Indian Hedgehog and its targets in human endometrium: menstrual cycle expression and response to CDB-2914.
BACKGROUND
Progesterone is critical for secretory endometrial differentiation in women, but its downstream mediators are poorly understood.


OBJECTIVE
Our objective was to investigate endometrial expression of Indian Hedgehog (IHH) and genes involved in its signaling [smoothened (SMO), patched-1 (PTCH1), glioma-associated oncogene homolog 1 (GLI1), and GLI2] during the menstrual cycle and the effects of the selective progesterone receptor modulator CDB-2914 on its expression.


METHODS
Comparisons between normally cycling volunteers and women with symptomatic fibroids who received CDB-2914 or placebo were made at a clinical research center.


METHODS
Endometrial biopsy was performed on 34 volunteers, 17 additional women with fibroids.


METHODS
Endometrial expression of IHH, SMO, PTCH1, GLI1, and GLI2 by in situ hybridization and/or RT-PCR and IHH, GLI1, and PTCH1 immunohistochemistry were evaluated.


RESULTS
RT-PCR showed expression of IHH, SMO, PTCH1, GLI1, and GLI2, with significant increases in IHH (5.2-fold) and GLI1 (3.6-fold) in endometrium exposed to CDB-2914 compared with placebo. In situ hybridization showed IHH mRNA expression in glands and stroma that was stronger in secretory samples. Among volunteers, IHH and GLI1 immunohistochemistry scores were higher in the secretory than proliferative phase in the nuclei and cytoplasm of glands and stroma (P=0.0002-0.04). Compared with follicular-phase controls, women exposed to CDB-2914 showed increased IHH expression in all compartments except stromal cytoplasm (P=0.0199-0.0423); GLI1 was up-regulated in glandular nuclei and cytoplasm compared with both volunteers and women receiving placebo (P≤0.0416).


CONCLUSIONS
The temporal increase in endometrial IHH and GLI1 during the secretory phase, and their modulation by CDB-2914, suggests progestin regulation and a potential role in endometrial differentiation and implantation.
Wei
Qingxiang
Q
Program in Reproductive and Adult Endocrinology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Building 10, CRC, Room 1-3140, 10 Center Drive, Bethesda, Maryland 20892, USA.
Levens
Eric D
ED
Stefansson
Lilja
L
Nieman
Lynnette K
LK
D001483:Base Sequence; D001706:Biopsy; D017931:DNA Primers; D004717:Endometrium / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D005260:Female; D053823:Hedgehog Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D007889:Leiomyoma / Q000235:genetics / Q000473:pathology; D037101:Luteinizing Hormone, beta Subunit / Q000187:drug effects / Q000378:metabolism; D008597:Menstrual Cycle / Q000187:drug effects / Q000502:physiology*; D009649:Norpregnadienes / Q000494:pharmacology*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D000072079:Patched Receptors; D000072081:Patched-1 Receptor; D017697:Premenopause; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D011980:Receptors, Progesterone / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D012016:Reference Values; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D000071676:Zinc Finger Protein GLI1; D000075746:Zinc Finger Protein Gli2
D017931:DNA Primers; C400551:GLI1 protein, human; C082593:GLI2 protein, human; D053823:Hedgehog Proteins; D051741:Kruppel-Like Transcription Factors; D037101:Luteinizing Hormone, beta Subunit; D009649:Norpregnadienes; D009687:Nuclear Proteins; C000606236:PTCH1 protein, human; D000072079:Patched Receptors; D000072081:Patched-1 Receptor; D011956:Receptors, Cell Surface; D011980:Receptors, Progesterone; D014157:Transcription Factors; D000071676:Zinc Finger Protein GLI1; D000075746:Zinc Finger Protein Gli2; C094854:ulipristal