title: FOXO1/3 and PTEN Depletion in Granulosa Cells Promotes Ovarian Granulosa Cell Tumor Development.
abstract: The forkhead box (FOX), FOXO1 and FOXO3, transcription factors regulate multiple functions in mammalian cells. Selective inactivation of the Foxo1 and Foxo3 genes in murine ovarian granulosa cells severely impairs follicular development and apoptosis causing infertility, and as shown here, granulosa cell tumor (GCT) formation. Coordinate depletion of the tumor suppressor Pten gene in the Foxo1/3 strain enhanced the penetrance and onset of GCT formation. Immunostaining and Western blot analyses confirmed FOXO1 and phosphatase and tensin homolog (PTEN) depletion, maintenance of globin transcription factor (GATA) 4 and nuclear localization of FOXL2 and phosphorylated small mothers against decapentaplegic (SMAD) 2/3 in the tumor cells, recapitulating results we observed in human adult GCTs. Microarray and quantitative PCR analyses of mouse GCTs further confirmed expression of specific genes (Foxl2, Gata4, and Wnt4) controlling granulosa cell fate specification and proliferation, whereas others (Emx2, Nr0b1, Rspo1, and Wt1) were suppressed. Key genes (Amh, Bmp2, and Fshr) controlling follicle growth, apoptosis, and differentiation were also suppressed. Inhbb and Grem1 were selectively elevated, whereas reduction of Inha provided additional evidence that activin signaling and small mothers against decapentaplegic (SMAD) 2/3 phosphorylation impact GCT formation. Unexpectedly, markers of Sertoli/epithelial cells (SRY [sex determining region Y]-box 9/keratin 8) and alternatively activated macrophages (chitinase 3-like 3) were elevated in discrete subpopulations within the mouse GCTs, indicating that Foxo1/3/Pten depletion not only leads to GCTs but also to altered granulosa cell fate decisions and immune responses. Thus, analyses of the Foxo1/3/Pten mouse GCTs and human adult GCTs provide strong evidence that impaired functions of the FOXO1/3/PTEN pathways lead to dramatic changes in the molecular program within granulosa cells, chronic activin signaling in the presence of FOXL2 and GATA4, and tumor formation.
authors.affiliation: Departments of Molecular and Cellular Biology (Z.L., Y.A.R., S.A.P., J.A., J.S.R.), Pathology and Immunology (S.A.P.), and Obstetrics and Gynecology (J.A.), Baylor College of Medicine, and Department of Experimental Radiation Oncology (W.Z.), The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030; Department of Pathology (D.H.C.), The University of Texas Southwestern Medical School, Dallas, Texas 75390; and Department of Anatomy and Developmental Biology (D.W.), Monash University, Clayton VIC 3800, Australia.
authors.forename: Zhilin
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Departments of Molecular and Cellular Biology (Z.L., Y.A.R., S.A.P., J.A., J.S.R.), Pathology and Immunology (S.A.P.), and Obstetrics and Gynecology (J.A.), Baylor College of Medicine, and Department of Experimental Radiation Oncology (W.Z.), The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030; Department of Pathology (D.H.C.), The University of Texas Southwestern Medical School, Dallas, Texas 75390; and Department of Anatomy and Developmental Biology (D.W.), Monash University, Clayton VIC 3800, Australia.
authors.forename: Yi A
authors.initials: YA
authors.lastname: Ren
authors.affiliation: Departments of Molecular and Cellular Biology (Z.L., Y.A.R., S.A.P., J.A., J.S.R.), Pathology and Immunology (S.A.P.), and Obstetrics and Gynecology (J.A.), Baylor College of Medicine, and Department of Experimental Radiation Oncology (W.Z.), The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030; Department of Pathology (D.H.C.), The University of Texas Southwestern Medical School, Dallas, Texas 75390; and Department of Anatomy and Developmental Biology (D.W.), Monash University, Clayton VIC 3800, Australia.
