Jing Cai

Efferocytosis‐Driven Polyamine Metabolism in Macrophages Enhances Cancer Stem Cell Enrichment after Chemotherapy in Ovarian Cancer

Abstract Chemotherapy‐induced enrichment of cancer stem cells (CSCs) is a key mechanism underlying acquired chemoresistance and recurrence of epithelial ovarian cancer (OC). Although chemotherapy may enrich CSCs through selection or by inducing dedifferentiation, the dynamic changes in the tumor niche and their impact on CSCs during chemotherapy remain unclear. In this study, single‐cell sequencing and multiplex immunohistochemical analysis are used to define microenvironmental changes, and a post‐chemotherapy increase in efferocytotic macrophages that phagocytosed chemotherapy‐induced apoptotic tumor cells is identified. Efferocytotic macrophages are associated with poor prognosis and CSCs in OC. Their conditioned medium facilitates OC stemness in vitro. Meanwhile, targeting efferocytosis suppresses CSC enrichment, chemoresistance, and regrowth in vivo. Mechanistically, it is demonstrated that enhanced expression of ODC1 driven by efferocytosis increases polyamine flux, particularly putrescine, by integrating metabolomics and transcriptomics. The increase in putrescine content leads to the SPP1 and OPN overexpression in macrophages, conferring cancer stemness to OC cells through the OPN‐CD44 axis. Treatment with an ODC1 selector inhibitor mitigates CSC enrichment, sensitizes tumors to cisplatin, and restricts tumor regrowth. Together, the study shows that efferocytosis and associated polyamine metabolic reprogramming support the chemotherapy‐induced enrichment of CSCs, providing new targets for addressing chemoresistance and recurrence of OC.

LEF1 confers resistance to DNA-damaging chemotherapies through upregulation of PARP1 and NUMA1 in ovarian cancer

Resistance to platinum-based drugs and PARP inhibitors (PARPi) is the leading cause of treatment failure in epithelial ovarian cancer (EOC). This study aimed to identify resistance mechanisms shared by both. Using bioinformatic analyses, EOC tissues, primary tumor cells and organoids, and chemoresistant cell lines, we identified lymphoid enhancer-binding factor 1 (LEF1) as a candidate, whose expression was increased in both platinum-resistant and PARPi-resistant tumors. Moreover, LEF1 deficiency increased EOC cell sensitivity to cisplatin and PARPi in vitro and in vivo. Mechanistically, LEF1 knockdown promoted double-strand breaks and significantly impaired both homologous recombination and nonhomologous end joining by directly downregulating the transcription of PARP1 and NUMA1. In addition, the LEF1 inhibitor niclosamide increased ovarian cancer sensitivity to Cisplatin and PARPi in patient-derived organoids and Niraparib-resistant cell lines. These findings indicate that LEF1 is a potential therapeutic target for overcoming resistance to chemotherapy based on platinum and PARPi in EOC.

Extracellular vesicles from ovarian cancer cells induce senescent lipid-laden macrophages to facilitate omental metastasis

Ovarian cancer exhibits striking metastatic tropism for the omentum, where lipid-laden macrophages are key mediators that fuel disease progression. However, the mechanisms governing their formation and pro-metastatic functions remain poorly understood. As extracellular vesicles (EVs) have as critical regulators of tumor-stroma crosstalk in metastatic niches, we sought to define how ovarian cancer-derived EVs orchestrate macrophages and adipocytes, and their impact on omental metastasis, aiming to explore potential therapeutic interventions. Single-cell transcriptomics of ovarian cancer revealed a distinct lipid-laden macrophage population in omentum, whose abundance correlated with metastatic burden and poor survival. Proteomics revealed that EVs from highly metastatic ovarian cancer cells were enriched in lipid metabolism regulators. In vivo experiments demonstrated that these tumor-derived vesicles mediated macrophage reprogramming, driving the acquisition of a pro-metastatic phenotype. Quantitative lipidomic profiling and lipid staining approaches confirmed the progressive lipid-laden in EV-treated macrophages. Using a patient-derived omentum-macrophage co-culture system, we demonstrated that tumor-derived EVs stimulate lipid release from omental adipocytes, which macrophages subsequently internalize through CD36-dependent uptake to drive lipid accumulation. This metabolic reprogramming culminated in cellular senescence, as evidenced by classical biomarkers including SA-β-galactosidase activity, elevated p16-INK4A and p53 levels, and the development of a matrix metalloproteinase-enriched senescence-associated secretory phenotype. Immunohistochemistry of clinical specimens demonstrated overexpression of CD36 correlated with omental metastasis and poor survival in ovarian cancer. In vivo experiments demonstrated that CD36 inhibition and senolytic therapy attenuated omental metastasis. This study unveils an EV-driven mechanism of adipose tropism in ovarian cancer metastasis, where EVs promote the formation of senescent lipid-laden macrophages via CD36-mediated lipid uptake, remodeling the metastatic niche. Targeting CD36 and senescent cells offers a promising therapeutic strategy against omental metastasis.

