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Designed angiopoietin-1 variant, COMP-angiopoietin-1, rescues erectile function through healthy cavernous angiogenesis in a hypercholesterolemic mouse.

Ryu JK, Kim WJ, Koh YJ, Piao S, Jin HR, Lee SW, Choi MJ, Shin HY, Kwon MH, Jung K, Koh GY, Suh JK - Sci Rep (2015)

Bottom Line: Therefore, neovascularization is a promising strategy for curing ED.Angiopoietin-1 (Ang1) is an angiogenic growth factor that promotes the generation of stable and functional vasculature.These findings constitute a new paradigm toward curative treatment of both cavernous angiopathy and ED.

View Article: PubMed Central - PubMed

Affiliation: 1] National Research Center for Sexual Medicine and Department of Urology, Inha University School of Medicine, Incheon 400-711, Republic of Korea [2] Inha Research Institute for Medical Sciences, Inha University School of Medicine, Incheon 400-711, Republic of Korea.

ABSTRACT
Despite the advent of oral phosphodiesterase-5 inhibitors, curative treatment for erectile dysfunction (ED) remains unavailable. Recently, the link between ED and cardiovascular disease was unveiled and the main etiology of ED was found to be vasculogenic. Therefore, neovascularization is a promising strategy for curing ED. Angiopoietin-1 (Ang1) is an angiogenic growth factor that promotes the generation of stable and functional vasculature. Here, we demonstrate that local delivery of the soluble, stable, and potent Ang1 variant, COMP-Ang1 gene or protein, into the penises of hypercholesterolemic mice increases cavernous angiogenesis, eNOS phosphorylation, and cGMP expression, resulting in full recovery of erectile function and cavernous blood flow up to 8 weeks after treatment. COMP-Ang1-induced promotion of cavernous angiogenesis and erectile function was abolished in Nos3(-/-) mice and in the presence of the NOS inhibitor, L-NAME. COMP-Ang1 also restored the integrity of endothelial cell-cell junction by down-regulating the expression of histone deacetylase 2 in the penis of hypercholesterolemic mice and in primary cultured mouse cavernous endothelial cells. These findings constitute a new paradigm toward curative treatment of both cavernous angiopathy and ED.

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In vivo expression of COMP-Ang1 gene and protein.(a) Detection of COMP-Ang1-specific mRNA in the corpus cavernosum from hypercholesterolemic mice. PCR was performed with primers specific for COMP-Ang1. RNA was extracted from the corpus cavernosum 3, 7, 14, and 21 days after intracavernous administration of ad-COMP-Ang1 (2 × 108 parts/20 μl). Densitometric data are presented as the relative ratio of COMP-Ang1 mRNA to β-actin mRNA. The relative ratio measured 3 days after injection of ad-COMP-Ang1 was arbitrarily set equivalent to 1. Bars represent the mean ± SEM of four independent experiments. M, marker. (b) Western blot analysis of the expression of COMP-Ang1 protein in cavernous tissues from hypercholesterolemic mice 1, 6, and 24 hours and 3, 7, 14, and 21 days after intracavernous injection of ad-COMP-Ang1 (left) or COMP-Ang1 protein (right). The relative ratio of COMP-Ang1 to β-actin measured 3 days after injection of ad-COMP-Ang1 or 1 hour after injection of COMP-Ang1 protein was arbitrarily set equivalent to 1. Bars represent the mean ± SEM of four independent experiments. (c) Anti-FLAG staining of cavernous tissue from hypercholesterolemic mice 7 days after intracavernous injection of ad-LacZ (2 × 108 parts/20 μl) or ad-COMP-Ang1 (2 × 108 parts/20 μl) and 1 hour after intracavernous injection of BSA (5.88 μg/20 μl) or COMP-Ang1 protein (5.88 μg/20 μl). 2nd Ab, secondary antibody control; arrowheads, endothelial cells; arrows, smooth muscle cells. Scale bar = 50 μm. (d) Immunohistochemical staining of cavernous tissue using antibodies to FLAG (red), PECAM-1 (CD31; green), and α-actin (green) in each group. Scale bar = 50 μm.
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f1: In vivo expression of COMP-Ang1 gene and protein.(a) Detection of COMP-Ang1-specific mRNA in the corpus cavernosum from hypercholesterolemic mice. PCR was performed with primers specific for COMP-Ang1. RNA was extracted from the corpus cavernosum 3, 7, 14, and 21 days after intracavernous administration of ad-COMP-Ang1 (2 × 108 parts/20 μl). Densitometric data are presented as the relative ratio of COMP-Ang1 mRNA to β-actin mRNA. The relative ratio measured 3 days after injection of ad-COMP-Ang1 was arbitrarily set equivalent to 1. Bars represent the mean ± SEM of four independent experiments. M, marker. (b) Western blot analysis of the expression of COMP-Ang1 protein in cavernous tissues from hypercholesterolemic mice 1, 6, and 24 hours and 3, 7, 14, and 21 days after intracavernous injection of ad-COMP-Ang1 (left) or COMP-Ang1 protein (right). The relative ratio of COMP-Ang1 to β-actin measured 3 days after injection of ad-COMP-Ang1 or 1 hour after injection of COMP-Ang1 protein was arbitrarily set equivalent to 1. Bars represent the mean ± SEM of four independent experiments. (c) Anti-FLAG staining of cavernous tissue from hypercholesterolemic mice 7 days after intracavernous injection of ad-LacZ (2 × 108 parts/20 μl) or ad-COMP-Ang1 (2 × 108 parts/20 μl) and 1 hour after intracavernous injection of BSA (5.88 μg/20 μl) or COMP-Ang1 protein (5.88 μg/20 μl). 2nd Ab, secondary antibody control; arrowheads, endothelial cells; arrows, smooth muscle cells. Scale bar = 50 μm. (d) Immunohistochemical staining of cavernous tissue using antibodies to FLAG (red), PECAM-1 (CD31; green), and α-actin (green) in each group. Scale bar = 50 μm.

