High Expression Levels Research Articles

Modulation of the malignant behavior of tongue squamous cell carcinoma cells by matrix metallopeptidase 25 through the NF-κB pathway.

Accumulating evidence has implicated matrix metalloproteinases (MMPs) in the progression of human cancers. Matrix metallopeptidase 25 (MMP25) is a membrane-type MMP whose role in tumorigenesis and cancer development is not well understood. Here, we investigated the functions of MMP25 in tongue squamous cell carcinoma (TSCC). Gene expression was measured using real-time PCR and western blot. CCK-8 and Transwell assays were used to determine the proliferation, migration, and invasion of TSCC cells. An NK cell co-culture experiment was performed to evaluate the killing of TSCC cells by NK cells. MMP25 had higher expression levels in TSCC tissues than in adjacent non-cancerous tissues. MMP25-overexpressing and MMP25-silenced TSCC cell lines were established by lentiviral transduction. Overexpression of MMP25 promoted proliferation, migration, and invasion of TSCC cells, whereas knockdown of MMP25 had opposite effects. MMP25 modulated the levels of proliferation- and apoptosis-related proteins (PCNA, cyclin D, cyclin B1, p27, and cleaved caspase 3 and 9) and upregulated two invasion-related MMPs (mature MMP2 and MMP9). Additionally, MMP25 promoted tumor growth of TSCC cells in athymic nude mice. Notably, MMP25 upregulated PD-L1 in TSCC cells, attenuated NK cell killing of TSCC cells, and inhibited the secretion of anti-tumor cytokines (TNF-α and IFN-γ). Furthermore, MMP25 promoted the nuclear translocation of NF-κB p65, suggesting that activation of NF-κB signaling may mediate the pro-tumor functions of MMP25 in TSCC. This study revealed a novel role for MMP25 in TSCC, highlighting the potential of MMP25 as a therapeutic target in TSCC.

Validation of the biological function and prognostic significance of AURKA in neuroblastoma.

Neuroblastoma (NB) is the most common extracranial solid tumor in children, and the AURKA gene encodes a protein kinase involved in cell cycle regulation that plays an oncogenic role in a variety of human cancers. The aim of this study was to validate the biological function and prognostic significance of AURKA in NB using basic experiments and bioinformatics. Data obtained from Target and GEO databases were analyzed using various bioinformatic techniques. The expression of AURKA in 77 NB samples was detected by immunohistochemistry (IHC) method. The lentiviral RNA interference technique was employed to downregulate AURKA gene expression in NB cell lines. Additionally, cell counting kit-8 and flow cytometry analysis were conducted to investigate the impact of AURKA expression on cell proliferation, cell cycle progression, and apoptosis. A bioinformatic analysis showed that patients with NB in the AURKA-high-expression group had shorter OS (Overall Survival). Immune cell infiltration analysis showed that only activated CD4 T cell and type 2 T helper cell infiltration levels were higher in the AURKA-high-expression group than in the AURKA-low-expression group, with the infiltration levels of most other immune cells and cytokines lower in the high-expression group. Furthermore, the enhanced infiltration of activated CD4 T cells was associated with worse OS in patients with NB. IHC results showed that the AURKA expression was correlated with MYCN status and INSS stage. Log-rank test showed that pathological type, MYCN status, INSS stage, COG risk group, and AURKA expression was related to PFS (Progression-free survival) of NB patients, but COX regression analysis showed that none of the above factors were independently prognostic for PFS. In vitro, shRNA delivered via an AURKA-specific lentivirus significantly and consistently silenced endogenous AURKA expression in the human NB cell line SK-N-AS. This inhibited tumor cell proliferation, induced apoptosis, and caused G2/M-phase cell cycle arrest. Moreover, western blot assay showed significant reductions in the levels of mTOR, p70S6K, and 4E-BP1 phosphorylation in the AURKA-knockdown group. I found in subsequent experiments that NFYB can bind to the AURKA promoter and thus promote AURKA expression. High-level AURKA expression in NB is associated with poor patient prognosis. Silencing AURKA inhibited tumor cell proliferation, induced tumor cell apoptosis, and led to cell cycle arrest in the G2/M phase. Mechanistically, AURKA knockdown inhibited the phosphorylation and the activation of the mTOR1/p70S6K/4E-BP1 signaling pathway. In addition, AURKA was observed to regulate the infiltration levels of various immune cells in the NB tumor microenvironment, resulting in remodeling of the immunosuppressive tumor microenvironment.

