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Experimental Therapeutics, Preclinical Pharmacology |
Gene Therapy Center, University of Alabama at Birmingham, Birmingham, Alabama 35294 [K. S., V. K., P. N. R., D. T. C., R. A.], and Department of Neuro-Oncology, University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030 [J. F.]
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ABSTRACT |
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 Top ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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INTRODUCTION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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However, realization of the full utility of CRAds in cancer
therapeutics depends on their ability to infect human tumors. Previous
studies on adenovirus-mediated gene delivery to human tumor cells have
pointed out the highly variable expression of primary adenoviral
receptor, CAR, in neoplastic cells (7
, 8)
, and this
variation may curtail the initial infection and lateral propagation of
CRAds (9)
. On the basis of these data, it has been
proposed that gene delivery via CAR-independent pathways is required to
overcome this aspect of tumor biology (10
, 11)
. We have
focused on 
V integrins as enhancers of
adenoviral infection according to a previous report on the correlation
of the levels of V integrins expressed by
tumor cells with the efficiency of adenovirus-mediated gene transfer
(12)
. Furthermore, previous studies demonstrate that
V integrins are aberrantly expressed in
several types of cancer (13
, 14)
and are present in tumor
blood vessels of breast cancer and malignant melanoma
(15)
.
Modifications of capsid proteins responsible for adenovirus
binding to target cells can alter its tissue tropism. These data favor
the incorporation of an Arg-Gly-Asp (RGD) sequence, known to interact
with V integrins, into the adenovirus fiber to
enhance tumor infection. Recently, we developed an approach based on
the genetic incorporation of a sequence encoding an RGD peptide into
the HI loop of the fiber knob. The addition of RGD-integrin
interactions on primary CAR binding confers an expanded tropism to the
fiber-modified adenovirus, and this effect has been demonstrated in
previous studies (16
, 17)
.
In this study, we combined the fiber knob modification strategy with a CRAd based on a partial deletion of the E1A gene, which synthesizes a defective protein unable to bind host cell Rb protein. The selectivity of this mutant adenovirus has been previously demonstrated by Fueyo et al. (18) and recently by another group that uses a virus with the same deletion (19) . Our results demonstrated that the incorporation of the RGD motif into the fiber of a CRAd enhances its oncolytic potency in vitro and in vivo.
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MATERIALS AND METHODS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Virus Construction
Ad5-
24 Mutant.
The replication-competent Ad5-24 adenovirus was provided by
J. F. (The University of Texas M. D. Anderson Cancer Center,
Houston, TX). This virus contains a 24-nucleotide deletion, from Ad5 bp
923 to 946 (both included), corresponding to the amino acid sequence
L122TCHEAGF129 of the E1A
protein known to be necessary for Rb protein binding (23)
.
Details of the tumor-specific replication of this virus are presented
elsewhere (18
, 19)
.
RGD Modification of Ad5luc and Ad5-24.
Ad5lucRGD is an E1-deleted virus containing the recombinant RGD fiber
and expressing the firefly luciferase. This vector was constructed by
homologous recombination of the E1 region containing the
luciferase gene into the plasmid pVK503 that contains the
modified fiber (15)
. A similar procedure was followed to
construct the RGD-modified version of Ad5-24. Briefly, an E1
fragment containing the 24-bp deletion was isolated from the plasmid
pXC1-24, originally used to construct Ad5-24 (18)
,
and cloned by homologous recombination into the
ClaI–digested plasmid pVK503 containing the RGD fiber
(15)
. The genome of the new virus was released from the
plasmid backbone by digestion with PacI, and the resulting
fragment was used to transfect 293 cells to rescue the Ad5-24RGD.
The presence of the RGD motif in Ad5-24RGD and Ad5lucRGD was
confirmed by PCR with the fiber primers FiberUp
(5'-CAAACGCTGTTGGATTTATG-3') and FiberDown
(5'-GTGTAAGAGGATGTGGCAAAT-3'). The 24 deletion was analyzed by
PCR with primers E1a-1 (5'-ATTACCGAAGAAATGGCCGC-3') and E1a-2
(5'-CCATTTAACACGCCATGCA–3') followed by BstXI digestion.
Virus DNA Replication.
