Next Article in Journal
Mechanisms and Neuroimaging Patterns of Hypereosinophilia-Related Ischemic Stroke: A Narrative Review through Three Cases
Next Article in Special Issue
Association between Endometriosis, Irritable Bowel Syndrome and Eating Disorders: ENDONUT Pilot Study
Previous Article in Journal
Current and Future Insights for Optimizing Antithrombotic Therapy to Reduce the Burden of Cardiovascular Ischemic Events in Patients with Acute Coronary Syndrome
Previous Article in Special Issue
Factors Associated with Increased Analgesic Use in German Women with Endometriosis during the COVID-19 Pandemic
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
JCM
Volume 11
Issue 19
10.3390/jcm11195606
jcm-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Expression of Fucosyltransferase 4 (FUT4) mRNA Is Increased in Endometrium from Women with Endometriosis

by
Marta Żeberkiewicz
1,2,
Anna Hyc
1,
Anna Iwan
1,
Aneta Zwierzchowska
3,4,
Aneta Ścieżyńska
1,
Ilona Kalaszczyńska
1,5,
Ewa Barcz
3,4 and
Jacek Malejczyk
1,5,*
1
Department of Histology and Embryology, Medical University of Warsaw, Chałubińskiego 5, 02-004 Warsaw, Poland
2
Postgraduate School of Molecular Medicine, Medical University of Warsaw, 02-091 Warsaw, Poland
3
Chair and Clinic of Gynecology and Obstetrics, Faculty of Medicine, Collegium Medicum, Cardinale Stefan Wyszyński University, 04-749 Warsaw, Poland
4
Department of Obstetrics and Gynecology, Multidisciplinary Hospital Warsaw-Miedzylesie, 04-749 Warsaw, Poland
5
Diagendo Ltd., 05-504 Bobrowiec, Poland
*
Author to whom correspondence should be addressed.
J. Clin. Med. 2022, 11(19), 5606; https://doi.org/10.3390/jcm11195606
Submission received: 9 August 2022 / Revised: 14 September 2022 / Accepted: 19 September 2022 / Published: 23 September 2022
(This article belongs to the Special Issue Endometriosis: Current Perspectives on Diagnosis and Treatment)
Browse Figures
Versions Notes

Abstract

:
Endometriosis is a common gynecological disorder defined as the presence of endometrial-like tissue (glands and stroma) outside the uterus. The etiopathogenesis of endometriosis is still poorly recognized. It is speculated that stage-specific embryonic antigen 1 (SSEA-1)-positive stem-like glandular epithelial cells may contribute to the development of the disease. The synthesis of SSEA-1 is mediated by fucosyltransferase 4 encoded by the FUT4 gene. Therefore, this study aimed to evaluate the specific expression of FUT4 mRNA in biopsies of the endometrium from women with and without endometriosis. FUT4 mRNA levels were examined in 49 women with laparoscopically confirmed endometriosis and 28 controls by means of quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The expression of FUT4 mRNA was significantly increased in the endometrium of patients with endometriosis when compared to the controls (p < 0.0001). Expression of FUT4 mRNA in the endometrium was correlated with the severity of endometriosis (rs = 0.5579, p < 0.0001); however, there were no differences in endometrial FUT4 mRNA expression when comparing endometriotic lesions from various locations. The discriminatory ability of FUT4 mRNA expression was evaluated by receiver-operating characteristics (ROC), which showed high statistical significance (AUC = 0.90, p < 0.0001), thus indicating that an increased level of endometrial FUT4 mRNA may serve as a specific marker for endometriosis.

1. Introduction

Endometriosis is a common estrogen-dependent gynecological disorder affecting approximately 10% of women of reproductive age. The disease is defined as the presence of endometrial glands and stroma outside the uterus, most frequently in the ovaries and peritoneal cavity. Clinical manifestations may include dysmenorrhea, dyspareunia, chronic pelvic pain and infertility. Endometriosis is associated with chronic pelvic inflammation and may account for up to 50% of female infertility [1,2,3,4,5]. The disease may significantly affect one’s quality of life and, therefore, constitutes an important social, socioeconomic and clinical problem.
The etiopathogenesis of endometriosis is still poorly understood [5,6,7,8]. According to the most commonly accepted theory, Sampson’s retrograde menstruation theory, endometriosis results from shed endometrial cells that survive in the peritoneal cavity, undergo implantation and form ectopic foci of endometrial-like tissue [9]. The mechanisms leading to the survival, implantation and growth of the ectopic endometrial cells may include decreased apoptosis, resistance to eradication by the local immune system, increased adhesiveness, invasiveness, proangiogenic potential and epithelial-to-mesenchymal transition [6,10,11,12,13,14].
There is a growing bulk of evidence that the stem-like precursor cells may contribute to the development of endometriotic lesions [15,16,17,18,19,20,21]. These may include mesenchymal stem-like cells that have been identified and characterized among endometrial stromal cells [19,20,22,23,24,25]. Of special interest, however, are epithelial precursor cells expressing N-cadherin (NCAD/CDH2) and/or stage-specific embryonic antigen 1 (SSEA-1/CD15) that were recently described in the basal glandular endometrial epithelium, where they seem to be responsible for the regeneration of endometrial glands following menstruation [26,27]. The NCAD+ cells are present in the most basal parts of the glands and gradually change their phenotype to the SSEA-1+ cells [15]. The SSEA-1+ cells display higher activity of telomerase and express nuclear SOX9 and β-catenin, thus suggesting activation of the WNT pathway [27]. SSEA-1 and SOX9 expression were also identified in ectopic endometrial cells. Most recently, it has been reported that the numbers of SSEA-1+ and SOX9+ glandular epithelial cells are significantly increased in the eutopic endometrium of women with endometriosis, indicating that they may contribute to the formation of ectopic lesions [28].
SSEA-1, better known as Sialyl-LewisX (SLeX) or CD15, is a tetrasaccharide carbohydrate composed of sialic acid, fucose and N-acetyllactosamine, which is usually attached to the cell surface by O-glycans [29]. SLeX is involved in embryonic development [30,31], and in adult humans, SLeX plays an essential role in oocyte fertilization [32,33], leukocyte interactions and homing, as well as in the induction of inflammation [34]. Additionally, SLeX is involved in cancer progression, metastasis and angiogenesis [35]. The synthesis of SLeX is mediated by fucosyltransferases 4 and 9 (FUT4, 9), which transfer fucose to N-acetyllactosamine polysaccharides [36]. The FUT4 gene, encoding a 41 kDa protein with α(1,3)-fucosyltransferase activity [37], is located on the long arm of chromosome 11 in position 11q21 [38].
Although endometriosis appears to be associated with increased numbers of SSEA-1/SLeX+ cells in the eutopic endometrium, it is highly plausible that this phenomenon may be related to the upregulated expression of the FUT4 gene. Therefore, the present study was aimed at comparing the expression of specific FUT4 mRNA in the endometrium of control women and women with endometriosis.

