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Tumor Biology |
The Burnham Institute, La Jolla, California 92037; University of Helsinki, Division of Biochemistry, Helsinki, Finland [E. K.]; Institute for Clinical Pathology, Department of Ultrastructural Pathology and Cell Biology, University of Vienna/AKH Wien, A-1090 Vienna, Austria [R. K.]; Institute for Medical Microbiology and Immunology, Panum Institute 18.3.22, DK-2200 Copenhagen, Denmark [A. S.]; Department of Experimental Oncology, St. Jude Children’s Research Hospital, Memphis, Tennessee 38105-2794 [R. A. A.]; and Department of Pathology and Laboratory Medicine, St. Jude Children’s Research Hospital, Memphis, Tennessee 38105-2794 [L. H. S.]
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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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vß3 and vß5 integrins (4
, 5)
, certain receptors for vascular growth factors (6
, 7)
, matrix metalloproteases (8
, 9)
, and other types
of molecules, such as a high-molecular-weight proteoglycan
(10, 11, 12)
. The growth factor receptors and integrins are
not only markers for angiogenesis, they also play an important
functional role in this process (13
, 14)
.
We have previously probed tumor vasculature by screening random
peptide libraries displayed on phage for the ability of the phage to
home to tumors in vivo. Selection of phage from the
libraries in tumor-bearing mice yielded three peptide motifs capable of
homing to tumor vasculature: an RGD (arginine-glycine-aspartic acid)
motif embedded in a double-cyclic peptide (termed RGD-4C), an NGR
(asparagine-glycine-arginine) motif, and a GSL (glycine-serine-leucine)
motif. Coupling an anticancer drug or a proapoptotic peptide to the
RGD-4C or CNGRC peptides yielded compounds with increased efficacies
against tumors and lowered toxicity to normal tissues in mice
(15
, 16) . Of the 
25 known integrins, many recognize the
RGD motif as a central feature of their binding site in their
extracellular matrix ligands (17)
. The binding specificity
of an RGD peptide for the individual integrins depends on the sequence
surrounding the RGD motif and on the conformation of the peptide
(17)
. The RGD-4C peptide binds selectively to the vß3
and vß5 integrins (18)
, which are specifically
expressed in angiogenesis (3
, 4)
and serve as receptors
for RGD-4C (15
, 19)
. A cyclic NGR peptide, CNGRC, homes
into tumors more effectively than linear peptides containing the NGR
motif (15)
.
The NGR motif resembles RGD, and NGR peptides can bind to integrins,
but the affinity of NGR peptides to integrins is lower than the
affinity of RGD-integrin binding (18
, 20)
. Despite the
integrin binding by the NGR peptides, the RGD-4C peptide does not
effectively compete with the tumor homing of a phage displaying the NGR
motif (15)
, indicating that the receptor for NGR is
different from the receptors for the RGD-4C peptide, the v
integrins.
Here we show that peptides containing the NGR motif bind to APN5 (also known as CD13; Ref. 21 ). APN is a membrane-spanning, Mr 140,000 cell surface protein that is expressed in various epithelial cells and in macrophages (21, 22, 23) . APN is thought to play a role in chemokine processing and tumor invasion (24, 25, 26) . We also show that the only vascular structures with detectable APN are tumor blood vessels and other types of vessels undergoing angiogenesis. In addition, APN antagonists are antiangiogenic in vivo. These findings indicate that APN plays a functional role in angiogenesis.
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MATERIALS AND METHODS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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were from R&D Systems. MDA-MB-435
human breast carcinoma cells and Molt-4 human T cell leukemia cells
were from American Type Culture Collection. KS1767 human Kaposi’s
sarcoma cells were obtained from Dr. J. A. Levy (University of
California, San Francisco, CA).
Transfection of APN cDNA and Cell Surface Expression of APN.
MDA-MB-435 cells were transfected by the calcium phosphate method with
20 µg of full-length APN or CD20 cDNA in the retroviral vector
pZIP(SV)X-1 (a gift from Dr. Richard Mulligan, Children’s Hospital,
Boston, MA) or the vector alone, as described (29)
. Molt-4
cells were electroporated with the APN cDNA inserted in the RcRSV
mammalian expression vector (Invitrogen). Cells expressing high levels
of APN on the cell surface were selected by culturing in
G418-containing medium, followed by three sequential rounds of
fluorescence-activated cell sorter sorting with the WM15 anti-APN
antibody.
