Objective Non–high-grade epithelial cancers originating in the ovary, fallopian tube, or peritoneum are rare, and the role of hyperthermic intraperitoneal chemotherapy (HIPEC) in patients with these cancers is undefined. We evaluated whether HIPEC is associated with improved overall survival and, secondarily, in which histologic subgroup the association was strongest. Methods Adults with stage III–IV non–high-grade epithelial ovarian, fallopian tube, or primary peritoneal carcinoma undergoing primary or interval cytoreductive surgery in the National Cancer Database (2004–2022) were classified by HIPEC receipt. Overall survival was administratively censored at 10 years. The entire eligible cohort was analyzed with era-balanced 1:4 propensity-score matching, full-cohort inverse-probability and overlap weighting, multiple imputation, and E-values. Results Among 15,654 patients, 72 (0.5%) received HIPEC and were matched to 288 controls. In the matched cohort, 5- and 10-year overall survival were 64.5% and 48.4% with HIPEC versus 50.0% and 29.0% without. HIPEC was associated with improved 10-year overall survival across estimators (overlap-weighted HR 0.66, 95% CI 0.46–0.95, p=0.024; inverse-probability-weighted HR 0.66, 95% CI 0.47–0.95, p=0.024; matched-set HR 0.52, 95% CI 0.33–0.81, p=0.004; multiply-imputed HR 0.66, 95% CI 0.46–0.94, p=0.023), with a directionally consistent multivariable estimate (HR 0.72, 95% CI 0.49–1.04, p=0.079). In exploratory analyses, the association was strongest in the 17 HIPEC-treated patients with mucinous carcinoma (multivariable HR 0.39, 95% CI 0.17–0.88, p=0.024) and was near-null in non-mucinous histologies (HR 0.93, 95% CI 0.56–1.54); these estimates rest on small numbers. Conclusions In this national cohort, HIPEC was associated with improved 10-year overall survival after adjustment for measured confounders, although unmeasured factors such as completeness of cytoreduction and tumor burden cannot be excluded. The association was strongest among patients with mucinous carcinoma. Pending prospective, histology-specific evaluation, these data may inform individualized discussion of HIPEC with selected patients at experienced centers, with counseling that high-level evidence is lacking.

Uterine corpus endometrial carcinoma (UCEC) is the most common gynecologic malignancy, with poor prognoses in advanced, high-grade, or treatment-resistant cases. The transcription factor NR2F6 (nuclear receptor subfamily 2 group F member 6) has emerged as a key regulator of immune evasion, tumor progression, and therapy resistance, suppressing T-cell activation by regulating PD-L1 (programmed death-ligand 1) expression. Nucleus accumbens-associated protein 1 (NAC1), encoded by the NACC1 gene, is a transcriptional repressor critical for tumor growth, metastasis, and drug resistance. NOTCH3 (Notch homolog 3), a downstream regulator, contributes to therapy resistance and tumor aggressiveness. This study investigates the significant therapeutic potential of targeting this axis. Gene expression data from 529 UCEC patients were obtained from The Cancer Genome Atlas (TCGA), and survival analyses were performed using R Studio. Microsatellite instability (MSI) cell line HEC1A and microsatellite stable (MSS) cell line KLE were obtained from ATCC. Knockdowns of NR2F6 and NACC1 were performed using small interfering RNA (siRNA). Semi-quantitative real-time PCR (RT-qPCR) and Western blot were used to measure NR2F6, NACC1, NOTCH3, and PD-L1 expression. Cell proliferation was evaluated using Promega assays. NIC3, a small-molecule inhibitor that prevents NAC1 homodimerization and induces proteasomal degradation, was applied to validate findings from siRNA experiments. Statistical significance was determined using paired Student’s t-tests. High expression of NR2F6 and NACC1 was associated with poor survival in UCEC patients (NR2F6: p = 0.0017; NACC1: p = 0.019). Correlation analyses showed a strong relationship between NR2F6 and NACC1 (Pearson r = 0.54, p < 0.0001) and a moderate relationship between NACC1 and NOTCH3 (r = 0.45, p < 0.0001). RT-qPCR and Western blot analyses revealed that NR2F6 knockdown significantly reduced NACC1, NOTCH3, and PD-L1 expression. NACC1 knockdown similarly decreased NOTCH3 and PD-L1 expression, confirming its regulatory role in the axis. Notably, siRNA-mediated knockdowns and NIC3 treatment significantly reduced cellular proliferation across all treated cell lines (HEC1A, KLE, and Colo 684). NIC3 similarly demonstrated the ability to degrade NAC1 protein and reduce downstream NOTCH3 and PD-L1 expression. This study highlights the therapeutic potential of targeting the NR2F6-NACC1-NOTCH3 axis in UCEC. Validation at both mRNA and protein levels, combined with the observed reduction in cellular proliferation, underscores the promise of this axis as a critical target in high-grade or treatment-resistant tumors. The small-molecule inhibitor NIC3 offers a novel strategy to disrupt NAC1-mediated signaling, and potentially target UCEC. Xiaoqi Sun, Xiaofang Guo, Tram Le, Vaagn Andikyan, Thomas J. Rutherford, Eduardo Sotomayor, Matthew L. Anderson, Adrian Kohut. Targeting the NR2F6-NACC1-NOTCH3 axis: a promising therapeutic strategy to combat tumor progression and resistance in endometrial cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 5685.