2. Materials and Methods

2.1. Patients and Material Collection

The study group consisted of 77 women who were admitted to the 1st Department of Obstetrics and Gynecology, Medical University of Warsaw, Poland between January 2010 and December 2015. The endometriosis group consisted of 49 patients in whom the disease was confirmed by laparoscopic and histological examinations. The severity of endometriosis was classified according to the revised American Fertility Society (rAFS) criteria [39]. The control group consisted of 28 women without any clinical or sonographic symptoms or signs of endometriosis who were subjected to the removal of a cervical polyp or underwent cervical biopsy due to an abnormal Pap smear result. Women in whom histopathological examination revealed cervical dysplasia grade 2+ or dysplasia within the cervical polyp were not included. All patients and control subjects had normal menstrual cycles, none of them suffered from any other chronic endocrine or autoimmune disorder and they had not been subjected to hormonal pharmacological treatment for at least 3 months prior to the study. The phase of the menstrual cycle was established based on the date of the last menstrual bleed and the mean cycle duration, as well as being confirmed by pelvic ultrasound examination. Written informed consent was obtained from all participants according to the guidelines of the Declaration of Helsinki and the study was approved by the Institutional Bioethical Review Board of the Medical University of Warsaw, Poland (ref. no. KB/223/2009 and KB/59/2012). The demographic and clinical characteristics of the patients from the endometriosis and control groups are presented in Table 1. As can be seen, there were no significant differences between the endometriosis and control groups, except for the parity and infertility status.
Tissue samples were collected from the participants during the mid-proliferative and mid-secretory phase of the menstrual cycle directly before surgical intervention. Endometrial tissue samples were obtained by an aspiration biopsy with a Pipelle® catheter (Pipelle de Cornier, Laboratoire C.C.D., Paris, France). Collected specimens were immediately placed in 5 volumes of RNAlaterTM solution (TermoFisher Scientific, Waltham, MA, USA), stored at 4 °C for one day and then stored frozen at −70 °C until RNA isolation was performed.

2.2. RNA Isolation and Quantitative Reversed-Transcription Polymerase Chain Reaction (qRT-PCR) Analysis

The total RNAs from the endometrial tissue samples (5 mm3) were isolated and purified using a NucleoSpin® miRNA Kit with rDNase digestion for removal of genomic DNA (Macherey-Nagel GmbH and Co. KG, Düren, Germany) according to the detailed protocol provided by the manufacturer. The quantity and quality of the isolated RNA were evaluated spectrophotometrically using an ND-2000 Spectrophotometer NanoDrop2000 with software for nucleic acid analysis (ThermoFisher Scientific, Waltham, MA, USA).
A total of 2 μg of large RNA was reverse-transcribed using a High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Cheshire, UK) according to the manufacturer’s protocol. Briefly, 2 μL of 10 × RT buffer, 0.8 μL of 25 × dNTP Mix, 2 μL of 10 × Random Primers, 1 μL of the Multiscribe Reverse Transcriptase, 4.2 μL of nuclease-free water and 10 μL of large RNA sample were used per reaction. The reaction was run in an Eppendorf Mastercycler gradient with the following conditions: 25 °C for 10 min, 37 °C for 120 min and 85 °C for 5 s. The cDNA samples obtained were stored at −20 °C until qRT-PCR assays were performed.
qRT-PCR was performed in an ABI PRISM 7500 system (Applied Biosystems, Waltham, MA, USA) using 96-well optical plates. For mRNA expression analysis, Hs01106466_s1 for FUT4 and Hs99999905_m1 for GAPDH, FAM-labeled TaqMan probes (Applied Biosystems) were used. Reactions were run in a total volume of 20 μL with TaqMan Universal Master Mix, the appropriate specific primer set, MGB probe and 50 ng of cDNA template. All qRT-PCR assays were carried out under universal conditions with an initial denaturation at 95 °C for 10 min, followed by product amplification through 50 successive cycles of 95 °C for 15 sec and 60 °C for 1 min. Each sample was run in triplicate. The FUT4 mRNA data were expressed relative to the GAPDH mRNA. The data analysis was carried out with Sequence Detection System (SDS) v1.2 software (Applied Biosystems) using the ΔCt method, and the relative mRNA expression is shown as 2−ΔCt.

