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Chronic effects of an anti-angiogenic thrombospondin-1 mimetic peptide, ABT-898, on female mouse reproductive outcomes

View Article: PubMed Central - PubMed

ABSTRACT

Background: Angiogenesis is an essential process in endometriosis disease progression. Earlier, we demonstrated that anti-angiogenic peptide, ABT-898 prevents neoangiogenesis of human endometriotic lesions in a xenograft mouse model. Since angiogenesis is essential for normal ovarian and uterine function, we evaluated effects of ABT-898 on normal female reproductive processes in mice.

Methods: Cycling female C57BL/6N mice were dosed with ABT-898 (100 mg/kg) or 5 % dextrose control for 21 consecutive days to cover multiple estrous cycles (average estrous cycle 4 to 5 days in mice). Pregnant female mice were dosed with ABT-898 (100 mg/kg) or control on alternate days over the course of gestation, beginning at gestation day 7.5 to 17.5 (gestation length 21 days). Histological analysis along with CD31 and Vimentin immunohistochemistry were performed on ovaries and uteri obtained from treated and control mice. To understand the influence of ABT-898 on systemic angiogenic factors, a Pro Mouse Cytokine 9-plex assay was performed on plasma samples obtained from mice prior to treatment, during the second week of ABAT-898 or control treatment and on the last day of treatment.

Results: ABT-898 did not affect the number of estrous cycles over the 21 day treatment compared to control. Histological analysis of ovaries found no difference in the number of primordial, primary, secondary, and antral follicles between ABT-898 treated and control groups. Similarly, no difference was observed in the microvessel density between ABT-898 treated and control uteri, ovarian follicles or corpus luteum when assessed using CD31 or vimentin immunohistochemistry. Electron microscopy revealed similar capillary structure and appearance in both ABT-898 treated and control uteri. Although peripheral blood angiogenic cytokine profiles (IL-15, IL-18, M-CSF, b-FGF, PDGF-bb, MIG, MIP-2, LIF and VEGF) changed over the course of the intervention, there was no significant difference between ABT-898 and control groups at any of the studied time points. Treatment with ABT-898 during pregnancy had no effect on litter size at birth, pup weight at birth or pup weight at weaning.

Conclusion: Our findings suggest that ABT-898 may not alter angiogenesis dependent reproductive processes in female mice. However, an extensive reproductive toxicology screening is required to substantiate use of ABT-898 in future.

Electronic supplementary material: The online version of this article (doi:10.1186/s12958-016-0192-7) contains supplementary material, which is available to authorized users.

No MeSH data available.


ABT-898 does not alter the number of ovarian follicles. Ovaries from mice dosed with ABT-898 (a) or 5 % dextrose (b) were sectioned and the numbers of primordial (c, g), primary (d, h), secondary (e, i) and antral (f, j) follicles were counted. The number of primordial, primary, secondary and antral follicles did not significantly differ between ABT-898 and 5 % dextrose groups (k). Red arrow: primordial follicle, black arrow: primary follicle, green arrow: secondary follicle, yellow arrow: antral follicle. Scale bar represents 75 μm. P < 0.05
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Fig2: ABT-898 does not alter the number of ovarian follicles. Ovaries from mice dosed with ABT-898 (a) or 5 % dextrose (b) were sectioned and the numbers of primordial (c, g), primary (d, h), secondary (e, i) and antral (f, j) follicles were counted. The number of primordial, primary, secondary and antral follicles did not significantly differ between ABT-898 and 5 % dextrose groups (k). Red arrow: primordial follicle, black arrow: primary follicle, green arrow: secondary follicle, yellow arrow: antral follicle. Scale bar represents 75 μm. P < 0.05

Mentions: After the 21 day treatment period mice were sacrificed and reproductive tracts were harvested for histological analysis. No gross differences were seen at this point in the size, structure, or vascularity of the reproductive tracts between ABT-898 (Fig. 1l), and control (Fig. 1m) groups. In an attempt to assess the effect of ABT-898 on follicular development, an angiogenesis dependent process, we performed follicle counts on ovaries from ABT-898 and control mice. There were no observable differences in the microscopic anatomy of the ovaries between ABT-898 (Fig. 2a) and control (Fig. 2b) groups. Using a modified version of Pederson’s and Peter’s classification system we counted the number of primordial, primary, secondary and antral follicles (representative follicle histology Fig. 2c–j), and found no difference between ABT-898 and control groups (Fig. 2k). These results provide initial evidence that ABT-898 does not disrupt the estrous cycle or follicular development in mice after a chronic daily treatment for 21 days.Fig. 2


