Therapeutic effect of angiostatin gene transfer in a murine model of endometriosis - PubMed (original) (raw)
Therapeutic effect of angiostatin gene transfer in a murine model of endometriosis
Charlotta Dabrosin et al. Am J Pathol. 2002 Sep.
Abstract
Endometriosis, the growth of ectopic endometrial tissue, is a chronic recurrent disease affecting 10% of the female population causing dyspareunia, pelvic pain, dysmenorrhea, and infertility. Suppression of ovarian activity is the cornerstone of medical therapy with limited benefit and severe adverse effects. Angiogenesis plays a major role in the development of endometriosis suggesting that anti-angiogenic therapy would offer a new therapeutic approach. We report successful treatment of endometriosis in estrogen-supplemented ovariectomized mice by transient overexpression (6 to 10 days of duration) of the gene for a natural angiogenesis inhibitor angiostatin, delivered to the peritoneum by a replication-deficient adenovirus vector (AdAngiostatin). Established endometriosis was eradicated in 14 of 14 AdAngiostatin-treated animals, whereas 11 of 13 control animals showed full disease development. Administered to normal cycling mice for the same transient period, AdAngiostatin caused impaired ovarian function with suppressed corpus luteum development, decreased production of estradiol and progesterone, decreased ovarian and uterine weight, and increased body weight. AdAngiostatin treatment lowered the levels of sex steroids but did not induce total castration. Gene therapy with angiogenic inhibitors is a highly effective treatment for endometriosis, even in a host with preserved estrogen levels. However, local or targeted delivery of the gene must be considered to avoid prolonged systemic effects and impaired ovarian function.
Figures
Figure 1.
Morphology of experimental endometriosis. A: Formalin-fixed and paraffin-embedded endometriosis 7 days after inoculation of endometrium into the peritoneal cavity of mice demonstrates typical morphology of endometriosis with endometrial glandular epithelium surrounded by an intact basement membrane and adjacent endometrial stroma. Arrows indicate endometrial stroma (A and D) and endometrial glandular epithelium surrounding an endometrial cyst with secretion (A) (H&E staining; results shown are representative of n = 10). B: Immunohistochemical analysis with antibodies to Factor VIII demonstrates abundant microvessel staining in the 7-day (pretreatment) endometrial transplant. Arrows indicate intact vessel structures (C and F). C: Factor VIII staining at day 7 examined at higher power (×100). D: Five days after treatment with AdAngiostatin 1 × 109 pfu intraperitoneally there were no visible glands left and the stroma was less abundant (H&E staining). E: Microvessel staining with Factor VIII 5 days after AdAngiostatin treatment shows residual endometrial stroma with significantly reduced staining for microvessels (arrows). F: Factor VIII staining at day 5 of AdAngiostatin treatment at higher power (×100). G: Staining for apoptosis (Apoptag) at day 5 after AdAngiostatin treatment (×100) shows endothelial cells (closed arrowhead) and endometrial stromal cell (open arrowhead) undergoing apoptosis.
Figure 2.
Expression of angiostatin in recovered peritoneal lavage of AdAngiostatin-treated mice. Western blot was used to detect murine angiostatin protein. Angiostatin was detected only in the AdAngiostatin-treated mice and could be seen as two bands at ∼40 to 50 kd, representing the two glycoforms of murine angiostatin (lane 2). No angiostatin bands could be detected in peritoneal lavage of Ad_dl_70-3 control-treated mice (lane 3). Positive control (lane 1) is rabbit angiostatin derived from human urokinase-digested rabbit plasminogen showing two angiostatin bands. A 50-kd marker position is shown (arrow).
Figure 3.
Body weight in normal cycling mice and OVX mice after AdAngiostatin treatment. AdAngiostatin, 1 × 109 pfu/mouse was given intraperitoneally. After 16 to 18 days normal cycling untreated mice or normal cycling mice treated with AdAngiostatin, OVX, OVX + estradiol supplemented (E2), and OVX + E2 + AdAngiostatin mice were weighed. The OVX- and AdAngiostatin-treated normal cycling mice increased their body weight significantly compared to all other groups (P < 0.0001, n = 5 to 6).
Figure 4.
Ovarian weight after AdAngiostatin treatment. Eighteen days after intraperitoneal injections of AdAngiostatin, 1 × 109 or 2 × 109 pfu, ovaries from FVB mice were removed and weighed. The weight decreased significantly in a dose-response pattern. *, P < 0.001 versus control; +, P < 0.05 versus AdAngiostatin 1 × 109 pfu (n = 5).
Figure 5.
Morphology of ovaries after AdAngiostatin treatment. Eighteen days after intraperitoneal injections of AdAngiostatin, 1 × 109 or 2 × 109 pfu, ovaries from mice were removed and fixed in formalin and stained with H&E. A: Normal ovary with primary follicles and corpora lutea. B: The treated ovaries showed an overall decrease in corpus luteum development and a loss of stroma (results are representative of n = 5).
Figure 6.
Serum hormone levels in normal cycling mice and OVX mice after AdAngiostatin treatment. A: Serum estradiol levels after AdAngiostatin treatment. Eighteen days after intraperitoneal injections of AdAngiostatin, 1 × 109 or 2 × 109 pfu, or after OVX, serum estradiol levels in mice were measured. In the control animals the serum levels showed a normal wide physiological range. *, P < 0.05 versus controls; +, P < 0.05 versus OVX (n = 4 to 6). B: Serum progesterone levels after AdAngiostatin treatment. Eighteen days after intraperitoneal injections of AdAngiostatin, 1 × 109 or 2 × 109 pfu, or after OVX, serum progesterone levels in mice were measured. In the control animals the serum levels showed a normal wide physiological range. *, P < 0.01 versus controls; +, P < 0.05 versus OVX (n = 4 to 6).
Figure 7.
Uterine weight after AdAngiostatin treatment. Eighteen days after intraperitoneal injection of AdAngiostatin, 1 × 109 or 2 × 109 pfu, or OVX or estradiol supplement, uteri from FVB mice were removed and weighed. *, P < 0.05 versus controls; +, P < 0.05 versus OVX; ***, P < 0.001 versus all other groups (n = 5 to 10).
References
- Garry R, Clayton R, Hawe J: The effect of endometriosis and its radical laparoscopic excision on quality of life indicators. Br J Obstet Gynaecol 2000, 107:44-54 - PubMed
- Strathy JH, Molgaard CA, Coulam CB, Melton LJ: Endometriosis and infertility: a laparoscopic study of endometriosis among fertile and infertile women. Fertil Steril 1982, 38:667-672 - PubMed
- Laufer MR, Goitein L, Bush M, Cramer DW, Emans SJ: Prevalence of endometriosis in adolescent girls with chronic pelvic pain not responding to conventional therapy. J Pediatr Adolesc Gynecol 1997, 10:199-202 - PubMed
- Brinton LA, Gridley G, Persson I, Baron J, Bergqvist A: Cancer risk after a hospital discharge diagnosis of endometriosis. Am J Obstet Gynecol 1997, 176:572-579 - PubMed
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