A549 cells, cultured in 6-well plates, were infected with Ad5-24 or
Ad5-24RGD at a dose of 0.01 viral particles/cell. The cells were
maintained in DMEM-5% FBS with 1 µCi/ml BrdUrd (Amersham
Pharmacia Biotech Inc., Piscataway, NJ). Attached and detached cells
were harvested at 2, 4, 6, and 8 days after infection, and
encapsidated viral DNA was purified by the spermine-HCl method
(24)
. The DNA was digested with HindIII and
resolved in 1% agarose gel. The BrdUrd incorporated into the DNA
resulting from viral replication was detected by Southwestern blot
using mouse anti-BrdUrd IgG (DAKO, Carpinteria, CA) and
peroxidase-labeled antimouse antibody (Amersham). The membrane was
exposed to Kodak Biomax ML film and developed in an automated
processor.
Adenovirus Yield Assay. A549 cells cultured in 6-well plates
were infected with 0.01 particle/cell Ad5lucRGD, Ad5-24, or
Ad5-24RGD, and maintained in DMEM-5% FBS. After 8 days, cells and
media were harvested, and the titer was determined by plaque assay.
Oncolysis Assay. A549 and LNCaP cells cultured by triplicate in 6-well plates were infected with one of the three types of adenovirus at doses of 0.001 or 0.01 viral particles/cell. Eight (A549) and 10 (LNCaP) days after infection, the cells were fixed and stained with crystal violet solution.
In Vitro Cytotoxicity Assay (XTT).
A549 and LNCaP cells were seeded and infected in parallel with the ones
used for the oncolysis assay described above. Eight and 10 days after
infection, cell survival was determined using XTT (Sigma, St. Louis,
MO). The number of living cells was calculated from noninfected cells
cultured and treated with XTT in the same way as were the experimental
groups.
s.c. Tumor Xenograft Model in Nude Mice.
Female athymic nu/nu mice (Frederick Cancer Research, Frederick,
MD), 8–10 weeks old, were kept under pathogen-free conditions
according to the American Association for Accreditation of Laboratory
Animal Care guidelines. Eight million A549 cells were xenografted under
the skin of each flank in anesthetized mice. When the nodules reached
60–100 mm3
, a single dose of
109 viral particles (high-dose experiment;
n = 5) or 107 viral particles
(low-dose experiment; n = 4) of Ad5lucRGD, Ad5-24,
Ad5-24RGD, or PBS was administered i.t. Tumor size was monitored
twice a week, and fractional volume was calculated from the formula:
(length x width x depth) x 1/2. The mice were
euthanized 35 days after the treatment because of the size of the
tumors in the control group. Animal protocols were reviewed and
approved by the Institutional Animal Care and Use Committee of the
University of Alabama at Birmingham. Statistical differences among
groups were assessed with Student’s t tests.
Adenovirus Hexon Immunodetection.
The presence of adenovirus hexon in the treated tumor xenografts was
assessed by immunofluorescence. A549 tumor sections were treated with
goat antihexon (Chemicon Inc., Temecula, CA) and Alexa Fluor
488-labeled donkey antigoat (Molecular Probes, Eugene, OR) antibodies,
and were counterstained with Hoechst 33342 (Molecular Probes). The
slides were analyzed under a fluorescent microscope (Leitz Orthoplan).
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RESULTS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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24 deletion of E1A and the RGD insertion in the fiber knob were
combined into a unique viral genome by homologous recombination, and
the resulting Ad5-24RGD was propagated efficiently in A549 cells.
The 24-bp deletion in the E1A gene and the RGD-encoding
sequence in the fiber were verified by PCR (Fig. 1)
. Of note, no adenoviruses having wild-type E1 or wild-type fiber
appeared throughout the propagation of Ad5-24RGD, a finding that
confirms the lack of endogenous adenoviral sequences in A549 cells.
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24RGD
and Ad524 was compared. A549 cells were infected with 0.01 viral
particle per cell of each virus and were maintained in
medium with BrdUrd throughout the 8-day incubation period. The
encapsidated viral DNA was purified on days 2, 4, 6, and 8
postinfection, and the samples were analyzed by Southwestern blot as
described in "Materials and Methods." As indicated by the BrdUrd
incorporated into replicating viral DNA, Ad5-24RGD propagation was
more efficient than that of Ad5-24 (Fig. 2)
. The Ad5-24RGD DNA can be detected not only sooner (day 6) compared
with Ad5-24 DNA (day 8) but in greater amounts. Thus, the
infectivity advantage conferred by RGD incorporation into the fiber
knob increased adenovirus propagation in target cells.