Chronic effects of an anti-angiogenic thrombospondin-1 mimetic peptide, ABT-898, on female mouse reproductive outcomes
ABT-898 does not alter the number of ovarian follicles. Ovaries from mice dosed with ABT-898 (a) or 5 % dextrose (b) were sectioned and the numbers of primordial (c, g), primary (d, h), secondary (e, i) and antral (f, j) follicles were counted. The number of primordial, primary, secondary and antral follicles did not significantly differ between ABT-898 and 5 % dextrose groups (k). Red arrow: primordial follicle, black arrow: primary follicle, green arrow: secondary follicle, yellow arrow: antral follicle. Scale bar represents 75 μm. P < 0.05
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5015213&req=5

Fig2: ABT-898 does not alter the number of ovarian follicles. Ovaries from mice dosed with ABT-898 (a) or 5 % dextrose (b) were sectioned and the numbers of primordial (c, g), primary (d, h), secondary (e, i) and antral (f, j) follicles were counted. The number of primordial, primary, secondary and antral follicles did not significantly differ between ABT-898 and 5 % dextrose groups (k). Red arrow: primordial follicle, black arrow: primary follicle, green arrow: secondary follicle, yellow arrow: antral follicle. Scale bar represents 75 μm. P < 0.05
Mentions: After the 21 day treatment period mice were sacrificed and reproductive tracts were harvested for histological analysis. No gross differences were seen at this point in the size, structure, or vascularity of the reproductive tracts between ABT-898 (Fig. 1l), and control (Fig. 1m) groups. In an attempt to assess the effect of ABT-898 on follicular development, an angiogenesis dependent process, we performed follicle counts on ovaries from ABT-898 and control mice. There were no observable differences in the microscopic anatomy of the ovaries between ABT-898 (Fig. 2a) and control (Fig. 2b) groups. Using a modified version of Pederson’s and Peter’s classification system we counted the number of primordial, primary, secondary and antral follicles (representative follicle histology Fig. 2c–j), and found no difference between ABT-898 and control groups (Fig. 2k). These results provide initial evidence that ABT-898 does not disrupt the estrous cycle or follicular development in mice after a chronic daily treatment for 21 days.Fig. 2

View Article: PubMed Central - PubMed

ABSTRACT

Background: Angiogenesis is an essential process in endometriosis disease progression. Earlier, we demonstrated that anti-angiogenic peptide, ABT-898 prevents neoangiogenesis of human endometriotic lesions in a xenograft mouse model. Since angiogenesis is essential for normal ovarian and uterine function, we evaluated effects of ABT-898 on normal female reproductive processes in mice.

Methods: Cycling female C57BL/6N mice were dosed with ABT-898 (100&nbsp;mg/kg) or 5&nbsp;% dextrose control for 21 consecutive days to cover multiple estrous cycles (average estrous cycle 4 to 5&nbsp;days in mice). Pregnant female mice were dosed with ABT-898 (100&nbsp;mg/kg) or control on alternate days over the course of gestation, beginning at gestation day 7.5 to 17.5 (gestation length 21&nbsp;days). Histological analysis along with CD31 and Vimentin immunohistochemistry were performed on ovaries and uteri obtained from treated and control mice. To understand the influence of ABT-898 on systemic angiogenic factors, a Pro Mouse Cytokine 9-plex assay was performed on plasma samples obtained from mice prior to treatment, during the second week of ABAT-898 or control treatment and on the last day of treatment.

Results: ABT-898 did not affect the number of estrous cycles over the 21&nbsp;day treatment compared to control. Histological analysis of ovaries found no difference in the number of primordial, primary, secondary, and antral follicles between ABT-898 treated and control groups. Similarly, no difference was observed in the microvessel density between ABT-898 treated and control uteri, ovarian follicles or corpus luteum when assessed using CD31 or vimentin immunohistochemistry. Electron microscopy revealed similar capillary structure and appearance in both ABT-898 treated and control uteri. Although peripheral blood angiogenic cytokine profiles (IL-15, IL-18, M-CSF, b-FGF, PDGF-bb, MIG, MIP-2, LIF and VEGF) changed over the course of the intervention, there was no significant difference between ABT-898 and control groups at any of the studied time points. Treatment with ABT-898 during pregnancy had no effect on litter size at birth, pup weight at birth or pup weight at weaning.

Conclusion: Our findings suggest that ABT-898 may not alter angiogenesis dependent reproductive processes in female mice. However, an extensive reproductive toxicology screening is required to substantiate use of ABT-898 in future.

Electronic supplementary material: The online version of this article (doi:10.1186/s12958-016-0192-7) contains supplementary material, which is available to authorized users.

No MeSH data available.