Induced Tumor-Suppressing (iTS) Cell-Based Approach for Protecting the Bone from Advanced Prostate Cancer

Biomolecules · 2026-02-03

Advanced prostate cancer frequently metastasizes to bone, but no effective therapy exists. To seek a novel treatment option and identify a new drug target, we took an induced tumor-suppressing (iTS) cell-based approach and produced tumor-suppressing proteins and conditioned medium (CM). Notably, the overexpression of Lrp5 and β-catenin, as well as the pharmacological Wnt activator, converted osteocytes, Murine mesenchymal stem cells, mononuclear cells, and monocytes into iTS cells. While Lrp5 conditional knockout mice presented severe bone loss, Lrp5-overexpressing osteocyte-derived CM rescued tumor-induced bone damage. Whole-genome proteomics analysis revealed that Moesin (MSN), which acted as an oncogene in tumor cells, was enriched in CM as an extracellular tumor-suppressing protein. Its anti-tumor action was mediated primarily by the interaction with CD44. Consistently, FRET live-cell imaging demonstrated that extracellular MSN reduced Src tyrosine kinase activity and nuclear localization of β-catenin. Collectively, we demonstrated herein the iTS cell-based approach to protect bone from prostate cancer and showed MSN as a potent extracellular tumor-suppressing protein.

Immunomodulation of the Prostate Tumor Microenvironment Following Inorganic Arsenic Exposure

Journal of Applied Toxicology · 2026-02-23 · 1 citations

ABSTRACT The tumor microenvironment (TME) influences prostate cancer (PCa) progression through stromal and immune interactions. Adipose‐derived mesenchymal stromal cells (ASCs) modulate immune tone, while inorganic arsenic (iAs), a widespread toxicant, is linked to immune suppression and carcinogenesis. Their combined impact on PCa immunity has remained unclear. Using a Ras‐driven murine PCa model (TC2Ras, which mimics aggressive, immune‐interactive PCa through constitutive Ras signaling), we assessed ASC and chronic iAs exposure effects on tumor growth, immune infiltration, and transcriptomic remodeling via flow cytometry, RNA‐seq, and qPCR. ASC‐conditioned media increased TC2Ras viability by up to 82%, an effect reversed by iAs (300–1000 ppb). In vivo, ASC co‐implantation significantly elevated tumor weight in ASC + iAs tumors. ASC promoted approximately twofold macrophage and CD4 + T‐cell infiltration, while iAs suppressed macrophages and MDSCs. We performed RNA‐seq and qPCR, confirming that a sustained IFNγ‐IRF1 activation (approximately eightfold) in ASC tumors occurred alongside an iAs‐driven downregulation of adaptive immunity, as well as an upregulation of immune checkpoint genes (Pdcd1, Lag3). These findings demonstrate that ASC–iAs crosstalk remodels the TME toward immune tolerance and chronic IFNγ signaling, potentially facilitating tumor progression and revealing novel mechanisms by which environmental toxicants may influence cancer immunity through stromal cell interactions.

Transcriptomic profiling reveals prognostic gene signatures and immune landscape alterations in canine diffuse large B-cell lymphoma