2.3. Statistical Analysis

All statistical analyses and graphical presentations were generated using GraphPad Prism 8.2.0 (GraphPad Software, San Diego, CA, USA). The normality of the data distribution was checked by tests included in the GraphPad Prism package. Differences between groups were determined by Student’s t-test or the Mann–Whitney U-test when applicable. The Kruskal–Wallis test followed by Dunn’s multiple comparison test was used when comparing more than two groups. The two-tailed Spearman’s correlation coefficient (rs) was used for correlation analyses. Generation of a receiver-operating characteristics (ROC) curve and calculation of the area under the curve (AUC) and 95% confidence intervals (95% CI) were used to assess the predictive power of FUT4 mRNA evaluation.
The results are shown as the mean ± SD or median with the interquartile range. A p-value < 0.05 was considered statistically significant.

3. Results

The relative levels of FUT4 mRNA expression in eutopic endometrium samples from the control group and women with endometriosis were compared and analyzed with regard to the phase of the menstrual cycle, the stage of disease and the lesion localization.
qRT-PCR analysis revealed that the relative FUT4 mRNA expression was significantly higher in eutopic endometrium samples from women with endometriosis compared to the control group (p < 0.0001) (Figure 1). There were no significant differences in FUT4 mRNA expression regarding the phase of the menstrual cycle in the endometrium samples from control subjects and patients with endometriosis (data not shown).
Analysis of the association between FUT4 mRNA expression in the endometrium and the stage of endometriosis showed that FUT4 mRNA expression was significantly upregulated in samples collected from patients with both minimal/mild and moderate/severe stages of disease when compared with the control endometrium samples (Figure 2A). We did not discover any significant differences between patients with less or more advanced stages of endometriosis. Nevertheless, Spearman’s rank correlation analysis revealed that there was a statistically significant positive association (rs = 0.5579, p < 0.0001) between FUT4 mRNA levels and the disease severity (Figure 2B).
Localization of endometriotic lesions (ovarian, peritoneal or both) had no impact on the expression of FUT4 mRNA in the endometrium of women with endometriosis. Irrespective of localization in all cases, the FUT4 mRNA levels were significantly higher compared to the endometrium samples from the control group (Figure 3).
The discriminatory ability of FUT4 mRNA expression in the total population analyzed was evaluated by ROC analysis. As can be seen in Figure 4, the AUC value was very high and was associated with high statistical significance (AUC = 0.90, 95% CI = 0.81–0.99, p < 0.0001). The diagnostic value of FUT4 mRNA evaluation is summarized in Table 2. As can be seen, the relative expression of FUT4 mRNA with a cutoff value of 0.002793 units (2−ΔCt) conferred 94.12% sensitivity and 89.29% specificity. The positive predictive value (PPV) and the negative predictive value (NPV) for the diagnosis of endometriosis were 93.9% and 89.3%, respectively. Youden’s statistics revealed a value of 0.8341.

4. Discussion

This study is the first to present data showing that FUT4 mRNA levels are significantly upregulated in the eutopic endometrium of patients with endometriosis when compared to the endometrium of healthy control women. ROC analysis revealed that the expression of FUT4 mRNA in samples of the eutopic endometrium could serve as a suitable marker for the minimally invasive diagnosis of endometriosis. These data also suggested that FUT4 may play an important role in the etiopathogenesis of this disease.
The expression of FUT4 in mammalian endometrium was reported to be specific for glandular and luminal epithelium [27,28,40,41], and as previously mentioned, it characterizes a population of endometrial glandular precursor/stem cells [27,28]. It has been suggested that FUT4 expression in the endometrium may be upregulated by progesterone and thus related to the phase of the menstrual or estrus cycle [40,41,42,43]. Conversely, other studies in mice claimed that FUT4 expression was suppressed by progesterone and stimulated by estrogen, indicating that the effects of both hormones are antagonistic [44]. In the present study, we did not discover any significant difference in FUT4 mRNA expression in the endometrium when comparing control women to women with endometriosis in the mid-proliferating and mid-secretory phase. Similarly, no changes were observed in serum levels of SLeX, a product of FUT4 activity, during the menstrual cycle in healthy women [45]. Inconsistent results for FUT4 mRNA expression across the literature in the case of human material may be related to different methods of sample collection, sample origin, PCR method and other factors. Thus, the mechanisms that regulate FUT4 expression during the menstrual cycle require further investigation.
Despite an increased number of SSEA-1+ cells, Hapangama et al. were unable to show increased expression of FUT4 mRNA in the eutopic endometrium of women with endometriosis [28]. This may have been due to the limited number of patients since in the present study, we found that the expression of FUT4 mRNA was markedly upregulated with very high statistical significance. A relationship between increased serum concentrations of SLeX and endometriosis was previously reported by Iwanari et al. [45,46]. However, increased serum concentrations of SLeX were also found in patients with ovarian [45,46] and breast cancer [47], meaning its diagnostic specificity appears to be limited. Our present results show that this problem may be circumvented by tissue-specific evaluation of FUT4 mRNA. ROC analysis confirmed that the expression of FUT4 mRNA in the eutopic endometrium is a specific marker of endometriosis. Evaluation of FUT4 mRNA for the diagnosis of endometriosis fulfills the criteria of high accuracy, sensitivity and specificity with high positive and negative prediction values. Samples of endometrium may be easily obtained for mRNA evaluations by a minimally invasive aspiration biopsy during a visit to a gynecological office. Therefore, the evaluation of FUT4 mRNA appears to be a suitable method for a minimally invasive diagnosis of endometriosis. This, however, will require further verification by an independent replicate study using different cohorts of patients.
Increased expression of FUT4 mRNA in endometriosis was observed irrespective of the lesion type or location. Furthermore, there was a positive correlation between the levels of FUT4 mRNA and the disease severity. This may support the role of FUT4 activity and FUT4-mediated glycosylation in the etiopathogenesis of endometriosis.
It is plausible that higher levels of FUT4 mRNA in the eutopic endometriosis tissue simply reflect the persistence of increased numbers of SSEA-1+ precursor/stem cells. However, the levels may also be explained by increased expression of the FUT4 gene in endometriosis-associated SSEA-1+ epithelial glandular cells [28]. The expression of fucosyltransferases and their activity are significantly increased in various tumors [48,49]; however, the mechanisms responsible for this phenomenon remain obscure. Possible stimulatory factors may include hypoxia [50], proinflammatory cytokines [51] or cell stress [52]. These factors are considered to play a part in the etiopathogenesis of endometriosis; therefore, it is plausible that they may contribute to increased expression of FUT4 mRNA in the endometrium of women suffering from endometriosis. This hypothesis, however, needs to be further investigated.
In a variety of tumor cells, overexpression of FUT4 is attributed to their increased proliferation rate, decreased susceptibility to apoptosis as well as increased adhesiveness, invasiveness and metastasis, and is generally related to a poor prognosis [48,49]. Interestingly, fucosylation may also be responsible for the modification of a variety of signaling molecules including TGF-β receptors that, in turn, may contribute to an increased rate of epithelial-to-mesenchymal transition and matrix metalloproteinases-related invasiveness [48]. It is not known whether similar effects may be observed in the course of endometriosis. However, it is intriguing to speculate that, in addition to being a marker of endometriosis, overexpression of FUT4 may play an important role in the development and persistence of endometriotic lesions. In future research, a pathogenic role of FUT4 in endometriosis awaits extensive investigation.