Isolation of APN and Assay of Enzymatic Activity.
APN was extracted with 50 mM octylglucoside from KS1767
Kaposi’s sarcoma xenografts grown in nude mice (15)
. APN
activity was measured with Ala-pNA and Leu-pNA substrates
(29)
in the presence or absence of the metalloenzyme
inhibitor o-phenatroline (Sigma).
Phage Binding Assays.
Phage that expressed peptides fused to the p111 surface protein
(30)
were used in binding assays as described (18
, 31)
. In these studies, the NGR phage were CNGRCVSGCAGRC and
CVLNGRMEC, referred to as NGR-phage, (15)
. To test phage
binding to APN, APN was immunocaptured to microtiter wells coated with
10 µg/ml of purified anti-APN antibody WM15 or with BSA. APN was
bound to the antibody from an octylglucoside extract of KS1767
Kaposi’s sarcoma tumors grown in nude mice. These KS1767 cells express
APN on their surface (result not shown). The extract was incubated in
antibody-coated and control wells for 1 h, the wells were washed,
and 2 x 109 TUs of phage was
added. After a 2-h incubation at room temperature, the wells were
washed, and bound phage were quantitated by plating with bacteria
(31)
.
Cytotoxicity Assay in Vitro.
MDA-MB-435 cells and their APN-transfectants were exposed to dox
or a dox-CNGRC peptide conjugate (15)
in 96-well
microtiter plates at 10 µg/well of the drug. After 20 min of drug
exposure, unbound drug was removed through extensive washing with DMEM,
and fresh medium was added. Surviving cells were quantitated at 24 h as described (31)
.
In Vivo Tumor Studies.
MDA-MB-435 mammary fat pad tumors were grown to a diameter of 0.5–1
cm3 (15)
. The homing of i.v.
injected phage to tumors was assessed by coinjecting 250 µg of
affinity-purified R3-63 antibody or normal rat IgG with the phage. As
an internal control, phage with no insert and an ampicillin marker were
used (32)
. In tumor growth studies, mice with size-matched
MDA-MB-435 tumors were treated i.p. with APN inhibitors in 200 µl of
DMEM or DMEM alone, and tumor volume was monitored. Tissues were
processed for immunohistochemistry as described (15)
. The
animal procedures were carried out under Avertin (0.017 ml/g)
anesthesia. All animal experimentation was reviewed and approved by the
Animal Research Committee of The Burnham Institute.
Angiogenesis Assays.
In the retinal neovascularization model (33)
, 1-week-old
mice were exposed to 75% oxygen in air for 5 days. After 5 more days
in room air, the proliferative neovascular response in the retina was
quantified in H&E-stained sections by counting the nuclei of neovessels
extending from the retina into the vitreous in 6-µm cross-sections.
CAM assays were performed as described (34)
.
Immunohistology.
Mouse tissues were processed as described (15)
. Human
tissue samples were fixed in 4% paraformaldehyde, and sections were
stained with various antibodies by using immunoperoxidase detection
with the Vectastain ABC Elite kit (Vector Laboratories) and the
Metal-enhanced DAB substrate (Pierce). Alternatively, fluorescently
labeled antimouse IgG (Sigma) was used for detecting the primary
antibody.
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RESULTS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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We tested the binding of NGR phage to APN immunocaptured onto
microtiter wells. Two different NGR phage bound specifically to
APN-containing wells, whereas the tumor-homing RGD-4C phage and another
RGD phage showed no binding (Fig. 1A)
. The antibody used to capture APN did not bind the NGR
phage without bound APN (not shown). The specificity of the binding was
further examined by inhibiting the APN binding of one of the NGR phage
with soluble peptides. Soluble CNGRC peptide blocked the binding of NGR
phage to APN. Two other cyclic peptides had no effect (Fig. 1B)
.
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and data not shown). The binding of NGR phage to cells
expressing APN was blocked by the CNGRC peptide in a dose-dependent
manner but was not blocked by a control peptide of a similar general
structure (CARAC).
Homing of NGR-Phage.
The in vivo homing of the CNGRC phage to tumors was blocked
by coinjection of a rat antimouse APN antibody (R3-63; Fig. 1D
). This antibody is capable of inhibiting the enzymatic
activity of APN (27)
. Tumor homing of RGD-4C phage was not
affected by R3-63, and normal rat IgG had no effect on the homing of
either phage.