Veterinary oncology. · 2026-01-06

Canine diffuse large B-cell lymphoma (cDLBCL) is a frequent and highly aggressive hematopoietic malignancy that serves as a robust translational model for human non-Hodgkin lymphomas. However, the molecular determinants underlying disease progression and prognosis remain poorly defined, hindering precise therapeutic decisions. Clarifying these mechanisms is essential to support the development of prognostic tools and targeted interventions in veterinary oncology. This study aimed to identify prognostically relevant genes and immune alterations associated with cDLBCL. RNA-Seq was performed on lymph node samples from 15 dogs with multicentric DLBCL and two healthy controls. Differential gene expression was assessed to identify key molecular alterations. Functional annotation, pathway analysis, and Kaplan–Meier survival correlations were used to characterize the biological and clinical significance of target genes. Additionally, immune cell deconvolution was conducted to explore changes in the tumor microenvironment (TME). Transcriptomic profiling revealed 410 differentially expressed genes (DEGs), comprising 408 downregulated and 2 upregulated genes in tumor samples compared to normal lymph nodes. Among these, STOM, TBC1D8, HMOX1, ABCB1, and CTLA4 emerged as genes of interest due to their involvement in immune regulation, oxidative stress response, vesicular trafficking, drug resistance, and immune checkpoint signaling. Lower expression of these genes was significantly associated with reduced overall survival. Immune infiltration analysis demonstrated a dominance of neoplastic B cells and a marked reduction in cytotoxic T cells and macrophages, consistent with an immunosuppressive TME. This study identifies key molecular and immunological signatures associated with prognosis in cDLBCL, providing insight into mechanisms of immune suppression and therapeutic resistance. The findings reinforce the translational value of canine lymphoma as a model for human disease and support the integration of molecular biomarkers into veterinary oncologic practice to guide individualized treatment strategies.

Interleukin-27-adipose-derived mesenchymal stromal cell-based gene therapy attenuates inflammation in lipopolysaccharide-induced acute respiratory distress syndrome

Stem Cell Research & Therapy · 2025-09-29

Abstract Background Acute respiratory distress syndrome (ARDS) is a lung inflammatory condition associated with the accumulation of fluid edema and cell infiltrates into the alveolar space along with dysregulation of the immune response. Current therapeutics are limited to palliative care, i.e., mechanical ventilators, thus highlighting the need to develop targeted therapeutic for ARDS. Interleukin-27 (IL-27) is a multifunctional cytokine with the capability for immune modulation. Our interest lies in exploring the properties of IL-27, particularly as an anti-inflammatory cytokine that functions as an antagonist of IL-6 signaling, as an inducer of anti-viral genes, as a promoter of tissue repair, and as a regulator of both the innate and adaptive immune responses, possessing promising potential as a therapeutic for ARDS. Methods To overcome the challenge of repeated administration due to the short half-life of cytokines, we utilized a cell-based gene therapy approach. An IL-27-expressing plasmid was transfected into adipose mesenchymal stromal cells (ASC) that serve as the gene therapy carriers. For in vitro studies, we treated mono- and co-culture lung lipopolysaccharide (LPS)-induced lung epithelial and monocytes/macrophages cell line with IL-27-expressing ASC (IL-27 ASC) conditioned media (CM) to determine the effects on pro-inflammatory gene expression. For in vivo studies, male C57BL/6 mice were intratracheally injected with LPS (5 mg/kg) and treated either PBS, ASC, or IL-27 ASC (5 × 10 5 cells/mouse) 24 h after LPS instillation. Measurements of gene expression, histopathological changes, cell infiltrations, and protein content in the bronchoalveolar lavage fluid and RNA-seq analysis were performed for the in vivo studies. Results IL-27 ASC CM reduced pro-inflammatory gene expression of lung epithelial and macrophages cultured in both mono- and co-culture systems. Additionally, IL-27 ASC were able to reduce pro-inflammatory markers, decrease cell infiltration into the lungs, promote genes and immune cells involved in tissue repair, and rebalance innate and adaptive immunity in an LPS-induced in vivo model. Conclusions Collectively, our in vitro and in vivo results show promising potential for IL-27 cell-based gene therapy as a treatment for ARDS.

