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Proteolytic exposure of a cryptic site within collagen type IV is required for angiogenesis and tumor growth in vivo.

Xu J, Rodriguez D, Petitclerc E, Kim JJ, Hangai M, Moon YS, Davis GE, Brooks PC, Yuen SM - J. Cell Biol. (2001)

Bottom Line: Exposure of this cryptic site was associated with angiogenic, but not quiescent, blood vessels and was required for angiogenesis in vivo.A monoclonal antibody (HUIV26) directed to this site disrupts integrin-dependent endothelial cell interactions and potently inhibits angiogenesis and tumor growth.Together, these studies suggest a novel mechanism by which proteolysis contributes to angiogenesis by exposing hidden regulatory elements within matrix-immobilized collagen type IV.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiation Oncology, Kaplan Cancer Center, New York University School of Medicine, New York, NY 10016, USA.

ABSTRACT
Evidence is provided that proteolytic cleavage of collagen type IV results in the exposure of a functionally important cryptic site hidden within its triple helical structure. Exposure of this cryptic site was associated with angiogenic, but not quiescent, blood vessels and was required for angiogenesis in vivo. Exposure of the HUIV26 epitope was associated with a loss of alpha1beta1 integrin binding and the gain of alphavbeta3 binding. A monoclonal antibody (HUIV26) directed to this site disrupts integrin-dependent endothelial cell interactions and potently inhibits angiogenesis and tumor growth. Together, these studies suggest a novel mechanism by which proteolysis contributes to angiogenesis by exposing hidden regulatory elements within matrix-immobilized collagen type IV.

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Effects of systemic administration of purified Mab HUIV26 on tumor growth in vivo. The effects of Mab HUIV26 on tumor growth was assessed in two independent models, including the chick embryo (A and B) and the SCID Mouse (C). HT1080 human fibrosarcoma cells (4 × 105) or CS-1 melanoma tumor cells (5 × 106) were inoculated on the CAMs of 10-d-old chick embryos. 24 h later, the embryos received a single intravenous injection of 100 μg of Mab HUIV26 or isotype-matched control. (A) Quantitation of HT1080 tumor growth in the chick embryo. (B) Quantitation of CS-1 tumor growth within the chick embryo. Data bars represent the mean tumor weights ± the standard errors from 5 to 10 embryos per condition. (C) SCID mice were injected subcutaneously with 2 × 106 M21 human melanoma cells. 3 d later mice were treated i.p. daily for 24 d with 100 μg of either Mab HUIV26 or an isotype-matched control antibody. Tumor size was monitored with calipers and tumor volumes were determined. Data represents the mean ± standard errors of the tumor volumes. All experiments were conducted 3 to 4 times with 5 to 10 animals per condition.
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fig5: Effects of systemic administration of purified Mab HUIV26 on tumor growth in vivo. The effects of Mab HUIV26 on tumor growth was assessed in two independent models, including the chick embryo (A and B) and the SCID Mouse (C). HT1080 human fibrosarcoma cells (4 × 105) or CS-1 melanoma tumor cells (5 × 106) were inoculated on the CAMs of 10-d-old chick embryos. 24 h later, the embryos received a single intravenous injection of 100 μg of Mab HUIV26 or isotype-matched control. (A) Quantitation of HT1080 tumor growth in the chick embryo. (B) Quantitation of CS-1 tumor growth within the chick embryo. Data bars represent the mean tumor weights ± the standard errors from 5 to 10 embryos per condition. (C) SCID mice were injected subcutaneously with 2 × 106 M21 human melanoma cells. 3 d later mice were treated i.p. daily for 24 d with 100 μg of either Mab HUIV26 or an isotype-matched control antibody. Tumor size was monitored with calipers and tumor volumes were determined. Data represents the mean ± standard errors of the tumor volumes. All experiments were conducted 3 to 4 times with 5 to 10 animals per condition.

Mentions: The growth of most all solid tumors is thought to depend on angiogenesis (Weidner et al., 1991, 1992). Therefore, we evaluated its effects on the growth of tumors of distinct histological origin within two independent animal models. First, CS1 melanoma or HT1080 human fibrosarcoma cells were applied to the CAMs of 10-d-old chick embryos (Brooks et al., 1996). 24 h latter, the embryos were treated systemically with a single injection (100 μg/embryo) of either Mab HUIV26 or an isotype-matched control antibody. As shown in Fig. 5, A and B , Mab HUIV26 inhibited HT1080 and CS1 tumor growth by 50 and 80%, respectively. Treatment of these embryos with either an irrelevant isotype-matched control antibody or an antibody directed to the ECM protein fibronectin showed little if any effect.


