Inflammatory Tumor Research Articles

Inhibitory Effect of Evodiamine on Psoriasis Lesions and Itching in Mice.

This study seeks to investigate the effect of evodiamine on psoriasis and psoriatic pruritus. Imiquimod-induced psoriasiform dermatitis in mice was used as a model, and evodiamine was topically applied for seven days. The mice were observed daily for skin damage on the back, clinical score and their scratching behavior was recorded. Blood samples were collected on the final day of the experiment, and the serum levels of pruritus-associated inflammatory cytokines tumor necrosis factor (TNF) -α, interleukin (IL) -23, and IL-17A were measured using enzyme-linked immunosorbent assay. Histopathological changes were observed in Hematoxylin and Eosin-stained skin specimens. The expression levels of transient receptor potential vanilloid (TRPV) 1, TRPV3, TRPV4, and the pruritus-related mediators Substance P (SP), nerve growth factor (NGF), and calcitonin gene-related peptide (CGRP) in the skin lesions were analyzed using Western blot and qRT-PCR. The effect of evodiamine on the exploratory behavior, motor, and coordination abilities of mice was assessed using open field, suspension, and Rota-Rod experiments. Molecular docking was utilized to verify the binding of evodiamine to the residues of TRPV1, TRPV3, and TRPV4. Evodiamine reduced pruritus and inhibited inflammation by decreasing the levels of inflammatory mediators TNF-α, IL-23, and IL-17A in the serum of Imiquimod-induced mice and attenuated the mRNA and protein expression levels of SP, NGF, CGRP, TRPV1, TRPV3, and TRPV4 in the skin. Evodiamine is an effective treatment for psoriasis and pruritus, due to its ability to inhibit immune inflammation and pruritic mediators.

Clinical utility of ctDNA as a tool to detect triple-negative breast cancer relapses: The CUPCAKE trial.

TPS1139 Background: The current follow-up of triple-negative breast cancer (TNBC) patients at high risk of relapse does not involve any blood or imaging monitoring to detect metastases, because of the low sensitivity and specificity of blood and imaging biomarkers in that context. In contrast, ctDNA detection techniques are highly specific, sensitive and can detect a molecular relapse several months before the occurrence of symptoms (1). We hypothesize that the early detection of molecular relapse by ctDNA combined with whole-body mapping of tumor recurrence sites by 68Ga-FAPI-46-PET/CT will enable the detection of relapses when they are still limited in size and number of sites. Detecting oligometastatic relapses could demonstrate a clinical utility by (i) allowing the use of local treatments in addition to systemic treatments; (ii) taking advantage of the higher efficacy of immunotherapy in patients with limited tumor burden and inflammation. Methods: CUPCAKE is a randomized, non-comparative, multicenter trial promoted by Institut Curie (Paris, France), which design is inspired from the Trials WIthin CohortS approach (2). N=450 non-metastatic TNBC patients at high risk of relapse will be included after having signed a written informed consent in a cohort allowing them to be followed by ctDNA monitoring. For each included patient, a ctDNA detection assay (FoundationOne Tracker) will be performed every 3 months, with a turnaround time of less than 3 weeks. When negative, ctDNA detection results will not be disclosed to patients nor clinicians. If, at any timepoint, ctDNA is detected (molecular relapse), patients will be randomized in a 1:1 ratio between the experimental arm (ctDNA-based intervention) and the control arm (no intervention). In the experimental arm, the positive ctDNA result will be disclosed. To locate metastatic foci, patients will be offered to undergo whole-body imaging with 18F-FDG and 68Ga-FAPI-46-PET/CT, in addition to any other workup considered as relevant by their treating physician. If/when a clinical/radiological relapse is observed, systemic or local treatments will be decided by physicians (standard of care). In the control arm, patients and their treating physician will not be made aware of the molecular relapse and will continue the standard surveillance with repeated ctDNA test every 3 months (blinded). At the time of the clinical/radiological diagnosis of relapse, similar procedures will be performed (18F-FDG & 68Ga-FAPI-46-PET/CT). The primary endpoint is overall survival rate at 24 months after the randomization in the experimental arm while the control arm will serve as reference. Secondary endpoints include efficacy criteria like progression-free survival but also clinical features upon radiological/radiological relapse, or performance criteria of 68Ga-FAPI-46 versus 18F-FDG PET/CT. 1. Coombes, Clin Cancer Res. 2019. 2. Relton, BMJ. 2010. Clinical trial information: 06225505.

Association between post-CART terminal complement complex (TCC) levels and clinically significant immune effector cell–associated neurotoxicity syndrome (ICANS).

