Objective: We investigated the clinical value of a novel circulating tumor cell (CTC) detection method-subtraction enrichment combined with immunostaining and fluorescence in situ hybridization (SEiFISH)-in ovarian cancer (OC). This study evaluated the diagnostic and prognostic significance of chromosome 8 aneuploidy in CTCs and circulating tumor endothelial cells (CTECs) for preoperative diagnosis, treatment efficacy assessment, and recurrence monitoring. Methods: A total of 331 patients were enrolled, including 56 with newly diagnosed primary OC, 265 with benign ovarian tumors, and 10 with borderline tumors. Peripheral blood CTCs and CTECs were detected using SEiFISH; their quantity and ploidy characteristics were analyzed in relation to clinical indicators. To assess dynamic CTC changes during disease progression and treatment response, 72 patients were followed longitudinally, of whom 19 experienced recurrence. Results: The CTC detection rate in OC patients was 92.9%, with significantly higher counts than that in the benign tumor group (median 5 vs. 2). Receiver operating characteristic analysis demonstrated good diagnostic performance for total CTCs [area under the curve (AUC)=0.699], with triploid CTCs achieving the highest efficacy (AUC=0.792), surpassing carbohydrate antigen 125 (CA125) (AUC=0.702). Postoperative follow-up showed that 70% of patients exhibited concurrent decreases in CTCs and CA125 levels, indicating disease improvement. In 30% of patients, CTC levels did not correlate with changes in CA125 levels. Individual case evidence suggests that CTC alterations may serve as an early indicator of recurrence or metastasis. Among the 19 recurrent cases, 73.7% showed elevated CTCs at recurrence that decreased following treatment. In four patients, CTCs reflected disease progression earlier than CA125, indicating higher sensitivity for recurrence monitoring. Conclusions: CTCs with chromosome 8 aneuploidy demonstrate significant clinical value in the preoperative diagnosis, treatment efficacy evaluation, and recurrence monitoring of OC. Dynamic CTC changes may serve as a more sensitive indicator than CA125 for disease surveillance, supporting the translational potential of CTC-based biomarkers in OC.

Background/Objectives: The diagnosis of endometriosis (EM) remains challenging due to the lack of a perfect diagnostic standard and the poor concordance between clinical symptoms and lesion severity. Although laparoscopy is widely used in clinical practice, it is invasive and associated with a non-negligible false-negative rate, while serum CA125 has limited diagnostic accuracy. In our previous studies, circulating endometrial cells (CECs) were identified in the peripheral blood of patients with EM, suggesting their potential as a non-invasive biomarker. Building on this finding, the present study aimed to systematically evaluate the clinical value of CECs in the diagnosis and stratified diagnosis of EM in the absence of a perfect diagnostic reference standard. Methods: Female patients treated at the Department of Obstetrics and Gynecology, Peking University People’s Hospital, between June 2022 and June 2024 were enrolled. Participants were clinically classified according to laparoscopic evaluation into an EM group and a non-EM group. However, laparoscopy was not treated as a definitive diagnostic gold standard in the statistical analysis. Instead, given the absence of a perfect reference standard, nonparametric latent class analysis was applied to jointly estimate disease status and the diagnostic performance of CECs, CA125, and laparoscopy. Patients with EM were further stratified according to dysmenorrhea severity (mild, moderate, and severe), lesion activity status (active or dormant), and menstrual cycle phase. Peripheral blood samples were collected from all participants, and CECs were detected using subtraction enrichment combined with immunofluorescence and fluorescence in situ hybridization (SE-iFISH). Serum CA125 levels were measured concurrently. Results: A total of 302 participants were included. The primary analysis focused on 133 surgically confirmed EM patients and 146 non-EM controls. After adjustment for an imperfect diagnostic reference standard, CECs demonstrated superior diagnostic performance compared with serum CA125 in the overall cohort, with higher sensitivity (0.58 vs. 0.37) and specificity (0.81 vs. 0.75). Under laparoscopic assessment in patients with severe dysmenorrhea (VAS ≥ 7), where the sensitivity and specificity were 0.759 and 1.00, respectively, CECs demonstrated superior diagnostic performance compared with serum CA125, with higher sensitivity (0.694 vs. 0.355) and specificity (0.946 vs. 0.429). Similarly, in patients with active EM, where laparoscopy showed a sensitivity of 0.79 and a specificity of 1.00, CECs again demonstrated superior diagnostic performance compared with CA125 (sensitivity 0.73 vs. 0.35; specificity 0.96 vs. 0.31), showing high concordance with laparoscopic diagnosis. When stratified by menstrual cycle phase, CECs maintained superior diagnostic performance over CA125 during both the proliferative and menstrual phases, with higher sensitivity (0.84 vs. 0.44) and specificity (0.83 vs. 0.65). Conclusions: Circulating endometrial cells (CECs) demonstrate high diagnostic accuracy for EM, significantly outperforming serum CA125, and show high concordance with laparoscopic diagnosis across clinically relevant stratified conditions in the absence of a perfect diagnostic gold standard.

