Current Applications and Challenges of Next-Generation Sequencing in Plasma Circulating Tumour DNA of Ovarian Cancer.

Circulating tumour DNA (ctDNA) facilitates longitudinal study of the tumour genome, which, unlike tumour tissue biopsies, globally reflects intratumor and intermetastatis heterogeneity. Despite its costs, next-generation sequencing (NGS) has revolutionised the study of ctDNA, ensuring a more comprehensive and multimodal approach, increasing data collection, and introducing new variables that can be correlated with clinical outcomes. Current NGS strategies can comprise a tumour-informed set of genes or the entire genome and detect a tumour fraction as low as 10-5. Despite some conflicting studies, there is evidence that ctDNA levels can predict the worse outcomes of ovarian cancer (OC) in both early and advanced disease. Changes in those levels can also be informative regarding treatment efficacy and tumour recurrence, capable of outperforming CA-125, currently the only universally utilised plasma biomarker in high-grade serous OC (HGSOC). Qualitative evaluation of sequencing shows that increasing copy number alterations and gene variants during treatment may correlate with a worse prognosis in HGSOC. However, following tumour clonality and emerging variants during treatment poses a more unique opportunity to define treatment response, select patients based on their emerging resistance mechanisms, like BRCA secondary mutations, and discover potential targetable variants. Sequencing of tumour biopsies and ctDNA is not always concordant, likely as a result of clonal heterogeneity, which is better captured in the plasma samples than it is in a large number of biopsies. These incoherences may reflect tumour clonality and reveal the acquired alterations that cause treatment resistance. Cell-free DNA methylation profiles can be used to distinguish OC from healthy individuals, and NGS methylation panels have been shown to have excellent diagnostic capabilities. Also, methylation signatures showed promise in explaining treatment responses, including BRCA dysfunction. ctDNA is evolving as a promising new biomarker to track tumour evolution and clonality through the treatment of early and advanced ovarian cancer, with potential applicability in prognostic prediction and treatment selection. While its role in HGSOC paves the way to clinical applicability, its potential interest in other histological subtypes of OC remains unknown.

The Concept behind the Suitability of Menstrual Blood-Derived Stem Cells for the Management of Vaginal Atrophy among BRCA Mutation Carriers after RRSO.

Risk-reducing bilateral salpingo-oophorectomy (RRSO) is recommended for breast cancer gene 1 (BRCA1) and 2 (BRCA2) mutation carriers. A major consequence of RRSO is surgical menopause associated with severe menopausal symptoms, mostly genitourinary complaints. Due to the inherent breast cancer risk, estrogen-based therapies are generally avoided in these patients. So far, the non-hormonal approaches available are not efficient to successfully treat the disabling vaginal atrophy-related symptoms. In regenerative medicine, mesenchymal stem cells (MSC) are the most frequently used cell type due to their remarkable and regenerative characteristics. Therapies based on MSC have revealed positive outcomes regarding symptoms and signs associated with vaginal atrophy by promoting angiogenesis, vaginal restoration, and the proliferation of vaginal mucosa cells. Menstrual blood-derived stem cells (MenSC) are a novel source of MSC, with promising therapeutic potential directly linked to their high proliferative rates; low immunogenicity; non-invasive, easy, and periodic acquisition; and almost no associated ethical issues. In this review, we update the current knowledge and research regarding the potential value of previously preserved MenSC in the therapy of vaginal atrophy among BRCA mutation carriers subjected to RRSO.

New Insights into Photobiomodulation of the Vaginal Microbiome-A Critical Review.

