmoter (pCMV-p). Transfeded or labeled SMC's (5 
t o. canr.,anim~ worn de~e~d to syngenec rec~ent rats by ~ect~n 
rote the jugular veto, the a~mals were sacnfce~ at tnf.e~s 15 n-~m.,'tes to 2 
weeks later, a~d the kings excised and examined. 
ResuRs: At 15 minutes post transplantation, injected cells were detected 
mainly m the lumen of small putmona~ a.,tenes and anenoles, often in greups 
of 3 ~r more ceils. After 24 hours, labeled SMC's were found incorporated 
[,~.~ ~ vascular wall o~ pulmonary adenoles, and transgene e~pres~on 
persmtad in situ for 14 days. ~ no e'.~lence ol {mmune response and 
minimal attrition. Using se1~ole geomett'K: assumptions, d was calculated that 
al~roximately 45 ± t 6% of the trans/e~ed cells r~ntroctuced ,nto the venous 
circulation could be identified in the lungs alter 1 hour, 25 ± 9% at 24 hours, 
13 ~: 6% at 1 week and 16 ± 6%at2 weeks (n = 6-8 for each group). 
Conc/us~on: These results suggest that a cell-based strategy of ex-tm~ 
transfedmn may provKte an effectwe r~wiral gene transfer approach for the 
treatment el pu l rnonan/~tar  d~eases. 
~T0"~ Adenoviral  of  Human Renel Gene Irramder V= 
Vasopressln Receptors Improves ContractilRy in 
Cardlomyocytes 
K.-L Laugwffz, M. Ungerer. H.-J. Weig. K Kmnsbein, 1-. Sch~eberg ~ ,
K. Hoffmann, G. Schutlz t, A. Sch0mig. Deutsches Herzzentrurn and 1 
Medizinische Klinik, Technische Unn/e/~Pat Munich. Germarry; rInst~tut f~r 
Pharmakologie, FU Berlin, Germany 
BackgrourK/: In heart failure, increased concentrations of arglnine vast> 
pressin (AVP) contribute fo vasoconstriction and reduced cardiac contractil- 
ity. These effects are mediated via the V1 subtype of vasopressin receptors 
(VIR) which stimulates intracellutar IP3 accumulation. 1"he V2 subtype (V2R) 
is not expressed in the myocardium physiologically. V2R expression is re- 
smcted to the kidney, where V2R activation stimulates adenylyl cyctase. 
Overexpression of V2R in the myocardium might allow to use AVP for an 
inotropic effect via a heterologous, cAMP.coupling receptor. 
Methods: We constructed a replication-deficient edenovirus for the hue 
man V2 vasopressin receptor (AdV2R). Cardiomyoblasts were infected w~,h 
10-1000 ptu AdV2R/cefl. Expression of the recombinant V2R was assessed 
by means of 3H-AVP binding in combination with specific V1/V2 antago- 
nists. We detected a maximum V2R density of 792 • 105 fmoVmg pro*.ein, 
whereas Control virus-infected cells only showed V1R binding. Maximum 
V2R expression was reached 72 h after infection. Confocal immunofluores- 
cence laser microscopy showed V2R expression in >90% of infected cells. 
AVP inducad a 30.told cAMP increase w=th an ECS0 of 25 ~.  Reo~'~r~t.n 
V2R-expmssing, spontaneously beating m~tatat c~rd~Wocytes howed 
a 4~ increase ~n co~lractillty aflor AVP ~imulatmo 8*, an EC~0.valu~ of 
10-100 pM. 
Concfusmns. A functionally active a0~vtrus for the htlmlm V2 mceip 
tor was O:mstru~ed, After mfect~on of ¢ 8 ~ ,  AVP ~! f~ of 
V2R-expfl~lsing ~IlSl I~1 10 an if~ft~s~ o~ ifl|fa(~tl!U~lf cAMP ~t~l ip  
t,:m The heterotogous exprffil0n 0.! lh~ cAMP~mmg r~¢AC~' 
(~)rttractde mstx~n~e to AVP in cardiomyoo/t~, 
~Xlgh  FmClUen W Ultmsouncl Sntmna~ TmnmCHon 
of Pomlne Vuculaf Smooth ~ Cells/n V/~ 
A L41wne, A,F. IBftskefl t , 5,S, Francis, 0,l, TItyl~, O.C, Cfotmm~m 
DC C ~ ,  CM. Newman. Unn~emtyotSheRte~, UK: 
'P~arma~h~-s Ine. Ca, USA 
Ba¢*~nd.- Progress in va~t~klr gene II~r=Loy ha= been ~',nde~d ~ me 
tim.'ted e l i l~  alXltor 10xl¢ity Of,:, f f~t  tra~sfe¢~ll~:l leChnlqu~ qt,~l~loul'~ 
iS safe and has been shown to facd~te tran~ite¢1,,on : ?d~'ObtaLt,. , v~fa We 
I~ve mves~galed the e f t~s  of of h~lh heqmmcy ~m~,-,~__,nd on t ra rm~ 
of porane agree vascular smooth muscle celia (VSMCs) m 
~ :  SuPconfluent VSMCa (pas~ges 2-7) were t ran~ w~ 
naked or l ip (~ (Promega T~,50~vec l  luo/efatm pla=ml0 ONA 
(7.5 l,g/ml) in medium corltainmg 10% (v/v) pomme se~u~; (m~tmuou= wave 
u~t~ at 0 (control). I .  3 or 10 MHz was ~ fo~ 60 s; ~ 
actn~y wa,~ aS~lyed at 48 h and expmsse~l ~ tight un~s per ,g  ~ cell 
protein (LUIpo); (:ell cmmts ~ pe~m~ed at 0. 3. 18 and 48 h. 