Although poly (ADP-ribose) polymerase inhibitors (PARPi) have been established to enhance ovarian cancer outcomes, the emergence of drug resistance poses considerable clinical challenges. In this study, we constructed a Hi-C atlas to systematically characterize the effect of olaparib on chromatin organization at multiple hierarchical scales, namely, chromosomes, A/B compartments, topologically associating domains, and chromatin loops. To investigate the effects of PARPi on expression of the cohesion subunit RAD21, we established olaparib-resistant ovarian cancer cell line. Furthermore, we examined the effects of RAD21 on the functions of ovarian cancer cells and spheroids based on cell proliferation, apoptosis, and comet assays. In addition, by performing integrated analyses using ChIP-seq datasets, ChIP-qPCR, and chromosome conformation capture assays, we assessed the influence of RAD21 on the enhancer–promoter interactions of a homologous recombination repair gene. Moreover, on the basis of our findings in previous studies using clinical samples, we further evaluated the clinical value of RAD21 in multiple databases. Genome-wide Hi-C heatmap analysis revealed that olaparib led to a reduction in the genome-wide contact frequency for long distance interactions, altered the degree of chromatin compartmentalization, and promoted compartment switching in ovarian cancer. Differences between the olaparib-treated and control cells with respect to topologically associating domain boundaries and chromatin loops were found to be associated with key cellular functions, such as DNA repair and transcriptional mis-regulation in cancer. Furthermore, PARPi treatment was observed to induce the expression of RAD21, whereas an upregulation of RAD21 promoted proliferation and inhibited apoptosis in ovarian cancer spheroids. Mechanistically, we obtained evidence to indicate that by maintaining enhancer–promoter interactions within chromatin conformation, RAD21 regulates the transcription of RAD51, thereby mediating olaparib resistance in ovarian cancer. The high expression of RAD21 was found to show a significant association with poor overall and progression-free survival in patients with ovarian cancer. Our findings in this study indicate that RAD21 could serve as a potential therapeutic target for overcoming olaparib resistance in ovarian cancer, and provide new insights into the mechanisms underlying the resistance to PARPi from the perspective of chromatin organization.

Endometriosis is a common gynecological disorder characterized by the ectopic growth of endometrium-like tissue outside the uterine cavity and is associated with pelvic pain, infertility, and reduced quality of life. Although several theories have been proposed, the mechanisms by which endometrial cells disseminate and establish ectopic lesions remain incompletely understood. Here, we developed a dual-color pseudo-menstrual mouse model that enables real-time in vivo tracking of circulating endometrial cells (CECs) while distinguishing their tissue origin as uterine-derived or lesion-derived. Using in vivo flow cytometry and two-photon intravital microscopy, we found that both uterine-derived and lesion-derived CECs can enter the circulation and are detected at higher levels during menstruation-like breakdown, potentially in association with local vascular remodeling as suggested by intravital imaging observations. These cells could localize to distant organs, including the lungs and other extrapelvic organs. In addition, both CEC populations exhibited three distinct daily peak periods, and CEC dynamics were significantly associated with fluctuations in serum prolactin levels. In conclusion, these findings provide in vivo support for hematogenous dissemination in endometriosis and suggest that hormonal rhythmicity may regulate CEC dynamics, with potential diagnostic and therapeutic implications.

