Aminopeptidase N (APN/CD13) is selectively expressed in vascular endothelial cells and plays multiple roles in angiogenesis

Fukasawa, Kazuteru; Fujii, H.; Saitoh, Yurika; Koizumi, Keiichi; Aozuka, Yasushi; Sekine, Keiko; Yamada, Masatoshi; Saiki, Ikuo; Nishikawa, Kiyohiro

doi:10.1016/j.canlet.2005.11.051

Cited by 124 publications

(99 citation statements)

References 25 publications

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“…Given that bestatin and G10T inhibited tube formation to similar levels, and in the absence of any cumulative or synergistic effects, data support the hypothesis that NGR-containing G10T can bind to APN. In addition, these results are consistent with functional analysis showing that knockdown of APN resulted in a similar inhibition of capillary tube formation using HUVECs (32). Also, present data somewhat shed new light on apparently perplexing results regarding anti-proliferative versus anti-migratory effects of endostatin and/or its hydrolysis products, given that APN participates in cell motility and invasion processes (35).…”

Section: Gene Silencing By Specific Sirnas Of Cathepsins L and S Impasupporting

confidence: 88%

“…This sequence has been used to improve the efficacy of anti-angiogenic and anti-tumorigenic agents through targeting to tumor sites; this effect is facilitated through the ability of the NGR sequence to target and bind APN (CD13) (30,31). APN is a zinc-binding type 2 transmembrane ectopeptidase of 150 kDa (Clan MA, family M1; Merops, the peptidase database) that is expressed selectively in vascular endothelial cells, including HUVECs and aortic endothelial cells (32). Previously, it has been shown that a synthetic NGR-containing peptide derived from endostatin (corresponding to the Ser-1451-Thr-1464 moiety and encompassing the sequence of presently identified peptides) binds to and inhibits APN more strongly than bestatin, which is its pharmacological inhibitor isolated from Streptomyces olivoreticuli and currently used as an anticancer drug (33,34).…”

Section: Gene Silencing By Specific Sirnas Of Cathepsins L and S Impamentioning

confidence: 99%

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Cysteine Cathepsins S and L Modulate Anti-angiogenic Activities of Human Endostatin

Veillard

Saidi

Burden

et al. 2011

Journal of Biological Chemistry

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Background: Cathepsins participate to the release of endostatin, a potent anti-angiogenic protein.Results: Both cathepsins L and S generate two peptides from human endostatin with increased angiostatic properties. Conclusion: Endostatin-derived peptides reduce tube formation of endothelial cells. Significance: Endostatin-derived peptides may represent novel molecular links between cysteine cathepsins and aminopeptidase N in the regulation of angiogenesis.

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Section: Gene Silencing By Specific Sirnas Of Cathepsins L and S Impasupporting

confidence: 88%

Section: Gene Silencing By Specific Sirnas Of Cathepsins L and S Impamentioning

confidence: 99%

Cysteine Cathepsins S and L Modulate Anti-angiogenic Activities of Human Endostatin

Veillard

Saidi

Burden

et al. 2011

Journal of Biological Chemistry

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“…That J1, in addition to its direct antitumoral effects, is able to significantly reduce the number of blood vessels in neuroblastoma, is a novel finding. The metalloproteinase aminopeptidase N is highly expressed in vascular endothelial cells and has been shown to play multiple roles in angiogenesis (25). Preliminary findings in our lab show that aminopeptidase N is a target of J1, 8 which may 50 Amol/kg of J1, melphalan, or no drug on days 0 and 6.…”

Section: Discussionmentioning

confidence: 81%

The novel melphalan prodrug J1 inhibits neuroblastoma growthin vitroandin vivo

Wickström

Johnsen²,

Ponthan³

et al. 2007

Molecular Cancer Therapeutics

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Neuroblastoma is the most common extracranial solid tumor of childhood. The activity of J1 (L-melphalanyl-p-Lfluorophenylalanine ethyl ester), an enzymatically activated melphalan prodrug, was evaluated in neuroblastoma models in vitro and in vivo. Seven neuroblastoma cell lines with various levels of drug resistance were screened for cytotoxicity of J1 alone or in combination with standard cytotoxic drugs, using a fluorometric cytotoxicity assay. J1 displayed high cytotoxic activity in vitro against all neuroblastoma cell lines, with IC 50 values in the submicromolar range, significantly more potent than melphalan. The cytotoxicity of J1, but not melphalan, could be significantly inhibited by the aminopeptidase inhibitor bestatin. J1 induced caspase-3 cleavage and apoptotic morphology, had additive effects in combination with doxorubicin, cyclophosphamide, carboplatin, and vincristine, and synergistically killed otherwise drugresistant cells when combined with etoposide. Athymic rats and mice carrying neuroblastoma xenografts [SH-SY5Y, SK-N-BE(2)] were treated with equimolar doses of melphalan, J1, or no drug, and effects on tumor growth and tissue morphology were analyzed. Tumor growth in vivo was significantly inhibited by J1 compared with untreated controls. Compared with melphalan, J1 more effectively inhibited the growth of mice with SH-SY5Y xenografts, was associated with higher caspase-3 activation, fewer proliferating tumor cells, and significantly decreased mean vascular density. In conclusion, the melphalan prodrug J1 is highly active in models of neuroblastoma in vitro and in vivo, encouraging further clinical development in this patient group.