Cytotoxicity of NGR Peptide Conjugate.
A conjugate of the CNGRC peptide and the cytotoxic drug dox was
selectively toxic to APN-expressing cells. Under conditions where the
exposure of APN-transfected and control MDA-MB-435 cells to the
conjugate was limited to 20 min by washing away the conjugate, the
conjugate killed a significant fraction of the APN-expressing cells.
APN-negative cells were essentially unaffected (Fig. 2)
. Free dox and dox coupled to the CARAC control peptide showed no
significant toxicity upon this short incubation. The doxorubicin-RGD-4C
conjugate was toxic to cells, regardless of their APN expression. This
outcome is in agreement with the expression of the vß3 integrin by
the MDA-MB-435 cells (15)
. The selectivity of the CNGRC
conjugate was lost when the incubation with the drug was prolonged; all
dox compounds were highly toxic to both types of cells. These results
provide further evidence for the binding of NGR to APN at cell
surfaces.
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. An antibody specific for human APN did not stain these
tumors (Fig. 3B)
, which agrees with the lack of APN
expression by these cells in vitro, as shown. The blood
vessels in all normal mouse tissues examined were negative for APN
(Fig. 3, C and D
show liver and spleen,
respectively).
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shows vascular APN staining in a human breast carcinoma.
The vasculature in human malignant gliomas and lymph node metastases
from multiple tumor types was also positive for APN (data not shown).
The blood vessels in various normal human tissues were essentially
negative for APN. Faint staining was sometimes seen in the endothelial
cells of arteries but not in capillaries; Fig. 4B
shows such
staining for normal breast tissue. Blood vessels in corpus luteum
expressed APN (Fig. 4C)
. As the blood vessels in this tissue
undergo angiogenesis (35)
, this finding suggests that the
expression of APN in blood vessels is related to angiogenesis.
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. The subendothelial APN staining colocalized with
staining for APA, a marker of pericytes and smooth muscle cells
(28)
in blood vessels (not shown). In the described tissue localization studies, two different antibodies, one against human APN (WM15) and the other against mouse APN (R3-63), were used. They both showed similar localization of APN in human and mouse tissues. Overlay of tissue sections with the CNGRC phage (detected by an anti-M13 antibody) revealed a similar staining pattern as anti-APN. Moreover, preincubating human tumor tissue sections with NGR phage blocked staining with the WM15 anti-APN antibody, whereas staining with the RC38 anti-APA was not affected. Conversely, preincubation of tumor tissue sections with the WM15 anti-APN antibody blocked the binding of the NGR phage to tissue sections (results not shown).
Functional Role of APN in Angiogenesis.
To test the functional role of the APN enzymatic activity in
angiogenesis, we studied the effects of APN inhibitors in angiogenesis
models. Two antibodies capable of inhibiting the enzymatic activity of
APN, antimouse APN R3-63 and 2M-7, both inhibited hypoxia-induced
retinal neovascularization (33)
upon systemic treatment of
mice (Fig. 5A)
. A chemical inhibitor, bestatin, had a similar effect.
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. Leupeptin, which does not inhibit APN activity, had no
effect.
Systemic treatment of mice with two rat antimouse APN antibodies
inhibited the growth of breast carcinoma xenografts derived from
MDA-MB-435 cells, whereas normal rat IgG had no effect (Fig. 5C)
. Because these cells do not express APN in
vitro or in vivo (Fig. 3B)
, the anti-APN
effects are attributable to APN expressed in tumor blood vessels.
Treatment with bestatin also inhibited tumor growth (Fig. 5C)
.
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DISCUSSION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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The expression pattern of APN agrees with its proposed role as the
receptor for the NGR peptides in tumor vasculature; APN is specifically
expressed in endothelial and subendothelial cells in angiogenesis.
Various types of tumors in two species, analyzed with two monoclonal
anti-APN antibodies and with an NGR phage overlay, consistently
revealed APN expression in tumor vasculature. The vascular APN
expression was independent of whether the tumor cells expressed APN. We
also found strong APN expression in the blood vessels of corpus luteum
and have shown in other work that retinal neovasculature expresses
APN.6
In each case, tumors, corpus luteum, and retinal neovascularization,
the vasculature is undergoing angiogenesis (1, 2, 3
, 33
, 35)
.
