Screening of Variants in the Transcript Profile of Eutopic Endometrium from Infertile Women with Endometriosis during the Implantation Window / Rastreio de variantes no perfil de tanscritos do endométrio eutópico de mulheres inférteis com endometriose durante a janela de implantação

Abstract Objective Abnormalities in the eutopic endometrium of women with endometriosis may be related to disease-associated infertility. Although previous RNA-sequencing analysis did not show differential expression in endometrial transcripts of endometriosis patients, other molecular alterations could impact protein synthesis and endometrial receptivity. Our aim was to screen for functional mutations in the transcripts of eutopic endometria of infertile women with endometriosis and controls during the implantation window. Methods Data from RNA-Sequencing of endometrial biopsies collected during the implantation window from 17 patients (6 infertile women with endometriosis, 6 infertile controls, 5 fertile controls) were analyzed for variant discovery and identification of functional mutations. A targeted study of the alterations found was performed to understand the data into disease's context. Results None of the variants identified was common to other samples within the same group, and no mutation was repeated among patients with endometriosis, infertile and fertile controls. In the endometriosis group, nine predicted deleterious mutations were identified, but only one was previously associated to a clinical condition with no endometrial impact. When crossing the mutated genes with the descriptors endometriosis and/or endometrium, the gene CMKLR1 was associated either with inflammatory response in endometriosis or with endometrial processes for pregnancy establishment. Conclusion Despite no pattern of mutation having been found, we ponder the small sample size and the analysis on RNA-sequencing data. Considering the purpose of the study of screening and the importance of the CMKLR1 gene on endometrial

Resumo Objetivo Anormalidades no endométrio eutópico de mulheres com endometriose podem estar relacionadas à infertilidade associada à doença. Embora a análise prévia de sequenciamento de RNA não tenha evidenciado expressão diferencial em transcritos endometriais de pacientes com endometriose, outras alterações moleculares poderiam afetar a síntese de proteínas e a receptividade endometrial. Nosso objetivo foi rastrear mutações funcionais em transcritos de endométrios eutópicos de mulheres inférteis com endometriose e de controles durante a janela de implantação. Métodos Os dados do sequenciamento de RNA de biópsias endometriais coletados durante a janela de implantação de 17 pacientes (6 mulheres inférteis com endometriose, 6 controles inférteis, 5 controles férteis) foram analisados para a descoberta de variantes e a identificação de mutações funcionais. Um estudo direcionado das alterações encontradas foi realizado para compreender os dados no contexto da doença. Resultados Nenhuma das variantes identificadas foi comuma outras amostras dentro do mesmo grupo, assim como nenhuma mutação se repetiu entre pacientes com endometriose, controles inférteis e férteis. No grupo de endometriose, foram identificadas nove mutações deletérias preditas, mas apenas uma foi previamente associada a uma condição clínica sem impacto endometrial. Ao cruzar os genes mutados com os descritores endometriose e/ou endométrio, o gene CMKLR1 foi associado a resposta inflamatória na endometriose e a processos endometriais para estabelecimento da gravidez. Conclusão Apesar de nenhum padrão de mutação ter sido encontrado, ponderamos o pequeno tamanho da amostra e a análise dos dados de sequenciamento de RNA. Considerando o objetivo do estudo de triagem e a importância do gene CMKLR1 na modulação endometrial, este poderia ser um gene candidato para estudos adicionais que avaliem mutações no endométrio eutópico de pacientes com endometriose.

Differential expression of upstream stimulatory factor (USF) 2 variants in eutopic endometria from women with endometriosis: estradiol regulation

BACKGROUND: Endometriosis, pro-inflammatory and invasive benign disease estrogen dependent, abnormally express in endometria the enzyme P450Arom, positively regulated by steroid factor-1 (SF-1). Our objective was to study the nuclear protein contents of upstream stimulating factor 2 (USF2a and USF2b), a positive regulator of SF-1, throughout the menstrual cycle in eutopic endometria from women with and without (control) endometriosis and the involvement of nuclear estrogen receptors (ER) and G-coupled protein estrogen receptor (GPER)-1. RESULTS: Upstream stimulating factor 2 protein contents were higher in mid (USF2b) and late (USF2a and USF2b) secretory phase in eutopic endometria from endometriosis than control (p < 0.05). In isolated control epithelial cells incubated with E2 and PGE2, to resemble the endometriosis condition, the data showed: (a) significant increase of USF2a and USF2b nuclear protein contents when treated with E2, PPT (specific agonist for ERa) or G1 (specific agonist for GPER1); (b) no increase in USF2 binding to SF-1 E-Box/DNA consensus sequence in E2-treated cells; (c) USF2 variants protein contents were not modified by PGE2; (d) SF-1 nuclear protein content was significantly higher than basal when treated with PGE2, E2 or G1, stimulation unaffected by ICI (nuclear ER antagonist); and (e) increased (p < 0.05) cytosolic protein contents of P450Arom when treated with PGE2, E2, PPT or G1 compared to basal, effect that was additive with E2 + PGE2 together. Nevertheless, in endometriosis cells, the high USF2, SF-1 and P450Arom protein contents in basal condition were unmodified. CONCLUSION: These data strongly suggest that USF2 variants and P450Arom are regulated by E2 through ERa and GPER1, whereas SF-1 through GPER1, visualized by the response of the cells obtained from control endometria, being unaffected the endogenously stimulated cells from endometriosis origin. The lack of E2 stimulation on USF2/SF-1 E-Box/DNA-sequence binding and the absence of PGE2 effect on USF2 variants opposite to the strong induction that they exert on SF1 and P450 proteins suggest different mechanisms and indirect regulations. The sustained USF2 variants protein expression during the secretory phase in eutopic endometria from women with endometriosis may participate in the pathophysiology of this disease strongly associated with infertility and its characteristic endometrial invasion to ectopic sites in the pelvic cavity.

Assessment of changes in eutopic endometrial vascularity of endometriosis using three-dimensional power Doppler ultrasound / 中华超声影像学杂志

Objective To evaluate the changes in eutopic endometrial vascularity of endometriosis using three-dimensional power Doppler ultrasound.Methods Forty-four women with endometriosis and 51 normal controls were studied by three-dimensional power Doppler ultrasound.Eutopic endometrial volume,vascularization index (VI),flow index (FI) and vascularization flow index (VFI) were calculated using VOCAL software.Results In proliferative phase,the mean VI was (5.499 ± 5.153)％ in patients with endometriosis and ( 1.135 ± 1.333) ％ in normal controls ( P ＜0.001 ),the mean FI was (28.832 ± 4.279)dB in patients with endometriosis and (25.544 ± 3.465)dB in normal controls ( P =0.006),the VFI was (1.769 ± 1.981 )dB in patients with endometriosis and (0.321 ± 0.397)dB in normal controls( P =0.002).In secretory phase,the mean VI was (8.693 ± 5.940) ％ in patients with endometriosis and ( 1.014±0.968)％ in normal controls ( P ＜0.001); the mean FI was (30.689 ± 3.632)dB in patients with endometriosis and (24.748 ± 3.811 )dB in normal controls( P ＜0.001 ),the VFI was (2.753 ± 2.044)dB in patients with endometriosis and (0.27 ± 0.293)dB in normal controls( P =0.001 ).Both in proliferative and secretory phase,no significant difference in endometrial volume between the two groups was observed ( P =0.108 and 0.068,respectively).Conclusions Three-dimensional power ultrasound provides a useful tool to investigate eutopic endometrial vascularization of endimetriosis.

Nerve Fibres Distribution in Eutopic Endometrium of Women with Endometriosis / 实用妇产科杂志

Objective:Nerve fibres distribution in the functional layer of endometrium of women with endometdosis was investigated.Methods:Histological sections of endometrial tissue were prepared from endometrialcurettings and hysterectomies performed on women with endometnosis(n=25)and without endometriosis(n=40).Immunohistochemistry was used to detect nerve fibres by highly specific polyclonal rabbit antibody PGP 9.5.The assessment of nerve fibre density was performed bv Image Pro Plus Discovery.Results:Nerve fibres were identified throughout the functional layers of the endometrium in all endometriosis patients,but not found in the functional layer of the endometrium in women without endometriosis(P<0.01).Conclusions:Nerve fibres detectad in the functional layer in all women with endometriosis may have important implications for understanding the generation of pain in these patients.

Evaluation of the relative efficacy of endometrial curettage for diagnosis of endometriosis by the detection of eutopic endometrial nerve fibers / 中国综合临床

Objective To evaluate endometrial biopsy and curettage in detecting small nerve fibers in eutopic endometrium for diagnosis of endometriosis.Methods Endometrial biopsies and curettings were taken in 65 women with menorrhalgia.Endometrial nerve fibers were immunohistochemically detected using the pan-neuronal marker PGP 9.5.All patients underwent laparoseopie approach.Results The specificity,sensitivity,positive and negative predictive value were 100% of endometrial biopsy and curettings for diagnosis of endometriosis.Condusions Careful endometrial biopsy combined with immunohistochemical staining for nerve fibers may be a reliable means of diagnosing or excluding endometriosis.

Survivin and Bcl-2 expression in eutopic endometrium with and without endometriosis / 대한산부인과학회지

OBJECTIVE: To examine expression of survivin and Bcl-2 in eutopic endometrium with endometriosis and without endometriosis by immunohistochemical stain. METHODS: Expression of survivin and Bcl-2 was immunohistochemically investigated in eutopic endometrium with endometriosis (n=30, study group I; laparoscopy-proved endometriosis, study group II; adenomyosis) or without endometriosis (n=34, control group). The score of immunohistochemical staining was evaluated semi-quantitative method modified by Sinicrope and Lu. Statistical analyses was carried out using one-way ANOVA with Turkey test. RESULTS: Survivin expression was significantly higher in nucleus of glandular epithelium of eutopic endometrium with endomtriosis compared to that without endometriosis (p<0.05). Increased expression of Bcl-2 was found in glandular epithelium of eutopic endometrium with endometriosis than without endometriosis, but there was no significantly difference. CONCLUSION: Our findings suggest that increased survivin expression may contribute to survival of endometriosis. Moreover, survivin could play an important role in pathogenesis of endometriosis.

Investigation of integrin ?_5 and integrin ?_3 in endometriosis / 医学研究生学报

Objective:To study the expression of integrin ?_5,?_3 in eutopic endometrium and ectopic endometrium of endometriosis. Methods:Reverse transcription polymerase chain reaction(RT-PCR) method was utilized to detect and quantitate integrin ?_5,?_3 mRNA expression levels in eutopic and ectopic endometrial tissues from 18 patients with endometriosis and the endometrial tissues from 18 non-endometriosis women.Serum concentration of estradiol(E_2),progesterone(P) and CA125 were measured by radioimmunoassay.AFS-r marking was used to describe the state of pelvic cavity during operation. Results:Various levels of positive expression of integrin subunits ?_5,?_3 were shown in both eutopic and ectopic endometrial tissues of endometriosis and the endometrium of non-endometriosis.The expressions of integrin ?_5 mRNA in eutopic endometrium,ectopic endometrium of endometriosis group and endometrium of control group were as 0.773?0.113,1.077?0.032,0.924?0.120,respectively.(Among) the groups,the differences were statistically significant(P0.05).Both CA125 level and AFS-r were positively correlated with integrin ?_5,?_3(P

Transcription characteristics of chemokine receptors in endometrium and endometrial tissues / 中国免疫学杂志

Objective:To investigate the expression of chemokine receptor in eupotic and ectopic endometrial tissues of women with endometriosis, and in endometrium of women without endometriosis.Methods:Normal endometrium, eutopic endometrium and endometriotic tissues were obtained from patients with endometriosis at laparoscopy. Total RNA was then extracted using the TRIzol reagent. The expression of chemokine receptors in these tissues were analyzed by way of semi-quantitative reverse transcriptase-polymerase chain reaction.Results:Compared to normal endometrium, the eutopic endometrium expressed significantly more CCR6, CCR8, CCR9 and CX3CR1(P