Mentions: Exogenous COMP-Ang1 mRNA or protein expression was detected in the corpus cavernosa of hypercholesterolemic mice 3, 7, 14, and 21 days after injection of ad-COMP-Ang1. The level of exogenous COMP-Ang1 transcript or protein expression was highest 7 days after injection and expression was still detectable at 21 days (Fig. 1a and Fig. 1b [left]).


Designed angiopoietin-1 variant, COMP-angiopoietin-1, rescues erectile function through healthy cavernous angiogenesis in a hypercholesterolemic mouse.

Ryu JK, Kim WJ, Koh YJ, Piao S, Jin HR, Lee SW, Choi MJ, Shin HY, Kwon MH, Jung K, Koh GY, Suh JK - Sci Rep (2015)

In vivo expression of COMP-Ang1 gene and protein.(a) Detection of COMP-Ang1-specific mRNA in the corpus cavernosum from hypercholesterolemic mice. PCR was performed with primers specific for COMP-Ang1. RNA was extracted from the corpus cavernosum 3, 7, 14, and 21 days after intracavernous administration of ad-COMP-Ang1 (2 × 108 parts/20 μl). Densitometric data are presented as the relative ratio of COMP-Ang1 mRNA to β-actin mRNA. The relative ratio measured 3 days after injection of ad-COMP-Ang1 was arbitrarily set equivalent to 1. Bars represent the mean ± SEM of four independent experiments. M, marker. (b) Western blot analysis of the expression of COMP-Ang1 protein in cavernous tissues from hypercholesterolemic mice 1, 6, and 24 hours and 3, 7, 14, and 21 days after intracavernous injection of ad-COMP-Ang1 (left) or COMP-Ang1 protein (right). The relative ratio of COMP-Ang1 to β-actin measured 3 days after injection of ad-COMP-Ang1 or 1 hour after injection of COMP-Ang1 protein was arbitrarily set equivalent to 1. Bars represent the mean ± SEM of four independent experiments. (c) Anti-FLAG staining of cavernous tissue from hypercholesterolemic mice 7 days after intracavernous injection of ad-LacZ (2 × 108 parts/20 μl) or ad-COMP-Ang1 (2 × 108 parts/20 μl) and 1 hour after intracavernous injection of BSA (5.88 μg/20 μl) or COMP-Ang1 protein (5.88 μg/20 μl). 2nd Ab, secondary antibody control; arrowheads, endothelial cells; arrows, smooth muscle cells. Scale bar = 50 μm. (d) Immunohistochemical staining of cavernous tissue using antibodies to FLAG (red), PECAM-1 (CD31; green), and α-actin (green) in each group. Scale bar = 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4363832&req=5