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24, or
Ad5-24RGD in A549 cells at 8 days after infection by plaque assay.
Ad5-24RGD produced a viral yield of 3.75 x
109 plaque-forming units/ml which was 43 times
higher than that of its unmodified Ad5-24 counterpart (8.75 x
107 plaque-forming units/ml). No virus was
obtained from the nonreplicative control Ad5lucRGD-infected cells.
These results are consistent with the fact that modifying the fiber
knob with an RGD motif led to enhancement of viral infectivity and an
increase in the production of infectious adenovirus.
Increased Oncolytic Potency of Infectivity-enhanced CRAd in
Vitro.
To demonstrate the increased lytic potency of Ad5-24RGD, we infected
A549 and LNCaP cells with small amounts of each virus to allow multiple
cycles of viral replication over the ensuing 8 days, then stained the
attached cells with crystal violet and counted viable cells by XTT
assay. In both cell lines, the fewest viable cells were detected in the
Ad5-24RGD-infected group (Fig. 3
, A and B). The cell lysis capacity of
Ad5-24RGD is 7 times higher in A549, and 3.5 times higher in LNCaP
compared with Ad5-24. These results demonstrate that the fiber knob
modification enhanced adenoviral lytic potency over that of the
Ad5-24 virus.
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24, P <
0.05; Ad5-24RGD, P < 0.01 compared with PBS group;
Fig. 4
A). Given these results, we conducted another experiment in
which we administered a 100-fold lower dose (107
viral particles) of the viruses. At this dose, Ad5-24 treatment did
not show a statistically significant difference compared with either
PBS or AdlucRGD. However, it demonstrated that the oncolytic effect of
Ad5-24RGD is maintained (Ad5-24RGD versus PBS,
P < 0.01; Ad5-24RGD versus Ad5-24,
P < 0.05). These variations observed between high-dose
and low-dose experiments suggest that a threshold dose over
107 viral particles of Ad5-24 is required to
obtain an oncolytic effect in tumor nodules (Fig. 4
B). To
confirm that the CRAds were present in the tumor tissue, we used
immunofluorescence to detect the virus hexon in tumor samples collected
after the low-dose experiment (35 days postinjection). Ad5-24RGD was
present in the tumor nodules, as was Ad5-24 to a lesser extent. PBS-
and Ad5lucRGD-treated nodules showed no hexon signal (Fig. 4
C). These results corroborated the observation that the
partial reduction of tumor mass was attributable to virus replication
and that the RGD modification of the fiber knob conferred infectivity
and oncolysis advantage to a CRAd in vivo.
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DISCUSSION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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In this report, we have demonstrated that the genetic introduction of
an RGD sequence in the fiber of a CRAd, such as previously
characterized Ad5-24 (18)
, allows CAR-independent
infection that leads to the enhancement of viral propagation and
oncolytic effect in vitro and in vivo. The
increased initial virus entry into the cells rendered by the RGD
modification results in earlier detection and augmented yields of
encapsidated DNA of Ad5-24RGD compared with the unmodified Ad5-24
(Fig. 2)
. Because this tropism modification is not anticipated to alter
fundamental aspects of the viral replication cycle, this effect was
likely attributable to the infectivity enhancement allowed by
delivering the virus through CAR-independent pathways. Subsequently, we
studied the oncolytic potency of CRAds in two cell lines and concluded
that Ad5-24RGD potency is higher than that of the unmodified virus.
Although the XTT assay was not sensitive enough to demonstrate the
lytic effect of Ad5-24 compared with the nonreplicative Ad5lucRGD,
the crystal violet showed early comet-like cytopathic areas in
Ad5-24-treated A549 and LNCaP cells, which indicated the presence of
an incipient lytic effect, whereas Ad5lucRGD-treated cells were intact
(Fig. 3
A). The less notable difference between Ad5-24RGD
and Ad5-24 seen in LNCaP cells is explained by the absence of the
Vß3 integrins
(25)
, compensated by the presence of other types of
RGD-binding integrins
(3ß1 and
5ß1; Ref.
26
) that were rapidly saturated (Fig. 3)
.