Objective:The C-reactive protein-albumin-lymphocyte (CALLY) index serves as an integrated indicator, reflecting the interplay between systemic inflammation, immune competence, and nutritional health. While validated in other chronic diseases, its relevance to endometriosis remains unexplored. We aimed to assess its association with endometriosis using nationally representative data. Methods:This cross-sectional analysis used data from four cycles of the National Health and Nutrition Examination Survey (NHANES, 1999-2006), including 4,602 US adults. The CALLY index was derived by integrating measurements of C-reactive protein (CRP) levels, serum albumin concentrations, and lymphocyte counts. Associations with endometriosis were estimated using weighted multivariable logistic regression across three progressively adjusted models. The discriminatory ability of CALLY was assessed using receiver operating characteristic (ROC) analyses and compared against that of other commonly used inflammatory biomarkers. Smoothed curve fitting and subgroup/interaction analyses assessed pattern and heterogeneity. Results:Log-transformed CALLY values (ln CALLY) levels showed a statistically significant inverse relationship with endometriosis prevalence (OR = 0.87; 95% CI: 0.79-0.96). Individuals in the top quartile of ln CALLY (Q4) had a 40% lower risk of endometriosis than those in the bottom quartile (Q1) (OR = 0.60, 95% CI: 0.41-0.87). Compared to other commonly used indices, CALLY exhibited superior discriminatory performance, achieving an AUC of 0.7219 (95% CI: 0.6970-0.7468) in the fully adjusted model. Curve fitting indicated that higher CALLY index levels were linked to a reduced likelihood of endometriosis. Subgroup analysis further confirmed the consistency and independence of this relationship (all p values for interactions > 0.05). Conclusion:A significant inverse association was observed between the CALLY index and endometriosis prevalence among American adults, supporting its potential as a novel inflammatory marker for identifying individuals at elevated risk. However, as a cross-sectional study, causality cannot be inferred. Future prospective cohorts across diverse populations are needed to evaluate its clinical utility for risk prediction and early identification.

Ovarian endometriosis is characterized by the growth of endometrial tissue within the ovary, causing infertility and chronic pain. However, its pathophysiology remains unclear. Utilizing high-precision single-cell RNA sequencing, we profile the normal, eutopic, and ectopic endometrium from 34 individuals across proliferative and secretory phases. We observe an increased proportion of ciliated cells in both eutopic and ectopic endometrium, characterized by a diminished expression of estrogen sulfotransferase, which likely confers apoptosis resistance. After translocating to ectopic lesions, endometrial epithelium upregulates nicotinamide N-methyltransferase expression that inhibits apoptosis by promoting deacetylation and subsequent nuclear exclusion of transcription factor forkhead box protein O1, thereby leading to the downregulation of the apoptotic gene BIM. Moreover, epithelial cells in ectopic lesions elevate HLA class II complex expression, which stimulates CD4+ T cells and consequently contributes to chronic inflammation. Altogether, our study provides a comprehensive atlas of ovarian endometriosis and highlights potential therapeutic targets for modulating apoptosis and inflammation.

