Abstract 5994: Mechanism of oncogenicity induced by overexpression of the PRL phosphatases

Cancer Research · 2026-04-03

Abstract The phosphatases of regenerating liver (PRL) phosphatases are frequently overexpressed in a wide variety of human cancers and are correlated with cancer progression, metastasis, and poor patient outcomes. Although PRLs appear to be linked to oncogenesis, it is not yet fully understood whether PRL overexpression is sufficient to drive spontaneous tumorigenesis in vivo or the mechanisms by which PRLs contribute to cancer. To address this gap, we developed a novel genetically modified mouse model that conditionally overexpresses PRL2 in the prostate epithelium, mimicking the onset of human cancers. Our findings indicate that transgenic overexpression of PRL2 leads to a multifocal low-grade prostatic intraepithelial neoplasia (LGPIN) phenotype, with a rare occurrence of malignancy in older mice. Furthermore, elevated PRL2 promotes significant acceleration and progression from high-grade prostatic intraepithelial neoplasia (HGPIN) to prostatic adenocarcinoma mediated by PTEN heterozygosity, whereas PRL2 overexpression is dispensable for PTEN-loss-mediated transformation. The initiation and progression of prostate cancer following PRL2 overexpression correlate with decreased PTEN levels and upregulation of AKT/mTOR pathways. Taken together, these findings elucidate the pivotal role of proto-oncogenic PRL2 in promoting tumorigenesis through the downregulation of PTEN. Therefore, PRLs are compelling therapeutic targets for cancer drug discovery, and PRL2 inhibition may be a novel approach for cancer treatment through PTEN augmentation in both PTEN-deficient and wild-type backgrounds. Citation Format: Jingmei Yu, Frederick Nguele Meke, Yunpeng Bai, Meaghan Maureen Broman, Haoran Zhang, Abhinanda Kar, Zhong-Yin Zhang, . Mechanism of oncogenicity induced by overexpression of the PRL phosphatases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 5994.

PD18-14 CHRONIC INFLAMMATION DRIVES EPITHELIAL REMODELING AND THERAPY-RESISTANT PROSTATE HYPERPLASIA

The Journal of Urology · 2026-04-27

AnnotateAnyCell: Open-Source AI Framework for Efficient Annotation in Digital Pathology

bioRxiv (Cold Spring Harbor Laboratory) · 2025-11-03

A bstract Manual annotation of histopathological whole slide images remains a critical bottleneck for computational pathology and clinical AI deployment, requiring prohibitive expert time at scale. Here we present an open-source semi-supervised framework combining active contrastive learning with iterative human-in-the-loop feedback for efficient cellular annotation and classification. The pipeline integrates Cellpose segmentation, UMAP-based latent space visualization, and contrastive learning with pseudolabel propagation, evaluated on five whole slide images of canine invasive urothelial carcinoma across low, intermediate, and high histological grades at 40× magnification. Latent space clustering-guided annotation required 47 minutes compared to 63 minutes for sequential annotation, a 25% reduction (95% CI 18–32%). Classification accuracy reached 96.3% ± 1.2% for mitotic figures and 98.3% ± 1.4% for nucleoli using 1,075 labeled samples, with nucleoli classification achieving 95.5% ± 1.5% accuracy from only 215 samples. Inter-annotator agreement was high for chromatin ( κ = 1.00) and nucleoli ( κ = 0.95) but moderate for mitotic figures ( κ = 0.58) and nuclear shape ( κ = 0.36), reflecting intrinsic morphological ambiguity in these categories. This framework substantially reduces annotation burden while achieving expert-level accuracy for well-defined morphological features, providing a scalable path toward AI-assisted diagnostics in resource-constrained pathology settings.

Immune dysregulation in the prostates of C57BL/6 ^Aire-/- mice mirrors that seen in human benign prostatic hyperplasia

bioRxiv (Cold Spring Harbor Laboratory) · 2025-08-15

Abstract Benign prostatic hyperplasia (BPH) is the most common urologic disease in aging men, resulting in significant morbidity. The etiologies of BPH are unknown, though chronic prostatic inflammation is known to promote hyperplasia, fibrotic remodeling, and therapeutic resistance in BPH. BPH is highly complex and heterogeneous, presenting with varying degrees of stromal and epithelial proliferation, fibrosis, inflammation and associated lower urinary tract symptoms. This complexity presents challenges in developing models. Here, we characterize an Aire transcription factor-deficient non-resolving (chronic) inflammation model in a C57BL/6J background for the study of the prostatic inflammation present in BPH. This chronic inflammatory model exhibits a lack of central tolerance but retains an otherwise intact and functional immune system. C57BL/6 Aire-/- mice were subcutaneously injected with prostate homogenate protein and Freund’s Complete Adjuvant and boosted after 10 days. After 21 and 35 days, whole prostates were collected for histology, flow cytometry, and scRNA-seq, which were then compared to human BPH scRNA-seq data Inflammation was confined to the prostate in C57BL/6 Aire-/- mice. Histological and scRNA-seq data show that the dominant leukocyte phenotypes in the prostates of C57BL/6 Aire-/- mice are B and T lymphocytes. Macrophages in C57BL/6 Aire-/- mouse prostates express signatures associated with an array of phenotypes as also seen in BPH. We identify a Trem2 + population of macrophages, and aging-associated Cd8 + GZMK hi GZMB low T cells in C57BL/6 Aire-/- prostates similar to those seen in human BPH. Further, fibroblast clusters in C57BL/6 Aire-/- are similar to fibroblasts identified from the prostates of patients diagnosed with BPH, and these clusters also express markers associated with aging. Overall, the inflammation and predicted interactions between leukocytes and stromal cells observed in the prostates of C57BL/6 Aire-/- mice resemble human BPH, making this model useful for studying the impact of inflammation-driven prostatic hyperplasia.