The autophagy-related gene PEA15 is a potential prognostic biomarker for early-stage endometrial carcinoma

Background The Cancer Genome Atlas (TCGA) molecular classification has advanced risk stratification for endometrial carcinoma but has demonstrated comparable survival outcomes between the microsatellite instability (MSI) and copy-number low (CN-L) subtypes. In this study, we aimed to identify potential autophagy-related molecular signatures to increase the precision of TCGA-based prognostic stratification in early-stage endometrial carcinoma. Methods Univariate Cox regression analysis of the TCGA-Uterine Corpus Endometrial Carcinoma cohort was used to identify autophagy-related genes associated with survival outcomes in patients with endometrial carcinoma. The candidates were analyzed by the Kaplan–Meier method. Multivariate Cox regression was used to assess whether PEA15 served as an independent prognostic factor, especially for the MSI and CN-L subtypes. We examined the correlation between PEA15 protein expression and patient survival through immunohistochemical analysis of tissue microarrays from our institutional cohort of stage I endometrial cancer patients. Results Univariate analysis revealed that NRG3, PEA15, DNAJB1, BAK1, DRAM1, KLHL24, ATF6, CDKN2A, MBTPS2, and UVRAG were significantly associated with survival outcomes in early-stage endometrial carcinoma patients. Multivariate analysis established PEA15 as an independent prognostic factor. Immunohistochemical analysis of tissue microarrays revealed that elevated PEA15 expression was significantly correlated with poorer overall survival and disease-free survival. Both univariate and multivariate Cox regression confirmed high PEA15 expression as an independent prognostic factor for recurrence in patients with stage I endometrioid adenocarcinoma. Conclusions The autophagy-related gene PEA15 is an independent prognostic biomarker in early-stage endometrial carcinoma, improving risk stratification between the MSI and CN-L subtypes. Immunohistochemical detection has clinical potential for molecular classification, offering opportunities for personalized postoperative management strategies.

AI-assisted cervical cytology precancerous screening for high-risk population in resource-limited regions using a compact microscope

Insufficient coverage of cervical cytology screening in resource-limited areas remains a major bottleneck for women's health, as traditional centralized methods require significant investment and many qualified pathologists. Using consumer-grade electronic hardware and aspherical lenses, we design an ultra-low-cost and compact microscope. Given the microscope's low resolution, which hinders accurate identification of lesion cells in cervical samples, we train a coarse instance classifier to screen and extract feature sequences of the top 200 instances containing potential lesions from a slide. We further develop Att-Transformer to focus on and integrate the sparse lesion information from these sequences, enabling slide grading. Our model is trained and validated using 3510 low-resolution slides from female patients at four hospitals, and subsequently evaluated on four independent datasets. The system achieves area under the receiver operating characteristic curve values of 0.87 and 0.89 for detecting squamous intraepithelial lesions on 364 slides from female patients at two external primary hospitals, 0.89 on 391 newly collected slides from female patients at the original four hospitals, and 0.85 on 570 human papillomavirus positive slides from female patients. These findings demonstrate the feasibility of our AI-assisted approach for effective detection of high-risk cervical precancer among women in resource-limited regions.

Risk assessment in a Chinese cohort of 96 318 females undergoing opportunistic cervical cancer screening