Resu/ts: ~ to ultrasound at 1 MHz yras ~ w~ a ? ~  
increase in transfeofiort wd~n l~aked DNA (DNA alone 0.4 ± 0.2 LU/.o; wlffl 1 
MHz tmatmef~t 3.0 ± 20  LU/.g, p = 0.028) a~l a 3 fold enhanceme~ wflh 
l i ~ ~ m d  plasm~d (tiposome/DNA 27.6 ± 6.9 LUIpg; w~lh t MHz 
treatme~ 72.8 ± 17 LUI.9, p = 0.007). Tm-.atme~ with 3 or 10 MHz had 
no eftecl on lransleclion efficmno/. Tra~Mment with t MHz mlultad in a 41% 
reduction in celt ~ r  at 1me 48 h t~nt  comparsd with corltr~ IP < 
0,051. 
~ -  H~ freque~y ultrasound enhances translect,m of VSMCs 
~n v~'ro and may also relam VSMC pmkreralton, both des~ propaff~s m 
the co~e~d of the preve~on of ~ s .  
~ Direct DNA Injection of Vascular Endothelial 
Growth Factor Induces Angiogemmis in 
Non-P.chemtc Myocam'iurn 
A.IM. Campt>ell, J.C Monge, D.A Latler. MA. Kul~,zewskl, D.J. Stewart. 
Terrence Donnel~ Heart Centre, St. Mc.ttaal~ t .~ l ,  Unn~rsfy of 
T~ont¢X Toronta Canada 
Background. Pre~ous work has ~monstrated the ability of vascular en- 
(K~ehal gro~d'n factor (VEGF} to induce ar~oganems after direct mlectma 
of naked ptasm¢l ONA in per i l  muscle. We determined d this tech~clue 
could prmm~ an effectwe method of inwnm myocardial germ transfer. 
Meffmds: Ptasmid DNA for the 165 an~ao-acid isoform of VEGF under the 
control of the cytorne~dowus enhancer/pmmomr (pVEGF~) was inieofed 
rote the left ventncular free wall of Spragu~Dawfey rats aged 10-12 weeks. 
Contrel animals were miectad w~th eith~ plasn'~l DNA ¢x:x3tng for the fe13offer 
gene beta-gatacios~tase (pCMV-~) or wifh buffered saline. Anin~ls 
sacrificed at intervals from 48 hours fe 2 weeks and [he hearts examined. 
Results: S~gnn~cant beta-gal stamin'3 was detected 7 days after in)eCt~. 
with approximately 3-5% of the volume of the left ventrficle xpmssJrlg the 
transgene. The hears in.lacteal with pCMV-p or buffered saline demortslrated 
a 3 fold increase in the numl0er of vessels along ~ course of ff~ needle tract 
and a 6 fold increase in the total vessel area as compared to non-lnteofed 
myocardium. Hearts iniected with pVEGF~e~ demonstrated marked neova~- 
cutanzatton w~th a 4 fold increase in the fetal number of veSSels and a 15 
fold increase in the total vessel area, as ~;o~-r~.,ared to the pCMV-# injected 
hearts. By 14 dayS after miec~on the number and total area ot vessels had 
decreased to non-injected control evels in e q animals, 
ConcluSiOn: These results indicate that direct myocardial in lucti¢,n of pies- 
mid DNA is an effective method of in.wvocjene transfer, and transfection with 
VEGF induCeS signdtcant but transient neovescularization of non-ischemic 
myocardium, greater than that seen w~th needle trauma alone. 


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Direct DNA injection of vascular endothelial growth factor induces angiogenesis in non-ischemic myocardium

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