Near-infrared fluorescence (NIRF) imaging is an excellent choice for image-guided surgery due to its simple operation and non-invasiveness. Developing tumor-specific fluorescent molecular probes is key to fluorescence imaging-guided surgery. EGFR (epidermal growth factor receptor) is closely related to the proliferation and growth of tumor cells and is highly expressed in epithelial ovarian cancer (EOC). The study aims to construct a NIR fluorescent molecular probe using cetuximab (an EGFR monoclonal antibody) and investigate its feasibility for targeting EOC in vivo through fluorescence imaging. We determined the expression of EGFR in EOC. NIR fluorescent molecular probe with cetuximab (cetuximab-Cy7) was chemically engineered and identified. The subcutaneous xenografted tumor model of EOC was induced using SKOV3-Luc cell line with positive expression of EGFR. Cetuximab-Cy7 was used for in vivo fluorescence imaging, and phosphate-buffered saline, free Cy7 dye and mouse isotype immunoglobulin G-Cy7 were used as controls. NIRF imaging system was performed to study the distribution and targeting of the probes. Tumors were imaged in situ and ex vivo, and fluorescent intensity was quantified. Resected specimens were analyzed to confirm diagnosis, and immunohistochemical (IHC) staining was used to identify EGFR expression. EGFR expression was increased in EOC tissues than fallopian tube tissues. The high expression of EGFR was significantly correlated with well-differentiation, residual lesions ≤ 1 cm, no recurrence and increased survival. NIRF imaging showed that the cetuximab-Cy7 enabled detection of tumor lesions in EOC-bearing mice with the optimal dose of 30 µg. The suitable imaging time window may be 24–96 h post-injection. Ex vivo fluorescence imaging indicated that fluorescent signal was mainly detected in the tumor and the lung. IHC results confirmed that xenografts were EGFR positive. Cetuximab-Cy7 can specifically target the tumors of EOC xenografted nude mice. This research lays the foundation for future studies on EOC surgery navigation.

Extracellular matrix protein 1 (ECM1) is a glycoprotein that may be a key player in tumorigenesis and tumor progression. However, knowledge regarding the role of ECM1 in endometriosis (EM) is still lacking. Microarray analyses were performed to compare the mRNA expression patterns between paired EU tissues and ectopic endometrial (EC) tissues (n = 4) from EM patients. ECM1 expression was significantly increased in the eutopic endometrial (EU) tissues than paired EC tissues of endometriotic patients and normal endometrial (NE) tissues of controls without EM. Blocking ECM1 with siRNA attenuated the migration and invasion of hEM15A cells and modified the distribution of the F-actin cytoskeleton. We conducted microarray analyses and bioinformatics analyses to investigate the differentially expressed genes (DEGs) and related pathways regulated by ECM1. A total of 161 DEGs between the siECM1 and the negative control (siNC) treatments were identified, consisting of 79 downregulated genes and 82 upregulated genes. Enriched DEGs were associated with 9 gene ontology (GO) terms. Moreover, a protein-protein interaction (PPI) network was constructed for the hub genes and modules. Radixin (RDX) was the second most downregulated gene in the siECM1 group compared with the siNC group. ECM1 knockdown significantly decreased the expression of RDX, RhoC, ROCK1, N-cadherin and β-catenin but not ROCK2. ECM1 showed high tissue-specific expression in EU tissues from EM patients, and may contribute to the migration, invasion and reorganization of the F-actin cytoskeleton in eutopic endometrial stromal cells via the RhoC/ROCK1 signaling pathway in EM.

BACKGROUND:Extracellular adenosine triphosphate (eATP) is an important inflammatory mediator that can boost the antitumour immune response, but its role in endometriosis remains unknown. We hypothesized that eATP could inhibit endometriosis cell function both directly and indirectly through macrophages.METHODS:Peritoneal and cyst fluid from endometriosis patients and non-endometriosis controls was collected to measure eATP levels. The addition of eATP was performed to explore its effects on endometriotic cell and macrophage functions, including cell proliferation, apoptosis, pyroptosis, mitochondrial membrane potential, phagocytosis, and the production of inflammatory cytokines and reactive oxygen species. A coculture of endometriotic epithelial cells and U937 macrophages was established, followed by P2X7 antagonist and eATP treatment. Endometriosis model eATP-treated rats were used to evaluate in situ cell death and macrophage marker expression.RESULTS:The pelvic microenvironment of endometriosis patients shows high eATP levels, which could induce endometriotic epithelial cell apoptosis and pyroptosis and significantly inhibit cell growth via the MAPK/JNK/Akt pathway. eATP treatment ameliorated endometriosis-related macrophage dysfunction and promoted macrophage recruitment. eATP treatment in the presence of macrophages exerted a stronger cytotoxic effect on endometriotic epithelial cells by regulating P2X7. eATP treatment effectively induced cell death in an endometriosis rat model and prominently increased the macrophage number without affecting the eutopic endometrium.CONCLUSION:eATP induces endometriotic epithelial cell death and enhances the immune function of macrophages to inhibit the progression of endometriosis, while eutopic endometrium is not affected. eATP treatment may serve as a nonhormonal therapeutic strategy for endometriosis.

