Direct regulation of HOXA10 by 1,25-(OH)\u3csub\u3e2\u3c/sub\u3eD\u3csub\u3e3\u3c/sub\u3e in human myelomonocytic cells and human endometrial stromal cells

Vitamin D receptor (VDR) and the functionally active form of its ligand, 1,25-(OH)2D3, have been implicated in female reproduction function and myeloid leukemic cell differentiation. HOXA10 is necessary for embryo implantation and fertility, as well as hematopoeitic development. In this study, we identified a direct role of vitamin D in the regulation of HOXA10 in primary human endometrial stromal cells, the human endometrial stromal cell line (HESC), and in the human myelomonocytic cell line, U937. Treatment of primary endometrial stromal cells, or the cell lines HESC and U937 with 1,25-(OH) 2D3 increased HOXA10 mRNA and protein expression. VDR mRNA and protein were detected in primary uterine stromal cells as well as HESC and U937 cells. We cloned the HOXA10 upstream regulatory sequence and two putative vitamin D response elements (VDRE) into luciferase reporter constructs and transfected primary stromal cells and HESC. One putative VDRE (P1: -385 to -434 bp upstream of HOXA10) drove reporter gene expression in response to treatment with 1,25-(OH)2D3. In EMSA, VDR demonstrated binding to the HOXA10 VDRE in the presence of 1,25-(OH)2D3. 1,25-(OH)2D3 up-regulates HOXA10 expression by binding VDR and interacting with a VDRE in the HOXA10 regulatory region. Direct regulation of HOXA10 by vitamin D has implications for fertility and myeloid differentiation. Copyright © 2005 by The Endocrine Society

Fertility and assisted reproductive technologies outcomes of women with non-surgically managed inflammatory bowel diseases: a systematic review.

peer reviewed[en] BACKGROUND AND AIM: In view of their frequent onset during childbearing years, the impact of inflammatory bowel diseases (IBD) on reproductive health is of important concern to young women and to the IBD physician. This study aims to assess the fertility and assisted reproductive technologies outcomes in non-surgically treated IBD female. METHODS: A systematic review was conducted using MEDLINE, SCOPUS and EMBASE (until March 2022) to identify studies assessing fertility and assisted reproductive technologies outcomes in women with non-operated IBD, compared to non-IBD patients. Two reviewers independently selected studies, assessed risk of bias and extracted study data. RESULTS: A total of 14 studies encompassing 18 012 patients with ulcerative colitis (UC) and 14 353 patients with Crohn's disease (CD) were included for analysis. The fertility rate in UC patients and in the general population was comparable, but UC patients tended to have fewer children, mainly by choice. On the contrary, the fertility of CD patients appeared to be reduced. Although a deliberate component cannot be not excluded, the disease itself could affect fertility. Disease activity was associated with reduced fertility in both UC and CD patients. In CD, the colonic involvement of the disease and perianal damage could be associated with subfertility, but data are less consistent. According to the only study reporting the assisted reproductive technologies outcomes, pregnancy rates after in vitro fertilization in subfertile non-operated UC patients and non-IBD patients were similar. CONCLUSION: There is low-quality evidence from observational studies that patients with CD and relapsing UC may have impaired fertility. After assisted reproductive technologies, pregnancy rates of subfertile nonoperated UC patients were similar to those of the general population, although this observation requires further scrutiny in larger studies that should include UC and CD patients

A novel role of the Sp/KLF transcription factor KLF11 in arresting progression of endometriosis.

Endometriosis affects approximately 10% of young, reproductive-aged women. Disease associated pelvic pain; infertility and sexual dysfunction have a significant adverse clinical, social and financial impact. As precise disease etiology has remained elusive, current therapeutic strategies are empiric, unfocused and often unsatisfactory. Lack of a suitable genetic model has impaired further translational research in the field. In this study, we evaluated the role of the Sp/KLF transcription factor KLF11/Klf11 in the pathogenesis of endometriosis. KLF11, a human disease-associated gene is etiologically implicated in diabetes, uterine fibroids and cancer. We found that KLF11 expression was diminished in human endometriosis implants and further investigated its pathogenic role in Klf11-/- knockout mice with surgically induced endometriotic lesions. Lesions in Klf11-/- animals were large and associated with prolific fibrotic adhesions resembling advanced human disease in contrast to wildtype controls. To determine phenotype-specificity, endometriosis was also generated in Klf9-/- animals. Unlike in Klf11-/- mice, lesions in Klf9-/- animals were neither large, nor associated with a significant fibrotic response. KLF11 also bound to specific elements located in the promoter regions of key fibrosis-related genes from the Collagen, MMP and TGF-β families in endometrial stromal cells. KLF11 binding resulted in transcriptional repression of these genes. In summary, we identify a novel pathogenic role for KLF11 in preventing de novo disease-associated fibrosis in endometriosis. Our model validates in vivo the phenotypic consequences of dysregulated Klf11 signaling. Additionally, it provides a robust means not only for further detailed mechanistic investigation but also the ability to test any emergent translational ramifications thereof, so as to expand the scope and capability for treatment of endometriosis