The development of new technologies such as sequencing has greatly enhanced our understanding of the human microbiome. The interactions between the human microbiome and the development of several diseases have been the subject of recent research. In-depth knowledge about the vaginal microbiome (VMB) has shown that dysbiosis is closely related to the development of gynecologic and obstetric disorders. To date, the progress in treating or modulating the VMB has lagged far behind research efforts. Photobiomodulation (PBM) uses low levels of light, usually red or near-infrared, to treat a diversity of conditions. Several studies have demonstrated that PBM can control the microbiome and improve the activity of the immune system. In recent years, increasing attention has been paid to the microbiome, mostly to the gut microbiome and its connections with many diseases, such as metabolic disorders, obesity, cardiovascular disorders, autoimmunity, and neurological disorders. The applicability of PBM therapeutics to treat gut dysbiosis has been studied, with promising results. The possible cellular and molecular effects of PBM on the vaginal microbiome constitute a theoretical and promising field that is starting to take its first steps. In this review, we will discuss the potential mechanisms and effects of photobiomodulation in the VMB.

Seminal human papillomavirus infection: a narrative review.

BACKGROUND: Human papillomavirus (HPV) infection is one of the most prevalent sexually transmitted infections that can lead to malignant pathologies as well as fertility problems. The aim of this study was to evaluate the prevalence of HPV infection in males, its impact on semen parameters, and reproductive consequences. We also evaluated potential measures that could prevent negative outcomes of HPV infection in males. METHODS: A systematic literature search using PubMed/Medline and Embase databases was performed to search for English articles published until July 2023. We explored three different aspects: (1) prevalence of HPV semen infection and its impact on seminal parameters; (2) the relationship between HPV semen infection and infertility risk and reproductive outcomes; and (3) potential measures that could prevent the negative outcomes related to HPV seminal infection. The identified studies were first screened and assessed independently by one author, and then validated by two additional authors. Data were extracted from 19 studies. RESULTS: The prevalence of seminal HPV infection was higher among infertile males. In addition to controversies about the real interference of seminal HPV infection on sperm parameters, a growing number of studies have demonstrated a correlation between unexplained infertility and seminal HPV infection. Semen HPV infection is also associated with lower rates of pregnancy and higher rates of miscarriage. Prevention measures such as HPV vaccination seem promising. CONCLUSION: Further studies are required to confirm not only the association between HPV infection and reproductive outcomes but also the benefit of preventive measures.

Marking Techniques for Targeted Axillary Dissection Among Patients With Node-Positive Breast Cancer Treated With Neoadjuvant Chemotherapy.

Invasive breast cancer with axillary lymph node (LN) invasion is a continuing problem worldwide. The morbidity associated with axillary LN dissection along with the high rate of nodal downstaging after neoadjuvant chemotherapy (NACT) made the standard treatment shift towards less invasive surgery. Sentinel lymph node biopsy (SLNB) after NACT is associated with high false-negative rates (13%-14%). To overcome this problem, it was concluded that the positive nodes should first be indicated with image-detectable markers and then removed together with SLNB: targeted axillary dissection (TAD). This review aims to describe and evaluate the different marking techniques for TAD in patients with node-positive breast cancer treated with NACT, namely: clip placement and guidewire localization; clip placement and 125I-labelled radioactive seed localization; clip placement and skin mark; clip placement and intraoperative ultrasound; tattooing with a sterile black carbon suspension; magnetic seeds; radar and infrared light technology localization. Targeted axillary dissection techniques have shown false-negative rates below 9% and identification rates above 95%. The most studied technique is guidewire localization, as it is also the oldest one. However, according to data gathered from this review, some newer techniques have shown to be very promising due to their statistical results and management factors.

The Psychosocial Impact of the Decision to Undergo Risk-Reducing Salpingo-Oophorectomy Surgery in BRCA Mutation Carriers and the Role of Physician-Patient Communication.

Risk-reducing bilateral salpingo-oophorectomy (RRSO) is an effective prophylactic surgery provided to premenopausal women carrying BRCA1 or BRCA2 mutations and presenting an increased risk of developing breast or ovarian cancer. This procedure is related to physiological, sexual, and psychosocial distress, which altogether increase uncertainty and complexity in the clinical decision-making process and post-surgery adaptation. Physician-patient communication (PPC) has been pointed out as a determinant factor in the decision-making to undergo RRSO, and the subsequent adjustment of women. However, studies examining the psychosocial impact of the decision-making process have been scarce and often lack clear theoretical frameworks. While the role of PPC in such processes has been highlighted in a few qualitative studies, there is a paucity of quantitative research addressing this question. Therefore, this narrative review, conducted using a multidisciplinary approach, was planned to: (1) present an updated medical background for RRSO; (2) analyze the psychosocial impact of the decision-making process within a theoretical framework of the Health Belief Model; and (3) discuss the role of PPC in such a decision-making process and in post-surgery. The collected research also enabled the recommendation of some additions to the existing clinical guidelines and the outlining of future research directions.