BACKGROUND:Endometriosis (EM) is a complex benign gynecological disease, but it has malignant biological behavior and can invade any part of the body. Clinical manifestations include pelvic pain, dysmenorrhea, infertility, pelvic nodules, and masses. Our previous study successfully detected circulating endometrial cells (CECs) in the peripheral blood of patients with EM. The purpose of this study is to overcome the limitation of cell size in the previous microfluidic chip method, to further accurately capture CECs, understand the characteristics of these cells, and explore the relationship between CECs and the clinical course characteristics of patients with EM. METHODS:Human peripheral venous blood used to detect CECs and circulating vascular endothelial cells (CVECs) was taken from EM patients ( n = 34) hospitalized in the Peking University People's Hospital. We used the subtraction enrichment and immunostaining fluorescence in situ hybridization (SE-iFISH) method to exclude the interference of red blood cells, white blood cells, and CVECs, so as to accurately capture the CECs in the peripheral blood of patients with EM. Then we clarified the size and ploidy number of chromosome 8 of CECs, and a second grouping of patients was performed based on clinical characteristics to determine the relationship between CECs and clinical course characteristics. RESULTS:The peripheral blood of 34 EM patients and 12 non-EM patients was evaluated by SE-iFISH. Overall, 34 eligible EM patients were enrolled. The results showed that the detection rates of CECs were 58.8% in EM patients and 16.7% in the control group. However, after classification according to clinical characteristics, more CECs could be detected in the peripheral blood of patients with rapidly progressive EM, with a detection rate of 94.4% (17/18). In total, 63.5% (40/63) of these cells were small cells with diameters below 5 μm, and 44.4% (28/63) were aneuploid cells. No significant association was found between the number of CECs and EM stage. CONCLUSION:The number and characteristics of CECs are related to the clinical course characteristics of patients with EM, such as pain and changes in lesion size, and may be used as biomarkers for personalized treatment and management of EM in the future.

High malignancy is a prominent characteristic of epithelial ovarian cancer (EOC), emphasizing the necessity for further elucidation of the potential mechanisms underlying cancer progression. Aneuploidy and copy number variation (CNV) partially contribute to the heightened malignancy observed in EOC; however, the precise features of aneuploidy and their underlying molecular patterns, as well as the relationship between CNV and aneuploidy in EOC, remain unclear. In this study, we employed single-cell sequencing data along with The Cancer Genome Atlas (TCGA) to investigate aneuploidy and CNV in EOC. The technique of fluorescence in situ hybridization (FISH) was employed using specific probes. The copy number variation within the genomic region of chromosome 8 (42754568-47889815) was assessed and utilized as a representative measure for the ploidy status of individual cells in chromosome 8. Differential expression analysis was performed between different subgroups based on chromosome 8 ploidy. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), protein–protein interaction (PPI), and hub–gene analyses were subsequently utilized to identify crucial genes involved. By classifying enriched tumor cells into distinct subtypes based on chromosome 8 ploidy combined with TCGA data integration, we identified key genes driving chromosome 8 aneuploidy in EOC, revealing that PRKDC gene involvement through the mediated non-homologous end-joining pathway may play a pivotal role in disease progression. Further validation through analysis of the GEO and TCGA database and survival assessment, considering both mRNA expression levels and CNV status of PRKDC, has confirmed its involvement in the progression of EOC. Further functional analysis revealed an upregulation of PRKDC in both ovarian EOC cells and tissues, with its expression showing a significant correlation with the extent of copy number variation (CNV) on chromosome 8. Taken together, CNV amplification and aneuploidy of chromosome 8 are important characteristics of EOC. PRKDC and the mediated NHEJ pathway may play a crucial role in driving aneuploidy on chromosome 8 during the progression of EOC.