Infiltrating lipid-rich macrophage subpopulations identified as a regulator of increasing prostate size in human benign prostatic hyperplasia

Frontiers in Immunology · 2025-01-10 · 8 citations

Introduction Macrophages exhibit marked phenotypic heterogeneity within and across disease states, with lipid metabolic reprogramming contributing to macrophage activation and heterogeneity. Chronic inflammation has been observed in human benign prostatic hyperplasia (BPH) tissues, however macrophage activation states and their contributions to this hyperplastic disease have not been defined. We postulated that a shift in macrophage phenotypes with increasing prostate size could involve metabolic alterations resulting in prostatic epithelial or stromal hyperplasia. Methods Single-cell RNA-seq of CD45 + transition zone leukocytes from 10 large (>90 grams) and 10 small (<40 grams) human prostates was conducted. Macrophage subpopulations were defined using marker genes and evaluated by flow cytometry. Results BPH macrophages do not distinctly categorize into M1 and M2 phenotypes. Instead, macrophages with neither polarization signature preferentially accumulate in large versus small prostates. Specifically, macrophage subpopulations with altered lipid metabolism pathways, demarcated by TREM2 and MARCO expression, accumulate with increased prostate volume. TREM2 high and MARCO high macrophage abundance positively correlates with patient body mass index and urinary symptom scores. TREM2 high macrophages have a statistically significant increase in neutral lipid compared to TREM2 low macrophages from BPH tissues. Lipid-rich macrophages were observed to localize within the stroma in BPH tissues. In vitro studies indicate that lipid-loaded macrophages increase prostate epithelial and stromal cell proliferation compared to control macrophages. Discussion These data define two new BPH immune subpopulations, TREM2 high and MARCO high macrophages, and suggest that lipid-rich macrophages may exacerbate lower urinary tract symptoms in patients with large prostates. Further investigation is needed to evaluate the therapeutic benefit of targeting these cells in BPH.

Corrigendum to “Identification of LIMK2 as a therapeutic target in castration resistant prostate cancer” [Cancer Lett 448 (2019)182–196]

Cancer Letters · 2025-08-26

Validation of protein biomarkers for early detection of cervical cancer and squamous intraepithelial lesions.

Journal of Clinical Oncology · 2025-05-28

e17506 Background: Human papillomavirus infections often resolve on their own; however, ongoing high risk HPV infections can progress to cervical cancer, a significant global health issue responsible for approximately 660,000 new cases and 350,000 deaths in 2022. Current screening methods emphasize cytology or HPV DNA detection, with a limited focus on protein biomarkers. Thus, identifying sensitive and specific cervical cancer protein biomarkers is essential, as this analysis provides opportunities for developing affordable point-of-care screening tests. Methods: This study optimized a unified protocol for cell lysis and protein extraction to validate biomarkers using cervical cell lines and cervical swab samples. Four proteins—Topoisomerase II Alpha (TOP2A), Minichromosome Maintenance Complex Component 2 (MCM2), Valosin Containing Protein (VCP), and Cyclin-Dependent Kinase Inhibitor 2A (p16INK4a)—were quantified using enzyme-linked immunosorbent assays. Furthermore, the expression of target biomarkers in cervical tumors (G1-G3) and squamous intraepithelial lesions (SIL) was evaluated using immunohistochemistry. Cytological evaluation of swab samples categorized them as normal, reactive cellular changes (RCC), atypical squamous cells of undetermined significance (ASC-US), low grade SIL (LSIL), or high grade SIL (HSIL). The study utilized cervical cancer cell lines HeLa, Ca Ski, HT-3, C-33A, and PCS. Statistical analysis was performed using a gamma-generalized linear mixed model, and imaging tools (Aperio and Visiopharm) were employed for tissue analysis. Results: RIPA buffer and acetone precipitation achieved optimal cell isolation and high-yield protein recovery. Biomarker protein concentrations were normalized to Beta-actin. Compared to normal samples, HSIL, LSIL, and ASC-US samples showed overexpression of p16INK4a, while HSIL samples exhibited higher levels of VCP. VCP and p16INK4a were overexpressed across all cancer cell lines compared to primary cells. MCM2 was overexpressed in HT-3 and HeLa, while TOP2A was underexpressed in Ca Ski. Further tissue analysis confirmed strong staining of all target biomarkers in precancerous and cancerous tissues, with significant differences in VCP, TOP2A, and p16INK4a expression observed across tumor grades and SIL. Conclusions: Through a simplified sample preparation and protein extraction protocol, four protein biomarkers were effectively identified and validated for cervical cancer screening. This research emphasizes the importance of detecting a panel of proteins expressed in both HPV-positive and HPV-negative cervical cancers. The results highlight the promise of these target proteins in recognizing precancerous lesions and the potential benefits of incorporating protein biomarker-based screening for improving early detection and cervical cancer outcomes, especially in resource limited settings.