One Step Nucleic Acid Amplification (OSNA) Lysate Samples Are Suitable to Establish a Transcriptional Metastatic Signature in Patients with Early Stage Hormone Receptors-Positive Breast Cancer.

The One Step Nucleic Acid Amplification (OSNA) is being adopted worldwide for sentinel lymph nodes (SLNs) staging in breast cancer (BC). As major disadvantage, OSNA precludes prognostic information based on structural evaluation of SLNs. Our aim is to identify biomarkers related to tumor-microenvironment interplay exploring gene expression data from the OSNA remaining lysate. This study included 32 patients with early stage hormone receptors-positive BC. Remaining OSNA lysates were prepared for targeted RNA-sequencing analysis. Identification of differentially expressed genes (DEGs) was performed by DESeq2 in R and data analysis in STATA. The results show that, in metastatic SLNs, several genes were upregulated: KRT7, VTCN1, CD44, GATA3, ALOX15B, RORC, NECTIN2, LRG1, CD276, FOXM1 and IGF1R. Hierarchical clustering analysis revealed three different clusters. The identified DEGs codify proteins mainly involved in cancer aggressiveness and with impact in immune response. The overexpression of the immune suppressive genes VTCN1 and CD276 may explain that no direct evidence of activation of immune response in metastatic SLNs was found. We show that OSNA results may be improved incorporating microenvironment-related biomarkers that may be useful in the future for prognosis stratification and immunotherapy selection. As OSNA assay is being implemented for SLNs staging in other cancers, this approach could also have a wider utility.

The accuracy of magnetic resonance imaging in predicting the size of pure ductal carcinoma in situ: a systematic review and meta-analysis.

Ductal carcinoma in situ (DCIS) is a putative precursor of invasive breast cancer and MRI is considered the most sensitive imaging technique for its detection. This study aims to evaluate the accuracy of MRI measuring the pure DCIS size, against pathology, to better understand the role of MRI in the management of this intraductal neoplasm.Potentially eligible studies in MEDLINE, Embase and Google Scholar, up to January 2021 were considered, and a systematic review and meta-analysis according to the published protocol (Prospero-CRD42021232228) was performed. Outcomes of mean differences and accuracy rates were analysed using IBM® SPSS® v26 and random-effect models in platform R v3.3.Twenty-two cross-sectional studies were selected and 15 proceeded to meta-analysis. MRI accurately predicted 55% of the tumours' sizes and, according to Bland-Altman plots, concordance between MRI and pathology was greater for smaller tumours. In the meta-analysis, difference of the means between MRI and pathology was 3.85 mm (CI 95% [-0.92;8.60]) with considerable heterogeneity (I2 = 96.7%). Subgroup analysis showed similar results for sizes between different MRI fields, temporal resolution, slice thickness and acquisition times, but lower heterogeneity in studies using 3-T MRI (I2 = 57.2%). Results were concordant with low risk of bias studies (2.46, CI 95% [0.57-4.36]), without heterogeneity (I2 = 0%).Therefore, MRI is shown to be an accurate method in pure DCIS size assessment. Once the best MRI protocol is established, evaluation of the impact of pure DCIS size in predicting treatment outcomes will contribute to clarifying current issues related to intraductal breast carcinoma.

The Emerging Role of Menstrual-Blood-Derived Stem Cells in Endometriosis.