Thus, APN expression correlates with angiogenesis. The expression of
APN in angiogenesis may depend on growth factors and cytokines, because
tumor necrosis factor- and bFGF up-regulate APN in cultured
endothelial cells
(36)
.7
We did not find APN expression in the vasculature of any normal tissues, including the blood vessels in the brain. However, others have found APN in the pericytes associated with the blood-brain barrier (37) . Although we are uncertain as to the cause, this discrepancy may relate to expression levels, which appear to be far higher in angiogenesis than in resting blood vessels. Importantly, both studies found the endothelial cells in the brain to lack APN.
Despite the presence of APN in various epithelial cells (22) , NGR phage injected i.v. home specifically to tumors and other sites of angiogenesis, presumably because phage are not able to traverse blood vessels to the epithelia. Low-molecular-weight drugs have been targeted previously to tumors as NGR peptide conjugates without apparent epithelial toxicity (15 , 16) . It may be that tumor vessels take up much of the conjugate before it diffuses into tissues. In addition, the homing peptide moiety may be proteolytically destroyed after the conjugate leaves the circulation, preventing specific uptake of the drug by APN-positive cells.
Our data show that APN is not only a previously unrecognized marker of angiogenic endothelial cells but is also functionally important in angiogenesis. We find that APN inhibitors, such as inhibitory antibodies, bestatin and actinonin, suppressed angiogenesis. In accord with earlier studies (25 , 26) the APN inhibitors also suppressed tumor growth. Our results suggest that inhibition of angiogenesis is likely to be a factor in the antitumor activity of these compounds.
The role of APN in angiogenesis may be to facilitate endothelial cell invasion of tissue, which is an essential component of angiogenesis (38 , 39) . Experiments with tumor cells have shown that APN expression can increase invasiveness (26) . Because compounds that are enzymatic inhibitors of APN inhibit angiogenesis, this putative invasion-promoting effect would appear to be related to the enzymatic activity of APN. Another possibility is that APN could modulate the activity of a growth factor or cytokine. Such activities are common among aminopeptidases, including APN (24 , 36 , 40 , 41) .
NGR-containing ligands show promise for targeting phage, drugs, and peptides to tumor vasculature (15 , 16) . The identification of APN as the vascular receptor responsible for the homing of NGR peptides to tumors will open new possibilities for refining such targeting.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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1 Supported by Grants DAMD-17-98-1-8041(to R. P.)
and DAMD-17-99-1-8164 (to W. A.) from the Department of Defense and
Grant CA74238 (to E. R.) and Cancer Center Support Grant CA30199 from
the National Cancer Institute. 
2 Present address: Box 13, M. D. Anderson Cancer
Center, 1515 Holcombe Boulevard, Houston, TX 77030.
3 Present address: Pathology Division, National
Cancer Center Research Institute, Tsukiji 5-1-1, Chuo-ku, Tokyo 104,
Japan.
4 To whom requests for reprints should be
addressed, at M. D. Anderson Cancer Center, Box 13, 1515 Holcombe
Boulevard, Houston, TX 77030. E-mail: rpasqual{at}notes.mdacc.tcm.edu
5 The abbreviations used are: APN, aminopeptidase
N; APA, aminopeptidase A; bFGF, basic fibroblast growth factor; TU,
transforming unit; dox, doxorubicin; CAM, chorioallantoic membrane.
6 W. Arap, M. Hagedorn, R. Pasqualini, and E.
Ruoslahti, unpublished results.
7 R. Pasqualini, unpublished results.
Received 10/ 5/99. Accepted 11/19/99.