7036 Background: The cornerstone of ICANS pathogenesis is endothelial dysfunction leading to blood-brain barrier disruption driven by inflammatory cytokines. Endothelial dysfunction is often associated with complement activation as in thrombotic microangiopathy, but the association between complement and ICANS has not yet been investigated. Herein, we describe the association between TCC levels (sC5b-9) and ICANS after CART. Methods: We retrospectively included 42 patients (pts) with B-cell non-Hodgkin lymphoma treated with axi-cel or brexu-cel from 03/2022 - 09/2023 at Ohio State University who had had sC5b-9 levels available pre-lymphodepletion (pre-LD), pre-CART and post-CART infusion. sC5b-9 levels were measured once in each of the timepoints: pre-LD (between D-7 and D-4), pre-CART (between D-3 and D0) and post-CART (between D2 and D8). We defined clinically significant ICANS as peak grade ≥2 (G≥2). Median and interquartile range (IQR) were used to describe the biomarkers levels, and Wilcoxon-rank sum test was used to compare values between groups. Results: Median age was 64 years (26-76), 30 (71%) pts had large B-cell lymphoma and 6 (14%) had untransformed follicular lymphoma who received axi-cel (36, 86%), and 6 (14%) had mantle cell lymphoma who received brexu-cel. Following CART, cytokine-release syndrome (CRS) peak grade was 2 and 1 in 16 (38%) and 22 (52%) of the pts, respectively, and 4 pts (10%) had no CRS. The ICANS peak grade was 4, 3, 2 and 1 in 4 (10%), 2 (5%), 6 (14%) and 6 (14%) of the pts, respectively, and 24 (57%) of the pts had no ICANS. The median time from CART to ICANS peak was 6 days (2-12). Tocilizumab and steroids were used in 28 (67%) and 20 (48%) of the pts, respectively. We compared sC5b-9 levels pre-LD, pre-CART and post-CART between pts that had ICANS peak G≥2 (N = 12) vs. grade 1 or did not have ICANS (G0-1, N = 30). We found that the post-CART sC5b-9 levels were significantly higher among pts who had ICANS G≥2 [median (IQR) 254 (129) vs. 183 (82) ng/mL, p = 0.013]. There was no statistical difference in the levels pre-LD and pre-CART between pts who had ICANS G≥2 vs. G0-1, median (IQR) 176 (103) vs. 159 (94) ng/mL, p = 0.37, and 152 (64) vs. 143 (69) ng/mL, p = 0.89, respectively. We also compared levels of other biomarkers associated with tumor burden, inflammation, and endothelial dysfunction at the same time points sC5b-9 was measured between pts with ICANS G≥2 vs. G0-1 and there were no statistically significant differences: median (IQR) pre-LD LDH 171 (73) vs. 180 (115) U/L, p =0.54, post-CART ferritin 262 (546) vs. 297 (333) ng/mL, p = 0.92 and post-CART fibrinogen 302 (221) vs. 405 (146) mg/dL, p = 0.29. Conclusions: Our findings support an association between post-CART sC5b-9 levels and clinically significant ICANS warranting further investigation of the role of complement in the ICANS pathogenesis and as a therapeutic target.

Neuroinflammation in Glioblastoma: The Role of the Microenvironment in Tumour Progression.

Glioblastoma (GBM) stands as the most aggressive and lethal among the main types of primary brain tumors. It exhibits malignant growth, infiltrating the brain tissue, and displaying resistance toward treatment. GBM is a complex disease characterized by high degrees of heterogeneity. During tumour growth, microglia and astrocytes, among other cells, infiltrate the tumour microenvironment and contribute extensively to gliomagenesis. Tumour-associated macrophages (TAMs), either of peripheral origin or representing brain-intrinsic microglia, are the most numerous nonneoplastic populations in the tumour microenvironment in GBM. The complex heterogeneous nature of GBM cells is facilitated by the local inflammatory tumour microenvironment, which mostly induces tumour aggressiveness and drug resistance. The immunosuppressive tumour microenvironment of GBM provides multiple pathways for tumour immune evasion, contributing to tumour progression. Additionally, TAMs and astrocytes can contribute to tumour progression through the release of cytokines and activation of signalling pathways. In this review, we summarize the role of the microenvironment in GBM progression, focusing on neuroinflammation. These recent advancements in research of the microenvironment hold the potential to offer a promising approach to the treatment of GBM in the coming times.

Interleukin-17A educated hepatic stellate cells promote hepatocellular carcinoma occurrence through fibroblast activation protein expression.

Liver cancer is typified by a complex inflammatory tumor microenvironment, where an array of cytokines and stromal cells orchestrate a milieu that significantly influences tumorigenesis. Interleukin-17A (IL-17A), a pivotal pro-inflammatory cytokine predominantly secreted by Th17 cells, is known to play a substantial role in the etiology and progression of liver cancer. However, the precise mechanism by which IL-17A engages with hepatic stellate cells (HSCs) to facilitate the development of hepatocellular carcinoma (HCC) remains to be fully elucidated. This investigation seeks to unravel the interplay between IL-17A and HSCs in the context of HCC. An HCC model was established in male Sprague-Dawley rats using diethylnitrosamine to explore the roles of IL-17A and HSCs in HCC pathogenesis. In vivo overexpression of Il17a was achieved using adeno-associated virus. A suite of molecular techniques, including RT-qPCR, enzyme-linked immunosorbent assays, Western blotting, cell counting kit-8 assays and colony formation assays, was employed for in vitro analyses. The study findings indicate that IL-17A is a key mediator in HCC promotion, primarily through the activation of hepatic progenitor cells (HPCs). This pro-tumorigenic influence appears to be mediated by HSCs, rather than through a direct effect on HPCs. Notably, IL-17A-induced expression of fibroblast activation protein (FAP) in HSCs emerged as a critical factor in HCC progression. Silencing Fap in IL-17A-stimulated HSCs was observed to reverse the HCC-promoting effects of HSCs. The collective evidence from this study implicates the IL-17A/FAP signaling axis within HSCs as a contributor to HCC development by enhancing HPC activation. These findings bolster the potential of IL-17A as a diagnostic and preventative target for HCC, offering new avenues for therapeutic intervention.