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“…Shapiro and colleagues later showed that APN is transcriptionally activated by angiogenic signals, mediated by the renin-ang system /MAPK/Ets-2 signaling pathway, and essential for endothelial morphogenesis and capillary tube formation (26)(27)(28). Recently, by using RNA interference, Fukasawa et al (29) confirmed that APN is selectively expressed in vascular endothelial cells, and that the enzyme plays multiple roles in angiogenesis. These results (21,(26)(27)(28)(29) are in agreement with our findings in the APN-null mouse.…”

Section: Discussionmentioning

confidence: 99%

Impaired angiogenesis in aminopeptidase N-null mice

Rangel

Sun

Guzman-Rojas

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

114

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Aminopeptidase N (APN, CD13; EC 3.4.11.2) is a transmembrane metalloprotease with several functions, depending on the cell type and tissue environment. In tumor vasculature, APN is overexpressed in the endothelium and promotes angiogenesis. However, there have been no reports of in vivo inactivation of the APN gene to validate these findings. Here we evaluated, by targeted disruption of the APN gene, whether APN participates in blood vessel formation and function under normal conditions. Surprisingly, APN-null mice developed with no gross or histological abnormalities. Standard neurological, cardiovascular, metabolic, locomotor, and hematological studies revealed no alterations. Nonetheless, in oxygen-induced retinopathy experiments, APN-deficient mice had a marked and dose-dependent deficiency of the expected retinal neovascularization. Moreover, gelfoams embedded with growth factors failed to induce functional blood vessel formation in APNnull mice. These findings establish that APN-null mice develop normally without physiological alterations and can undergo physiological angiogenesis but show a severely impaired angiogenic response under pathological conditions. Finally, in addition to vascular biology research, APN-null mice may be useful reagents in other medical fields such as malignant, cardiovascular, immunological, or infectious diseases.CD13 ͉ knockout mice ͉ retinopathy ͉ vasculogenesis T he aminopeptidases are a large family of proteolytic enzymes that affect protein maturation, degradation, and regulation (1, 2). Aminopeptidase N (APN) is a membrane-bound zincdependent metalloprotease originally identified as a surface marker in myeloid cells (3,4). APN is widely distributed in many cell types, and its role in hydrolyzing unsubstituted N-terminal residues with neutral side chains varies in different locations. In the epithelium of the renal proximal tubule, APN cleaves its only known natural substrate, angiotensin (ang) III, to ang IV; in synaptic membranes, APN metabolizes enkephalins and endorphins; in the heart, it is an integral component of cardiac remodeling postmyocardial infarction (5-10); and in the respiratory system, APN is the cell surface receptor for certain human coronaviruses and potentially for the severe acute respiratory syndrome (SARS) virus (11-13). Additionally, APN functions in signal transduction, cell cycle control, and differentiation (14, 15).We have developed an in vivo system by using ligand peptides displayed on the surface of phage to study organ-and tumorspecific vascular homing; this methodology enables the identification of vascular markers (16,17). We have isolated phage displaying an asparagine-glycine-arginine (NGR)-containing peptide in a tumor-homing selection and have shown that these phage bind selectively to angiogenic blood vessels. When coupled to a cytotoxic drug (18) or fused to a proapoptotic peptide (19) or to tumor necrosis factor (20), NGR-targeted compounds were more effective and less toxic than the respective controls. The cell surface receptor fo...

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Aminopeptidase N (APN/CD13) is selectively expressed in vascular endothelial cells and plays multiple roles in angiogenesis

Cited by 124 publications

References 25 publications

Cysteine Cathepsins S and L Modulate Anti-angiogenic Activities of Human Endostatin

Cysteine Cathepsins S and L Modulate Anti-angiogenic Activities of Human Endostatin

The novel melphalan prodrug J1 inhibits neuroblastoma growthin vitroandin vivo

Impaired angiogenesis in aminopeptidase N-null mice

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