f1: In vivo expression of COMP-Ang1 gene and protein.(a) Detection of COMP-Ang1-specific mRNA in the corpus cavernosum from hypercholesterolemic mice. PCR was performed with primers specific for COMP-Ang1. RNA was extracted from the corpus cavernosum 3, 7, 14, and 21 days after intracavernous administration of ad-COMP-Ang1 (2 × 108 parts/20 μl). Densitometric data are presented as the relative ratio of COMP-Ang1 mRNA to β-actin mRNA. The relative ratio measured 3 days after injection of ad-COMP-Ang1 was arbitrarily set equivalent to 1. Bars represent the mean ± SEM of four independent experiments. M, marker. (b) Western blot analysis of the expression of COMP-Ang1 protein in cavernous tissues from hypercholesterolemic mice 1, 6, and 24 hours and 3, 7, 14, and 21 days after intracavernous injection of ad-COMP-Ang1 (left) or COMP-Ang1 protein (right). The relative ratio of COMP-Ang1 to β-actin measured 3 days after injection of ad-COMP-Ang1 or 1 hour after injection of COMP-Ang1 protein was arbitrarily set equivalent to 1. Bars represent the mean ± SEM of four independent experiments. (c) Anti-FLAG staining of cavernous tissue from hypercholesterolemic mice 7 days after intracavernous injection of ad-LacZ (2 × 108 parts/20 μl) or ad-COMP-Ang1 (2 × 108 parts/20 μl) and 1 hour after intracavernous injection of BSA (5.88 μg/20 μl) or COMP-Ang1 protein (5.88 μg/20 μl). 2nd Ab, secondary antibody control; arrowheads, endothelial cells; arrows, smooth muscle cells. Scale bar = 50 μm. (d) Immunohistochemical staining of cavernous tissue using antibodies to FLAG (red), PECAM-1 (CD31; green), and α-actin (green) in each group. Scale bar = 50 μm.
Mentions: Exogenous COMP-Ang1 mRNA or protein expression was detected in the corpus cavernosa of hypercholesterolemic mice 3, 7, 14, and 21 days after injection of ad-COMP-Ang1. The level of exogenous COMP-Ang1 transcript or protein expression was highest 7 days after injection and expression was still detectable at 21 days (Fig. 1a and Fig. 1b [left]).

Bottom Line: Therefore, neovascularization is a promising strategy for curing ED.Angiopoietin-1 (Ang1) is an angiogenic growth factor that promotes the generation of stable and functional vasculature.These findings constitute a new paradigm toward curative treatment of both cavernous angiopathy and ED.

View Article: PubMed Central - PubMed

Affiliation: 1] National Research Center for Sexual Medicine and Department of Urology, Inha University School of Medicine, Incheon 400-711, Republic of Korea [2] Inha Research Institute for Medical Sciences, Inha University School of Medicine, Incheon 400-711, Republic of Korea.

ABSTRACT
Despite the advent of oral phosphodiesterase-5 inhibitors, curative treatment for erectile dysfunction (ED) remains unavailable. Recently, the link between ED and cardiovascular disease was unveiled and the main etiology of ED was found to be vasculogenic. Therefore, neovascularization is a promising strategy for curing ED. Angiopoietin-1 (Ang1) is an angiogenic growth factor that promotes the generation of stable and functional vasculature. Here, we demonstrate that local delivery of the soluble, stable, and potent Ang1 variant, COMP-Ang1 gene or protein, into the penises of hypercholesterolemic mice increases cavernous angiogenesis, eNOS phosphorylation, and cGMP expression, resulting in full recovery of erectile function and cavernous blood flow up to 8 weeks after treatment. COMP-Ang1-induced promotion of cavernous angiogenesis and erectile function was abolished in Nos3(-/-) mice and in the presence of the NOS inhibitor, L-NAME. COMP-Ang1 also restored the integrity of endothelial cell-cell junction by down-regulating the expression of histone deacetylase 2 in the penis of hypercholesterolemic mice and in primary cultured mouse cavernous endothelial cells. These findings constitute a new paradigm toward curative treatment of both cavernous angiopathy and ED.

Show MeSH
Related in: MedlinePlus