Biological subgroups in melanoma: A correlation of tumor genomic and transcriptomic analysis with spatial immunophenotyping.

e14667 Background: A prominent challenge advancing immunotherapy is the heterogeneity of resistance mechanisms leading to drug development failure despite efficacy in subgroup of patients. To address these shortcomings, precision medicine platforms aimed at prospectively stratifying patients into treatment groups based on tumor mutational burden (TMB) and tumor inflammation scores by gene expression profiling (GEP) are now being explored. Whether these gene-based variables reflect the true immunologic state of the tumor remains an open question. Here, we sought to evaluate TMB and GEP as markers of intratumoral inflammatory state by single-cell spatial analysis. Methods: Under an IRB approved protocol through the Huntsman Cancer Institute, 25 melanoma tumors with available clinical, WES, and RNAseq data were stained with a multiplex immunofluorescence-based protocol using Opal dye and Leica Bond Rx (CD8, CD68, PD1, PD-L1, Ki67, SOX10, DAPI) and scanned with Akoya Polaris whole slide scanner. Images were analyzed using HALO. Results: Melanoma tumors were stratified into 4 subgroups based on TMB (<10 vs ≥10 m/Mb) by WES, and inflammatory (Inflm) gene signature expression (GSEA >0 vs ≤0) by RNAseq, denoted as S1 (TMBhi/Inflmhi; n=4), S2 (TMBhi/Inflmlo; n=4), S3 (TMBlo/Inflmhi; n=8), and S4 (TMBlo/Inflmlo; n=9). The rate of disease progression was 0% (S1), 25% (S2), 25% (S3), and 56% (S4). Infiltration analysis of CD8 T cells showed elevated intratumoral CD8 density in S1 (249/mm2) and S3 (157/mm2) vs S2 (65/mm2) and S4 (65/mm2; p=0.0002), consistent with GEP findings. Spatial analysis showed a similar pattern of CD8 distribution across subgroups with peak CD8 density in the tumor periphery. Proximity analyses showed average distance of CD8 T cell to the nearest tumor cell was not different across subgroups. Furthermore, we found Inflmhi tumors had a higher frequency of PD1+ CD8 T cells relative to Inflmlo tumors (12.9 vs 2.8 cells/HPF; p=0.0002), suggesting GEP also reflects CD8 activation and functional status. Increased frequency of PD1+ CD8 T cells was correlated with a much improved overall survival (HR=3.3; p=0.017). A similar analysis performed on CD68+ macrophages showed significantly increased peri-tumoral macrophage recruitment in S4 (445/mm2) vs S1 (209/mm2; p<0.0001), S2 (91/mm2; p<0.0001), and S3 (267/mm2; p=0.0003) subgroups. Interestingly, Inflmhi tumors were associated with an increased proportion of PDL1+ macrophages regardless of TMB status (43% vs 3%; p<0.0001), suggestive of an upregulated adaptive immune response in tumors with active inflammation. Conclusions: Our findings illustrate that biological groupings based on TMB and GEP biomarkers correlate with CD8 spatial location and activation state. Each subgroup has unique tumor immune features that can be related to immune resistance and warrant further investigation.

Risk predictors as companion diagnostics for ER positive, HER2 negative early staged breast cancers from India.

