Unique Molecular Features in High-Risk Histology Endometrial Cancers

Endometrial cancer is the most common gynecologic malignancy in the United States and the sixth most common cancer in women worldwide. Fortunately, most women who develop endometrial cancer have low-grade early-stage endometrioid carcinomas, and simple hysterectomy is curative. Unfortunately, 15% of women with endometrial cancer will develop high-risk histologic tumors including uterine carcinosarcoma or high-grade endometrioid, clear cell, or serous carcinomas. These high-risk histologic tumors account for more than 50% of deaths from this disease. In this review, we will highlight the biologic differences between low- and high-risk carcinomas with a focus on the cell of origin, early precursor lesions including atrophic and proliferative endometrium, and the potential role of stem cells. We will discuss treatment, including standard of care therapy, hormonal therapy, and precision medicine-based or targeted molecular therapies. We will also discuss the impact and need for model systems. The molecular underpinnings behind this high death to incidence ratio are important to understand and improve outcomes

Transcriptomic analyses of ovarian clear-cell carcinoma with concurrent endometriosis

IntroductionEndometriosis, a benign inflammatory disease whereby endometrial-like tissue grows outside the uterus, is a risk factor for endometriosis-associated ovarian cancers. In particular, ovarian endometriomas, cystic lesions of deeply invasive endometriosis, are considered the precursor lesion for ovarian clear-cell carcinoma (OCCC).MethodsTo explore this transcriptomic landscape, OCCC from women with pathology-proven concurrent endometriosis (n = 4) were compared to benign endometriomas (n = 4) by bulk RNA and small-RNA sequencing.ResultsAnalysis of protein-coding genes identified 2449 upregulated and 3131 downregulated protein-coding genes (DESeq2, P< 0.05, log2 fold-change > |1|) in OCCC with concurrent endometriosis compared to endometriomas. Gene set enrichment analysis showed upregulation of pathways involved in cell cycle regulation and DNA replication and downregulation of pathways involved in cytokine receptor signaling and matrisome. Comparison of pathway activation scores between the clinical samples and publicly-available datasets for OCCC cell lines revealed significant molecular similarities between OCCC with concurrent endometriosis and OVTOKO, OVISE, RMG1, OVMANA, TOV21G, IGROV1, and JHOC5 cell lines. Analysis of miRNAs revealed 64 upregulated and 61 downregulated mature miRNA molecules (DESeq2, P< 0.05, log2 fold-change > |1|). MiR-10a-5p represented over 21% of the miRNA molecules in OCCC with endometriosis and was significantly upregulated (NGS: log2fold change = 4.37, P = 2.43e-18; QPCR: 8.1-fold change, P< 0.05). Correlation between miR-10a expression level in OCCC cell lines and IC50 (50% inhibitory concentration) of carboplatin in vitro revealed a positive correlation (R2 = 0.93). MiR-10a overexpression in vitro resulted in a significant decrease in proliferation (n = 6; P< 0.05) compared to transfection with a non-targeting control miRNA. Similarly, the cell-cycle analysis revealed a significant shift in cells from S and G2 to G1 (n = 6; P< 0.0001). Bioinformatic analysis predicted that miR-10a-5p target genes that were downregulated in OCCC with endometriosis were involved in receptor signaling pathways, proliferation, and cell cycle progression. MiR-10a overexpression in vitro was correlated with decreased expression of predicted miR-10a target genes critical for proliferation, cell-cycle regulation, and cell survival including [SERPINE1 (3-fold downregulated; P< 0.05), CDK6 (2.4-fold downregulated; P< 0.05), and RAP2A (2-3-fold downregulated; P< 0.05)].DiscussionThese studies in OCCC suggest that miR-10a-5p is an impactful, potentially oncogenic molecule, which warrants further studies

Building a Systematic Online Living Evidence Summary of COVID-19 Research

Throughout the global coronavirus pandemic, we have seen an unprecedented volume of COVID-19 researchpublications. This vast body of evidence continues to grow, making it difficult for research users to keep up with the pace of evolving research findings. To enable the synthesis of this evidence for timely use by researchers, policymakers, and other stakeholders, we developed an automated workflow to collect, categorise, and visualise the evidence from primary COVID-19 research studies. We trained a crowd of volunteer reviewers to annotate studies by relevance to COVID-19, study objectives, and methodological approaches. Using these human decisions, we are training machine learning classifiers and applying text-mining tools to continually categorise the findings and evaluate the quality of COVID-19 evidence

Behavioral stress alters corticolimbic microglia in a sex- and brain region-specific manner

<div><p>Women are more susceptible to numerous stress-linked psychological disorders (e.g., depression) characterized by dysfunction of corticolimbic brain regions critical for emotion regulation and cognitive function. Although sparsely investigated, a number of studies indicate sex differences in stress effects on neuronal structure, function, and behaviors associated with these regions. We recently demonstrated a basal sex difference in- and differential effects of stress on- microglial activation in medial prefrontal cortex (mPFC). The resident immune cells of the brain, microglia are implicated in synaptic and dendritic plasticity, and cognitive-behavioral function. Here, we examined the effects of acute (3h/day, 1 day) and chronic (3h/day, 10 days) restraint stress on microglial density and morphology, as well as immune factor expression in orbitofrontal cortex (OFC), basolateral amygdala (BLA), and dorsal hippocampus (DHC) in male and female rats. Microglia were visualized, classified based on their morphology, and stereologically counted. Microglia-associated transcripts (CD40, iNOS, Arg1, CX3CL1, CX3CR1, CD200, and CD200R) were assessed in brain punches from each region. Expression of genes linked with cellular stress, neuroimmune state, and neuron-microglia communication varied between unstressed male and female rats in a region-specific manner. In OFC, chronic stress upregulated a wider variety of immune factors in females than in males. Acute stress increased microglia-associated transcripts in BLA in males, whereas chronic stress altered immune factor expression in BLA more broadly in females. In DHC, chronic stress increased immune factor expression in males but not females. Moreover, acute and chronic stress differentially affected microglial morphological activation state in male and female rats across all brain regions investigated. In males, chronic stress altered microglial activation in a pattern consistent with microglial involvement in stress-induced dendritic remodeling across OFC, BLA, and DHC. Together, these data suggest the potential for microglia-mediated sex differences in stress effects on neural structure, function, and behavior.</p></div

Corticolimbic sampling and microglial cell morphology.

