National Cancer Institute

Our previous preclinical studies demonstrated that non-steroidal anti-inflammatory drugs (NSAIDs) such as aspirin and naproxen significantly inhibited prostate tumorigenesis in a TMPRSS2-ERG fusion-driven model compared to non-fusion models. Since TMPRSS2-ERG fusion-positive tumors display heightened inflammatory signaling and substantial immune infiltration, we hypothesized that the differential efficacy of NSAIDs may arise from their ability to remodel the tumor immune microenvironment. Accordingly, in the present study, we systematically profiled innate and adaptive immune-cell populations: F4/80⁺ macrophages, mast cells, neutrophils, CD3⁺ T cells, CD8⁺ cytotoxic T cells, FoxP3⁺ regulatory T cells, CD20⁺ B cells, IgKC⁺ plasma cells, and Granzyme B⁺ effector cells in highly infiltrated dorso-lateral prostate regions of TMPRSS2-ERG fusion-driven and non-fusion PCa models, with and without NSAID intervention. Our analyses revealed pronounced macrophage infiltration in TMPRSS2-ERG. Ptenflox/flox and Hi-Myc+/ - model, which was further augmented by NSAIDs. Importantly, NSAID intervention shifted macrophage polarization toward an M1-like, pro-inflammatory state, contrasting with the M2-dominant phenotype characteristic of untreated tumors. NSAID treatment reduced mast cell density within the stromal compartment, suggesting suppression of mast cell-mediated tumor-promoting signals. In the fusion model, infiltration of total T cells and CD8⁺ cytotoxic T cells decreased following NSAID exposure, whereas FoxP3⁺ Tregs remained largely unaffected. Both models showed increased B-cell infiltration independent of NSAID efficacy, and no clear correlation was observed between plasma-cell presence and treatment response. Collectively, our findings offer new insight into NSAID-mediated immunomodulation in TMPRSS2-ERG fusion-driven PCa; however, further in-depth immune subtyping and spatial mapping could fully delineate the immunological mechanisms driving NSAID responsiveness.

VEGF-A exerts complex immunomodulatory effects that foster an immunosuppressive tumour microenvironment, including impaired dendritic cell (DC) maturation, expansion of regulatory T cells (Tregs), and induction of T-cell exhaustion, while transiently enhancing effector T-cell responses. This dual activity underscores the need to clarify the immunosuppressive versus immunostimulatory consequences of VEGF-A receptor activation to optimise selective inhibition. This proof-of-concept study evaluated whether selective VEGFR-1 blockade can counteract VEGF-A-driven immunosuppression in human immune cells using the monoclonal antibody (mAb) D16F7, which inhibits membrane-bound VEGFR-1 while preserving the decoy and anti-angiogenic activity of its soluble form. The impact of VEGFR-1 blockade by D16F7 mAb on VEGF-A-mediated immune modulation was assessed by multiparametric flow cytometry using primary immune cells from healthy donors. Human DCs were generated from CD14+ monocytes and cultured with VEGF-A; tolerogenic and pro-exhaustion activity was evaluated via maturation markers (CD80, CD83, HLA-DR) and PD-1 expression on DC-stimulated T cells after 14days. Short-term effects on effector T cells, stratified by PD-1 and CD28 expression, were analysed by intracellular cytokine staining at 5-6 and 18h. Purified Tregs, isolated via a two-step magnetic separation, were cultured for 1week to assess the acquisition of an immunosuppressive phenotype by measuring the expression of checkpoint markers (PD-1, ICOS, TIM-3). D16F7 reversed VEGF-A-induced DC maturation defects, reduced DC-mediated T-cell tolerance, and attenuated the emergence of a highly immunosuppressive Treg phenotype, while largely preserving short-term effector T-cell function. Despite the limitations of an in vitro PBMC-based system, selective VEGFR-1 blockade by D16F7 mAb alleviates VEGF-A-driven immune tolerance while preserving T-cell activation, supporting its translational potential as a complementary immunomodulatory approach.

There are no cures for neurodegenerative diseases. The biggest hurdle to treating these disorders is that their clinical manifestation is rooted in multiple physiological processes. Therefore, efficacious pharmaceutical options will likely require two or more agents with different mechanisms of action. However, drug combinations have significant drawbacks, including overlapping toxicities and unique pharmacokinetic properties, particularly the rate and extent of central nervous system (CNS) penetration. A single agent with multiple mechanisms of action could overcome these drawbacks. We have recently discovered first-in-class novel single agents (compounds 1 and 2) that mildly inhibit clinically important kinases and subtly favor microtubule stability at concentrations that show no evidence of neuronal toxicity in primary neurons, while maintaining their ability to penetrate the CNS in vivo. It is important to note that the effects of these analogs are mild and are predicated on avoiding neurotoxicity. These multitargeting single agents provide a new structural modality with the potential to influence treatments for Parkinson's and Alzheimer's disease and serve as lead compounds for further optimization.