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REFERENCES |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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v integrins. Science (Washington DC), 270: 1500-1502, 1995.vß3 for angiogenesis. Science (Washington DC), 264: 569-571, 1994.v integrins as receptors for tumor targeting by circulating ligands. Nat. Biotechnol., 15: 542-546, 1997.[Medline]
vß3 selected from random phage display libraries. Biochemistry, 34: 3948-3955, 1995.[Medline]
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P. Valadon, J. D. Garnett, J. E. Testa, M. Bauerle, P. Oh, and J. E. Schnitzer Screening phage display libraries for organ-specific vascular immunotargeting in vivo PNAS, January 10, 2006; 103(2): 407 - 412. [Abstract] [Full Text] [PDF]
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M. G. Kolonin, L. Bover, J. Sun, A. J. Zurita, K.-A. Do, J. Lahdenranta, M. Cardo-Vila, R. J. Giordano, D. E. Jaalouk, M. G. Ozawa, et al. Ligand-Directed Surface Profiling of Human Cancer Cells with Combinatorial Peptide Libraries Cancer Res., January 1, 2006; 66(1): 34 - 40. [Abstract] [Full Text] [PDF]
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A. Sacchi, A. Gasparri, C. Gallo-Stampino, S. Toma, F. Curnis, and A. Corti Synergistic Antitumor Activity of Cisplatin, Paclitaxel, and Gemcitabine with Tumor Vasculature-Targeted Tumor Necrosis Factor-{alpha} Clin. Cancer Res., January 1, 2006; 12(1): 175 - 182. [Abstract] [Full Text] [PDF]
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H. Murakami, A. Yokoyama, K. Kondo, S. Nakanishi, N. Kohno, and M. Miyake Circulating Aminopeptidase N/CD13 Is an Independent Prognostic Factor in Patients with Non-Small Cell Lung Cancer Clin. Cancer Res., December 15, 2005; 11(24): 8674 - 8679. [Abstract] [Full Text] [PDF]
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 M. Tan and X. Jiang The P Domain of Norovirus Capsid Protein Forms a Subviral Particle That Binds to Histo-Blood Group Antigen Receptors J. Virol., November 15, 2005; 79(22): 14017 - 14030. [Abstract] [Full Text] [PDF]
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 M. S. Lionakis, J. Lahdenranta, J. Sun, W. Liu, R. E. Lewis, N. D. Albert, R. Pasqualini, W. Arap, and D. P. Kontoyiannis Development of a Ligand-Directed Approach To Study the Pathogenesis of Invasive Aspergillosis Infect. Immun., November 1, 2005; 73(11): 7747 - 7758. [Abstract] [Full Text] [PDF]
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V. Askoxylakis, S. Zitzmann, W. Mier, K. Graham, S. Kramer, F. von Wegner, R. H.A. Fink, M. Schwab, M. Eisenhut, and U. Haberkorn Preclinical Evaluation of the Breast Cancer Cell-Binding Peptide, p160 Clin. Cancer Res., September 15, 2005; 11(18): 6705 - 6712. [Abstract] [Full Text] [PDF]
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 L. Zhang, J. A. Hoffman, and E. Ruoslahti Molecular Profiling of Heart Endothelial Cells Circulation, September 13, 2005; 112(11): 1601 - 1611. [Abstract] [Full Text] [PDF]
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N. Ferrari, U. Pfeffer, R. Dell'Eva, C. Ambrosini, D. M. Noonan, and A. Albini The Transforming Growth Factor-{beta} Family Members Bone Morphogenetic Protein-2 and Macrophage Inhibitory Cytokine-1 as Mediators of the Antiangiogenic Activity of N-(4-Hydroxyphenyl)Retinamide Clin. Cancer Res., June 15, 2005; 11(12): 4610 - 4619. [Abstract] [Full Text] [PDF]
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O. H. Aina, J. Marik, R. Liu, D. H. Lau, and K. S. Lam Identification of novel targeting peptides for human ovarian cancer cells using "one-bead one-compound" combinatorial libraries Mol. Cancer Ther., May 1, 2005; 4(5): 806 - 813. [Abstract] [Full Text] [PDF]
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A. Diaz-Perales, V. Quesada, L. M. Sanchez, A. P. Ugalde, M. F. Suarez, A. Fueyo, and C. Lopez-Otin Identification of Human Aminopeptidase O, a Novel Metalloprotease with Structural Similarity to Aminopeptidase B and Leukotriene A4 Hydrolase J. Biol. Chem., April 8, 2005; 280(14): 14310 - 14317. [Abstract] [Full Text] [PDF]
