BackgroundAneurysmal subarachnoid hemorrhage (SAH) is a catastrophic disease with devastating consequences, including a high mortality rate and severe disabilities among survivors. Inflammation is induced following SAH, but the exact role and phenotype of innate immune cells remain poorly characterized. We investigated the inflammatory components of the early brain injury in an animal model and in SAH patients.MethodSAH was induced through injection of blood in the subarachnoid space of C57Bl/6 J wild-type mice. Prospective blood collections were obtained at 12 h, days 1, 2, and 7 to evaluate the systemic inflammatory consequences of SAH by flow cytometry and enzyme-linked immunosorbent-assay (ELISA). Brains were collected, enzymatically digested, or fixed to characterize infiltrating inflammatory cells and neuronal death using flow cytometry and immunofluorescence. Phenotypic evaluation was performed at day 7 using the holding time and footprint tests. We then compared the identified inflammatory proteins to the profiles obtained from the plasma of 13 human SAH patients.ResultsFollowing SAH, systemic IL-6 levels increased rapidly, whereas IL-10 levels were reduced. Neutrophils were increased both in the brain and in the blood reflecting local and peripheral inflammation following SAH. More intracerebral pro-inflammatory monocytes were found at early time points. Astrocyte and microglia activation were also increased, and mice had severe motor deficits, which were associated with an increase in the percentage of caspase-3-positive apoptotic neurons. Similarly, we found that IL-6 levels in patients were rapidly increased following SAH. ICAM-1, bFGF, IL-7, IL-12p40, and MCP-4 variations over time were different between SAH patients with good versus bad outcomes. Moreover, high levels of Flt-1 and VEGF at admission were associated with worse outcomes.ConclusionSAH induces an early intracerebral infiltration and peripheral activation of innate immune cells. Furthermore, microglia and astrocytic activation are present at later time points. Our human and mouse data illustrate that SAH is a systemic inflammatory disease and that immune cells represent potential therapeutic targets to help this population of patients in need of new treatments.
Within the prostate tumor microenvironment (TME) there are complex multi-faceted and dynamic communication occurring between cancer cells and immune cells. Macrophages are key cells which infiltrate and surround tumor cells and are recognized to significantly contribute to tumor resistance and metastases. Our understanding of their function in the TME is commonly based on in vitro and in vivo models, with limited research to confirm these model observations in human prostates. Macrophage infiltration was evaluated within the TME of human prostates after 72 h culture of fresh biopsies samples in the presence of control or enzalutamide. In addition to immunohistochemistry, an optimized protocol for multi-parametric evaluation of cellular surface markers was developed using flow cytometry. Flow cytometry parameters were compared to clinicopathological features. Immunohistochemistry staining for 19 patients with paired samples suggested enzalutamide increased the expression of CD163 relative to CD68 staining. Techniques to validate these results using flow cytometry of dissociated biopsies after 72 h of culture are described. In a second cohort of patients with Gleason grade group ≥ 3 prostate cancer, global macrophage expression of CD163 was unchanged with enzalutamide treatment. However, exploratory analyses of our results using multi-parametric flow cytometry for multiple immunosuppressive macrophage markers suggest subgroup changes as well as novel associations between circulating biomarkers like the neutrophil to lymphocyte ratio (NLR) and immune cell phenotype composition in the prostate TME. Further, we observed an association between B7–H3 expressing tumor-associated macrophages and the presence of intraductal carcinoma. The use of flow cytometry to evaluate ex vivo cultured prostate biopsies fills an important gap in our ability to understand the immune cell composition of the prostate TME. Our results highlight novel associations for further investigation.
Emerging evidence associates translation factors and regulators to tumorigenesis. However, our understanding of translational changes in cancer resistance is still limited. Here, we generated an enzalutamide-resistant prostate cancer (PCa) model, which recapitulated key features of clinical enzalutamide-resistant PCa. Using this model and poly(ribo)some profiling, we investigated global translation changes that occur during acquisition of PCa resistance. We found that enzalutamide-resistant cells exhibit an overall decrease in mRNA translation with a specific deregulation in the abundance of proteins involved in mitochondrial processes and in translational regulation. However, several mRNAs escape this translational downregulation and are nonetheless bound to heavy polysomes in enzalutamide-resistant cells suggesting active translation. Moreover, expressing these corresponding genes in enzalutamide-sensitive cells promotes resistance to enzalutamide treatment. We also found increased association of long non-coding RNAs (lncRNAs) with heavy polysomes in enzalutamide-resistant cells, suggesting that some lncRNAs are actively translated during enzalutamide resistance. Consistent with these findings, expressing the predicted coding sequences of known lncRNAs JPX, CRNDE and LINC00467 in enzalutamide-sensitive cells drove resistance to enzalutamide. Taken together, this suggests that aberrant translation of specific mRNAs and lncRNAs is a strong indicator of PCa enzalutamide resistance, which points towards novel therapeutic avenues that may target enzalutamide-resistant PCa.
INTRODUCTION AND OBJECTIVE: Men are 3-4 times more likely to be diagnosed with bladder cancer (BCa) than women, who often have more aggressive tumors. Intravesical instillation of BCG is one of the first immunotherapies in widespread use and is given to patients with non-muscle invasive bladder cancer (NMIBC). Newer PD-1/PD-L1 checkpoint inhibitors now also have proven efficacy against BCa. Nonetheless, approximately 35% of patients fail to respond to BCG, with response rates to checkpoint immunotherapy much lower. Based on prior work suggesting the importance of the androgen receptor (AR) in BCa, we investigated AR-targeting as a novel strategy to improve the response to BCa immunotherapy in the MBT-2 murine BCa model.METHODS: Subcutaneous injection of 10 6 MBT-2 cells into male and female C3H mice was performed, with tumors harvested at tumor size limits. The AR antagonist enzalutamide (ENZ) was given in alone or combination with anti-PD-1 or intra-tumoral BCG treatments as previously described. Selected human NMIBC were collected following transurethral resection. After digestion of fresh human and murine tumors, flow cytometry analyses characterized the immune cells present. The anti-proliferative effect of AR antagonists against human and murine BCa cell lines was tested in vitro.RESULTS: The CD45 þ /CD45ratio, extent of tumor infiltrating lymphocytes, and expression of immune checkpoint markers from the MBT-2 model is comparable to the immune composition of NMIBC tumors resected from patients. Moreover, the model also recapitulates previously observed human tumor immune cell gender differences. AR antagonist ENZ demonstrated anti-proliferative effects in lower grade human and murine BCa cell lines, while limited differences were seen in human high grade BCa cell lines. While ENZ alone had no effect in vivo, its use in combination with either anti-PD-1 or BCG treatment in male mice synergized to improve response rates. Complete response was confirmed by re-challenge of mice, demonstrating immune memory. Notably, the proportion of complete responses in male mice treated with the combination treatment is similar to that observed in female mice with either immunotherapy alone. Flow cytometry analyses showed that these therapeutic benefits in male mice are related to an increase in pro-inflammatory tumor infiltrating dendritic cells as well as a decrease in anti-inflammatory tumor infiltrating dendritic cells and myeloid-derived suppressor cells.CONCLUSIONS: Our results suggest that combining AR antagonism with immunotherapy in male BCa patients may potentiate the antitumor immune response and increase response rates. Accordingly, a Phase II trial to evaluate the efficacy of AR antagonism in combination with BCG to decrease NMIBC tumor recurrences in men is planned. Further, the MBT-2 murine model appears relevant to investigate immunologic BCa sex differences.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.