Exploring tumor dynamics and responses of prostate cancer to IL-27 based treatment combinations through biodynamic imaging and RNA sequencing analyses

Scientific Reports · 2025-11-26

The tumor microenvironment (TME) is a highly dynamic network shaped by immune, vascular, and stromal interactions, further modulated by extracellular factors. The high tumor heterogeneity complicates treatment due to evolving resistance. Biodynamic imaging (BDI), a 3D coherence-gated holography technique, quantifies intracellular motion to assess phenotypic responses to therapy. We applied BDI to prostate cancer (PCa) treated with IL-27-based combinations to evaluate the dynamic responses of immunotherapy. This study is the first to integrate BDI with RNA-seq to correlate physiological changes with gene expression. Using a subcutaneous PCa model, tumors were treated ex vivo with chemotherapy and immunomodulatory agents. BDI enabled capturing motion frequency shifts (0.01-12.5 Hz), while RNA-seq revealed the molecular changes in the tumors. IL-27 showed potential to modulate the immunologically "cold" TME and enhance therapeutic efficacy. This integrative approach offers a novel platform for evaluating combination strategies in PCa and may inform more effective treatment decisions.

Abstract 5693: Diagnostic and predictive biomarkers for canine prostatic carcinoma: translational therapeutic insights

Cancer Research · 2025-04-21

The prostate is an organ prone to diseases typical of human aging, with benign and malignant prostatic disorders being among the most common diseases that affect men. There are a wide variety of treatment options for diagnosing prostate cancer in humans, depending on the risk category the disease falls into. The dog is the only mammal, besides man, that spontaneously develops canine prostatic hyperplasia (PH), prostatic atrophy and prostatic carcinoma (PCa), which are associated with age and androgenic hormones. However, unlike PCa in humans, PCa in dogs is not androgen dependent, therefore androgen deprivation therapy is not effective, and animals develop castration-resistant prostatic carcinoma (CRPC). Many treatment modalities usually for human PCa cannot be applied in CRPC. And regarding dogs, unfortunately, there is still no effective therapy for the treatment of these tumors. Available pharmacological treatments are scarce, based primarily on the prescription of anti-inflammatory drugs, with a low degree of survival. Therefore, we intend to carry out new translational studies on human and canine PCa samples, aiming to identify predictive markers and potential common therapeutic targets, to subsequently elucidate the antitumor effect of existing drugs, aiming to develop targeted therapies. The study included 20 animals, with 4 in the control group and 16 in the neoplasia group, under CEUA approval. Samples underwent proteomic analysis, RT-qPCR, RNA sequencing (RNAseq), and Next-Generation Sequencing (NGS) to evaluate gene expression patterns, tumor microenvironment interactions, and immune cell infiltrate estimates. Cross-validation with independent data from canine and human prostate carcinoma databases was performed. Biological triplicates were analyzed, and differential protein and gene expression were assessed using Student’s t-test (p < 0.05, expression change > 1.5x). A total of 44 proteins, including significantly elevated levels of Vimentin and Peptidase S1, were identified in the neoplasia group. Additionally, 1,412 differentially expressed genes were enriched in prostate cancer-related pathways, with 536,200 SNPs identified across 291 genes in the canine exome. Thirteen genes co-expressed in RNAseq and NGS analyses emerged as potential direct therapy targets. Increased secretion of Vimentin and Peptidase S1 in the neoplasia group highlighted their potential as tumor biomarkers. These findings reveal a significant overlap in tumor progression and biomarker expression between canine and human PCa, offering promising avenues for translational studies to develop effective diagnostic tools and targeted therapies for both species. Citation Format: Alexandre Matheus Baesso Cavalca, Marxa Leao Figueiredo, Maricy Apparício Ferreira, Carlos Eduardo Fonseca Alves. Diagnostic and predictive biomarkers for canine prostatic carcinoma: translational therapeutic insights [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 5693.