Leiomyomas are benign proliferations of uterine smooth muscle found in 60% of women. A spatial redistribution of ecto-5'-nucleotidase (CD73, NT5E) that results in reduced extracellular concentrations of adenosine has recently been described in leiomyomas. However, the mechanisms by which altered extracellular adenosine levels contribute to leiomyoma growth remain poorly understood. To address this deficiency, a series of tissue specimens and primary cultures generated from matched specimens of myometrium and leiomyoma were used. Overexpression of Type 1 adenosine receptors (ADORA1) was observed when matched specimens and primary cultures were interrogated by RT-qPCR and western blot. By immunohistochemistry, ADORA1 expression was diffusely observed in myocytes in the leiomyoma complex, with only limited expression in vascular and other structures. Overexpression of ADORA1 was also observed in fibroblasts and multiple smooth muscle subtypes in the leiomyoma complex when single-cell transcriptomics data were interrogated. Incubation with N6-cyclopentyladenosine (CPA), a selective ADORA1 agonist, resulted in decreased proliferation of primary leiomyoma cultures, accompanied by decreased intracellular cAMP and enhanced cyclin D1 and phospho-AKT1 expression. To confirm the specificity of this observation, ADORA1 expression was directly targeted by siRNA, resulting in decreased proliferation, increased intracellular cAMP, and lower levels of cyclin D1 and phospho-AKT1. Collectively, these data indicate that overexpression of the ADORA1 receptor is a robust feature of uterine leiomyomas, where its activation by residual levels of extracellular adenosine potentially contributes to tumor growth by regulating AKT1-mediated signaling.

Hyperthermic intraperitoneal chemotherapy (HIPEC) is used to eliminate minimal residual disease in patients with peritoneal surface malignancies, including advanced epithelial ovarian cancer (EOC). While some trials suggest potential benefits, the role of HIPEC during cytoreductive surgery (CRS) in EOC remains uncertain. This study aimed to evaluate outcomes for patients undergoing HIPEC during CRS for advanced-stage EOC in the United States (US). This multicenter, retrospective cohort study included women with stage III–IV EOC who underwent CRS with or without HIPEC between 2006 and 2021 at Commission on Cancer-accredited US facilities. Propensity score matching was used to create a control group of patients who underwent CRS only. Overall survival (OS) was analyzed using the Kaplan–Meier log-rank method and adjusted for confounding factors with Cox proportional hazards regression. Among 1400 patients identified, 700 underwent CRS with HIPEC and 700 underwent CRS only. Of these 1400 patients, 932 underwent interval CRS and 468 underwent primary CRS. No significant difference in median OS was observed between the overall CRS+HIPEC and CRS-only groups (57.6 vs. 47.6 months; p = 0.105). However, interval CRS+HIPEC was associated with significantly improved median OS compared with interval CRS-only (57.6 vs. 45.7 months; p = 0.003). After adjustment, HIPEC remained significantly associated with improved survival (hazard ratio 0.77, 95

Primers used to clone CARG-2020, VLV-12 and VLV-GFP

Diagram showing major immune phenotypes in each phase of ovarian tumor progression.