5. Conclusions

The results of the present study show that the expression of FUT4 mRNA in the endometrium of patients with endometriosis is significantly increased when compared to healthy women. Therefore, FUT4 mRNA from the endometrium may serve as a specific marker that can be used to diagnose endometriosis in a minimally invasive manner. Furthermore, it appears that FUT4-mediated fucosylation may play an important role in the development and persistence of endometriotic lesions and, therefore, may be considered a target for endometriosis therapies.

Author Contributions

M.Ż.: design of the project, sample collection, experiment conduction, data analysis, interpretation of data, critical revision and final approval of the manuscript. A.H.: design of the project, experiment conduction, data analysis, interpretation of data, critical revision and final approval of the manuscript. A.I.: design of the project, experiment conduction, data analysis, interpretation of data, critical revision and final approval of the manuscript. A.Z.: design of the project, patients’ enrollment, sample collection, clinical data acquisition, data analysis, interpretation of data, critical revision and final approval of the manuscript. A.Ś.: design of the project, data analysis, interpretation of data, critical revision and final approval of the manuscript. I.K.: interpretation of data, critical revision and final approval of the manuscript. E.B.: design of the project, patients’ enrollment, sample collection, clinical data acquisition, data analysis, interpretation of data, critical revision and final approval of the manuscript. J.M.: design of the project, data analysis, interpretation of data, writing of the manuscript, critical revision and final approval of the manuscript. All authors have read and agreed to the published version of the manuscript.

Funding

The study was supported by the 1st Faculty of Medicine, Warsaw Medical University intramural grants 1M15/18 and 1M15/19. M.Ż. was a fellow at the Postgraduate School of Molecular Medicine, Warsaw, Poland. We would like to thank Barbara Tomaszyńska for her excellent technical assistance. We are also indebted to the patients enrolled in the study.

Institutional Review Board Statement

The study was conducted in accordance with the Declaration of Helsinki and approved by the Institutional Bioethical Review Board of the Medical University of Warsaw, Poland (ref. no. KB/223/2009, 15 December 2009 and KB/59/2012, 13 March 2012).

Informed Consent Statement

Informed consent was obtained from all patients involved in the study.

Data Availability Statement

Specific data are available from the authors upon request.

Conflicts of Interest

M.Ż., A.H., A.I., A.Z., A.Ś., E.B. and J.M. are listed as co-inventors on the Polish (P.431605) and European (PCT/PL2020/050078) patent application. I.K. and J.M. are co-owners of Diagendo Ltd. The authors declare no other conflicts of interest.