Proteolytic exposure of a cryptic site within collagen type IV is required for angiogenesis and tumor growth in vivo.

Xu J, Rodriguez D, Petitclerc E, Kim JJ, Hangai M, Moon YS, Davis GE, Brooks PC, Yuen SM - J. Cell Biol. (2001)

Effects of systemic administration of purified Mab HUIV26 on tumor growth in vivo. The effects of Mab HUIV26 on tumor growth was assessed in two independent models, including the chick embryo (A and B) and the SCID Mouse (C). HT1080 human fibrosarcoma cells (4 × 105) or CS-1 melanoma tumor cells (5 × 106) were inoculated on the CAMs of 10-d-old chick embryos. 24 h later, the embryos received a single intravenous injection of 100 μg of Mab HUIV26 or isotype-matched control. (A) Quantitation of HT1080 tumor growth in the chick embryo. (B) Quantitation of CS-1 tumor growth within the chick embryo. Data bars represent the mean tumor weights ± the standard errors from 5 to 10 embryos per condition. (C) SCID mice were injected subcutaneously with 2 × 106 M21 human melanoma cells. 3 d later mice were treated i.p. daily for 24 d with 100 μg of either Mab HUIV26 or an isotype-matched control antibody. Tumor size was monitored with calipers and tumor volumes were determined. Data represents the mean ± standard errors of the tumor volumes. All experiments were conducted 3 to 4 times with 5 to 10 animals per condition.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196184&req=5

fig5: Effects of systemic administration of purified Mab HUIV26 on tumor growth in vivo. The effects of Mab HUIV26 on tumor growth was assessed in two independent models, including the chick embryo (A and B) and the SCID Mouse (C). HT1080 human fibrosarcoma cells (4 × 105) or CS-1 melanoma tumor cells (5 × 106) were inoculated on the CAMs of 10-d-old chick embryos. 24 h later, the embryos received a single intravenous injection of 100 μg of Mab HUIV26 or isotype-matched control. (A) Quantitation of HT1080 tumor growth in the chick embryo. (B) Quantitation of CS-1 tumor growth within the chick embryo. Data bars represent the mean tumor weights ± the standard errors from 5 to 10 embryos per condition. (C) SCID mice were injected subcutaneously with 2 × 106 M21 human melanoma cells. 3 d later mice were treated i.p. daily for 24 d with 100 μg of either Mab HUIV26 or an isotype-matched control antibody. Tumor size was monitored with calipers and tumor volumes were determined. Data represents the mean ± standard errors of the tumor volumes. All experiments were conducted 3 to 4 times with 5 to 10 animals per condition.
Mentions: The growth of most all solid tumors is thought to depend on angiogenesis (Weidner et al., 1991, 1992). Therefore, we evaluated its effects on the growth of tumors of distinct histological origin within two independent animal models. First, CS1 melanoma or HT1080 human fibrosarcoma cells were applied to the CAMs of 10-d-old chick embryos (Brooks et al., 1996). 24 h latter, the embryos were treated systemically with a single injection (100 μg/embryo) of either Mab HUIV26 or an isotype-matched control antibody. As shown in Fig. 5, A and B , Mab HUIV26 inhibited HT1080 and CS1 tumor growth by 50 and 80%, respectively. Treatment of these embryos with either an irrelevant isotype-matched control antibody or an antibody directed to the ECM protein fibronectin showed little if any effect.

Bottom Line: Exposure of this cryptic site was associated with angiogenic, but not quiescent, blood vessels and was required for angiogenesis in vivo.A monoclonal antibody (HUIV26) directed to this site disrupts integrin-dependent endothelial cell interactions and potently inhibits angiogenesis and tumor growth.Together, these studies suggest a novel mechanism by which proteolysis contributes to angiogenesis by exposing hidden regulatory elements within matrix-immobilized collagen type IV.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiation Oncology, Kaplan Cancer Center, New York University School of Medicine, New York, NY 10016, USA.

ABSTRACT
Evidence is provided that proteolytic cleavage of collagen type IV results in the exposure of a functionally important cryptic site hidden within its triple helical structure. Exposure of this cryptic site was associated with angiogenic, but not quiescent, blood vessels and was required for angiogenesis in vivo. Exposure of the HUIV26 epitope was associated with a loss of alpha1beta1 integrin binding and the gain of alphavbeta3 binding. A monoclonal antibody (HUIV26) directed to this site disrupts integrin-dependent endothelial cell interactions and potently inhibits angiogenesis and tumor growth. Together, these studies suggest a novel mechanism by which proteolysis contributes to angiogenesis by exposing hidden regulatory elements within matrix-immobilized collagen type IV.

Show MeSH
Related in: MedlinePlus