Overexpression of a DENND1A isoform produces a polycystic ovary syndrome theca phenotype.
Polycystic ovary syndrome (PCOS), characterized by increased ovarian androgen biosynthesis, anovulation, and infertility, affects 5-7% of reproductive-age women. Genome-wide association studies identified PCOS candidate loci that were replicated in subsequent reports, including DENND1A, which encodes a protein associated with clathrin-coated pits where cell-surface receptors reside. However, these studies provided no information about functional roles for DENND1A in the pathogenesis of PCOS. DENND1A protein was located in the cytoplasm as well as nuclei of theca cells, suggesting a possible role in gene regulation. DENND1A immunostaining was more intense in the theca of PCOS ovaries. Using theca cells isolated and propagated from normal cycling and PCOS women, we found that DENND1A variant 2 (DENND1A.V2) protein and mRNA levels are increased in PCOS theca cells. Exosomal DENND1A.V2 RNA was significantly elevated in urine from PCOS women compared with normal cycling women. Forced overexpression of DENND1A.V2 in normal theca cells resulted in a PCOS phenotype of augmented CYP17A1 and CYP11A1 gene transcription, mRNA abundance, and androgen biosynthesis. Knock-down of DENND1A.V2 in PCOS theca cells reduced androgen biosynthesis and CYP17A1 and CYP11A1 gene transcription. An IgG specific to DENND1A.V2 also reduced androgen biosynthesis and CYP17 and CYP11A1 mRNA when added to the medium of cultured PCOS theca cells. We conclude that the PCOS candidate gene, DENND1A, plays a key role in the hyperandrogenemia associated with PCOS. These observations have both diagnostic and therapeutic implications for this common disorder.
Departments of Pathology and Obstetrics and Gynecology, Pennsylvania State College of Medicine, Hershey, PA 17033.
Jan M
JM
McAllister
Bhavi
B
Modi
Bruce A
BA
Miller
Jessica
J
Biegler
Richard
R
Bruggeman
Richard S
RS
Legro
Jerome F
JF
Strauss
D000704:Analysis of Variance; D000728:Androgens / Q000096:biosynthesis; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000378:metabolism; D053418:Death Domain Receptor Signaling Adaptor Proteins / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D055106:Genome-Wide Association Study; D020662:Guanine Nucleotide Exchange Factors; D006801:Humans; D017588:Hyperandrogenism / Q000235:genetics* / Q000378:metabolism; D007074:Immunoglobulin G / Q000276:immunology; D007150:Immunohistochemistry; D011085:Polycystic Ovary Syndrome / Q000235:genetics* / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D013254:Steroid 17-alpha-Hydroxylase / Q000378:metabolism; D013799:Theca Cells / Q000378:metabolism
D000728:Androgens; C571984:DENND1A protein, human; D053418:Death Domain Receptor Signaling Adaptor Proteins; D020662:Guanine Nucleotide Exchange Factors; D007074:Immunoglobulin G; D020033:Protein Isoforms; C501892:CYP17A1 protein, human; D013254:Steroid 17-alpha-Hydroxylase; D002786:Cholesterol Side-Chain Cleavage Enzyme