Comparison of the role of Klf9 and Klf11 in an animal endometriosis model.

<p>(<b>A</b>) Endometriotic Lesions (circled) in <i>Klf9-/-</i> animals were either unchanged or had regressed in size from the time of initial peritoneal implantation, when evaluated at necropsy 3 weeks later. (<b>B</b>) Endometriotic lesions in these animals also did not elicit a progressive fibrotic response as seen in <i>Klf11-/-</i> animals. Any adhesions in <i>Klf9-/-</i> animals (black arrow) were flimsy, transparent and peri-lesional in extent. (<b>C</b>) Tissue planes were unaltered with preservation of intra-abdominal anatomy. Consequently, intestinal length was not foreshortened due to lack of mesenteric fibrosis (unraveled intestinal loops denoted by white arrows). (<b>D</b>): A composite adhesion score for each mouse was determined and compared between <i>Klf11-/-</i>, <i>Klf9-/-</i> and wildtype genotypes, based on the Murine Adhesion Scoring System. The adhesion score for <i>Klf11-/-</i> mice (81.7±4.8) was significantly different from that calculated in either <i>Klf9-/-</i> (12.3±1.8) or wildtype animals (9.17±0.8); (* = p<0.05, 14 lesions/genotype). The scores objectively reflected observed anatomical findings in these animals.</p

KLF11 expression in human urogenital tissues, uterine eutopic endometrium and endometriosis.

<p>(<b>A</b>) Pooled Human Organ-specific RNA (1 µg/reaction) was analyzed for <i>KLF11</i> mRNA expression by PCR. mRNA levels of the housekeeping gene <i>GAPDH</i> were simultaneously assessed as loading control. Relative organ-specific <i>KLF11</i> mRNA expression levels were determined by densitometric comparison to corresponding <i>GAPDH</i> levels. KLF11 mRNA expression levels were increased in urogenital tissues such as the kidney, ovary, placenta, testes and uterus (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060165#pone-0060165-g001" target="_blank">Figure 1A</a>, arrows) compared to other non-urogenital tissues. (<b>B</b>) Expression of KLF11 mRNA and protein were also assessed in two cell lines Ishikawa, a well-differentiated endometrial adenocarcinoma cell line and Human Endometrial Stromal Cells (T-HESC), which are frequently used as model endometrial epithelial and stromal cell lines respectively. <i>KLF11</i>/KLF11 were expressed in both cell-lines; <i>GAPDH/</i>α-TUBULIN were used as a reference controls. To qualitatively demonstrate KLF11 expression in each of these endometrial cell lines, lysate for RNA or protein extraction was obtained from one 10-cm cell culture dish of confluent cells. Due to much lower abundance of stromal compared to epithelial cells there appears to be differential <i>KLF11</i>/KLF11 expression between the cell-types. However, this difference was also reflected in expression of the loading controls <i>GAPDH</i>/α-TUBULIN between the two cell lines. (<b>C, D</b>) KLF11 expression was also evaluated by immunohistochemistry in eutopic endometrium as well as in ectopic endometrial implants (magnification: 200×: panel; 400×: inset). KLF11 was expressed in the nuclei and cytoplasm of epithelial as well as stromal cells in both eutopic endometrium as well as in endometriotic implants. KLF11 expression was diminished in endometriotic implants compared to eutopic endometrium. Representative samples shown. (<b>E</b>) KLF11 expression levels were compared and scored in paired eutopic endometrium and ectopic endometriotic implants obtained from the same patient (N = 28 paired samples). The expression was significantly reduced in the implants (epithelium: 103±2.6; stroma: 40.4±3.9) compared to that in eutopic uterine endometrium (epithelium: 278±2.5; stroma: 114±8.7). * and. = p<0.05 and represent comparisons between epithelial and stromal cells in eutopic and ectopic endometrium respectively.</p