Exploring the Relationship Between VMAT2 and DAT Expression, Psychotic Experiences, Craving, and Treatment Motivation in Male Patients with Methamphetamine Use Disorder.

Objectives: We aimed to examine the relationship of Dopamine transporter (DAT) and vesicular monoamine transporter (VMAT-2) gene and protein levels with psychic experiences and other clinical parameters in individuals with Methamphetamine Use Disorder (MUD). Methods: This study included 50 males diagnosed with MUD and 50 males as a smoking control (SC) and nonsmoking control (NSC). Community Assessment of Psychic Experiences (CAPE) was administered to patients and controls; Addiction Profile Index, Treatment Motivation Questionnaire, and Substance Craving Scale were administered only to the patient group. DAT and VMAT2 gene and protein levels were determined in blood obtained from the controls and patient groups. Results: CAPE positive, depressive, total, and distress scores were significantly higher in the patient group. DAT protein level and VMAT2 gene and protein levels were lower in the patient group compared to the controls. The DAT gene expression level was higher in the patient group compared to the controls. There was no correlation between any clinical variables and expression levels. A low VMAT2 gene expression level could diagnose MUD with a 5% probability when NSCs were used as a reference. A high DAT gene expression level could diagnose tobacco use disorder (TUD) with a 99.9% probability when NSCs were used as a reference. Conclusions: The patient group showed more psychic experiences than healthy people. The low expression of the VMAT2 gene was identified as a predictor of MUD, while the high expression of the DAT gene was predictive of TUD.

The Prognostic Significance of CD47, CD68, and CD163 Expression Levels and Their Relationship with MLR and MAR in Locally Advanced and Oligometastatic Nasopharyngeal Carcinoma.

This study aimed to assess the prognostic and predictive implications of CD47, CD68, and CD163, biomarkers of tumor-associated macrophages (TAMs), on the treatment efficacy and clinical outcomes of nasopharyngeal carcinoma (NPC). Additionally, the prognostic value of TAM-related indices, such as the monocyte-to-lymphocyte ratio (MLR) and monocyte-to-albumin ratio (MAR), was evaluated. A retrospective cohort of 54 patients with locally advanced or oligometastatic NPC treated with concurrent chemoradiotherapy (CCRT), with or without induction chemotherapy, was analyzed. Patients were categorized based on the cumulative expression scores for CD47, CD68, and CD163: negative/low (0-3 points) and high (4-6 points). MLR and MAR were also stratified as low MLR (<0.545) vs. high MLR (≥0.545) and low MAR (<16.145) vs. high MAR (≥16.145). The primary endpoint was overall survival (OS). High CD47, CD68, and CD163 expression levels were correlated with advanced clinical stage, reduced CCRT response, and elevated MLR and MAR. These TAM biomarkers were linearly correlated with each other and with established risk factors such as advanced age and elevated EBV-DNA levels. Kaplan-Meier analysis revealed that patients with low TAM expression had significantly longer OS and progression-free survival (PFS) than those with high TAM expression. Multivariate analysis identified high CD163, MLR, and MAR levels as independent adverse prognostic factors for OS. Elevated MLR is an independent risk factor for both OS and PFS in patients with NPC. CD47, CD68, and CD163 are significant prognostic markers in NPC, with higher levels being associated with poorer OS and PFS. Elevated MLR and MAR values also predict worse outcomes, underscoring their value as prognostic tools. CD163 and MLR are particularly strong predictors, highlighting the crucial role of TAMs in NPC management and suggesting that CD163 is a potential therapeutic target within the immune checkpoint pathway.