Our ultimate goal was to demonstrate the superior oncolytic
effect of Ad5-24RGD in an in vivo model. To this end,
A549 cells xenografted in nude mice were treated with single, high-dose
(109 viral particles), i.t. injections of
Ad5lucRGD, Ad5-24, Ad5-24RGD, or PBS, and the results showed that
both CRAds (modified and unmodified) yielded similar oncolysis (Fig. 4
A). However, when a 100-fold lower dose
(107 viral particles) was administered, it became
clear that the oncolytic effect of Ad5-24RGD was higher than that of
Ad5-24 (P < 0.05; Fig. 4
B). Furthermore,
we were able to correlate the observed oncolytic effect with the
presence of virus progeny in the tumor samples by immunofluorescent
detection of adenoviral hexon. Hexon was not detected in PBS- (not
shown) and Ad5lucRGD-treated nodules (Fig. 4
C,
a), whereas it was detected throughout the tumors treated
with CRAds. The comparison between the two CRAds showed that
fluorescence in Ad5-24RGD-treated tumors was stronger than the one
observed in Ad5-24-treated tumors (Fig. 4
C, b
and c, respectively). The lack of fluorescent staining in
tumors treated with the nonreplicative control Ad5lucRGD indicates that
the detected hexon belongs to the viral progeny of Ad5-24 and
Ad5-24RGD, and not to the initial inoculum. As regards the high
divergence of the volumes of PBS- and Ad5lucRGD-treated tumors, factors
such as highly heterogeneous cell replication rates and hypoxic and
necrotic areas are known to affect individual tumor volume after a
critical size is reached. These differences have been noted previously
when using oncolytic viruses (27
, 28) . Nevertheless, total
resolution of the tumors in the s.c. xenograft model was seen only in
some nodules treated with Ad5-24RGD, which indicated that
administration volume and schema adjustments, such as the ones
suggested recently by Heise et al. (29)
, might
be necessary to achieve complete oncolysis.
As presented here and elsewhere (30)
, the efficacy
of replication-competent viruses used as oncolytic agents can improved
at the level of infectivity. As other tumor-binding peptides are
isolated (30
, 31) , modifications in addition to the RGD
insertion can be considered as well. Of note, the RGD modification
described here does not preclude the binding of the fiber to CAR, and
the modified virus can enter the cells through
V integrins and CAR. One approach to improve
specific tumor infection/transduction would be the combination of CAR
ablation and tumor-specific ligands to redirect the virus tropism.
Recently, the adenovirus fiber amino acids crucial for CAR-binding
abrogation and new tumor-selective peptides have been defined
(15
, 31
, 32)
. This combination will generate truly
targeted viruses, but the efficiency of their propagation will depend
on the amount of the targeted receptor in the same way as the
propagation of the unmodified virus depends on CAR.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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1 This work was supported by NIH Grants RO1
CA68245-01, RO1 CA74242, RO1 HL-50255, T32CA75930, CA-98-008, and NO1
CO-97110; the United States Department of Defense Grants PC 970193 and
PC 991018; and by the Susan B. Komen Foundation. 
2 To whom requests for reprints should be
addressed, at Gene Therapy Center, The University of Alabama at
Birmingham, 1824 Sixth Avenue South, WTI-620, Birmingham, AL
35294-3300. Phone: (205) 934-8629; Fax: (205) 975-7949; E-mail: ramon.alemany@ccc.uab.edu, david.curiel{at}ccc.uab.edu
3 The abbreviations used are: CRAd, conditionally
replicative adenovirus; BrdUrd, bromodeoxyuridine; CAR, Coxsackievirus
and adenovirus receptor; i.t., intratumoral/intratumorally; Rb,
retinoblastoma; FBS, fetal bovine serum; XTT,
2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxyanilide.
Received 8/30/00; revised 10/25/00; accepted 10/31/00.
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REFERENCES |
|---|
|
TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
E1B adenovirus replication in human cells. Oncogene, 18: 5032-5043, 1999.[CrossRef][Medline]
vß5 may predict the susceptibility to adenovirus-mediated gene transfer in human lung cancer cells. Gene Ther., 5: 361-368, 1998.[CrossRef][Medline]
vß3 integrin. Adhesion mechanism for transformed glial cells. J. Clin. Investig., 88: 1924-1932, 1991.