A. Mean tumor burden over time of nude mice bearing OCSC1-F2 human ovarian tumors. mCherry fluorescent ROI was used to provide a measure of tumor burden. Data are presented as mean ± SEM. Treatments began on day 6 and continued through to day 12. B. Survival analysis using Kaplan-Meier.

Flow cytometry gating strategy CD8 T cells, Macrophages and MDSC.

A. Diagram of CARG-2020 vector and GFP control vector; B. TKO mouse ovarian cancer cells were treated in vitro with PBS Control or 1x105 PFU/cell of CARG-2020 for 24h. IL12 mRNA was quantified by qPCR and secreted IL-12p40 and p70 protein were quantified using xMAP technology; C. Expression of IL-12 and IL-17R ECD were detected by western blot analysis in total cell lysate. D. Secretion of IL-17 RA-ECD was detected by western blot analysis in the supernatant of cells treated with CARG-2020. Control cells were treated with VLV-GFP vector. Representative images of 3 independent experiments. E. Expression of PDL-1 following treatment with CARG-2020. Note the time dependent decrease on PDL-1 expression on cells treated with CARG-2020. Control=VLV-GFP. Representative imagesof 3 independent experiments.

Abstract Current immunotherapies have proven effective in strengthening antitumor immune responses, but constant opposing signals from tumor cells and the surrounding microenvironment eventually lead to immune escape. We hypothesized that in situ release of antigens and regulation of both the innate and adaptive arms of the immune system would provide a robust and long-term antitumor effect by creating immunologic memory against tumors. To achieve this, we developed CARG-2020, a genetically modified virus-like vesicle (VLV) that is a self-amplifying RNA with oncolytic capacity and encodes immune regulatory genes. CARG-2020 carries three immune modulators: (i) the pleiotropic antitumor cytokine IL12, in which the subunits (p35 and p40) are tethered together; (ii) the extracellular domain (ECD) of the protumor IL17RA, which serves as a dominant-negative antagonist; and (iii) a shRNA targeting PD-L1. Using a mouse model of ovarian cancer, we demonstrated the oncolytic effect and immune-modulatory capacities of CARG-2020. By enhancing IL12 and blocking IL17 and PD-L1, CARG-2020 successfully reactivated immune surveillance by promoting M1, instead of M2, macrophage differentiation, inhibiting MDSC expansion and establishing a potent CD8+ T cell–mediated antitumoral response. Furthermore, we demonstrated that this therapeutic approach provided tumor-specific and long-term protection against the establishment of new tumors. Our results provide a rationale for the further development of this platform as a therapeutic modality for ovarian cancer patients to enhance antitumor responses and prevent a recurrence.