authors.forename: Stephanie A
authors.initials: SA
authors.lastname: Pangas
authors.affiliation: Departments of Molecular and Cellular Biology (Z.L., Y.A.R., S.A.P., J.A., J.S.R.), Pathology and Immunology (S.A.P.), and Obstetrics and Gynecology (J.A.), Baylor College of Medicine, and Department of Experimental Radiation Oncology (W.Z.), The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030; Department of Pathology (D.H.C.), The University of Texas Southwestern Medical School, Dallas, Texas 75390; and Department of Anatomy and Developmental Biology (D.W.), Monash University, Clayton VIC 3800, Australia.
authors.forename: Jaye
authors.initials: J
authors.lastname: Adams
authors.affiliation: Departments of Molecular and Cellular Biology (Z.L., Y.A.R., S.A.P., J.A., J.S.R.), Pathology and Immunology (S.A.P.), and Obstetrics and Gynecology (J.A.), Baylor College of Medicine, and Department of Experimental Radiation Oncology (W.Z.), The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030; Department of Pathology (D.H.C.), The University of Texas Southwestern Medical School, Dallas, Texas 75390; and Department of Anatomy and Developmental Biology (D.W.), Monash University, Clayton VIC 3800, Australia.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhou
authors.affiliation: Departments of Molecular and Cellular Biology (Z.L., Y.A.R., S.A.P., J.A., J.S.R.), Pathology and Immunology (S.A.P.), and Obstetrics and Gynecology (J.A.), Baylor College of Medicine, and Department of Experimental Radiation Oncology (W.Z.), The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030; Department of Pathology (D.H.C.), The University of Texas Southwestern Medical School, Dallas, Texas 75390; and Department of Anatomy and Developmental Biology (D.W.), Monash University, Clayton VIC 3800, Australia.
authors.forename: Diego H
authors.initials: DH
authors.lastname: Castrillon
authors.affiliation: Departments of Molecular and Cellular Biology (Z.L., Y.A.R., S.A.P., J.A., J.S.R.), Pathology and Immunology (S.A.P.), and Obstetrics and Gynecology (J.A.), Baylor College of Medicine, and Department of Experimental Radiation Oncology (W.Z.), The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030; Department of Pathology (D.H.C.), The University of Texas Southwestern Medical School, Dallas, Texas 75390; and Department of Anatomy and Developmental Biology (D.W.), Monash University, Clayton VIC 3800, Australia.
authors.forename: Dagmar
authors.initials: D
authors.lastname: Wilhelm
authors.affiliation: Departments of Molecular and Cellular Biology (Z.L., Y.A.R., S.A.P., J.A., J.S.R.), Pathology and Immunology (S.A.P.), and Obstetrics and Gynecology (J.A.), Baylor College of Medicine, and Department of Experimental Radiation Oncology (W.Z.), The University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030; Department of Pathology (D.H.C.), The University of Texas Southwestern Medical School, Dallas, Texas 75390; and Department of Anatomy and Developmental Biology (D.W.), Monash University, Clayton VIC 3800, Australia.
authors.forename: JoAnne S
authors.initials: JS
authors.lastname: Richards
mesh_terms: D000328:Adult; D000818:Animals; D063646:Carcinogenesis / Q000378:metabolism / Q000473:pathology*; D005260:Female; D000071161:Forkhead Box Protein O1; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors / Q000378:metabolism*; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006062:Gonadotropins / Q000494:pharmacology; D006106:Granulosa Cell Tumor / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D019426:Integrases / Q000378:metabolism; D053533:Keratin-8 / Q000378:metabolism; D037102:Lectins / Q000378:metabolism; D018345:Mice, Knockout; D008875:Middle Aged; D008954:Models, Biological; D051059:PTEN Phosphohydrolase / Q000378:metabolism*; D055758:SOX9 Transcription Factor / Q000378:metabolism; D051785:Smad Proteins / Q000378:metabolism; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
chemical_list: C085126:FOXO1 protein, human; C499517:FOXO3 protein, human; D000071161:Forkhead Box Protein O1; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors; C474033:FoxO3 protein, mouse; C467200:Foxo1 protein, mouse; D006062:Gonadotropins; D053533:Keratin-8; D037102:Lectins; D055758:SOX9 Transcription Factor; D051785:Smad Proteins; C045073:Cre recombinase; D019426:Integrases; D051059:PTEN Phosphohydrolase; C429541:Chil3 protein, mouse; D001619:beta-N-Acetylhexosaminidases