The human endometrium has a complex cellular composition that is capable of promoting cyclic regeneration, where endometrial stem cells play a critical role. Menstrual blood-derived stem cells (MenSC) were first discovered in 2007 and described as exhibiting mesenchymal stem cell properties, setting them in the spotlight for endometriosis research. The stem cell theory for endometriosis pathogenesis, supported by the consensual mechanism of retrograde menstruation, highlights the recognized importance that MenSC have gained by potentially being directly related to the genesis, development and maintenance of ectopic endometriotic lesions. Meanwhile, the differences observed between MenSC in patients with endometriosis and in healthy women underlines the applicability of these cells as a putative biomarker for the early diagnosis of endometriosis, as well as for the development of targeted therapies. It is expected that in the near future MenSC will have the potential to change the way we manage this complex disease, once their long-term safety and effectiveness are assessed.

Providing new insights into the endometriosisassociated cancer arising in episiotomy scars.

Endometriosis-associated malignancy in an episiotomy scar is rare. The predictive factors are poorly understood as are the mechanisms and pathways associated with implantation and malignant transformation. In this study we describe the cases reported in the literature of malignancies arising in endometriosis foci of an episiotomy scar. We identified 5 cases described between 1990 and 2016. These cases represent recurrence of endometriotic lesions in an episiotomy scar after previous diagnosis of endometriosis, 3 to 25 months before. Histology revealed clear cell tumours in 4 cases and a serous papillary carcinoma. The approach encompassed surgical removal for diagnosis and as part of the therapeutic strategy. Adjuvant treatment was performed depending on classical prognostic factors. Mechanisms of endometriosis implantation in scars include the influence of estrogens in the healing process and activation of COX-2, aromatase and matrix metalloproteinases. Nevertheless, for malignant transformation, other pathways seem to play a role, namely inflammation, immune response and oxidative stress, induced by iron deposits due to haemorrhage. Further studies are needed to allow the establishment of a predictive model for malignant transformation of endometriosis in episiotomy scars.

Epithelial-mesenchymal interconversions in ovarian cancer: The levels and functions of E-cadherin in intraabdominal dissemination.

The metastatic process of ovarian cancer (OC) is almost exclusively defined by direct shedding of tumor cells into the abdominal cavity, followed by clustering into multicellular aggregates and posterior peritoneal anchorage. This process relies on dynamic intercellular interactions which are modified by epithelial- mesenchymal interconversions and, therefore, E-cadherin expression variability. Although widely accepted as a tumor suppressor in many types of cancer, E-cadherin is currently known to have a dynamic expression and a much more complex role in OC. First, high E-cadherin expression is considered a sign of metaplasia in the normal ovarian epithelium, due to its association with epithelial growth factor receptor (EGFR) mediated cell proliferation. Subsequently, it is the decreased expression of E-cadherin that allows the acquisition of a more invasive phenotype, leading to the spread of primary tumor cells into the peritoneal fluid. This downregulation seems to depend on complex regulatory mechanisms, from molecular proteolysis to microenvironment interference and epigenetic regulation. E-cadherin cleavage and its resulting fragments appear to be essential to the process of dissemination and even to the formation of multicellular aggregates. Paradoxically, the maintenance of some E-cadherin expression seems to promote intercellular adhesion, resistance, and survival while decreasing cancer response to chemotherapy. Multiple studies have shown that reversing epithelial-mesenchymal transaction (EMT) and increasing E-cadherin expression prevents OC intraperitoneal dissemination, but findings that simultaneously correlate E-cadherin downregulation to higher chemotherapy sensitivity should not be ignored. Nevertheless, EMT and E-cadherin seem to have a potential interest as therapeutic targets in novel approaches to OC treatment.

Prevention of Human Papillomavirus Infection. Beyond Cervical Cancer: A Brief Review.