References

  1. Broi, M.G.D.; Ferriani, R.A.; Navarro, P.A. Ethiopathogenic mechanisms of endometriosis-related infertility. JBRA Assist. Reprod. 2019, 23, 273–280. [Google Scholar] [CrossRef]
  2. Giudice, L.C.; Kao, L.C. Endometriosis. Lancet 2004, 364, 1789–1799. [Google Scholar] [CrossRef]
  3. Practice Committee of the American Society for Reproductive Medicine. Endometriosis and infertility. Fertil. Steril. 2006, 86, S156–S160. [Google Scholar] [CrossRef] [PubMed]
  4. Zondervan, K.T.; Becker, C.M.; Koga, K.; Missmer, S.A.; Taylor, R.N.; Vigano, P. Endometriosis. Nat. Rev. Dis. Primers 2018, 4, 9. [Google Scholar] [CrossRef] [PubMed]
  5. Vercellini, P.; Vigano, P.; Somigliana, E.; Fedele, L. Endometriosis: Pathogenesis and treatment. Nat. Rev. Endocrinol. 2014, 10, 261–275. [Google Scholar] [CrossRef] [PubMed]
  6. Klemmt, P.A.B.; Starzinski-Powitz, A. Molecular and Cellular Pathogenesis of Endometriosis. Curr. Womens Health Rev. 2018, 14, 106–116. [Google Scholar] [CrossRef] [PubMed]
  7. Koninckx, P.R.; Ussia, A.; Adamyan, L.; Wattiez, A.; Gomel, V.; Martin, D.C. Pathogenesis of endometriosis: The genetic/epigenetic theory. Fertil. Steril. 2019, 111, 327–340. [Google Scholar] [CrossRef]
  8. Patel, B.G.; Lenk, E.E.; Lebovic, D.I.; Shu, Y.; Yu, J.; Taylor, R.N. Pathogenesis of endometriosis: Interaction between Endocrine and inflammatory pathways. Best Pract. Res. Clin. Obstet. Gynaecol. 2018, 50, 50–60. [Google Scholar] [CrossRef]
  9. Sampson, J.A. Metastatic or Embolic Endometriosis, due to the Menstrual Dissemination of Endometrial Tissue into the Venous Circulation. Am. J. Pathol. 1927, 3, 93–110 143. [Google Scholar]
  10. Gazvani, R.; Templeton, A. Peritoneal environment, cytokines and angiogenesis in the pathophysiology of endometriosis. Reproduction 2002, 123, 217–226. [Google Scholar] [CrossRef]
  11. Klemmt, P.A.; Carver, J.G.; Koninckx, P.; McVeigh, E.J.; Mardon, H.J. Endometrial cells from women with endometriosis have increased adhesion and proliferative capacity in response to extracellular matrix components: Towards a mechanistic model for endometriosis progression. Hum. Reprod. 2007, 22, 3139–3147. [Google Scholar] [CrossRef] [Green Version]
  12. Matarese, G.; De Placido, G.; Nikas, Y.; Alviggi, C. Pathogenesis of endometriosis: Natural immunity dysfunction or autoimmune disease? Trends Mol. Med. 2003, 9, 223–228. [Google Scholar] [CrossRef]
  13. Yang, Y.M.; Yang, W.X. Epithelial-to-mesenchymal transition in the development of endometriosis. Oncotarget 2017, 8, 41679–41689. [Google Scholar] [CrossRef] [PubMed]
  14. Sciezynska, A.; Komorowski, M.; Soszynska, M.; Malejczyk, J. NK Cells as Potential Targets for Immunotherapy in Endometriosis. J. Clin. Med. 2019, 8, 1468. [Google Scholar] [CrossRef] [PubMed]
  15. Cousins, F.L.; Dorien, F.O.; Gargett, C.E. Endometrial stem/progenitor cells and their role in the pathogenesis of endometriosis. Best Pract. Res. Clin. Obstet. Gynaecol. 2018, 50, 27–38. [Google Scholar] [CrossRef]
  16. Gargett, C.E.; Schwab, K.E.; Deane, J.A. Endometrial stem/progenitor cells: The first 10 years. Hum. Reprod. Update 2016, 22, 137–163. [Google Scholar] [CrossRef]
  17. Nikoo, S.; Ebtekar, M.; Jeddi-Tehrani, M.; Shervin, A.; Bozorgmehr, M.; Vafaei, S.; Kazemnejad, S.; Zarnani, A.-H. Menstrual blood-derived stromal stem cells from women with and without endometriosis reveal different phenotypic and functional characteristics. Mol. Hum. Reprod. 2014, 20, 905–918. [Google Scholar] [CrossRef]
  18. Gargett, C.E.; Schwab, K.E.; Brosens, J.J.; Puttemans, P.; Benagiano, G.; Brosens, I. Potential role of endometrial stem/progenitor cells in the pathogenesis of early-onset endometriosis. Mol. Hum. Reprod. 2014, 20, 591–598. [Google Scholar] [CrossRef] [PubMed]
  19. Gotte, M.; Wolf, M.; Staebler, A.; Buchweitz, O.; Kiesel, L.; Schuring, A.N. Aberrant expression of the pluripotency marker SOX-2 in endometriosis. Fertil. Steril. 2011, 95, 338–341. [Google Scholar] [CrossRef]
  20. Gotte, M.; Wolf, M.; Staebler, A.; Buchweitz, O.; Kelsch, R.; Schuring, A.N.; Kiesel, L. Increased expression of the adult stem cell marker Musashi-1 in endometriosis and endometrial carcinoma. J. Pathol. 2008, 215, 317–329. [Google Scholar] [CrossRef]
  21. Schuring, A.N.; Dahlhues, B.; Korte, A.; Kiesel, L.; Titze, U.; Heitkotter, B.; Ruckert, C.; Gotte, M. The endometrial stem cell markers notch-1 and numb are associated with endometriosis. Reprod. Biomed. Online 2018, 36, 294–301. [Google Scholar] [CrossRef] [PubMed]
  22. Masuda, H.; Anwar, S.S.; Buhring, H.J.; Rao, J.R.; Gargett, C.E. A novel marker of human endometrial mesenchymal stem-like cells. Cell Transplant 2012, 21, 2201–2214. [Google Scholar] [CrossRef] [PubMed]
  23. Schuring, A.N.; Schulte, N.; Kelsch, R.; Ropke, A.; Kiesel, L.; Gotte, M. Characterization of endometrial mesenchymal stem-like cells obtained by endometrial biopsy during routine diagnostics. Fertil. Steril. 2011, 95, 423–426. [Google Scholar] [CrossRef] [PubMed]