Oligodendroglioma is genetically defined by mutations in isocitrate dehydrogenase 1 or 2 (IDH1/IDH2) and 1p/19q codeletion. We previously showed that in IDH1-mutant oligodendroglioma, the oncometabolite D-2-hydroxyglutarate biases the sphingosine-1-phosphate–to–ceramide rheostat toward ceramides. Taking advantage of this intrinsic metabolic vulnerability, we investigated whether further elevating ceramide levels through inhibition of acid ceramidase could exacerbate this imbalance and promote apoptotic cell death.Analysis of patient datasets demonstrated that acid ceramidase is expressed at higher levels in both low- and high-grade gliomas compared with normal tissue. Pharmacologic inhibition of acid ceramidase with SABRAC preferentially reduced viability in human IDH1-mutant oligodendroglioma cell lines. In these sensitive models, acid ceramidase inhibition markedly increased ceramide levels and induced coordinated sphingolipid remodeling. Subcellular imaging using a fluorescent ceramide analogue demonstrated increased ceramide localization to lysosomes and mitochondria following acid ceramidase inhibition. This was accompanied by cytochrome c redistribution, executioner caspase activation, and caspase-dependent apoptotic cell death, consistent with engagement of intrinsic mitochondrial apoptosis. Transcriptomic and biochemical analyses further revealed activation of endoplasmic reticulum stress and unfolded protein response signaling, including PERK- and IRE1α-associated programs, suggesting coordinated multi-organelle stress responses under sustained ceramide elevation. These mechanistic effects translated into a survival benefit in oligodendroglioma xenograft-bearing mice.Together, these findings suggest that IDH1-mutant oligodendroglioma harbors a pre-existing heightened sensitivity to ceramide stress and identify acid ceramidase as a therapeutically actionable target in this disease.

Cutaneous leishmaniasis (CL) remains a clinically and immunologically heterogeneous disease influenced by parasite species and host immune responses. In this study, seventeen CL cases were assessed through lesion characteristics, histopathological analysis and Kinetoplast DNA (kDNA) nested Polymerase Chain Reaction (PCR)figure. Lesions mainly appeared on exposed body parts, especially the face (64.70%) and most patients had a single lesion. kDNA PCR confirmed CL in 58.82% (n=10) of cases, identifying Leishmania donovani (720 bp) in 5 cases and L. major (560–590 bp) 5 cases as the circulating species. Immunological analysis showed reduction in T-lymphocytes in CL patients compared to Healthy Controls (HC), largely due to a significant decrease in CD8⁺ T cells (p=0.036), while CD4⁺ T cell levels remained stable. Further stratification based on CL PCR positivity showed that reductions in T-lymphocytes (p=0.045) were confined to patients with confirmed Leishmania infection, again attributed mainly to decreased CD8⁺ T cells (p=0.040). No notable changes were observed in B cells, NK cells, or NKT cells. These findings highlight CL infection positivity status-dependent variations in host immune responses and emphasize the importance of CD8⁺ T-lymphocytes in CL pathogenesis and disease outcome.

Bacteriophage-based display has been utilized for a variety of purposes, such as to assemble protein libraries and conduct biopanning. We have created a modified lambda (λ) bacteriophage platform, ideal for the display and delivery of proteins. Our system utilizes counter-selection recombineering for versatile modification, temperature-sensitive induction for timely lysate production, and an arabinose-inducible mechanism for high-titer, stable yield. Here, we investigated the ability of this specialized λ phage display platform to stimulate highly specific antibodies in mice against the displayed cancer-variant cell-surface receptor EGFRvIII, demonstrating its potential in cancer immunotherapy and broader vaccine development. λ display immunogenicity was explored by generating fusion proteins between the λ head protein D and a 13-mer peptide from the N terminus of glioblastoma variant cell-surface receptor, EGFRvIII. The 13-mer peptide was fused to either the N or C terminus of the λD protein while λ remained a dormant lysogen in the bacterial host chromosome. Recombinant phage lysates were then generated with ~420 displayed fusion proteins per phage particle. Mice were injected with purified recombinant λ phage without an adjuvant via both intraperitoneal and intramuscular routes, and sera harvested at various timepoints were profiled for immunogenicity. Analysis of serum samples by ELISA and Western blotting demonstrated the ability of the λD~EGFRvIII phage display, especially in the C-terminal fusion construction, to elicit a robust anti-EGFRvIII humoral response by either injection route. Notably, the antibody response was highly specific to EGFRvIII without exhibiting cross-reactivity to wild-type EGFR. The data generated in this study demonstrate the λ system's immunotherapeutic potential as a high-titer, stable, self-adjuvanting vector for the stimulation of robust antibody titers with defined specificity.