INTRODUCTION: Human papillomavirus is responsible for almost all cases of cervical cancer, an important portion of anogenital and oropharyngeal invasive and preinvasive lesions, as well as genital warts (condyloma acuminatum) and recurrent respiratory papillomatosis. Currently, three prophylactic vaccines against high-risk Human papillomavirus are commercialized in many countries worldwide. METHODS: To this non-systematic review the authors searched in MEDLINE/PubMed for systematic reviews, meta-analysis and randomized controlled trials, published in the last six years, using the terms "HPV", "non-cervical cancer" and "vaccine". Non-cervical cancers caused by human papillomavirus are less common lesions. However, its incidence has been increasing, while cervical cancer has declined, due mainly to highly effective screening programs. There are no formal screening programs for non-cervical cancers, so universal vaccination could have an important impact. The preventive effect of the vaccine is mainly studied and established in relation to cervical cancer, although it has also been demonstrated in the development of vulvar and vaginal lesions. To date, the efficacy in preventing anal and oropharyngeal diseases related with human papillomavirus is uncertain due to scarce supporting data and low vaccination coverage in men. The prevalence of injuries and subsequent absolute benefit of vaccination is lower in men, but it provides an additional benefit to the herd immunity achieved with the vaccination of women. CONCLUSION: The total fraction of malignant and pre-malignant lesions attributed to Human papillomavirus genotypes contained in the nonavalent vaccine is significant in both women and men, which turns this vaccine into a great asset in terms of Public Health.

Introdução: O vírus do papiloma humano é responsável por quase todos os casos de cancro do colo do útero, de uma importante fração de lesões anogenitais e orofaríngeas pré-invasivas e invasivas bem como de condilomas genitais e da papilomatose respiratória recorrente. Atualmente existem três vacinas profiláticas contra o vírus do papiloma humano de alto risco comercializadas em vários países do mundo. Métodos: Para esta revisão não-sistemática, os autores pesquisaram na MEDLINE/PubMed revisões sistemáticas, metanálises e ensaios clínicos randomizados, publicados nos últimos seis anos, utilizando os termos "HPV", "cancro não cervical" e "vacina". Os cancros não cervicais causados pelo vírus do papiloma humano são lesões menos comuns. Contudo, a sua incidência tem aumentado, a par de uma diminuição do cancro do colo do útero, devido principalmente à implementação de programas de rastreio altamente eficazes. Uma vez que não existem programas oficiais de rastreio para cancros não cervicais, a vacinação universal pode ter um impacto importante. O efeito preventivo da vacina é principalmente estudado e estabelecido em relação ao cancro do colo do útero, embora também tenha sido demonstrado no desenvolvimento de lesões vulvares e vaginais. Até ao momento, a eficácia na prevenção de doenças anais e orofaríngeas relacionadas com o vírus do papiloma humano é incerta, devido à escassez de dados na literatura e baixa cobertura de vacinação em homens. A prevalência de lesões e o consequente benefício absoluto da vacinação é inferior nos homens, porém proporciona um benefício adicional à imunidade de grupo alcançada com a vacinação de mulheres. Conclusão: A fração total de lesões malignas e pré-malignas atribuídas aos genótipos de vírus do papiloma humano contidos na vacina nonavalente é significativa tanto em mulheres quanto em homens, o que confere a essa vacina um grande potencial em termos de Saúde Pública.

Controversial association between polycystic ovary syndrome and breast cancer.

Polycystic ovary syndrome (PCOS) risk factors overlap with breast cancer, and the hormonal profile may be implicated in breast cancer pathogenesis. This study aims to report a literature review considering epidemiological and molecular mechanisms that correlate PCOS and breast cancer, as well as the influence of PCOS treatment on the incidence of breast cancer. Epidemiological studies failed to adjust potential variables that affect the risk and have thus provided inconclusive results. Molecular effects of androgenic pathways in breast cancer have been studied and androgens seem to have an inhibitory effect on mammary epithelial proliferation. However, increased bioavailable androgens were associated with recurrence of breast cancer due to conversion to oestrogens. Sex hormone-binding globulin has a role in hormone-dependent cancers and can be considered a marker for PCOS; a gene profile has already been linked to breast cancer risk in these patients. PCOS medical treatment is a promising tool for stratifying breast cancer risk due to the metabolic influence and hormonal environment. Clinical reports are inconsistent, emphasizing the need for further studies with a prospective design. In the future, the role of pharmacological interventions in PCOS will increase knowledge and awareness of breast cancer pathogenesis and will help to refine breast cancer risk stratification.