  24. Gurung, S.; Werkmeister, J.A.; Gargett, C.E. Inhibition of Transforming Growth Factor-β Receptor signaling promotes culture expansion of undifferentiated human Endometrial Mesenchymal Stem/stromal Cells. Sci. Rep. 2015, 5, 15042. [Google Scholar] [CrossRef] [PubMed]
  25. Gargett, C.E.; Gurung, S. Endometrial Mesenchymal Stem/Stromal Cells, Their Fibroblast Progeny in Endometriosis, and More. Biol. Reprod. 2016, 94, 129. [Google Scholar] [CrossRef]
  26. Nguyen, H.P.T.; Xiao, L.; Deane, J.A.; Tan, K.-S.; Cousins, F.L.; Masuda, H.; Sprung, C.N.; Rosamilia, A.; Gargett, C.E. N-cadherin identifies human endometrial epithelial progenitor cells by in vitro stem cell assays. Hum. Reprod. 2017, 32, 2254–2268. [Google Scholar] [CrossRef]
  27. Valentijn, A.J.; Palial, K.; Al-Lamee, H.; Tempest, N.; Drury, J.; Von Zglinicki, T.; Saretzki, G.; Murray, P.; Gargett, C.E.; Hapangama, D.K. SSEA-1 isolates human endometrial basal glandular epithelial cells: Phenotypic and functional characterization and implications in the pathogenesis of endometriosis. Hum. Reprod. 2013, 28, 2695–2708. [Google Scholar] [CrossRef]
  28. Hapangama, D.K.; Drury, J.; Da Silva, L.; Al-Lamee, H.; Earp, A.; Valentijn, A.J.; Edirisinghe, D.P.; Murray, P.A.; Fazleabas, A.T.; Gargett, C.E. Abnormally located SSEA1+/SOX9+ endometrial epithelial cells with a basalis-like phenotype in the eutopic functionalis layer may play a role in the pathogenesis of endometriosis. Hum. Reprod. 2019, 34, 56–68. [Google Scholar] [CrossRef]
  29. Stanley, P.; Cummings, R.D. Structures Common to Different Glycans. In Essentials of Glycobiology, 3rd ed.; Varki, A., Cummings, R.D., Esko, J.D., Stanley, P., Hart, G.W., Aebi, M., Darvill, A.G., Kinoshita, T., Packer, N.H., Prestegard, J.H., Eds.; Cold Spring Harbor Laboratory Press: Cold Spring Harbor, NY, USA, 2015; pp. 161–178. [Google Scholar] [CrossRef]
  30. Fenderson, B.A.; Eddy, E.M.; Hakomori, S. Glycoconjugate expression during embryogenesis and its biological significance. Bioessays 1990, 12, 173–179. [Google Scholar] [CrossRef]
  31. Handa, K.; Hakomori, S.I. Changes of glycoconjugate expression profiles during early development. Glycoconj. J. 2017, 34, 693–699. [Google Scholar] [CrossRef]
  32. Pang, P.C.; Chiu, P.C.; Lee, C.L.; Chang, L.Y.; Panico, M.; Morris, H.R.; Haslam, S.M.; Khoo, K.H.; Clark, G.F.; Yeung, W.S.; et al. Human sperm binding is mediated by the sialyl-Lewis(x) oligosaccharide on the zona pellucida. Science 2011, 333, 1761–1764. [Google Scholar] [CrossRef] [PubMed]
  33. Clark, G.F. The role of carbohydrate recognition during human sperm-egg binding. Hum. Reprod. 2013, 28, 566–577. [Google Scholar] [CrossRef] [PubMed]
  34. Kawashima, H. Roles of sulfated glycans in lymphocyte homing. Biol. Pharm. Bull. 2006, 29, 2343–2349. [Google Scholar] [CrossRef] [PubMed]
  35. Kannagi, R.; Izawa, M.; Koike, T.; Miyazaki, K.; Kimura, N. Carbohydrate-mediated cell adhesion in cancer metastasis and angiogenesis. Cancer Sci. 2004, 95, 377–384. [Google Scholar] [CrossRef]
  36. Nakayama, F.; Nishihara, S.; Iwasaki, H.; Kudo, T.; Okubo, R.; Kaneko, M.; Nakamura, M.; Karube, M.; Sasaki, K.; Narimatsu, H. CD15 expression in mature granulocytes is determined by α1,3-fucosyltransferase IX, but in promyelocytes and monocytes by α1,3-fucosyltransferase IV. J. Biol. Chem. 2001, 276, 16100–16106. [Google Scholar] [CrossRef]
  37. Goelz, S.E.; Hession, C.; Goff, D.; Griffiths, B.; Tizard, R.; Newman, B.; Chi-Rosso, G.; Lobb, R. ELFT: A gene that directs the expression of an ELAM-1 ligand. Cell 1990, 63, 1349–1356. [Google Scholar] [CrossRef]
  38. Reguigne, I.; James, M.R.; Richard, C.W., 3rd; Mollicone, R.; Seawright, A.; Lowe, J.B.; Oriol, R.; Couillin, P. The gene encoding myeloid alpha-3-fucosyl-transferase (FUT4) is located between D1 1S388 and D11S919 on 11q21. Cytogenet. Cell Genet. 1994, 66, 104–106. [Google Scholar] [CrossRef]
  39. Society, A.F. Revised American Fertility Society classification of endometriosis: 1985. Fertil. Steril. 1985, 43, 351–352. [Google Scholar] [CrossRef]
  40. Ponnampalam, A.P.; Rogers, P.A. Expression and regulation of fucosyltransferase 4 in human endometrium. Reproduction 2008, 136, 117–123. [Google Scholar] [CrossRef]
  41. Zhang, Y.-M.; Zhang, Y.-Y.; Bulbul, A.; Shan, X.; Wang, X.-Q.; Yan, Q. Baicalin promotes embryo adhesion and implantation by upregulating fucosyltransferase IV (FUT4) via Wnt/beta-catenin signaling pathway. FEBS Lett. 2015, 589, 1225–1233. [Google Scholar] [CrossRef] [Green Version]
  42. Ponnampalam, A.P.; Weston, G.C.; Trajstman, A.C.; Susil, B.; Rogers, P.A. Molecular classification of human endometrial cycle stages by transcriptional profiling. Mol. Hum. Reprod. 2004, 10, 879–893. [Google Scholar] [CrossRef] [PubMed]
  43. Lee, S.-H.; Song, E.-J.; Hwangbo, Y.; Lee, S.; Park, C.K. Change of uterine histroph proteins during follicular and luteal phase in pigs. Anim. Reprod. Sci. 2016, 168, 26–33. [Google Scholar] [CrossRef] [PubMed]