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F. Curnis, A. Gasparri, A. Sacchi, A. Cattaneo, F. Magni, and A. Corti Targeted Delivery of IFN{gamma} to Tumor Vessels Uncouples Antitumor from Counterregulatory Mechanisms Cancer Res., April 1, 2005; 65(7): 2906 - 2913. [Abstract] [Full Text] [PDF]
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 V. J. Yao, M. G. Ozawa, M. Trepel, W. Arap, D. M. McDonald, and R. Pasqualini Targeting Pancreatic Islets with Phage Display Assisted by Laser Pressure Catapult Microdissection Am. J. Pathol., February 1, 2005; 166(2): 625 - 636. [Abstract] [Full Text] [PDF]
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G. E.R. Weller, M. K.K. Wong, R. A. Modzelewski, E. Lu, A. L. Klibanov, W. R. Wagner, and F. S. Villanueva Ultrasonic Imaging of Tumor Angiogenesis Using Contrast Microbubbles Targeted via the Tumor-Binding Peptide Arginine-Arginine-Leucine Cancer Res., January 15, 2005; 65(2): 533 - 539. [Abstract] [Full Text] [PDF]
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Y. van Hensbergen, H. J. Broxterman, S. Rana, P. J. van Diest, M. C. A. Duyndam, K. Hoekman, H. M. Pinedo, and E. Boven Reduced Growth, Increased Vascular Area, and Reduced Response to Cisplatin in CD13-Overexpressing Human Ovarian Cancer Xenografts Clin. Cancer Res., February 1, 2004; 10(3): 1180 - 1191. [Abstract] [Full Text] [PDF]
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F. Curnis, A. Gasparri, A. Sacchi, R. Longhi, and A. Corti Coupling Tumor Necrosis Factor-{alpha} with {alpha}V Integrin Ligands Improves Its Antineoplastic Activity Cancer Res., January 15, 2004; 64(2): 565 - 571. [Abstract] [Full Text] [PDF]
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N. Petrovic, S. V. Bhagwat, W. J. Ratzan, M. C. Ostrowski, and L. H. Shapiro CD13/APN Transcription Is Induced by RAS/MAPK-mediated Phosphorylation of Ets-2 in Activated Endothelial Cells J. Biol. Chem., December 5, 2003; 278(49): 49358 - 49368. [Abstract] [Full Text] [PDF]
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A. Kehlen, U. Lendeckel, H. Dralle, J. Langner, and C. Hoang-Vu Biological Significance of Aminopeptidase N/CD13 in Thyroid Carcinomas Cancer Res., December 1, 2003; 63(23): 8500 - 8506. [Abstract] [Full Text] [PDF]
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 S. Christian, J. Pilch, M. E. Akerman, K. Porkka, P. Laakkonen, and E. Ruoslahti Nucleolin expressed at the cell surface is a marker of endothelial cells in angiogenic blood vessels J. Cell Biol., November 24, 2003; 163(4): 871 - 878. [Abstract] [Full Text] [PDF]
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F. Pastorino, C. Brignole, D. Marimpietri, M. Cilli, C. Gambini, D. Ribatti, R. Longhi, T. M. Allen, A. Corti, and M. Ponzoni Vascular Damage and Anti-angiogenic Effects of Tumor Vessel-Targeted Liposomal Chemotherapy Cancer Res., November 1, 2003; 63(21): 7400 - 7409. [Abstract] [Full Text] [PDF]
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 Y. Ichinose, K. Genka, T. Koike, H. Kato, Y. Watanabe, T. Mori, S. Iioka, A. Sakuma, and M. Ohta Randomized Double-Blind Placebo-Controlled Trial of Bestatin in Patients With Resected Stage I Squamous-Cell Lung Carcinoma J Natl Cancer Inst, April 16, 2003; 95(8): 605 - 610. [Abstract] [Full Text] [PDF]
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N. Ikeda, Y. Nakajima, T. Tokuhara, N. Hattori, M. Sho, H. Kanehiro, and M. Miyake Clinical Significance of Aminopeptidase N/CD13 Expression in Human Pancreatic Carcinoma Clin. Cancer Res., April 1, 2003; 9(4): 1503 - 1508. [Abstract] [Full Text] [PDF]
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S. V. Bhagwat, N. Petrovic, Y. Okamoto, and L. H. Shapiro The angiogenic regulator CD13/APN is a transcriptional target of Ras signaling pathways in endothelial morphogenesis Blood, March 1, 2003; 101(5): 1818 - 1826. [Abstract] [Full Text] [PDF]