e13049 Background: Gene expression profiling tests can identify women with breast cancer most likely to benefit from chemotherapy providing accurate prognostic information than other non-genetic clinicopathological parameters. These derive specific molecular features of a tumour indicating an increased likelihood of rapid growth, metastasis risk, and response to chemotherapy. The tests are typically performed after surgery, in conjunction with other available information such as tumour size and grade. The objective of the current study was to evaluate the risk prediction as a companion diagnostic assay for women with early staged breast cancers (ER+,HER2-ve). Methods: The risk prediction was done evaluating quantitative gene expression profiling on Nanostring nCounter Flex platform using total RNA extracted from FFPE tissues (n = 81) obtained from trucut biopsy tissues and post MRM specimens after ensuring the tumour representation. The gene quantitations were derived after normalisation and using nSolver and Roselind software based statistical analysis. Results: We found 55.5% (45/81) and 44.5% (36/81) to be of intrinsic subtype Luminal A and Luminal B among our breast tumours respectively. The risk of recurrence score was high for luminal B (36.1% ; 13/36) compared to luminal A (13.4% ; 6/45). There was no difference pertaining to the Tumour Inflammation score (TIS score) among the luminal A and B that depicted good response to immunotherapy with 80% (36/45) among the luminal A and 72.3% (26/36) among the luminal B. We have identified top 38 differentially expressed genes that drive the high breast cancer proliferation score among the breast cancers in Indian women. The top 5 genes influencing the risk of recurrence with high BCP among the luminal A tumours were GNLY, CHRNA5, CDC20, MELK, RRM2 and among luminal B we got FGFR2, AURKA, MYBL2, CCNE2 and UBE2C. The top genes combining both types influencing the genomic risk of recurrence were FGFR2, MYBL2, AURKA, TTK, UBE2c and ORC6. The genomic risk of recurrence is based only on the gene expression unlike the risk of recurrence which includes the tumour size and nodal involvement. The top genes influencing a poor response to immunotherapy among luminal A was CNTFR, HAPLN1, FGF12, COL4A6 and CDH2 and the top genes influencing poor response to immunotherapy among luminal B was FGFR2, MYCN, AURKA and FAM198B. Conclusions: This is the first study from India using risk prediction test as a companion diagnostics assay for ER+ HER2-ve early staged breast cancer. The salient genes important to drive the breast cancer proliferation increasing the risk of recurrence scores in both luminal A and Luminal B intrinsic subtypes have been derived. We can validate these gene signatures as smaller panels of tests that can be low cost and more suitable for LMIC like India. The BCP panel of genes were relevant driving the recurrence.

The In Vitro Antioxidant and Anti-Inflammatory Activities of Selected Australian Seagrasses.

Recent studies have shown that seagrasses could possess potential applications in the treatment of inflammatory disorders. Five seagrass species (Zostera muelleri, Halodule uninervis, Cymodocea rotundata, Syringodium isoetifolium, and Thalassia hemprichii) from the Great Barrier Reef (QLD, Australia) were thus collected, and their preliminary antioxidant and anti-inflammatory activities were evaluated. From the acetone extracts of five seagrass species subjected to 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging antioxidant assay, the extract of Z. muelleri had the highest activity (half minimal concentration of inhibition (IC50) = 138 µg/mL), with the aerial parts (IC50 = 119 µg/mL) possessing significantly higher antioxidant activity than the roots (IC50 ≥ 500 µg/mL). A human peripheral blood mononuclear cells (PBMCs) assay with bacterial lipopolysaccharide (LPS) activation and LEGENDplex cytokine analysis showed that the aerial extract of Z. muelleri significantly reduced the levels of inflammatory cytokines tumour necrosis factor alpha (TNF-α), interleukin (IL)-1β, and IL-6 by 29%, 74%, and 90%, respectively, relative to the LPS treatment group. The aerial extract was thus fractionated with methanol (MeOH) and hexane fraction, and purification of the MeOH fraction by HPLC led to the isolation of 4-hydroxybenzoic acid (1), luteolin (2), and apigenin (3) as its major constituents. These compounds have been previously shown to reduce levels of TNF-α, IL-1β, and IL-6 and represent some of the major bioactive components of Z. muelleri aerial parts. This investigation represents the first study of the antioxidant and anti-inflammatory properties of Z. muelleri and the first isolation of small molecules from this species. These results highlight the potential for using seagrasses in treating inflammation and the need for further investigation.

Immune cell infiltration and inflammatory landscape in primary brain tumours

BackgroundPrimary malignant brain tumours are more than one-third of all brain tumours and despite the molecular investigation to identify cancer driver mutations, the current therapeutic options available are challenging due to high intratumour heterogeneity. In addition, an immunosuppressive and inflammatory tumour microenvironment strengthens cancer progression. Therefore, we defined an immune and inflammatory profiling of meningioma and glial tumours to elucidate the role of the immune infiltration in these cancer types.MethodsUsing tissue microarrays of 158 brain tumour samples, we assessed CD3, CD4, CD8, CD20, CD138, Granzyme B (GzmB), 5-Lipoxygenase (5-LOX), Programmed Death-Ligand 1 (PD-L1), O-6-Methylguanine-DNA Methyltransferase (MGMT) and Transglutaminase 2 (TG2) expression by immunohistochemistry (IHC). IHC results were correlated using a Spearman correlation matrix. Transcript expression, correlation, and overall survival (OS) analyses were evaluated using public datasets available on GEPIA2 in Glioblastoma (GBM) and Lower Grade Glioma (LGG) cohorts.ResultsSeven out of ten markers showed a significantly different IHC expression in at least one of the evaluated cohorts whereas CD3, CD4 and 5-LOX were differentially expressed between GBMs and astrocytomas. Correlation matrix analysis revealed that 5-LOX and GzmB expression were associated in both meningiomas and GBMs, whereas 5-LOX expression was significantly and positively correlated to TG2 in both meningioma and astrocytoma cohorts. These findings were confirmed with the correlation analysis of TCGA-GBM and LGG datasets. Profiling of mRNA levels indicated a significant increase in CD3 (CD3D, CD3E), and CD138 (SDC1) expression in GBM compared to control tissues. CD4 and 5-LOX (ALOX5) mRNA levels were significantly more expressed in tumour samples than in normal tissues in both GBM and LGG. In GBM cohort, GzmB (GZMB), SDC1 and MGMT gene expression predicted a poor overall survival (OS). Moreover, in LGG cohort, an increased expression of CD3 (CD3D, CD3E, CD3G), CD8 (CD8A), GZMB, CD20 (MS4A1), SDC1, PD-L1, ALOX5, and TG2 (TGM2) genes was associated with worse OS.ConclusionsOur data have revealed that there is a positive and significant correlation between the expression of 5-LOX and GzmB, both at RNA and protein level. Further evaluation is needed to understand the interplay of 5-LOX and immune infiltration in glioma progression.