[Pregnancy after Breast Cancer: State of the Art]. / Gravidez após o Cancro da Mama: Estado da Arte.

Breast cancer survivors have given rise to several issues of major relevance from a clinical and scientific point of view. In fact, breast cancer is the most prevalent malignancy in women of reproductive age. The effect of pregnancy on overall survival and in the recurrence after treatment of breast cancer, as well as the questions related to heredity continue to be matter of the highest timeliness and scientific interest. Most recent studies seem to agree in admitting that pregnancy after breast cancer appears to be potentially safe to both the women and her offspring, although this issue remains complex. Heredity and genetics seem to play an important role in this subject, but the conclusions lack absolute and unequivocal consistency. There is a need for meta-analysis, cohort and case-control studies, translational and prospective studies extended in time, in order to obtain greater safety in the establishment of strategies and guidelines for clinicians and adequate objective information for young breast cancer patients.

As sobreviventes de cancro da mama têm originado um conjunto de questões da maior relevância sob o ponto de vista clínico e científico. De facto, o cancro da mama é a neoplasia mais prevalente em mulheres em idade reprodutiva. O efeito da gravidez na sobrevida global e na recorrência após tratamento de cancro da mama, bem como as questões relacionadas com a hereditariedade continuam a ser matéria da maior atualidade e do mais elevado interesse científico. Os estudos mais recentes são consensuais em admitir que a gravidez após o cancro da mama é potencialmente segura para a doente e sua descendência, embora a questão permaneça complexa. A hereditariedade e a genética parecem desempenhar um papel importante nesta temática mas, também aqui, as conclusões carecem de absoluta e inequívoca consistência. Verifica-se a necessidade de meta-análises, estudos coorte e caso-controlo, translacionais e prospetivos, alargados e prolongados no tempo, no sentido de obter uma maior segurança no estabelecimento de estratégias e linhas de orientação para os clínicos e informação adequada e objetiva para as jovens doentes com cancro da mama.

Intracystic Primary Squamous Cell Carcinoma of the Breast: A Challenging Diagnosis.

We report a case of a 36-year-old woman that presented with a painful mass in the outer quadrants of the left breast that had grown rapidly. Physical examination revealed a well circumscribed elastic mass and breast ultrasound showed a cyst measuring 26 mm with vegetation growing on the inner wall. Microscopic evaluation, after fine needle aspiration cytology (FNAC), suggested benign lesion. Tumorectomy was performed and the final diagnosis was a pure squamous cell carcinoma (SCC) of the breast. A simple mastectomy with sentinel node biopsy was performed. The histological study of the specimen revealed residual SCC and the sentinel lymph node was negative. The patient received 6 cycles of adjuvant chemotherapy and adjuvant radiotherapy. Four years later, the patient is free of disease.

A Scary Onset of a Rare and Aggressive Type of Primary Breast Sarcoma: A Case Report.

Primary breast sarcoma, arising from connective tissue within the breast, is extremely rare, accounting for less than 1% of all primary breast malignancies and no more than 5% of all sarcomas. The rarity of this pathology limits most studies to case reports and small retrospective studies, which has led to a lack of consensus on the clinical management. We report a clinical case of a 52-year-old woman, perimenopausal, previously healthy, with regular breast surveillance, who presented with a large (>20 cm) and rapidly expanding hypervascularized tumor of the left breast developed over 10 days, with a very thin preulcerative skin over the last 4 days. There was no systemic dissemination. The patient was submitted to total mastectomy and excision of axillary adenopathy. The tumor was diagnosed histologically as malignant phyllodes tumor associated with areas of high-grade sarcoma. Due to rapid growth and aggressive histological characteristics, adjuvant chemotherapy and radiotherapy were performed. There is a lot of evidence that tumors larger than 5 cm are associated with a poor prognosis. Despite the poor prognosis associated with this aggressive entity, the patient had no recurrence during 5 years of follow-up. We review the relevant literature about primary breast sarcomas.