Protein Biomarkers Enable Sensitive and Specific Cervical Intraepithelial Neoplasia (CIN) II/III+ Detection: One Step Closer to Universal Cervical Cancer Screening

Cancers · 2025-05-24 · 1 citations

BACKGROUND/OBJECTIVES: Cervical cancer (CC) is a significant global health challenge, particularly in low- and middle-income countries (LMICs), where limited access to human papillomavirus (HPV) vaccination and effective CC screening results in a majority of cases and fatalities among women. Moreover, existing vaccines do not target HPV-independent cancers. Current screening methods are expensive and time-consuming, with a limited emphasis on CC protein biomarkers. Therefore, we aimed to validate critical markers that allow the development of affordable point-of-care screening tests for resource-limited settings. METHODS: This study first optimized a cell lysis and protein extraction protocol for CC cell lines and clinical cervical swabs. Subsequently, four proteins-topoisomerase II alpha (TOP2A), minichromosome maintenance complex component 2 (MCM2), valosin-containing protein (VCP), and cyclin-dependent kinase inhibitor 2A (p16INK4a)-were quantified in the resulting lysates using enzyme-linked immunosorbent assays, as well as in cervical tumors and squamous intraepithelial lesions (SILs) using immunohistochemistry for further validation. RESULTS: Acetone precipitation allowed for efficient cell isolation, and radioimmunoprecipitation assay buffer yielded the highest protein recovery. VCP and p16INK4a were overexpressed across all cancer cell lines compared to primary cells. All four biomarkers were overexpressed in high-grade SIL (HSIL) swab specimens and tumor samples, including CC subtypes, G1-G3 tumor grades, and HSILs. Lastly, we showed that the proteins could accurately classify swabs and tissue specimens into clinically relevant groups. CONCLUSIONS: The quantitative analysis of these biomarkers, along with the subsequent sensitive and specific clinical classification, highlights their potential application in SIL early detection and CC prevention, particularly in LMICs.

Immune cell single-cell RNA sequencing analyses link an age-associated T cell subset to symptomatic benign prostatic hyperplasia

Frontiers in Immunology · 2025-07-07 · 3 citations

Introduction: Benign prostatic hyperplasia (BPH) is among the most common age-associated diseases in men. Prostatic immune cell infiltration is frequently observed with aging coincident with BPH; however, the contribution of age-related changes in immune cells to BPH is not clear. As T cells are the predominate immune cell in aged prostates, it is hypothesized that age-associated alterations in T cell subsets contribute to BPH symptoms. Methods: scRNA-seq data from immune cells isolated from small (≤40g) and large (≥90g) prostates from aged men (>50 years) were combined with previously published scRNA-seq data from three young organ donor prostates to compare young to aged prostate T cells and small to large aged prostate T cells. Cycling and senescent BPH patient-derived fibroblasts were treated with granzyme K and senescence-associated secretory phenotype (SASP)-associated cytokines were measured by ELISA. Results: T cell differentiation is altered in large BPH prostates compared to small age-matched prostates, favoring Taa accumulation. In vitro granzyme K treatment of human BPH patient-derived large prostate fibroblasts increased secretion of pro-inflammatory senescence-associated secretory phenotype (SASP)-associated cytokines. Discussion: These data suggest that granzyme K-mediated stimulation of prostate stromal fibroblast SASP cytokine and chemokine production promotes prostate immune cell recruitment and activation. Overall, these results connect symptomatic BPH with immune aging.

Immune cell single-cell RNA sequencing analyses link an age-associated T cell subset to symptomatic benign prostatic hyperplasia

bioRxiv (Cold Spring Harbor Laboratory) · 2024 · 3 citations

• Biology
• Computational biology
• Medicine

T cell differentiation is altered in large BPH prostates compared to small age-matched prostates, favoring Taa accumulation. In vitro granzyme K treatment of human BPH patient-derived large prostate fibroblasts increased secretion of pro-inflammatory senescence-associated secretory phenotype (SASP)-associated cytokines. These data suggest that granzyme K-mediated stimulation of prostate stromal fibroblast SASP cytokine and chemokine production promotes prostate immune cell recruitment and activation. Overall, these results connect symptomatic BPH with immune aging.