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Y. van Hensbergen, H. J. Broxterman, R. Hanemaaijer, A. S. Jorna, N. A. van Lent, H. M. W. Verheul, H. M. Pinedo, and K. Hoekman Soluble Aminopeptidase N/CD13 in Malignant and Nonmalignant Effusions and Intratumoral Fluid Clin. Cancer Res., December 1, 2002; 8(12): 3747 - 3754. [Abstract] [Full Text] [PDF]
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M. G. Kolonin, R. Pasqualini, and W. Arap Teratogenicity induced by targeting a placental immunoglobulin transporter PNAS, October 1, 2002; 99(20): 13055 - 13060. [Abstract] [Full Text] [PDF]
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S. Zitzmann, V. Ehemann, and M. Schwab Arginine-Glycine-Aspartic Acid (RGD)-Peptide Binds to Both Tumor and Tumor-Endothelial Cells in Vivo Cancer Res., September 15, 2002; 62(18): 5139 - 5143. [Abstract] [Full Text] [PDF]
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 F. A. Scappaticci Mechanisms and Future Directions for Angiogenesis-Based Cancer Therapies J. Clin. Oncol., September 15, 2002; 20(18): 3906 - 3927. [Abstract] [Full Text] [PDF]
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J. Renkonen, O. Tynninen, P. Hayry, T. Paavonen, and R. Renkonen Glycosylation Might Provide Endothelial Zip Codes for Organ-Specific Leukocyte Traffic into Inflammatory Sites Am. J. Pathol., August 1, 2002; 161(2): 543 - 550. [Abstract] [Full Text] [PDF]
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K. Porkka, P. Laakkonen, J. A. Hoffman, M. Bernasconi, and E. Ruoslahti A fragment of the HMGN2 protein homes to the nuclei of tumor cells and tumor endothelial cells in vivo PNAS, May 28, 2002; 99(11): 7444 - 7449. [Abstract] [Full Text] [PDF]
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H. Miyashita, T. Yamazaki, T. Akada, O. Niizeki, M. Ogawa, S.-i. Nishikawa, and Y. Sato A mouse orthologue of puromycin-insensitive leucyl-specific aminopeptidase is expressed in endothelial cells and plays an important role in angiogenesis Blood, May 1, 2002; 99(9): 3241 - 3249. [Abstract] [Full Text] [PDF]
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S. Morikawa, P. Baluk, T. Kaidoh, A. Haskell, R. K. Jain, and D. M. McDonald Abnormalities in Pericytes on Blood Vessels and Endothelial Sprouts in Tumors Am. J. Pathol., March 1, 2002; 160(3): 985 - 1000. [Abstract] [Full Text] [PDF]
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B. Carnemolla, L. Borsi, E. Balza, P. Castellani, R. Meazza, A. Berndt, S. Ferrini, H. Kosmehl, D. Neri, and L. Zardi Enhancement of the antitumor properties of interleukin-2 by its targeted delivery to the tumor blood vessel extracellular matrix Blood, March 1, 2002; 99(5): 1659 - 1665. [Abstract] [Full Text] [PDF]
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M. Essler and E. Ruoslahti Molecular specialization of breast vasculature: A breast-homing phage-displayed peptide binds to aminopeptidase P in breast vasculature PNAS, February 19, 2002; 99(4): 2252 - 2257. [Abstract] [Full Text] [PDF]
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F. Curnis, G. Arrigoni, A. Sacchi, L. Fischetti, W. Arap, R. Pasqualini, and A. Corti Differential Binding of Drugs Containing the NGR Motif to CD13 Isoforms in Tumor Vessels, Epithelia, and Myeloid Cells Cancer Res., February 1, 2002; 62(3): 867 - 874. [Abstract] [Full Text] [PDF]
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 R. PASQUALINI and W. ARAP Profiling the Molecular Diversity of Blood Vessels Cold Spring Harb Symp Quant Biol, January 1, 2002; 67(0): 223 - 226. [Abstract] [PDF]
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S. V. Bhagwat, J. Lahdenranta, R. Giordano, W. Arap, R. Pasqualini, and L. H. Shapiro CD13/APN is activated by angiogenic signals and is essential for capillary tube formation Blood, February 1, 2001; 97(3): 652 - 659. [Abstract] [Full Text] [PDF]
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F. D. Hong and G. L. Clayman Isolation of a Peptide for Targeted Drug Delivery into Human Head and Neck Solid Tumors Cancer Res., December 1, 2000; 60(23): 6551 - 6556. [Abstract] [Full Text]
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