Duhuo Jisheng Decoction in reduction of inflammatory response via Transforming growth factor-β/Smad signaling pathway for repairing rabbit articular cartilage Injury: A Randomized Controlled Trial

ObjectiveThis study aims to investigate the mechanism underlying the effect of Duhuo Jisheng Decoction on the repair of rabbit articular cartilage injury through a reduction in the inflammatory response mediated by the Transforming growth factor (TGF)-β/Smad signaling pathway. MethodsA rabbit articular cartilage injury model was constructed using a ring bone extraction drill. Twenty-four Japanese white rabbits were randomly divided into six groups, namely Sham operation, model, low-dose Duhuo Jisheng Decoction, medium-dose Duhuo Jisheng Decoction, high-dose Duhuo Jisheng Decoction, and positive control groups. The treatment lasted 12 weeks. Gross observation, International Cartilage Repair Society score, Wakitani score, and Micro-computed tomography analysis were used to evaluate the structural repair of cartilage injury. Histology and immunohistochemistry were used to observe the proteoglycan, P-TβRII, P-Smad2, and type II collagen expression levels. Enzyme-linked immunosorbent assay was used to analyze the concentrations of Matrix Metalloproteinase-13 and Syndecan-4 in the joint fluid; and RT-PCR and Western Blot were used to observe the mRNA and protein expressions of ALK5, Sox-9, P-Smad3, and TGF-β1 at the injury repair site. ResultsThe repair effect of cartilage injury, as seen through gross observation and quantitative scoring, was better in all the Duhuo Jisheng Decoction treatment groups than in the model group. The medium dose group of Duhuo Jisheng Decoction had the best repair effect. We observed remarkable structural restoration of cartilage injury in the medium-dose Duhuo Jisheng Decoction group, with the subchondral bone presenting a distinct hierarchy, and parameters such as bone volume fraction and trabecular separation/spacing being significantly augmented. We found high expression levels of proteoglycans, P-TβRII, P-Smad2, and type II collagen. The concentrations of Matrix Metalloproteinase-13 and Syndecan-4 in the joint fluid were significantly lower following treatment. The low gene expression levels of ALK5, Sox-9, P-Smad3, and TGF-β1 in the injury site of the model group could be reversed in the medium-dose Duhuo Jisheng Decoction group. ConclusionDuhuo Jisheng Decoction can repair rabbit cartilage injury and reverse the levels of inflammatory factors in the joint fluid. The mechanism underlying its therapeutic effect is related to the activation of the TGF-β/Smad signaling pathway. This study provides a reliable basis for using Duhuo Jisheng Decoction to treat cartilage injury following knee osteoarthritis.

Physiological and genomic comparisons revealed the selective breeding improved the oceanic-salinity adaptability of estuarine oysters (Crassostrea ariakensis)