SRS 16-86 promotes diabetic nephropathy recovery by regulating ferroptosis.

Diabetic nephropathy (DN) is a common complication of diabetes mellitus (DM), and cell death plays an important role. Ferroptosis is a recently discovered type of iron-dependent cell death and one that is different from other kinds of cell death including apoptosis and necrosis. However, ferroptosis has not been described in the context of DN. This study explored the role of ferroptosis in DN pathophysiology and aimed to confirm the efficacy of the ferroptosis inhibitor SRS 16-86 on DN. Streptozotocin injection was used to establish the DM and DN animal models. To investigate the presence or occurrence of ferroptosis in DN, we assessed the concentrations of iron, reactive oxygen species and specific markers associated with ferroptosis in a rat model of DN. Additionally, we performed haematoxylin-eosin staining, blood biochemistry, urine biochemistry and kidney function analysis to evaluate the efficacy of the ferroptosis inhibitor SRS 16-86 in ameliorating DN. We found that SRS 16-86 could improve the recovery of renal function after DN by upregulating glutathione peroxidase 4, glutathione and system xc -light chain and by downregulating the lipid peroxidation markers and 4-hydroxynonenal. SRS 16-86 treatment could improve renal organization after DN. The inflammatory cytokines interleukin 1β and tumour necrosis factor α and intercellular adhesion molecule 1 were significantly decreased following SRS 16-86 treatment after DN. The results indicate that there is a strong connection between ferroptosis and the pathological mechanism of DN. The efficacy of the ferroptosis inhibitor SRS 16-86 in DN repair supports its use as a new therapeutic treatment for DN.

In Situ Colonization of Trojan‐Yeast for Targeted Glucose Starvation and Reconstruction of Tumor Immune Environment

AbstractGlucose is an important energy source for living cells. Particularly, most tumor cells use glycolysis to harness energy for their fast growth, invasion and metastasis even under aerobic conditions. Thus, reduction of the glucose level in cancer can lead to suppression of tumor growth. Here, macrophage membrane dressed yeast, Trojan M‐yeast, is developed, via coating the surface of yeast with macrophage membrane, for targeted accumulation and suppression of tumor by glucose starvation. Due to the inflammatory homing effects of macrophage membrane, M‐yeast efficiently target and colonize in the inflammatory tumor site, where they compete with tumor cells for glucose consumption, thereby disrupting glycolysis of cancer cells, resulting in the reduction of lactate accumulation and reversal of the acidosis of tumor microenvironment. Subsequently, the apoptotic tumor cells and yeast promote dendritic cell maturation, increase tumor‐infiltrating cytotoxic T lymphocytes. Meanwhile, ethanol and acetaldehyde produced by yeast metabolism of glucose can re‐polarize tumor associated macrophages (TAMs) to the anti‐tumor phenotype (M1), thus reprogramming the tumor immunosuppressive microenvironment, leading to synergistic tumor suppression together with glucose starvation, which is different from traditional chemotherapy and immunotherapy. M‐yeast exhibits promising targeted cancer therapy with a decent safety profile. This work may provide new insights as well as a new paradigm for yeast‐based cancer therapy.

Olsalazine pretreatment augments chemosensitivity of gemcitabine in hepatocellular carcinoma.