Abstract Objective To assess CIN3+ risk in a Chinese cohort of outpatients undergoing contesting screening and to evaluate the portability of the American Society for Colposcopy and Cervical Pathology (ASCCP) risk-based management, which was primarily developed using data from the Kaiser Permanente of Northern California (KPNC) cohort. Methods Females aged 25-65 years who were screened with cytology and high-risk human papillomavirus (hrHPV) co-testing between 2011 and 2020 at Wuhan Union Hospital (WHUH) were retrospectively studied. The risks of immediate and 3-year CIN3+ were estimated via prevalence-incidence mixture models. Portability was evaluated via the ratio of the observation risk in the WHUH cohort to the expected risk in the KPNC cohort (O/E) and its 95% CI. Results A total of 96 318 females were included, and 16.83% of the women tested hrHPV-positive at initial screening, who had a CIN3+ immediate risk of 14.14%. The CIN3+ immediate risk varied between subgroups of positive HPV16 (34.09%), HPV18 (13.38%), other HPV types (6.71%), and negative hrHPV (0.12%). Compared to the KPNC cohort, our cohort exhibited a significantly higher CIN3+ immediate risk (1.42% vs 0.46%; O/E, 3.09; 95% CI, 2.92-3.26) and disproportionately increased cancer immediate risks in most subgroups requiring immediate colposcopy or treatment, as well as higher 3-year CIN3+ risks in women with hrHPV-negative ASC-US/NILM. Yet, the action threshold suggested by ASCCP, a CIN3+ immediate risk of 4%, showed good portability to our cohort. Conclusions Despite the higher risks in our cohort, the ASCCP clinical action threshold remains portable. For women with minimal abnormalities or normal results, shortened follow-up intervals should be considered.

Association between body mass index and lymph node metastasis among women with cervical cancer: a systematic review and network meta-analysis

Lymph node status is a determinant of survival in patients with early-stage cervical cancer. However, the relationship between obesity and lymph node status remains unclear. Therefore, this systematic review aims to evaluate the correlation between body mass index (BMI) and lymph node metastasis in cervical cancer. Cohort studies through six databases were reviewed until December 2021. Odds ratios (ORs) for lymphatic metastasis were estimated using random-effects models and network meta-analysis. BMI groups for lymph node metastasis were ranked. Heterogeneities were assessed using I No significant difference was found between obese (BMI ≥ 25) and non-obese patients (BMI < 25) (OR = 1.01; 95% CI 0.69-1.47; P = 0.97). In subgroup analyses, obesity was associated with higher risk among the Americans and advanced-stage patients. The grouping analysis based on BMI and the rankogram values revealed that the '35 ≤ BMI' group had the highest risk of lymph node metastasis. Although there were no significant differences in lymph node metastasis between obese and non-obese cervical cancer patients in overall analysis, patients with BMI ≥ 35 were at significantly higher risk of lymph node metastasis.

Tumor cell-derived exosomes deliver TIE2 protein to macrophages to promote angiogenesis in cervical cancer

Tyrosine kinase with immunoglobulin and epidermal growth factor homology domains 2 (TIE2)-expressing macrophages (TEMs) are an angiogenesis-promoting subset of tumor-associated macrophages that have been demonstrated to be increased in solid tumors and associated with the progression of cervical cancer. However, the induction mechanism of TEMs remains unclear. Here, based on multicolor immunofluorescence of 58 cervical cancer tissues and the GEPIA database, we found that TEMs were increased in TIE2-high cervical cancer and related to shorter survival. In vitro and in vivo experiments verified that exosomes derived from TIE2-high cervical cancer cells transferred TIE2 protein directly to macrophages, thereby inducing TEMs. Similar to primary TEMs, TEMs induced by tumor-derived exosomes promoted angiogenesis, could be induced by angiopoietin-2, and possessed an M2-like phenotype. In conclusion, exosomes derived from TIE2-high cervical cancer cells induce TEMs by directly transporting TIE2 to promote tumor angiogenesis.

CBX2 phase-separation contributes to homologous recombination repair and drug resistance in ovarian cancer

Abstract Drug resistance jeopardizes the prognosis of high-grade serous ovarian carcinoma (HGSOC) patients via DNA damage repair-coupled mechanism. The role of biomolecular phase separation in DNA damage repair has loomed. Here we find that CBX2 condensates are associated with drug resistance and contribute to DNA double-strand break (DSB) repair in HGSOC. Specifically, CBX2 condensates facilitate the recruitment of key DSB repair factors PARP1, 53BP1, and BRCA1 to chromatin. Patients with a CBX2-negative pattern exhibit the best prognosis, followed by those with non-condensate CBX2, while the worst outcomes are observed in patients with condensate CBX2. By drug screening, Ibrutinib is identified as an effective inhibitor of HGSOC cells and patient-derived organoids with CBX2 condensates. Overall, CBX2 phase separation enhances DSB repair-mediated drug resistance in HGSOC cells, and Ibrutinib may offer a viable therapeutic option for CBX2-positive HGSOC patients.