BACKGROUND:High-grade serous ovarian cancer (HGSOC) is the biggest cause of gynecological cancer-related mortality because of its extremely metastatic nature. This study aimed to explore and evaluate the characteristics of candidate factors associated with the metastasis and progression of HGSOC.METHODS:Transcriptomic data of HGSOC patients' samples collected from primary tumors and matched omental metastatic tumors were obtained from three independent studies in the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) were selected to evaluate the effects on the prognosis and progression of ovarian cancer using data from The Cancer Genome Atlas (TCGA) database. Hub genes' immune landscapes were estimated by the Tumor Immune Estimation Resource (TIMER) database. Finally, using 25 HGSOC patients' cancer tissues and 10 normal fallopian tube tissues, immunohistochemistry (IHC) was performed to quantify the expression levels of hub genes associated with International Federation of Gynecology and Obstetrics (FIGO) stages.RESULTS:Fourteen DEGs, ADIPOQ , ALPK2 , BARX1 , CD37 , CNR2 , COL5A3 , FABP4 , FAP , GPR68 , ITGBL1 , MOXD1 , PODNL1 , SFRP2 , and TRAF3IP3 , were upregulated in metastatic tumors in every database while CADPS , GATA4 , STAR , and TSPAN8 were downregulated. ALPK2 , FAP , SFRP2 , GATA4 , STAR , and TSPAN8 were selected as hub genes significantly associated with survival and recurrence. All hub genes were correlated with tumor microenvironment infiltration, especially cancer-associated fibroblasts and natural killer (NK) cells. Furthermore, the expression of FAP and SFRP2 was positively correlated with the International Federation of Gynecology and Obstetrics (FIGO) stage, and their increased protein expression levels in metastatic samples compared with primary tumor samples and normal tissues were confirmed by IHC ( P = 0.0002 and P = 0.0001, respectively).CONCLUSIONS:This study describes screening for DEGs in HGSOC primary tumors and matched metastasis tumors using integrated bioinformatics analyses. We identified six hub genes that were correlated with the progression of HGSOC, particularly FAP and SFRP2 , which might provide effective targets to predict prognosis and provide novel insights into individual therapeutic strategies for HGSOC.

Cancer stem cells (CSCs) express pluripotent markers and share many features with normal pluripotent stem cells. It is possible that immunity induced by embryonic stem cells (ESCs) and induced pluripotent stem cells- (IPSCs-) based vaccines may selectively target CSCs. In our study, cells expressing the pluripotent marker CD133 in the murine ovarian cancer cell-line ID8 were isolated and identified as CSCs. We investigated the preventive efficacy of ESCs and IPSCs-based vaccines against the development of ovarian cancer in vivo and evaluated the humoral and cellular immunities targeting CSCs in vitro. Our study showed that preimmunization with both mouse-derived embryonic stem cells (mESCs) and mouse-induced pluripotent stem cells (mIPSCs) lysates, combined with an immunostimulatory adjuvant CpG, elicited strong humoral and cellular responses. These responses effectively suppressed the development of CSC-derived tumors. Immune sera collected from mESCs and mIPSCs-vaccinated mice contained antibodies that were capable of selectively targeting CSCs, resulting in the lysis of CSCs in the presence of complement. Cytotoxic T-lymphocytes generated from splenocytes of mESCs and mIPSCs-vaccinated hosts could secrete interferon- (IFN-) γ in response to CSCs and kill CSCs in vitro. These findings indicate that vaccines based on mESCs and mIPSCs can elicit effective antitumor immunities. These immunities are related to the conferring of humoral and cellular responses that directly target CSCs.