  44. Wang, H.; Ge, C.H.; Kong, Y.; Xin, Y.; Zhu, Z.M. Ovary Hormonal Control of Le(y) Oligosaccharide Expression during Peri-implantation of Mouse Endometrium. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 2001, 33, 542–546. [Google Scholar] [PubMed]
  45. Iwanari, O.; Miyako, J.; Date, Y.; Nakayama, S.; Kijima, S.; Moriyama, M.; Takahashi, K.; Yoshino, N.; Karino, K.; Endoh, J.; et al. Differential diagnosis of ovarian cancer, benign ovarian tumor and endometriosis by a combination assay of serum sialyl SSEA-1 antigen and CA125 levels. Gynecol. Obstet. Investig. 1990, 29, 71–74. [Google Scholar] [CrossRef]
  46. Iwanari, O.; Miyako, J.; Date, Y.; Nakayama, S.; Kijima, S.; Moriyama, M.; Karino, K.; Endoh, J.; Kitao, M. Clinical evaluations of the tumor marker sialyl SSEA-1 antigen for clinical gynecological disease. Gynecol. Obstet. Investig. 1990, 29, 214–218. [Google Scholar] [CrossRef]
  47. Yan, X.; Lin, Y.; Liu, S.; Aziz, F.; Yan, Q. Fucosyltransferase IV (FUT4) as an effective biomarker for the diagnosis of breast cancer. Biomed. Pharmacother. 2015, 70, 299–304. [Google Scholar] [CrossRef]
  48. Shan, M.; Yang, D.; Dou, H.; Zhang, L. Fucosylation in cancer biology and its clinical applications. Prog. Mol. Biol. Transl. Sci. 2019, 162, 93–119. [Google Scholar] [CrossRef]
  49. Keeley, T.S.; Yang, S.; Lau, E. The Diverse Contributions of Fucose Linkages in Cancer. Cancers 2019, 11, 1241. [Google Scholar] [CrossRef]
  50. Koike, T.; Kimura, N.; Miyazaki, K.; Yabuta, T.; Kumamoto, K.; Takenoshita, S.; Chen, J.; Kobayashi, M.; Hosokawa, M.; Taniguchi, A.; et al. Hypoxia induces adhesion molecules on cancer cells: A missing link between Warburg effect and induction of selectin-ligand carbohydrates. Proc. Natl. Acad. Sci. USA 2004, 101, 8132–8137. [Google Scholar] [CrossRef]
  51. Schnyder-Candrian, S.; Borsig, L.; Moser, R.; Berger, E.G. Localization of α1,3-fucosyltransferase VI in Weibel-Palade bodies of human endothelial cells. Proc. Natl. Acad. Sci. USA 2000, 97, 8369–8374. [Google Scholar] [CrossRef] [Green Version]
  52. Yang, X.; Wang, J.; Liu, S.; Yan, Q. HSF1 and Sp1 regulate FUT4 gene expression and cell proliferation in breast cancer cells. J. Cell Biochem. 2014, 115, 168–178. [Google Scholar] [CrossRef] [PubMed]
Jcm 11 05606 g001 550
Figure 1. Expression of FUT4 mRNA in the endometrium of control women and women with endometriosis. Expression of FUT4 mRNA was evaluated by qRT-PCR in the endometrium of control women (n = 28) and the endometrium of women with endometriosis (n = 49). The data are shown as scatterplots with the median and interquartile range. Samples from the endometriosis group displaying relative expression values over 0.08 (n = 3) are not included in the graph. Statistically significant differences between groups were computed by the Mann–Whitney U-test.
Figure 1. Expression of FUT4 mRNA in the endometrium of control women and women with endometriosis. Expression of FUT4 mRNA was evaluated by qRT-PCR in the endometrium of control women (n = 28) and the endometrium of women with endometriosis (n = 49). The data are shown as scatterplots with the median and interquartile range. Samples from the endometriosis group displaying relative expression values over 0.08 (n = 3) are not included in the graph. Statistically significant differences between groups were computed by the Mann–Whitney U-test.
Jcm 11 05606 g001
Jcm 11 05606 g002 550
Figure 2. Expression of FUT4 mRNA in the endometrium relative to the stage of endometriosis. (A) Expression of FUT4 mRNA in the endometrium from patients with minimal/mild (I/II) (n = 14) and moderate/severe (III/IV) (n = 35) stages of endometriosis compared to the endometrium from control women (n = 28). Data are shown as scatterplots with the median and interquartile range. Statistical differences between groups were computed by the Kruskal–Wallis test and Dunn’s post hoc test. (B) Correlation between FUT4 mRNA and the rAFS score of the disease. The results are shown as scatterplots with a regression line. Samples from the endometriosis group displaying relative expression values over 0.08 (n = 3) are not included in the graph. Spearman’s correlation coefficients (rs), their p-values and the number of cases (n) are shown in an inset.
Figure 2. Expression of FUT4 mRNA in the endometrium relative to the stage of endometriosis. (A) Expression of FUT4 mRNA in the endometrium from patients with minimal/mild (I/II) (n = 14) and moderate/severe (III/IV) (n = 35) stages of endometriosis compared to the endometrium from control women (n = 28). Data are shown as scatterplots with the median and interquartile range. Statistical differences between groups were computed by the Kruskal–Wallis test and Dunn’s post hoc test. (B) Correlation between FUT4 mRNA and the rAFS score of the disease. The results are shown as scatterplots with a regression line. Samples from the endometriosis group displaying relative expression values over 0.08 (n = 3) are not included in the graph. Spearman’s correlation coefficients (rs), their p-values and the number of cases (n) are shown in an inset.