Recently, olsalazine a DNA hypomethylating agent was found to inhibit the growth of breast cancer cells. The present study was carried out to evaluate the effects of olsalazine pretreatment in the potentiation of chemosensitivity of gemcitabine for the treatment of hepatocellular carcinoma (HCC).In silico molecular docking was performed to analyze the interaction of olsalazine and gemcitabine with DNMT1 and DNA, respectively, using the AutoDock tools 1.5.6. Cytotoxicity of olsalazine, gemcitabine, and combination were measured on human HePG2 cells using MTT assay. Antiproliferative effects were assessed using animal model of N-nitrosodiethylamine and carbon tetrachloride-induced HCC. Treatment was initiated from 8th week of induction to 11th week and change in body weight, liver weight, and survival rate were measured. Following treatment, blood samples were collected for estimation serum biochemistry. Blood serum was used for the estimation of inflammatory cytokines tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6), C-reactive protein[CRP],lactate dehydrogenase (LDH), and P53 levels. Oxidative stress markers were measured in liver tissue homogenates. Histopathology and immunohistochemistry (IHC) were performed on liver sections to detect the morphological changes and P53 expression. Docking analysis revealed the interactions between olsalazine and DNMT1 with a binding energy score of -5.34 and gemcitabine and DNA with a binding energy score of -5.93. Olsalazine pretreatment potentiated the antiproliferative effect of gemcitabine in cell line study. In the group receiving olsalazine pretreatment showed significant reductions in relative liver weight and improved survival rate of gemcitabine treatment group. Serum biochemical markers:serum glutamate pyruvate transaminase,serum glutamic oxaloacetic transaminase, alkaline phosphatase, and bilirubin revealed improved liver functions. Olsalazine pretreatment also reduced the levels of inflammatory markers like CRP, LDH, TNF-α, and IL-6 and oxidative stress markers dose dependently. Histopathology and IHC showed improved liver morphology with potentiated the induction of P53 upon olsalazine pretreatment in combination with gemcitabine. In conclusion, sequential combination of olsalazine and gemcitabine improvedthe treatment outcomes during the progression of HCC.

SLC7A5 correlated with malignancies and immunotherapy response in bladder cancer

BackgroundMetabolic reprogramming contributes to bladder cancer development. This study aimed to understand the role of SLC7A5 in bladder cancer.MethodsWe systematically analyzed the correlation between SLC7A5 and bladder cancer through various approaches, including bioinformatics, western blotting, cell cycle analysis, cell proliferation assays, and invasion experiments. We also investigated the immunological features within the tumor microenvironment (TME), encompassing cancer immune cycles, immune modulators, immune checkpoints, tumor-infiltrating immune cells (TIIC), T cell inflammation scores, and treatment responses. Additionally, for a comprehensive assessment of the expression patterns and immunological roles of SLC7A5, pan-cancer analysis was performed using cancer genomics datasets.ResultsSLC7A5 was associated with adverse prognosis in bladder cancer patients, activating the Wnt pathway and promoting bladder cancer cell cycle progression, proliferation, migration, and invasion. Based on the evidence that SLC7A5 positively correlated with immunomodulators, TIIC, the cancer immune cycle, immune checkpoint and T cell inflammation scores, we also found that SLC7A5 was associated with the inflammatory tumor immune microenvironment. EGFR-targeted therapy, cancer immunotherapy, and radiation therapy were effective for patients with high SLC7A5 expression in bladder cancer. Low SLC7A5 patients were, however, sensitive to targeted therapies and anti-angiogenic therapy, such as blocking β-catenin network, PPAR-γ and FGFR3 signaling. Anti-SLC7A5 combined with cancer immunotherapy may have greater effectiveness than either therapy alone. Furthermore, we observed specific overexpression of SLC7A5 in TME of various cancers.ConclusionSLC7A5 can predict therapeutic response to immunotherapy, radiotherapy and chemotherapy in bladder cancer patients. Targeting SLC7A5 in combination with immunotherapy may be a potentially appropriate treatment option.

Cryo-EM reveals that iRhom2 restrains ADAM17 protease activity to control the release of growth factor and inflammatory signals

A disintegrin and metalloprotease 17 (ADAM17) is a membrane-tethered protease that triggers multiple signaling pathways. It releases active forms of the primary inflammatory cytokine tumor necrosis factor (TNF) and cancer-implicated epidermal growth factor (EGF) family growth factors. iRhom2, a rhomboid-like, membrane-embedded pseudoprotease, is an essential cofactor of ADAM17. Here, we present cryoelectron microscopy (cryo-EM) structures of the human ADAM17/iRhom2 complex in both inactive and active states. These reveal three regulatory mechanisms. First, exploiting the rhomboid-like hallmark of TMD recognition, iRhom2 interacts with the ADAM17 TMD to promote ADAM17 trafficking and enzyme maturation. Second, a unique iRhom2 extracellular domain unexpectedly retains the cleaved ADAM17 inhibitory prodomain, safeguarding against premature activation and dysregulated proteolysis. Finally, loss of the prodomain from the complex mobilizes the ADAM17 protease domain, contributing to its ability to engage substrates. Our results reveal how a rhomboid-like pseudoprotease has been repurposed during evolution to regulate a potent membrane-tethered enzyme, ADAM17, ensuring the fidelity of inflammatory and growth factor signaling.

Diabetes and Cancer: A Twisted Bond.