v integrins as receptors for tumor targeting by circulating ligands. Nat. Biotechnol., 15: 542-546, 1997.[CrossRef][Medline]
vß3 integrin is modulated by a focal adhesion kinase pathway. Cancer Res., 59: 1655-1664, 1999.This article has been cited by other articles:
|
|
 |
|
  A. M. Sonabend, I. V. Ulasov, M. A. Tyler, A. A. Rivera, J. M. Mathis, and M. S. Lesniak Mesenchymal Stem Cells Effectively Deliver an Oncolytic Adenovirus to Intracranial Glioma Stem Cells, March 1, 2008; 26(3): 831 - 841. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. M. Alonso, C. Gomez-Manzano, B. N. Bekele, W.K. A. Yung, and J. Fueyo Adenovirus-Based Strategies Overcome Temozolomide Resistance by Silencing the O6-Methylguanine-DNA Methyltransferase Promoter Cancer Res., December 15, 2007; 67(24): 11499 - 11504. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. L.M. Lamfers, S. Idema, L. Bosscher, S. Heukelom, S. Moeniralm, I. H. van der Meulen-Muileman, R. M. Overmeer, P. van der Valk, V. W. van Beusechem, W. R. Gerritsen, et al. Differential Effects of Combined Ad5-{Delta}24RGD and Radiation Therapy in In vitro versus In vivo Models of Malignant Glioma Clin. Cancer Res., December 15, 2007; 13(24): 7451 - 7458. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
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|
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|
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|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
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|
|
|
|
|
|
L. Kangasniemi, T. Kiviluoto, A. Kanerva, M. Raki, T. Ranki, M. Sarkioja, H. Wu, F. Marini, K. Hockerstedt, H. Isoniemi, et al. Infectivity-Enhanced Adenoviruses Deliver Efficacy in Clinical Samples and Orthotopic Models of Disseminated Gastric Cancer. Clin. Cancer Res., May 15, 2006; 12(10): 3137 - 3144. [Abstract] [Full Text] [PDF]
|
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|
|
|
|
|
|
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|
|
|
|
 |
|
 C. Gomez-Manzano, W.K. A. Yung, R. Alemany, and J. Fueyo Genetically modified adenoviruses against gliomas: From bench to bedside Neurology, August 10, 2004; 63(3): 418 - 426. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Davydova, L. P. Le, T. Gavrikova, M. Wang, V. Krasnykh, and M. Yamamoto Infectivity-Enhanced Cyclooxygenase-2-Based Conditionally Replicative Adenoviruses for Esophageal Adenocarcinoma Treatment Cancer Res., June 15, 2004; 64(12): 4319 - 4327. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. M. Witlox, V. W. van Beusechem, B. Molenaar, H. Bras, G. R. Schaap, R. Alemany, D. T. Curiel, H. M. Pinedo, P. I. J. M. Wuisman, and W. R. Gerritsen Conditionally Replicative Adenovirus with Tropism Expanded towards Integrins Inhibits Osteosarcoma Tumor Growth in Vitro and in Vivo Clin. Cancer Res., January 1, 2004; 10(1): 61 - 67. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. E. Goldsmith, M. Kitazono, P. Fok, T. Aikou, S. Bates, and T. Fojo The Histone Deacetylase Inhibitor FK228 Preferentially Enhances Adenovirus Transgene Expression in Malignant Cells Clin. Cancer Res., November 1, 2003; 9(14): 5394 - 5401. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Cascallo, G. Capella, A. Mazo, and R. Alemany Ras-dependent Oncolysis with an Adenovirus VAI Mutant Cancer Res., September 1, 2003; 63(17): 5544 - 5550. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Fueyo, R. Alemany, C. Gomez-Manzano, G. N. Fuller, A. Khan, C. A. Conrad, T.-J. Liu, H. Jiang, M. G. Lemoine, K. Suzuki, et al. Preclinical Characterization of the Antiglioma Activity of a Tropism-Enhanced Adenovirus Targeted to the Retinoblastoma Pathway J Natl Cancer Inst, May 7, 2003; 95(9): 652 - 660. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Kawakami, H. Li, J. T. Lam, V. Krasnykh, D. T. Curiel, and J. L. Blackwell Substitution of the Adenovirus Serotype 5 Knob with a Serotype 3 Knob Enhances Multiple Steps in Virus Replication Cancer Res., March 15, 2003; 63(6): 1262 - 1269. [Abstract] [Full Text] [PDF]
|
|
|
|
|