ABSTRACT Research question : What is the implication of aberrant CD24 expressions in endometriosis (EM) and the relevant mechanisms? Design : Thirty women with EM and twenty-seven women with other benign gynecological diseases, who underwent laparoscopy and hysteroscopy, were recruited in this study. The hEM15A cells were used in functional experiments in vitro. The microarray assay and bioinformatics analyses were performed to investigate the differentially expressed genes (DEGs) and signaling pathways regulated by CD24. Results : CD24 was expressed at significantly higher levels in eutopic endometrial (EU) tissues than in paired ectopic endometrial (EC) tissues from EM patients and normal endometrial (NE) tissues from controls. However, the CD24 expressions had no obvious relationships with clinical features such as age, infertility conditions, and menstrual cycle phase. Suppression of CD24 by small interfering RNAs in hEM15A cells impaired cell migration and reduced cytoskeletal filamentous actin (F-actin), but had no effects on cell apoptosis, cell cycle or cell viability. A total of 862 DEGs were identified between siCD24 and siNC, which consisted of 517 upregulated genes and 345 downregulated genes. The gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses for the DEGs mainly focused on the type I interferon signaling pathway and TGF-beta signaling pathway. Gene set enrichment analysis (GSEA) was also conducted. Furthermore, a protein-protein interaction (PPI) network and subnetwork were constructed to explore the hub modules and DEGs. Conclusions : This study provides a better understanding of the role of CD24 in EM.

Supplementary Table from Single-Cell Dissection of the Multiomic Landscape of High-Grade Serous Ovarian Cancer

BackgroundOur previous work revealed the high expression of fibrinogen alpha chain (FGA) in patients with endometriosis (EM) and that it could promote the migration and invasion of endometrial stromal cells. Angiogenesis is the key condition for the development of EM. This study was aimed to elucidate the role of FGA in endometrial stromal cells involved in angiogenesis in EM. MethodsImmunohistochemistry was used to detect the microvessel density (MVD) and VEGF expression in the eutopic endometrium samples from EM and non-EM. The conditioned medium (CM) of human primary eutopic endometrial stromal cells (EuESC) and immortalized endometrial stromal cell line hEM15A with FGA knockdown were collected and used to treat human umbilical vein endothelial cells (HUVECs). Then, tube formation assay, EdU assay, wound assay, transwell assay and flow cytometry assays were performed to assess the function of HUEVCs in vitro. The angiogenic capability of HUVECs was further measured using a matrigel plug assay with BALB/c nude mice in vivo. Immunofluorescence was used to detect the expression of F-actin and VE-cadherin. RT-PCR and western blotting were used to detect the expression of angiogenesis-related factors in endometrial stromal cells and downstream signalling pathways in HUVECs. ResultsMVD and VEGF expression in the eutopic endometrium of EM patients were significantly higher than those in the normal endometrium of non-EM patients, and the increased MVD in EM indicates an increased risk of recurrence. Functionally, we found that CM of endometrial stromal cells with FGA knockdown could inhibit HUEVCs migration and tube formation in vitro and in vivo, while having no significant effect on HUVECs proliferation, apoptosis and cell cycle. Mechanically, the expression of VEGFA, PDGF, FGF-B, VEGF, MMP-2 and MMP-9 was reduced in hEM15A cells with FGA knockdown. CM of hEM15A cells with FGA knockdown reduced the number of microfilaments and pseudopodia, as well as the expression of VE-cadherin, and inhibited the activity of VEGFR2 and the FAK signalling pathway in HUVECs. ConclusionOur study demonstrated FGA could enhance the interaction between endometrial stromal cells and HUVECs via the potential VEGA-VEGFR-FAK signalling axis and promote EM angiogenesis, revealing a promising therapeutic approach for EM.

