Mullerian Inhibiting Substance, a product of the fetal testic, is a large molecular weight glycoprotein which initiates regression of the Mullerian duct, the anlagan of the uterus, fallopian tube, and upper vagina. The underlying hypothesis of this proposal is that a fetal regressor which initiated morphogenic dissociation of an organ system in the embryo may produce the same effects against tumors derived from the cell types which respond to the fetal regressor in embryonic life. Mullerian Inhibiting Substance is detected by an embryonic organ culture assay and has been purified from the newborn calf testis. Using sequential ion exchange, carbohydrate affinity and dye affinity chromatography steps, we have achieved a 30,000-fold purification from starting testes and 2,000-fold from incubation media with 65% yield. Using hybridoma technology, we have developed monoclonal antibodies which both absorb out and block biological activity and from which we have eluted almost completely purified biologically active Mullerian Inhibiting Substance. Initial attempts at scale-up production (10 x) from 1,000 newborn calf testes were successful and will be continued from both bovine and avian sources. The availability of more purified material with MIS bioactivity makes broader oncological studies feasible. We propose to use purified MIS to study its effect against post-transplant, human ovarian tumor, as well as endometrial and cervical carcinomas, in nude mice and in the subrenal capsule assay. We continue to harvest fresh tumor from patients with ovarian and endometrial carcinoma to test their sensitivity to MIS. We have perfected PAP techniques and have localized MIS to the Sertoli cell endoplasmic reticulum and golgi apparatus. We will now attempt to localize MIS receptors with either monoclonal antibodies, antiidiotypic antibodies, polyclonal antibodies, or receptor antibodies. Phosphorylation events, which appear to be involved in Mullerian duct regression, will be studied both in the embryo and in the human ovarian cell line to investigate the mechanisms of MIS action and to propose newer chemotherapeutic strategies. We plan to study the ontogeny of steroid receptors both autoradiographically and biochemically during Mullerian duct regression and to correlate these events with the steroid modulation of MIS that we have observed. These findings will be applied to modulation of animal tumor hosts prior to treatment of Mullerian duct tumors with Mullerian Inhibiting Substance.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA017393-11
Application #
3164685
Study Section
(SSS)
Project Start
1978-06-01
Project End
1988-05-31
Budget Start
1985-06-01
Budget End
1986-05-31
Support Year
11
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
Chung, Youn Jee; Kim, Hyun Jung; Park, Sang Ho et al. (2015) Transcriptome analysis reveals that Müllerian inhibiting substance regulates signaling pathways that contribute to endometrial carcinogenesis. Int J Oncol 46:2039-46
Pépin, David; Sosulski, Amanda; Zhang, Lihua et al. (2015) AAV9 delivering a modified human Mullerian inhibiting substance as a gene therapy in patient-derived xenografts of ovarian cancer. Proc Natl Acad Sci U S A 112:E4418-27
Park, Joo Hyun; Tanaka, Yoshihiro; Arango, Nelson A et al. (2014) Induction of WNT inhibitory factor 1 expression by Müllerian inhibiting substance/antiMullerian hormone in the Müllerian duct mesenchyme is linked to Müllerian duct regression. Dev Biol 386:227-36
Arango, Nelson Alexander; Li, Li; Dabir, Deepa et al. (2013) Meiosis I arrest abnormalities lead to severe oligozoospermia in meiosis 1 arresting protein (M1ap)-deficient mice. Biol Reprod 88:76
Pépin, D; Hoang, M; Nicolaou, F et al. (2013) An albumin leader sequence coupled with a cleavage site modification enhances the yield of recombinant C-terminal Mullerian Inhibiting Substance. Technology 1:63-71
Song, Jae Yen; Jo, Hyun Hee; Kim, Mee Ran et al. (2012) Expression of Müllerian inhibiting substance type II receptor and antiproliferative effects of MIS on human cervical cancer. Int J Oncol 40:2013-21
Namkung, Jeong; Song, Jae Yen; Jo, Hyun Hee et al. (2012) Mullerian inhibiting substance induces apoptosis of human endometrial stromal cells in endometriosis. J Clin Endocrinol Metab 97:3224-30
Meirelles, Katia; Benedict, Leo Andrew; Dombkowski, David et al. (2012) Human ovarian cancer stem/progenitor cells are stimulated by doxorubicin but inhibited by Mullerian inhibiting substance. Proc Natl Acad Sci U S A 109:2358-63
Hwang, Seong Jin; Suh, Min Jung; Yoon, Joo Hee et al. (2011) Identification of characteristic molecular signature of Müllerian inhibiting substance in human HPV-related cervical cancer cells. Int J Oncol 39:811-20
Clarkson, Andrew N; Talbot, Caroline L; Wang, Pei-Yu et al. (2011) Müllerian inhibiting substance is anterogradely transported and does not attenuate avulsion-induced death of hypoglossal motor neurons. Exp Neurol 231:304-8

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