Jcm 11 05606 g002
Jcm 11 05606 g003 550
Figure 3. Expression of FUT4 mRNA in the eutopic endometrium from women with endometriosis in relation to the localization of ectopic endometriotic lesions.FUT4 mRNA levels were compared in patients with different lesion locations. The normal endometrium from control women served as a control. All data are shown as scatterplots with median and interquartile ranges. Samples from the endometriosis group displaying relative expression values over 0.08 (n = 3) are not included in the graph. Statistical differences between groups were computed by the Kruskal–Wallis test and Dunn’s post hoc test.
Figure 3. Expression of FUT4 mRNA in the eutopic endometrium from women with endometriosis in relation to the localization of ectopic endometriotic lesions.FUT4 mRNA levels were compared in patients with different lesion locations. The normal endometrium from control women served as a control. All data are shown as scatterplots with median and interquartile ranges. Samples from the endometriosis group displaying relative expression values over 0.08 (n = 3) are not included in the graph. Statistical differences between groups were computed by the Kruskal–Wallis test and Dunn’s post hoc test.
Jcm 11 05606 g003
Jcm 11 05606 g004 550
Figure 4. ROC analysis of FUT4 mRNA expression in the endometrium of women with endometriosis and control women. The area under the curve (AUC), standard error (SE), 95% confidence interval (95% CI), p-value and the total number of analyzed samples (n) are given in an inset.
Figure 4. ROC analysis of FUT4 mRNA expression in the endometrium of women with endometriosis and control women. The area under the curve (AUC), standard error (SE), 95% confidence interval (95% CI), p-value and the total number of analyzed samples (n) are given in an inset.
Jcm 11 05606 g004
Table
Table 1. Demographic and clinical characteristics of patients with endometriosis and control subjects.
Table 1. Demographic and clinical characteristics of patients with endometriosis and control subjects.
Control GroupEndometriosis Group
AllMid-Proliferative PhaseMid-Secretory PhaseAllMid-Proliferative PhaseMid-Secretory Phase
Number of cases (N)2812 (42.9%)16 (57.1%)4918 (36.7%)31 (63.3%)
Age, years (mean ± SD)35.2 ± 6.4036.0 ± 5.0234.5 ± 7.3731.9 ± 5.5329.4 ± 3.7333.4 ± 5.93
BMI, kg/m2 (mean ± SD)22.2 ± 4.7922.8 ± 6.0821.9 ± 3.9621.7 ± 3.1221.5 ± 2.4421.8 ± 3.50
Parity0.8 ± 0.961.0 ± 1.100.6 ± 0.840.35 ± 0.67 *0.18 ± 0.390.45 ± 0.78
Infertility00018 (36.7%) **9 (50.0%)9 (29.0%)
rAFSI (minimal)---8 (16.3%)2 (11.1%)6 (19.4%)
II (mild)---6 (12.2%)0 (0%)6 (19.4%)
III (moderate)---25 (51.0%)11 (61.1%)14 (45.1%)
IV (severe)---10 (20.5%)5 (27.8%)5 (16.1%)
Lesion localizationOvarian---37 (75.5%)16 (88.9%)21 (67.7%)
Peritoneal---35 (71.4%)13 (72.2%)22 (71.0%)
Both---23 (46.9%)11 (61.1%)12 (38.7%)
p-values were calculated by one-way ANOVA, Student’s t-test or the Chi-squared test when applicable. * Different from all control groups at p = 0.0200. ** Different from all control groups at p = 0.0058.
Table
Table 2. Diagnostic value of FUT4 mRNA expression in the eutopic endometrium to predict endometriosis.
Table 2. Diagnostic value of FUT4 mRNA expression in the eutopic endometrium to predict endometriosis.
Sensitivity (95%CI)94.12% (84.08–98.40%)
Specificity (95%CI)89.29% (72.80–96.29%)
Positive Predictive Value (PPV)93.9%
Negative Predictive Value (NPV)89.3%
Likelihood Ratio8.784
Accuracy92.2%
Youden’s Index0.8341
The threshold for relative FUT4 mRNA expression was established at 2−ΔCt = 0.002793; 95% confidence interval (95%CI) values are given in parentheses.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Żeberkiewicz, M.; Hyc, A.; Iwan, A.; Zwierzchowska, A.; Ścieżyńska, A.; Kalaszczyńska, I.; Barcz, E.; Malejczyk, J. Expression of Fucosyltransferase 4 (FUT4) mRNA Is Increased in Endometrium from Women with Endometriosis. J. Clin. Med. 2022, 11, 5606. https://doi.org/10.3390/jcm11195606

AMA Style

Żeberkiewicz M, Hyc A, Iwan A, Zwierzchowska A, Ścieżyńska A, Kalaszczyńska I, Barcz E, Malejczyk J. Expression of Fucosyltransferase 4 (FUT4) mRNA Is Increased in Endometrium from Women with Endometriosis. Journal of Clinical Medicine. 2022; 11(19):5606. https://doi.org/10.3390/jcm11195606

Chicago/Turabian Style

Żeberkiewicz, Marta, Anna Hyc, Anna Iwan, Aneta Zwierzchowska, Aneta Ścieżyńska, Ilona Kalaszczyńska, Ewa Barcz, and Jacek Malejczyk. 2022. "Expression of Fucosyltransferase 4 (FUT4) mRNA Is Increased in Endometrium from Women with Endometriosis" Journal of Clinical Medicine 11, no. 19: 5606. https://doi.org/10.3390/jcm11195606

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop