Ecto-nucleotidases activities in the contents of ovarian endometriomas: potential biomarkers of endometriosis

Endometriosis, defined as the growth of endometrial tissue outside the uterus, is a common gynecologic condition affecting millions of women worldwide. It is an inflammatory, estrogen-dependent complex disorder, with broad symptomatic variability, pelvic pain, and infertility being the main characteristics. Ovarian endometriomas are frequently developed in women with endometriosis. Late diagnosis is one of the main problems of endometriosis; thus, it is important to identify biomarkers for early diagnosis. The aim of the present work is to evaluate the ecto-nucleotidases activities in the contents of endometriomas. These enzymes, through the regulation of extracellular ATP and adenosine levels, are key enzymes in inflammatory processes, and their expression has been previously characterized in human endometrium. To achieve our objective, the echo-guided aspirated fluids of endometriomas were analyzed by evaluating the ecto-nucleotidases activities and compared with simple cysts. Our results show that enzyme activities are quantifiable in the ovarian cysts aspirates and that endometriomas show significantly higher ecto-nucleotidases activities than simple cysts (5.5-fold increase for ATPase and 20-fold for ADPase), thus being possible candidates for new endometriosis biomarkers. Moreover, we demonstrate the presence of ecto-nucleotidases bearing exosomes in these fluids. These results add up to the knowledge of the physiopathologic mechanisms underlying endometriosis and, open up a promising new field of study

Impaired expression of ectonucleotidases in ectopic and eutopic endometrial tissue is in favor of ATP accumulation in the tissue microenvironment in endometriosis

Endometriosis is a prevalent disease defined by the presence of endometrial tissue outside the uterus. Adenosine triphosphate (ATP), as a proinflammatory molecule, promotes and helps maintain the inflammatory state of endometriosis. Moreover, ATP has a direct influence on the two main symptoms of endometriosis: infertility and pain. Purinergic signaling, the group of biological responses to extracellular nucleotides such as ATP and nucleosides such as adenosine, is involved in the biology of reproduction and is impaired in pathologies with an inflammatory component such as endometriosis. We have previously demonstrated that ectonucleotidases, the enzymes regulating extracellular ATP levels, are active in non-pathological endometria, with hormone-dependent changes in expression throughout the cycle. In the present study we have focused on the expression of ectonucleotidases by means of immunohistochemistry and in situ activity in eutopic and ectopic endometrial tissue of women with endometriosis, and we compared the results with endometria of women without the disease. We have demonstrated that the axis CD39-CD73 is altered in endometriosis, with loss of CD39 and CD73 expression in deep infiltrating endometriosis, the most severe, and most recurring, endometriosis subtype. Our results indicate that this altered expression of ectonucleotidases in endometriosis boosts ATP accumulation in the tissue microenvironment. An important finding is the identification of the nucleotide pyrophophatase/phosphodiesterase 3 (NPP3) as a new histopathological marker of the disease since we have demonstrated its expression in the stroma only in endometriosis, in both eutopic and ectopic tissue. Therefore, targeting the proteins directly involved in ATP breakdown could be an appropriate approach to consider in the treatment of endometriosis

Human Papillomavirus Infection in HIV-1 Infected Women in Catalonia (Spain): Implications for Prevention of Cervical Cancer

Background High-risk human Papillomavirus infection is a necessary factor for cervical squamous intraepithelial lesions and invasive cervical cancer. In HIV-1-infected women, HPV infection is more prevalent and a higher risk of cervical cancer has been identified. We aimed to calculate the prevalence of infection by HR-HPV, determine the factors associated with this infection and abnormal cytology findings and to describe the history of cervical cancer screening in HIV-1-infected women. Methods We enrolled 479 HIV-1-infected women from the PISCIS cohort. Each patient underwent a gynecological check-up, PAP smear, HPV AND Hybrid capture, HPV genotyping, and colposcopy and biopsy, if necessary. We applied questionnaires to obtain information on sociodemographic, behavioral, clinical, and cervical screening variables. We present a cross-sectional analysis. Results Median age was 42 years. The prevalence of HR-HPV infection was 33.2% and that of high-grade squamous intraepithelial lesions (HSIL) was 3.8%. The most common genotypes were 16(23%), 53(20.3%), and 52(16.2%). The factor associated with HR-HPV infection was age 500cells/mm3 (OR,8.4; 95%CI,3.7-19.2), HIV-1 viral load >10,000copies/mL versus <400copies/mL (OR,2.1; 95%CI,1.0-4.4), and use of oral contraceptives (OR,2.0; 95%CI,1.0-3.9). Sixty percent of HIV-1-infected women had had one Pap smear within the last 2 years. Conclusions The high prevalence of HPV infection and cervical lesions in the HIV-1-infected population in Catalonia, as well as the low coverage and frequency of screening in this group, means that better preventive efforts are necessary and should include vaccination against HPV, better accessibility to screening programs, training of health care professionals, and specific health education for HIV-1-infected women

Long‐term predictors of residual or recurrent cervical intraepithelial neoplasia 2‐3 after treatment with a large loop excision of the transformation zone: a retrospective study

Objective: To assess the long-term risk factors predicting residual/recurrent cervical intraepithelial neoplasia (CIN 2–3) and time to recurrence after large loop excision of the transformation zone (LLETZ). Design: Retrospective study. Setting: Colposcopy clinic. Population: 242 women with CIN 2–3 treated between 1996 and 2006 and followed up until June 2016. Methods: Age, margins and high risk-human papillomavirus (HR-HPV) were estimated using Cox proportional hazard and unconditional logistic regression models. The cumulative probability of treatment failure was estimated by Kaplan-Meier analysis. Main Outcome measure: Histologically confirmed CIN 2-3, HR-HPV, margins, age. Results: CIN 2-3 was associated with HR-HPV (HR =30.5; 95% CI =3.80-246.20); age >35 years (HR =5.53; 95% CI =1.22-25.13); and margins (HR = 7.31; 95% CI = 1.60–33.44). HR-HPV showed a sensitivity of 88.8% and a specificity of 80%. Ecto(+)/endocervical(+)(16.7%), uncertain (19.4%) and ecto(-)/endocervical(+) margins (9.1%) showed a higher risk of recurrence (OR = 13.20(95% CI =1.02-170.96), 15.84(95% CI =3.02-83.01), and 6.60(95% CI =0.88-49.53)), respectively. Women with involved margins and/or HR-HPV positive had more treatment failure than those who were HR-HPV negative or had clear margins (P-log rank<0.001). Conclusions: HR-HPV and margins seem essential for stratifying post- LLETZ risk, and enable personalised management. Given that clear margins present a lower risk, a large excision may be indicated in older women in order to reduce the risk. Funding statement: This study has received no funding

Characterization of the endometrial MSC marker ectonucleoside triphosphate diphosphohydrolase-2 (NTPDase2/CD39L1) in low- and high-grade endometrial carcinomas: loss of stromal expression in the invasive phenotypes

Ectonucleoside triphosphate diphosphohydrolase-2 (NTPDase2/CD39L1) has been described in human non-pathological endometrium in both epithelial and stromal components without changes along the cycle. It was identified as a stromal marker of basalis. In the present study, we aimed to evaluate NTPDase2 distribution, using immunolabeling and in situ enzyme activity approaches, in endometrial carcinoma (EC) at different tumor grades. NTPDase2 was present in tumor epithelial EC cells, as in the non-pathological endometria, but the expression underwent changes in subcellular distribution and also tended to decrease with the tumor grade. In stroma, NTPDase2 was identified exclusively at the tumor-myometrial junction but this expression was lost in tumors of invasive phenotype. We have also identified in EC samples the presence of the perivascular population of endometrial mesenchymal stem cells (eMSCs) positive for sushi domain containing 2 (SUSD2) and for NTPDase2, already described in non-tumoral endometrium. Our results point to NTPDase2 as a histopathological marker of tumor invasion in EC, with diagnostic relevance especially in cases of EC coexisting with other endometrial disorders, such as adenomyosis, which occasionally hampers the assessment of tumor invasion parameters

Role of sentinel node biopsy in breast cancer: a review

Axillary lymph node involvement is still an important predictor of recurrence and survival in breast cancer. Axillary staging was classically done by axillary lymph node dissection (ALND), but the introduction of sentinel lymph node biopsy (SLNB) has led to a progressive and continuing de-escalation in its use. Therefore, SLNB can now be considered the standard procedure for axillary staging in clinically N0 patients. Different studies have also begun to report that a positive sentinel node does not always require ALND, reducing the morbidity derived from this technique. Fears that this sentinel node approach might not be accurate for neoadjuvant chemotherapy have been allayed by several studies showing that post-neoadjuvant SLNB in clinical N0 patients reduces the rate of ALND. This approach benefits from axillary pathological complete response with an acceptable false-negative rate. By contrast, however, cN1 disease still requires that we optimise the technique to reduce the rate of false negatives. Currently, SLNB is the best method for axillary staging in breast cancer, allowing patients to be treated according to risk of recurrence, and with good evidence that morbidity is lower than with other more radical techniques

Ecto-nucleotidases as potential biomarkers for diagnosis of endometriosis

Introduction: Endometriosis is an inflammatory disorder characterized by the growth of endometrial tissue in extrauterine locations. Ovarian endometriomas are a common form of endometriosis. Late diagnosis is the main problem in the pathology; thus, it is important to identify clinical biomarkers. Ecto-nucleotidases are enzymes that hydrolyze extracellular ATP to adenosine and are involved in many inflammatory processes. Alterations of ecto-nucleotidase activity are related to inflammatory disease states

Ecto-nucleotidases activities in the contents of ovarian endometriomas: potential biomarkers of endometriosis

Endometriosis, defined as the growth of endometrial tissue outside the uterus, is a common gynecologic condition affecting millions of women worldwide. It is an inflammatory, estrogen-dependent complex disorder, with broad symptomatic variability, pelvic pain, and infertility being the main characteristics. Ovarian endometriomas are frequently developed in women with endometriosis. Late diagnosis is one of the main problems of endometriosis; thus, it is important to identify biomarkers for early diagnosis. The aim of the present work is to evaluate the ecto-nucleotidases activities in the contents of endometriomas. These enzymes, through the regulation of extracellular ATP and adenosine levels, are key enzymes in inflammatory processes, and their expression has been previously characterized in human endometrium. To achieve our objective, the echo-guided aspirated fluids of endometriomas were analyzed by evaluating the ecto-nucleotidases activities and compared with simple cysts. Our results show that enzyme activities are quantifiable in the ovarian cysts aspirates and that endometriomas show significantly higher ecto-nucleotidases activities than simple cysts (5.5-fold increase for ATPase and 20-fold for ADPase), thus being possible candidates for new endometriosis biomarkers. Moreover, we demonstrate the presence of ecto-nucleotidases bearing exosomes in these fluids. These results add up to the knowledge of the physiopathologic mechanisms underlying endometriosis and, open up a promising new field of study

Role of ecto-nucleotidases in human endometrial pathologies

Abstract from the VII International Congress of Histology and Tissue Engineering - XIX Congress of the Spanish Society of Histology and Tissue Engineering - IV Iberoamerican Congress of Histology. 5-8 September, 2017, Santiago de Compostela, Spai