Estuarine oysters (Crassostrea ariakensis), a species with significant economic and ecological value, widely distrubted along the estuaries of East Asia. Overfishing, reduced runoff and shortage of fresh water volume in northern China have made the natural resources declined. We initiated a selective breeding program to improve the oceanic adaptability of estuarine oysters and now evaluated the selective breeding effect and the underlying mechanism. Comparisons of physiological and genetic characteristics were carried out under optimal salinity (20 ‰), ocean salinity (30 ‰), and hypersaline conditions (55 ‰) between fourth-generation oysters from the selected population and wild population. With respect to physiological characteristics, the selected population had a stronger energy supply and antioxidant capacities. As salinity increased, the selected population maintained a stable respiration rate, whereas the respiration rate of the wild population significantly decreased. The activities of pyruvate kinase (PK), Na+/K+-ATPase, and superoxide dismutase (SOD) exhibited higher levels in the selected population. In contrast, the phosphoenolpyruvate carboxykinase (PEPCK) activity, ratio of PEPCK to PK activities, and malonaldehyde (MDA) content were lower in the selected than in the wild population. Based on specific-locus amplified fragment sequencing, 4,821,296 SNPs from 347,280 polymorphic SLAF tags were identified and 269,174 highly consistent population SNPs were retained after filtering. Selected and wild populations exhibited significant genetic divergence. The selected population exhibited relatively large linkage disequilibrium blocks compared with the wild population. We identified twelve structurally differential genes with higher expression levels and amplitude of change under stressed conditions. These genes included nine hypersaline-stress responsive genes, three and six of which were upregulated (Gadd45a-1, Gadd45g, and Gadd45a-2) and downregulated (UN-Nat, Slc23a2, FucTA, UN-Pi3, UN-Pi5, and UN-Cpe), respectively. Our findings suggest that artificial selection significantly affects physiological and genomic responses to salinity change and these responses are important for improved resistance to salinity changes. Our data provide practical implications for oyster breeding programs.

Leveraging Bioinformatics and Machine Learning for Identifying Prognostic Biomarkers and Predicting Clinical Outcomes in Lung Adenocarcinoma.

Background/Objectives: There exist significant challenges for lung adenocarcinoma (LUAD) due to its poor prognosis and limited treatment options, particularly in the advanced stages. It is crucial to identify genetic biomarkers for improving outcome predictions and guiding personalized therapies. Methods: In this study, we utilize a multi-step approach that combines principled sure independence screening, penalized regression methods and information gain to identify the key genetic features of the ultra-high dimensional RNA-sequencing data from LUAD patients. We then evaluate three methods of survival analysis: the Cox model, survival tree, and random survival forests (RSFs), to compare their predictive performance. Additionally, a protein-protein interaction network is used to explore the biological significance of identified genes. Results:DKK1 and TNS4 are consistently selected as significant predictors across all feature selection methods. The Kaplan-Meier method shows that high expression levels of these genes are strongly correlated with poorer survival outcomes, suggesting their potential as prognostic biomarkers. RSF outperforms Cox and survival tree methods, showing higher AUC and C-index values. The protein-protein interaction network highlights key nodes such as VEGFC and LAMA3, which play central roles in LUAD progression. Conclusions: Our findings provide valuable insights into the genetic mechanisms of LUAD. These results contribute to the development of more accurate prognostic tools and personalized treatment strategies for LUAD.

Endocannabinoid receptor 2 is a potential biomarker and therapeutic target for the lysosomal storage disorders.

Herein, we studied the expression of endocannabinoid receptor 2 (CB2R), a known inflammation mediator, in several lysosomal storage disorder (LSD) animal models and evaluated it as a potential biomarker and therapeutic target for these diseases. CB2R was highly elevated in the plasma of Farber disease and mucopolysaccharidosis (MPS) type IIIA mice, followed by Fabry disease and MPS type I mice. Mice with acid sphingomyelinase-deficient Niemann-Pick disease (ASMD) and rats with MPS type VI exhibited little or no plasma CB2R elevation. High-level expression of CB2R was also observed in tissues of Farber and MPS IIIA mice. Treatment of MPS IIIIA patient cells with CB2R agonists led to a reduction of CB2R and monocyte chemoattractant protein-1 (MCP-1), a chemotactic factor that is elevated in this LSD. Treatment of MPS IIIA mice with one of these agonists (JWH133) led to a reduction of plasma and tissue CB2R and MCP-1, a reduction of glial fibrillary acidic protein (GFAP) in the brain, and an improvement in hanging test performance. JWH133 treatment of Farber disease mice also led to a reduction of MCP-1 in tissues and plasma, and treatment of these mice by enzyme replacement therapy (ERT) led to a reduction of plasma CB2R, indicating its potential to monitor treatment response. Overall, these findings suggest that CB2R should be further examined as a potential therapeutic target for the LSDs and may also be a useful biomarker to monitor the impact of therapies.