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K. Suzuki, R. Alemany, M. Yamamoto, and D. T. Curiel The Presence of the Adenovirus E3 Region Improves the Oncolytic Potency of Conditionally Replicative Adenoviruses Clin. Cancer Res., November 1, 2002; 8(11): 3348 - 3359. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. L. M. Lamfers, J. Grill, C. M. F. Dirven, V. W. van Beusechem, B. Geoerger, J. van den Berg, R. Alemany, J. Fueyo, D. T. Curiel, G. Vassal, et al. Potential of the Conditionally Replicative Adenovirus Ad5-{Delta}24RGD in the Treatment of Malignant Gliomas and Its Enhanced Effect with Radiotherapy Cancer Res., October 15, 2002; 62(20): 5736 - 5742. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Bernt, M. Liang, X. Ye, S. Ni, Z.-Y. Li, S. L. Ye, F. Hu, and A. Lieber A New Type of Adenovirus Vector That Utilizes Homologous Recombination To Achieve Tumor-Specific Replication J. Virol., October 2, 2002; 76(21): 10994 - 11002. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. S. Haviv, J. L. Blackwell, A. Kanerva, P. Nagi, V. Krasnykh, I. Dmitriev, M. Wang, S. Naito, X. Lei, A. Hemminki, et al. Adenoviral Gene Therapy for Renal Cancer Requires Retargeting to Alternative Cellular Receptors Cancer Res., August 1, 2002; 62(15): 4273 - 4281. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Tsukuda, R. Wiewrodt, K. Molnar-Kimber, V. P. Jovanovic, and K. M. Amin An E2F-responsive Replication-selective Adenovirus Targeted to the Defective Cell Cycle in Cancer Cells: Potent Antitumoral Efficacy but No Toxicity to Normal Cell Cancer Res., June 1, 2002; 62(12): 3438 - 3447. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. K. Bergsland and A. P. Venook Shedding Old Paradigms: Developing Viruses to Treat Cancer J. Clin. Oncol., May 1, 2002; 20(9): 2220 - 2222. [Full Text] [PDF]
|
|
|
|
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M. N. Barnes, C. J. Coolidge, A. Hemminki, R. D. Alvarez, and D. T. Curiel Conditionally Replicative Adenoviruses for Ovarian Cancer Therapy Mol. Cancer Ther., April 1, 2002; 1(6): 435 - 439. [Abstract] [Full Text] [PDF]
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 C. J. A. Ring Cytolytic viruses as potential anti-cancer agents J. Gen. Virol., March 1, 2002; 83(3): 491 - 502. [Abstract] [Full Text] [PDF]
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G. J. Bauerschmitz, J. T. Lam, A. Kanerva, K. Suzuki, D. M. Nettelbeck, I. Dmitriev, V. Krasnykh, G. V. Mikheeva, M. N. Barnes, R. D. Alvarez, et al. Treatment of Ovarian Cancer with a Tropism Modified Oncolytic Adenovirus Cancer Res., March 1, 2002; 62(5): 1266 - 1270. [Abstract] [Full Text] [PDF]
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J. Kim, T. Smith, N. Idamakanti, K. Mulgrew, M. Kaloss, H. Kylefjord, P. C. Ryan, M. Kaleko, and S. C. Stevenson Targeting Adenoviral Vectors by Using the Extracellular Domain of the Coxsackie-Adenovirus Receptor: Improved Potency via Trimerization J. Virol., February 15, 2002; 76(4): 1892 - 1903. [Abstract] [Full Text] [PDF]
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C. Balague, F. Noya, R. Alemany, L. T. Chow, and D. T. Curiel Human Papillomavirus E6E7-Mediated Adenovirus Cell Killing: Selectivity of Mutant Adenovirus Replication in Organotypic Cultures of Human Keratinocytes J. Virol., August 15, 2001; 75(16): 7602 - 7611. [Abstract] [Full Text] [PDF]
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T. P. Cripe, E. J. Dunphy, A. D. Holub, A. Saini, N. H. Vasi, Y. Y. Mahller, M. H. Collins, J. D. Snyder, V. Krasnykh, D. T. Curiel, et al. Fiber Knob Modifications Overcome Low, Heterogeneous Expression of the Coxsackievirus-Adenovirus Receptor That Limits Adenovirus Gene Transfer and Oncolysis for Human Rhabdomyosarcoma Cells Cancer Res., April 1, 2001; 61(7): 2953 - 2960. [Abstract] [Full Text]
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J. T. Douglas, M. Kim, L. A. Sumerel, D. E. Carey, and D. T. Curiel Efficient Oncolysis by a Replicating Adenovirus (Ad) in Vivo Is Critically Dependent on Tumor Expression of Primary Ad Receptors Cancer Res., February 1, 2001; 61(3): 813 - 817. [Abstract] [Full Text]
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