<p>Stereological estimates (shaded in grey) and micropunch samples (dashed circle, indicated in one hemisphere for simplicity) were derived from both hemispheres of OFC, BLA, and DHC (CA3 stratum radiatum, dark grey; CA3 stratum oriens, light grey) using standard gross morphology (micropunch) and cytoarchitecture (stereology) [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0187631#pone.0187631.ref045" target="_blank">45</a>]. Microglia were classified as A) surveillant (numerous thin processes, radial branching); B) primed (thickened processes, increased polarity, proliferation, some reduced secondary branching); C) reactive (thickened stout processes with highly reduced branching); or D) amoeboid (few-to-no processes, enlarged cell body). Scale bars = 10 μm. Arrowheads indicate exemplars of each type.</p

Sex and stress effects on microglial morphology and immune factor expression in orbitofrontal cortex.

<p>A. Total microglial density based on estimated volume. Chronic stress slightly decreased total microglial density in females. B. Chronic stress reduced the proportion of primed to surveillant microglia in males. C. There were no sex differences in- or stress effects on CD40 expression. D. Acute stress increased iNOS transcript in females. Chronic stress induced iNOS expression in males and females. E. Chronic stress increases Arg1 mRNA in females. F. CX3CL1 expression is lower in unstressed females compared to unstressed males. Chronic stress increased CX3CL1 mRNA in males. G. Expression of CX3CR1 is reduced in unstressed females compared to unstressed males. Acute stress increases CX3CR1 mRNA in males, whereas chronic stress increases CX3CR1 transcript in females. H. Acute and chronic stress increase CD200 expression in females. I. Chronic stress increases CD200R mRNA in males. ^†p < .05, unstressed males compared to unstressed females. ^#p < .05, acute stress group compared to same-sex unstressed group. *p < .05, chronic stress group compared to same-sex unstressed group. Error bars indicate SEM. For graphs C-I, data are expressed relative to unstressed males.</p

Chronic stress reduces daily weight gain and induces adrenal hypertrophy in male and female rats.

<p>A. Data are displayed as mean weight gain per day (end body weight–starting body weight/number of study days). B. Data are displayed as mean adrenal-weight-to-body-weight ratios. *p < .05 chronic stress group compared to same-sex unstressed group. Error bars indicate SEM.</p

Sex differences in- and stress effects on- microglial morphological activation across corticolimbic brain regions.

<p>Associations between microglial morphological activation states across OFC, mPFC, BLA, and DHC were examined using Pearson correlation coefficients. Increased strength and/or number of associations would suggest greater microglial coupling, or decreased heterogeneity in microglial morphological activation state across corticolimbic brain regions. Microglial activation state is strongly correlated across corticolimbic brain regions in unstressed male- but not unstressed female- rats. Acute stress altered patterns of microglial morphological associations in both male and female rats. Remarkably, chronic stress reduced associations in microglial morphological activation state across corticolimbic circuitry in males, but induced associations in females. Green brain region nodes indicate an increase in microglial morphological activation state- and red nodes indicate a decrease in microglial morphological activation state as compared to same-sex unstressed animals. The strength of each association is represented in line weight (, r = -1.00 –-.71; , r = -0.70 –-0.61; , r = -0.60 –-0.50; , <i>r</i> = 0.50–0.60; , <i>r</i> = 0.61–0.70; , r = 0.71–1.00;); positive correlations are indicated with a solid line; negative correlations are indicated with a dashed line.</p

Sex and stress effects on microglial morphology and immune factor expression in basolateral amygdala.

<p>A. Total microglial density based on estimated volume. There were no effects of sex or stress on microglial density. B. Acute stress reduced the proportion of primed to surveillant microglia in males and females. Chronic stress reduced this proportion in males only. C. Expression of CD40 is significantly higher in unstressed females compared to unstressed males. Acute stress induced CD40 expression in males. Chronic stress decreased CD40 transcript in females. D. Chronic stress increased iNOS mRNA in females only. E. Unstressed females exhibited greater Arg1 expression compared to unstressed males. Acute stress induced Arg1 expression in males, whereas acute and chronic stress reduced Arg1 transcript in females. F. CX3CL1 expression did not vary with sex or stress. G. Acute stress increased CX3CR1 transcript in males only. H. There were no effects of sex or stress on CD200 mRNA. I. CD200R expression is higher in unstressed females compared to unstressed males. Acute and chronic stress increase CD200R transcript in males only. ^†p < .05, unstressed males compared to unstressed females. ^#p < .05, acute stress group compared to same-sex unstressed group. *p < .05, chronic stress group compared to same-sex unstressed group. Error bars indicate SEM. For graphs C-I, data are expressed relative to unstressed males.</p

qPCR primer sequences.

<p>qPCR primer sequences.</p