Lapsi (Choerospondias axillaris), a plant native to Nepal, has been traditionally used in Asian countries to treat cardiovascular conditions. However, its effects on immune regulatory function remain largely unexplored. This study aimed to in vivo evaluate the immunoregulatory properties of Lapsi fruit extract in mice on immunotoxic responses with analysis on in vitro mechanism for immune suppression, oxidative stress, and inflammatory response. Male Balb/c mice were intragastrically administered various doses of the extract for 21 days. In some mice, immune suppression was induced with cyclophosphamide, and subsequent immune recovery was assessed. In addition, RAW264.7 cells and THP-1-derived macrophages were treated in vitro with lipopolysaccharide and different concentrations of the extract. Administration of extract increased the IgG2a/IgG1 ratio while reducing serum IgE and IgG1 level compared with control mice. Tumor necrosis factor (TNF)-α and interleukin (IL)-17 levels were lower in splenic culture supernatants of mice administered extract. Lapsi extract also effectively reversed cyclophosphamide (CP)-induced immunosuppression by enhancing serum levels of IgA and IgG2a, of interferon-γ and interleukin (IL)-4 secreted by splenic T cells, and of IgG1 and IgG2a secreted by B cells, as well as by increasing immune cell counts. In cell cultures, the extract decreased the levels of inflammation markers, including nitric oxide, reactive oxygen species, prostaglandin E2, and pro-inflammatory cytokines (IL-6, TNF-α, and IL-1β). Mechanistic analysis showed that Lapsi extract modulated the NF-κB p65, MAPK, and inflammasome pathways. Lapsi extract may act as both an immunostimulatory and anti-inflammatory agent, indicating its potential as a candidate immunomodulatory activity under polyclonal and CP-suppressed conditions; however, further disease-specific studies, along with isolation and characterization of active phytochemicals, are warranted to evaluate its therapeutic applicability.

The constituents of tobacco smoke that specifically contribute to lung cancer risk have yet to be fully identified. We evaluated associations between biomarkers of potentially harmful constituents-polycyclic aromatic hydrocarbons (PAHs), tobacco-specific nitrosamines (TSNAs), nicotine, and volatile organic compounds (VOCs)-and lung cancer incidence among US women. In a case-cohort study nested within the Sister Study (women aged 35 to 74 years at baseline, enrolled 2003 to 2009), data were obtained for a random subcohort and all remaining incident lung cancers through September 2017 (median follow-up 9.6 years), stratified by race and ethnicity (Hispanic, non-Hispanic Black, non-Hispanic White, others) and smoking status (current, former, never). The analytic sample included 356 cases and 433 non-cases. We quantified 30 biomarkers in baseline urine samples and calculated hazard ratios (HRs) for associations between one-unit increase in biomarker concentrations (log-scale) and lung cancer incidence using weighted Cox regression models adjusted for urinary creatinine and demographic, health, and lifestyle factors. Among women who were currently smoking at enrolment, positive associations were observed for biomarkers of PAHs (naphthalene, phenanthrene, pyrene, fluorene; HRs 1.4 to 5.3), TSNAs (particularly 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK); HRs : 1.3-2.2), and VOCs (xylene, acrylamide, acrylonitrile, 1,2-dibromoethane/vinyl-chloride/ethylene-oxide/acrylonitrile, acrolein, styrene/ethylbenzene, benzene, dimethylformamide/methylisocyanate, 1,3-butadiene, crotonaldehyde, isoprene; HRs : 1.6-4.4). Associations with biomarkers of most PAHs, NNK, xylene, and dimethylformamide/methylisocyanate remained after additional adjustment for smoking frequency, duration, and nicotine metabolites. In women who did not smoke, positive associations were observed for styrene/ethylbenzene and dimethylformamide/methylisocyanate biomarkers. Exposure to PAHs, TSNAs and several VOCs through tobacco smoking were associated with increased lung cancer risk among women.