Found in as many as 80% of women, uterine leiomyomas are a frequent cause of abnormal uterine bleeding, pelvic pain, and infertility. Despite their significant clinical impact, the mechanisms responsible for driving leiomyoma growth remain poorly understood. After obtaining IRB permission, expression of ecto-5'-nucleotidase (NT5E, CD73) was assessed in matched specimens of myometrium and leiomyoma by real-time qPCR, Western blot, and immunohistochemistry (IHC). Adenosine concentrations were measured by enzyme-linked assay. Primary cultures were used to assess the impact of adenosine and/or adenosine receptor agonists on proliferation, apoptosis, and patterns of intracellular signaling in vitro. When compared to matched specimens of healthy myometrium, uterine leiomyomas were characterized by reduced CD73 expression. Largely limited to thin-walled vascular structures and the pseudocapsule of leiomyomas despite diffuse myometrial distribution. Restricted intra-tumoral CD73 expression was accompanied by decreased levels of intra-tumoral adenosine. In vitro, incubation of primary leiomyoma cultures with adenosine or its hydrolysis-resistant analog 2-chloro-adenosine (2-CL-AD) inhibited proliferation, induced apoptosis, and reduced proportion of myocytes in S- and G2-M phases of the cell cycle. Decreased proliferation was accompanied by reduced expression of phospho-Akt, phospho-Cdk2-Tyr15, and phospho-Histone H3. Enforced expression of the A2B adenosine receptor (ADORA2B) and ADORA2B-selective agonists similarly suppressed proliferation and inhibited Akt phosphorylation. Collectively, these observations broadly implicate CD73 and reduced extracellular concentrations of adenosine as key regulators of leiomyoma growth and potentially identify novel strategies for clinically managing these common tumors.

VEGFR2 (Vascular endothelial growth factor receptor 2) is a central regulator of placental angiogenesis. The study of the VEGFR2 proteome of chorionic villi at term revealed its partners MDMX (Double minute 4 protein) and PICALM (Phosphatidylinositol-binding clathrin assembly protein). Subsequently, the oxytocin receptor (OT-R) and vasopressin V1aR receptor were detected in MDMX and PICALM immunoprecipitations. Immunogold electron microscopy showed VEGFR2 on endothelial cell (EC) nuclei, mitochondria, and Hofbauer cells (HC), tissue-resident macrophages of the placenta. MDMX, PICALM, and V1aR were located on EC plasma membranes, nuclei, and HC nuclei. Unexpectedly, PICALM and OT-R were detected on EC projections into the fetal lumen and OT-R on 20–150 nm clusters therein, prompting the hypothesis that placental exosomes transport OT-R to the fetus and across the blood–brain barrier. Insights on gestational complications were gained by univariable and multivariable regression analyses associating preeclampsia with lower MDMX protein levels in membrane extracts of chorionic villi, and lower MDMX, PICALM, OT-R, and V1aR with spontaneous vaginal deliveries compared to cesarean deliveries before the onset of labor. We found select associations between higher MDMX, PICALM, OT-R protein levels and either gravidity, diabetes, BMI, maternal age, or neonatal weight, and correlations only between PICALM-OT-R (p < 2.7 × 10–8), PICALM-V1aR (p < 0.006), and OT-R-V1aR (p < 0.001). These results offer for exploration new partnerships in metabolic networks, tissue-resident immunity, and labor, notably for HC that predominantly express MDMX.

A. Kaplan Meier curve comparing overall survival in mice treated with PBS Control, CARG-2020, VLV-IL-12, VLV-17R ECD, VLV-shRNA PD-L1, and VLV-GFP; p<0.0001. B. CARG-2020 but not VLV-IL-12 modulates the innate immune system. Peritoneal lavage from VLV-IL-12 and CARG-2020-treated mice (n=6) were analyzed by flow cytometry for macrophages using CD11b and CD206. VLV-IL-12 treatment is not able to increase the number of CD11blow/CD206- macrophages compared to CARG2020. Representative dot plots shown. Gating strategy is shown in Supp. Fig. 1. C. Decrease on Ly6C+/Ly6G+ MSC is observed only in the CARG2020 treated group but not in those treated with VLV-IL-12 group. Representative dot plots shown. Gating strategy is shown in Supp. Fig. 1.

Chromobox protein homolog 7 (CBX7), a member of the Polycomb repressor complex, is a potent epigenetic regulator and gene silencer. Our group previously reported that CBX7 functions as a tumor suppressor in ovarian cancer cells, and its loss accelerated the formation of carcinomatosis and drove tumor progression in an ovarian cancer mouse model. The goal of this study was to identify specific signaling pathways in the ovarian tumor microenvironment that can downregulate CBX7. Adipocytes are an integral component of the peritoneal cavity and the ovarian tumor microenvironment.