Tibial Biomechanics and Adaptive Response to Mechanical Stimuli in the Green Iguana

Integrative Organismal Biology · 2025-01-01

Synopsis Mechanical loading models are used to study adaptive skeletal mechanobiology mechanisms. However, most studies have used mammal models, leaving a knowledge gap regarding how these mechanisms differ among vertebrate groups. To address this gap, we evaluated the in vivo bone strain environment of the left tibia in green iguanas during locomotion, axial compressive loading, and with finite element analysis. Our study involved examining male green iguanas (n = 7) over a range of speeds (0.45–1.34 m/s) and axial load magnitudes (–25 to –100 N) to determine peak strains. Bone strains were measured using single-element strain gauges and rosette strain gauges, surgically attached to the tibial anterior, posterior, and medial surfaces. At a speed of 1.34 m/s, peak strains ± standard deviation observed were 645 ± 699 µε, –448 ± 464 µε, and 206 ± 168 µε at the anterior, posterior, and medial surfaces, respectively. Peak principal tensile and compressive strains on the medial surface were 199 ± 113 µε and –153 ± 98 µε at 1.34 m/s. During –100 N compressive loading, peak strains were 403 ± 277 µε, –506 ± 460 µε, and –52 ± 177 µε at the anterior, posterior, and medial surfaces, respectively. Our finite element model demonstrated a close correlation with experimentally measured strain values at the gauge sites (slope = 1.07, R = 0.8381). Using these foundational in vivo strain results and a daily strain stimulus formula, our objective was to develop a novel noninvasive axial compressive tibial loading model to induce a cortical bone adaptive response in the green iguana tibia (n = 9). However, following 3 weeks of daily applied compressive loading, no significant difference was detected in critical bone parameters at 37 and 50% (midshaft) volume of interests from the proximal tibia (P < 0.05). While this study did not yield significant differences in critical bone parameters following the application of daily compressive loading, it provided new knowledge regarding the bone strain environment and the potential for inducing adaptive responses in the green iguana tibia. Further research may refine our understanding of skeletal mechanobiology mechanisms across vertebrate groups and develop more effective loading models for studying bone adaptation. Overall, the findings of this study, although limited, contribute to the broader field of musculoskeletal mechanobiology, giving insights that may inform bone health and adaptation in diverse species.

Experimental models for developing oncolytic virotherapy for metastatic prostate cancer

Frontiers in Immunology · 2025-07-10 · 1 citations

Cancer has remained the second leading cause of death worldwide for over a century. Despite significant advances, effectively targeting cancer cells and overcoming therapeutic challenges remain critical goals. In this review, we focus on advanced metastatic prostate tumors, where the patients’ five-year survival rate is less than 35%. While standard androgen deprivation therapy (ADT) has been effective for most prostate cancer patients, recurrence of aggressive tumors is common, emphasizing an urgent need for new treatment strategies. Immunotherapy has gained attention for its potential to harness the immune system against cancer cells. Among these, oncolytic virotherapy stands out for its tumor-specific tropism, its ability to transform or convert the immune-suppressive tumor microenvironment by enhancing immune cell infiltration, and its capacity for therapeutic gene delivery. This review explores the background of commonly used viruses, evaluation models (including cell culture, animal models, ex vivo platforms, and clinical trials), and the anticipated outcomes and challenges of oncolytic virotherapy. By addressing these aspects, we aim to provide a comprehensive overview of the current state and future directions of oncolytic virotherapy models in the treatment of advanced prostate cancer.

Single-Cell RNA Sequencing Identifies CCR6-Driven Immune Landscape Changes in RM1 Prostate Cancer Bone Metastasis