Ovarian cancer is a leading cause of death among gynecological malignancies, and novel therapies are urgently needed. Here we report preliminary findings on the potential safety and efficacy of 6B11-OCIK, an adoptive cell therapy of autologous T cells induced by the humanized anti-idiotypic antibody 6B11 minibody plus dendritic cells and cytokines, against platinum-resistant recurrent or refractory ovarian cancer in three patients. We found that 6B11-OCIK treatment was safe and well tolerated after five cycles of intravenous infusion with an initial dose of 1–2×109 cells and a dose-climbing strategy. Hemoglobin, platelets, white cell count, creatinine or liver enzyme values, coagulation function, kidney and heart function were not significantly affected over the duration of therapy. Two of the three enrolled patients showed potentially drug-related grade 1 and 2 weakness, and no other adverse events were observed. Of the three enrolled patients, one had stable disease and two showed disease progression. The patient with favorable clinical efficacy had better immune response as measured by 6B11-OCIK proliferation capacity, activation ability of CD3+CD8+ tumor-specific cytotoxic T lymphocytes and CD3+CD56+ cytokine-induced killer cells, and tumor cell killing efficiency. Changes in circulating tumor cells after treatment were consistent with serum level CA125 in the patient with stable disease (both decreased), while differences were observed in the two patients with disease progression (increased CA125 in both and decreased CTC in the patient with better immune response), suggesting that variation of circulating tumor cells was more consistent with immune response and reflected efficacy directly. This preliminary study suggested that autologous 6B11-OCIK treatment was safe and had potential clinical efficacy against ovarian cancer. Patients with better immune response had more favorable efficacy. In addition to imaging, CA125 and immunophenotypes, CTC monitoring may represent a potential indicator of immunotherapy response.

细胞治疗作为新兴的现代生物医学技术,促进了以精准医学和系统生物学为代表的治疗变革,也不断地与中医关于生命认识的整体观和"天人合一"的思维接近.细胞治疗为肿瘤等多种疾病的治疗提供了新的策略,但其不良反应和应用局限制约了其在临床上的广泛应用.中医药与细胞治疗联合使用,发挥协同作用,在提高细胞治疗疗效、减轻毒副作用方面已经做了一些探索和尝试.概述中医药在免疫细胞治疗、干细胞治疗和基因修饰体细胞治疗中的应用现状,初步探讨中医药联合细胞治疗在未来的发展趋势.

Recent studies have shown the involvement of exosomes in intercellular communication during tumor progression. Circular RNAs (circRNAs) can be packaged into exosomes for extracellular communication, however, the possible effects of exosomal circRNAs in epithelial ovarian cancer (EOC) cells with high metastatic potential have been rarely studied. In this study, we identified exosomal circRNA051239 from high-metastatic ovarian cancer SKOV3.ip cells and subsequently analyzed circRNA051239 levels in both EOC tissues and exosomes derived from plasma and cells by qRT-PCR. A variety of in vitro assays were employed to observe the effects of exosomal circRNA051239 derived from high-metastatic ovarian cancer SKOV3.ip cells on low-metastatic ovarian cancer SKOV3 cells. Bioinformatics analysis and luciferase activity assays were further utilized to confirm the relationship between circRNA051239, miR-509-5p and PRSS3. As a result, circRNA051239 expression was increased in tissues and plasma exosomes from EOC patients. Moreover, si-circRNA051239-Exo (exosomes derived from circRNA051239 knockdown SKOV3.ip cells) inhibited the proliferation, migration as well as invasion of SKOV3 cells. Mechanistically, circRNA051239 functioned as a competitive endogenous RNA (ceRNA) by sponging miR-509-5p to facilitate PRSS3 expression. Exosomal circRNA051239 derived from high-metastatic ovarian cancer SKOV3.ip cells promoted the progression of low-metastatic ovarian cancer SKOV3 cells. Collectively, these outcomes implicated that higher metastatic EOC cells can confer this potential to lower metastatic potential via exosomal circRNA051239, causing enhanced proliferative, migratory and invasive capacities in recipient cells.

基因编辑技术可针对性增强体细胞的靶向性,一定程度上解决了传统过继性免疫疗法在肿瘤等多种疾病治疗中显露出的靶向性差等问题.最具代表性的是嵌合抗原受体T细胞(CAR-T)免疫疗法,其在血液系统肿瘤治疗中已取得良好疗效,此外,T细胞受体基因工程化的T细胞(TCR-T)、基因修饰的树突状细胞以及基因修饰的干细胞在多种疾病治疗的临床研究中也显现了良好的安全性和有效性.目前国内外多家研究机构和生物公司已在基因修饰体细胞行业加速布局,相关产业即将进入快速发展期,基因修饰体细胞疗法的临床转化应用具有广阔前景,对目前己经开展疾病临床研究的几种基因修饰免疫细胞疗法进行总结.