Enhanced purification and production of Müllerian inhibiting substance for therapeutic applications.
It is almost 60 years since Prof. Alfred Jost reported the seminal observations regarding Müllerian inhibiting substance (MIS). His experiments clearly showed that a testicular product other than testosterone, a Müllerian inhibitor, was responsible for Müllerian duct regression. Twenty-five years later Dr. Picon established an organ culture assay which paved the way for the initial studies into the biochemistry and biology of Müllerian inhibiting substance, also known as Anti-Müllerian hormone (AMH), undertaken first in Dr. Nathalie Josso's Laboratory in Paris then in our own laboratory in Boston. Purification of MIS led to cloning the human gene and production of recombinant human (rhMIS). MIS is a 140 kDa glycoprotein homodimer which is activated by a biosynthetic protease, cleaving MIS into an aminoterminus (110 kDa) and a carboxyterminus (25 kDa). The latter domain is sufficient for biological activities. MIS functions by interacting with two receptors; a type II binds the hormone and at type I that initiates downstream signaling. The MIS type II receptor has been cloned and functionally confirmed as distinct from that of other members of the TGFbeta superfamily. MIS can employ a number of type I receptors (ALK2, ALK3, ALK6) and BMP receptor specific SMADS 1, 5, and 8 in various tissue specific contexts. Cell lines derived from human ovarian, breast, and prostate tumors, and from rodent Leydig cell tumors, which respond to MIS in growth inhibition assays, all express the MIS type II receptor. A variety of signal transduction pathways are associated with the grown inhibition mediated by MIS. For example, breast and prostate cancer cell lines use a MIS-mediated NFkappaB pathway leading to G1 arrest and apoptosis. The ovarian cancer cell lines employ a pathway which enhances p16, modulates the E2Fs, and induces apoptosis. These signal transduction events can establish new rational treatment strategies to complement the growth inhibitory effects mediated by MIS. These combination strategies are being tested in vitro, and where appropriate will be tested in vivo using the highly purified MIS preparations, prior to use in early human clinical trials.
Donahoe
Patricia K
PK
Pediatric Surgical Research Laboratories, Massachusetts General Hospital and Harvard Medical School, Warren 11, 55 Fruit Street, Boston, MA 02114, USA. pdonahoe@partners.org
Clarke
Trent
T
Teixeira
Jose
J
Maheswaran
Shyamala
S
MacLaughlin
David T
DT
D030201:Activin Receptors, Type I / Q000502:physiology; D000818:Animals; D054304:Anti-Mullerian Hormone; D052005:Bone Morphogenetic Protein Receptors, Type I; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000502:physiology; D005260:Female; D005341:Fibrinolysin / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006023:Glycoproteins / Q000096:biosynthesis* / Q000494:pharmacology / Q000627:therapeutic use*; D006801:Humans; D051379:Mice; D009369:Neoplasms / Q000188:drug therapy; D010051:Ovarian Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000502:physiology; D017978:Receptors, Growth Factor / Q000502:physiology; D018000:Receptors, Peptide / Q000502:physiology; D018125:Receptors, Transforming Growth Factor beta; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000494:pharmacology / Q000627:therapeutic use; D012733:Sex Differentiation / Q000502:physiology; D013735:Testicular Hormones / Q000096:biosynthesis* / Q000494:pharmacology / Q000627:therapeutic use*
D019941:Cyclin-Dependent Kinase Inhibitor p16; D006023:Glycoproteins; D017978:Receptors, Growth Factor; D018000:Receptors, Peptide; D018125:Receptors, Transforming Growth Factor beta; D011994:Recombinant Proteins; D013735:Testicular Hormones; C084621:anti-Mullerian hormone receptor; D054304:Anti-Mullerian Hormone; D017346:Protein Serine-Threonine Kinases; D030201:Activin Receptors, Type I; C498214:BMPR1B protein, human; C498212:Bmpr1a protein, mouse; C498215:Bmpr1b protein, mouse; D052005:Bone Morphogenetic Protein Receptors, Type I; D005341:Fibrinolysin