DNA and cell biology reports. · 2025-03-13

Background: Prostate cancer commonly metastasizes to bone, creating an immunosuppressive microenvironment that supports tumor progression. The RM1 (Ras/Myc) mouse model is a valuable tool for studying interactions between prostate tumors and the bone marrow immune landscape. Methods: Single-cell RNA sequencing was employed to investigate how CCR6 deficiency affects immune cell comsmunication in bone marrow from RM1 prostate cancer bone metastasis. Immune responses were compared between wild-type and CCR6 knockout (CCR6ko) C57BL/6-mice injected with RM1-BoM3 cells. Results: Distinct immune cell dynamics were observed between the two groups. CCR6ko bone marrow showed enhanced signaling from macrophages, regulatory T cells, and myeloid-derived suppressor cell-like cells, with altered receptor activity in naïve T cells, NK cells, and conventional dendritic cells. Differential gene expression highlighted immune regulation pathways and transcriptional networks centered on JUN, JUNB, and FOSB, linked to immune suppression and tumor progression. Cell-cell communication analysis revealed changes in CCL, TGFb, MIF, and CXCL pathways, aligning with observations in human castration-resistant prostate cancer (CRPC) bone-metastasis. Profiling of immune cell populations showed increased tumor-infiltrating monocytic cells, M2 macrophages, and NKT-like CD8 cells in RM1 tumors compared to CCR6ko bone marrow. Pathway enrichment analysis identified upregulated pathways, including IL-17 signaling and osteoclast differentiation, associated with immune-modulation and inflammation. Gene regulatory networks in the RM1 bone metastasis microenvironment involved NFKB1, STAT1, IRF8, and JUN family proteins, emphasizing their roles in immune suppression and tumor progression. Conclusions: These results suggest that targeting CCR6 may enhance immunotherapy efficacy in prostate cancer bone metastasis by reshaping the immune microenvironment. Validation in human CRPC bone metastasis datasets revealed conserved immune interactions.

Proteomic profiling of adipose-derived mesenchymal stromal cell response to novel compounds targeting the Laminin Receptor – PEDF interaction

Biochemistry and Biophysics Reports · 2025-11-25

This study characterizes the proteomic responses of adipose-derived mesenchymal stromal cells (ASC) to novel compounds that target the interaction between pigment epithelium-derived factor (PEDF) and the 37 KDa Laminin Receptor (LR). Building on previous research, we introduced the second-generation analog, 02–09, for comparative analysis with C3, the original compound. C3 was identified through in silico screening and experimental validation, showing anti-inflammatory activity. Analog 02–09 was synthesized by modifying C3's aromatic ring, resulting in similar binding affinities to LR. The distinct molecular effects of C3 and 02–09, along with their potential as anti-inflammatory therapies from prior work, suggested their multi-functional utility. Proteomic profiling of ASC, under both undifferentiated and chondrogenic differentiation conditions, was performed using LC-MS/MS, with controls including the PEDF-derived peptide p18 and the cartilage-regenerative compound Kartogenin (Krt). Both C3 and 02–09 induced significant proteomic modulation compared to controls, with enrichment in pathways related to integrin β1 signaling, extracellular matrix (ECM) organization , and TGF-β signaling . C3 uniquely activated the nuclear receptors meta-pathway, whereas 02–09 was associated with laminin interactions . Protein-protein interaction and transcription factor enrichment analyses revealed distinct and overlapping regulatory networks for LR-targeting compounds. During chondrogenic differentiation, C3 primarily enhanced proteins linked to a chondrogenic phenotype with minimal fibrogenic activity, while 02–09 showed an influence on both chondrogenic and fibrogenic pathways. Comparative analysis of the proteomic response to C3 or 02–09 with a single-cell RNA-sequencing dataset confirmed that C3's modulated proteins primarily correlated with a chondrogenic differentiation phenotype, with minimal fibrogenic influence. The proteins modulated by 02–09 correlated with a dual profile (both chondrogenic and fibrogenic potential). Our results suggest that C3 has potential as a targeted chondrogenic agent, while 02–09 may serve as a multifaceted modulator of both cartilage repair and ECM protection. These findings indicate the utility of these compounds for regenerative medicine and the development of disease-modifying therapies for inflammatory and immune-related conditions. • Proteomics showed ASC responses to LR–PEDF-targeting molecules. • Analog 02-09 and C3 displayed distinct differentiation profiles. • C3 promoted chondrogenic phenotype with minimal fibrogenic activity. • 02–09 induced chondrogenic and fibrogenic signatures. • LR-targeting modulated ECM, integrin, and TGF-β signaling pathways.