This paper presents an overview of the interconnection between various factors related to both cancer and type 2 diabetes mellitus (T2DM). Hyperglycemia, hyperinsulinemia, chronic inflammation, and obesity are involved in the development and progression of both diseases but, strong evidence for a direct causal relationship between diabetes and cancer, is lacking. Several studies described a relationship between hyperglycemia and cancer at the cellular, tissular and organismic levels but at the same time recent Mendelian randomization studies proved a significant causal relationship only between hyperglycemia and breast cancer. On the other hand, the association between both hyperinsulinemia and obesity and several cancer types appears to be robust as demonstrated by Mendelian randomized studies. Metabolic alterations, including the Warburg effect and excessive glucose consumption by tumors, are discussed, highlighting the potential impact of dietary restrictions, such as fasting and low-carb diets, on tumor growth and inflammation. Recent data indicates that circulating branched-chain amino acids levels, may represent novel biomarkers that may contribute to both better diabetes control and early pancreatic cancer detection. Understanding the underlying mechanisms and shared risk factors between cancer and T2DM can provide valuable insights for cancer prevention, early detection, and management strategies.

TRPC absence induces pro-inflammatory macrophage polarization to promote obesity and exacerbate colorectal cancer.

During the past half-century, although numerous interventions for obesity have arisen, the condition's prevalence has relentlessly escalated annually. Obesity represents a substantial public health challenge, especially due to its robust correlation with co-morbidities, such as colorectal cancer (CRC), which often thrives in an inflammatory tumor milieu. Of note, individuals with obesity commonly present with calcium and vitamin D insufficiencies. Transient receptor potential canonical (TRPC) channels, a subclass within the broader TRP family, function as critical calcium transporters in calcium-mediated signaling pathways. However, the exact role of TRPC channels in both obesity and CRC pathogenesis remains poorly understood. This study set out to elucidate the part played by TRPC channels in obesity and CRC development using a mouse model lacking all seven TRPC proteins (TRPC HeptaKO mice). Relative to wild-type counterparts, TRPC HeptaKO mice manifested severe obesity, evidenced by significantly heightened body weights, augmented weights of epididymal white adipose tissue (eWAT) and inguinal white adipose tissue (iWAT), increased hepatic lipid deposition, and raised serum levels of total cholesterol (T-CHO) and low-density lipoprotein cholesterol (LDL-C). Moreover, TRPC deficiency was accompanied by an decrease in thermogenic molecules like PGC1-α and UCP1, alongside a upsurge in inflammatory factors within adipose tissue. Mechanistically, it was revealed that pro-inflammatory factors originating from inflammatory macrophages in adipose tissue triggered lipid accumulation and exacerbated obesity-related phenotypes. Intriguingly, considering the well-established connection between obesity and disrupted gut microbiota balance, substantial changes in the gut microbiota composition were detected in TRPC HeptaKO mice, contributing to CRC development. This study provides valuable insights into the role and underlying mechanisms of TRPC deficiency in obesity and its related complication, CRC. Our findings offer a theoretical foundation for the prevention of adverse effects associated with TRPC inhibitors, potentially leading to new therapeutic strategies for obesity and CRC prevention.

Alleviative effect of betaine against copper oxide nanoparticles-induced hepatotoxicity in adult male albino rats: histopathological, biochemical, and molecular studies

BackgroundCopper oxide nanoparticles (CuO-NPs) have gained interest due to their availability, efficiency, and their cost-effectiveness. Betaine is an essential methyl donor and takes part in various physiological activities inside the body; it is found to have protective and curative effects against various liver diseases. The present study aimed to evaluate the hepatotoxic effect of CuO-NPs on adult male albino rats and the ability of betaine to alleviate such hepatotoxicity.MethodsForty adult male albino Wister rats were grouped into 4 groups (10 rats/group): group I a negative control, group II (CuO-NPs) injected with CuO-NPs intra peritoneal by insulin needle (0.5 mg/kg/day), group III (betaine + CuO-NPs) administered betaine orally by gavage needle (250 mg/kg/day 1 h before CuO-NPs) and CuO-NPs (0.5 mg/kg/day) finally, group IV (betaine) administered betaine orally by gavage needle (250 mg/kg/day) for consecutive 28 days. Blood and liver samples were gathered and processed for biochemical, molecular, histopathological, and immunohistochemical investigations.ResultsGroup II displayed a marked rise in alanine aminotransferase (ALT), aspartate aminotransferase (AST), and malondialdehyde (MDA) levels. Furthermore, there is an excessive upregulation of the inflammatory biomarkers interleukin1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α). On the other hand, substantial reduction in glutathione (GSH) levels and significant downregulation at glutathione peroxidase (GPx) mRNA gene expression. Regarding the histopathological deviations, there were severe congestion, dilatation and hyalinization of blood vessels, steatosis, hydropic degeneration, hepatocytic necrosis, increased binucleation, degenerated bile ducts, hyperplasia of ducts epithelial lining, and inflammatory cells infiltration. Immunohistochemically, there was a pronounced immunoreactivity toward IL-1β. Luckily, the pre-administration of betaine was able to mitigate these changes. MDA was dramatically reduced, resulting in the downregulation of IL-1β and TNF-α. Additionally, there was a considerable rise in GSH levels and an upregulation of GPx. Histopathological deviations were substantially improved as diminished dilatation, hyalinization and congestion of blood vessels, hepatocytes, and bile ducts are normal to some extent. In addition, IL-1β immunohistochemical analysis revealed marked decreased intensity.ConclusionBetaine can effectively reduce the hepatotoxicity caused by CuO-NPs via its antioxidant properties and its ability to stimulate the cell redox system.

Immune profiling of dedifferentiated liposarcoma and identification of novel antigens for targeted immunotherapy

Dedifferentiated liposarcoma (DDLS) is an aggressive, recurring sarcoma with limited treatments. T-cell immunotherapies selectively target malignant cells, holding promise against DDLS. The development of successful immunotherapy for DDLS requires a thorough evaluation of the tumor immune microenvironment and the identification and characterization of targetable immunogenic tumor antigens. To assess the complexity of the human DDLS tumor immune microenvironment and to identify target antigens, we used the nCounter NanoString platform, analyzing gene expression profiles across 29 DDLS and 10 healthy adipose tissue samples. Hierarchical clustering of tumors based on expression of tumor inflammation signature genes revealed two distinct groups, consisting of 15 inflamed tumors and 14 non-inflamed tumors, demonstrating tumor heterogeneity within this sarcoma subtype. Among the identified antigens, PBK and TTK exhibited substantial upregulation in mRNA expression compared to healthy adipose tissue controls, further corroborated by positive protein expression by IHC. This data shows considerable inter-tumoral heterogeneity of inflammation, which should be taken into consideration when designing an immunotherapy for DDLS, and provides a novel targetable antigen in DDLS. The results of this study lay the groundwork for the development of a novel immunotherapy for this highly aggressive sarcoma.

Effects of dietary astaxanthin on growth performance, immunity, and tissue composition in largemouth bass, Micropterus salmoides

This study examined the effects of feeding largemouth bass (Micropterus salmoides) with diets containing different doses of astaxanthin (0, 50, 100, 150, and 200 mg/kg) for 8 weeks. The results showed that the values of weight gain significantly increased from 620.32 ± 50.38% to 826.14 ± 33.49% as dietary astaxanthin levels increased from 0 mg/kg to 100 mg/kg. When the astaxanthin level exceeded 150mg/kg, the weight gain rate showed a downward trend, but there was no significant difference among of the 100, 150 and 200 mg/kg groups. The feed conversion ratio (FCR) and protein efficiency ratio (PER) were also improved by adding astaxanthin to diets (P < 0.05). Meanwhile, adding astaxanthin to the feed increased the length and thickness of intestinal villus and muscle layer thickness (P < 0.05). The astaxanthin supplementation increased the expression of the NF-E2-related factor (Nrf2) gene and reduced malondialdehyde (MDA) content and the expression of apoptosis genes Caspase-9 and Caspase-3 (P < 0.05), indicating that it has a good antioxidant ability. Furthermore, adding astaxanthin increased the content of non-specific immune markers and decreased the expression levels of the inflammatory factors interleukin-15 (IL-15) and tumor necrosis factor-α (TNF-α). Moreover, fish fed diets with astaxanthin exhibited lower blood cortisol levels (P < 0.05). The proportions of C20:4n6 (ARA) and C20:5n3 (EPA) in the liver decreased with increasing dietary astaxanthin levels. Based on WGR and SGR values, the optimal addition level of astaxanthin in largemouth bass feed is 134.8 mg/kg ~ 135.75 mg/kg. In summary, the appropriate dietary astaxanthin enhanced the antioxidant capacity and immune response of largemouth bass and had a positive effect on its intestinal health.

Hsa_circ_0007482 Promotes Proliferation and Differentiation of Chondrocytes in Knee Osteoarthritis.

Knee osteoarthritis (KOA) is a complex degenerative joint disease and a major cause of joint dysfunction. This study aimed to explore the function of hsa_circ_0007482 on inflammation, proliferation, differentiation, and apoptosis in KOA. Real-time quantitative polymerase chain reaction (PCR) was performed to detect the expression of circ_0007482, inflammatory factors, and differentiation-related molecules in KOA chondrocytes and interleukin (IL)-1β-stimulated chondrocytes. The correlation between the circ_0007482 expression and inflammatory factors was analyzed by the Pearson method. KOA cell model was established using IL-1β for 24 hours. The proliferation activity of chondrocytes was evaluated by CCK-8 assay, and cell apoptosis rate was assessed by flow cytometry. The downstream miRNA of circ_0007482 was validated using dual-luciferase reporter assay. The circ_0007482 expression was elevated in both KOA cartilage tissues and IL-1β-treated chondrocytes and positively correlated with inflammatory factors expression. In comparison to the control group, IL-1β treatment diminished chondrocyte proliferation abilities and increased cell apoptosis and inflammatory factors IL-6, IL-8, and tumor necrosis factor (TNF)-α mRNA expression. Inhibition of circ_0007482 partially improved IL-1β-induced inflammatory reaction. Circ_0007482 could negatively regulate the expression of miR-558. Interfering of circ_0007482 might partially promote cell proliferation and differentiation, while inhibit cell apoptosis to improve joint injury by regulating miR-558 in IL-1β-treated chondrocyte cell model.