Prevalence, prognostic and clinicopathological value of HIF-1α in glioblastoma patients: a systematic review and meta-analysis.

Several studies have investigated the role of HIF-1α in predicting the prognosis of patients with glioblastoma, yielding contradictory results. Therefore, we performed a meta-analysis to document the correlation between HIF-1α and glioblastoma in individuals diagnosed with glioblastoma. We searched the PubMed, Cochrane Library, EMBASE, and Web of Science by January 25, 2024. Hazard Ratio (HR) was used to evaluate the relationship between HIF-1α and survival outcome, and Odds Ratio (OR) was adopted for tumor features.There was incorporation of nine observational studies with 607 individuals. The total prevalence of HIF-1α (higher than cut-off values) among individuals with glioblastoma was 0.72 (95% confidence interval (CI) = 0.68-0.75, I2 = 95.1%). There is a strong association between increased levels of HIF-1α in tumour tissues and shorter Overall Survival (OS) (HR = 1.82, 95% CI = 1.41-2.34, I2 = 13.7%). Subgroup analysis also indicated a correlation between higher levels of HIF-1α and reduced OS, specifically in the Asian population (HR = 1.48, 95% CI = 1.13-1.83, I2 = 41.5%). In addition, there was a correlation between HIF-1α and age (older vs. younger, OR = 2.19, 95% CI = 1.25-3.86, P = 0.260). High levels of HIF-1α expression were associated with poorer survival outcomes and other clinicopathological characteristics of glioblastoma. Integrating HIF-1α into prognostic tools for glioblastoma aids in predicting survival, categorising risk, and advising patients on suitable treatment regimens.

Clinicopathological Analysis of 14 Cases of Primary Pulmonary Lymphoepithelial Carcinoma

Primary pulmonary lymphoepithelial carcinoma (PPLEC) is a rare form of lung malignancy, accounting for only 0.7% of all lung cancers. It is currently classified as a distinct subtype within squamous cell carcinomas. This study aims to explore the clinicopathological characteristics of PPLEC and its subtypes, with the objective of enhancing understanding and improving diagnostic accuracy for this disease. A retrospective analysis was conducted on the clinical, pathological, imaging, and prognostic data of 14 patients diagnosed with PPLEC at the First Affiliated Hospital of Soochow University between February 2019 and June 2023. A total of 14 cases of PPLEC were identified, including 5 cases of the Regaud type, with ages ranging from 33 to 73 years, comprising 2 males and 3 females; and 9 cases of the Schmincke type, with ages ranging from 36 to 79 years, including 4 males and 5 females. Computed tomography (CT) scans consistently demonstrated soft tissue masses or nodular shadows. Reagud type mainly showed peripheral masses and Schmincke type mainly showed central masses. Pathological examination revealed tumor cells exhibiting syncytial-like growth, accompanied by lymphocytic infiltration and stromal fibrosis, with the Regaud type showing well-defined borders combined with granulomatous inflammation, while the Schmincke type exhibited indistinct tumor margins. Immunohistochemistry showed that CK, CK5/6, P40 and P63 were positive, and the Ki-67 index of Regaud type was lower than that of Schmincke type; notably, all 8 cases tested for programmed death-ligand 1 (PD-L1) were positive. Epstein-Barr virus-encoded RNA (EBER) in situ hybridization was positive in all instances. Among these cases, 6 underwent surgical treatment, and 8 received comprehensive therapy; by the end of the follow-up period, all 14 patients remained alive. PPLEC is a rare form of malignant lung tumor associated with Epstein-Barr virus (EBV) infection. The Regaud and Schmincke subtypes display distinct imaging and pathological characteristics. In the early stages of the disease, surgical intervention is the primary treatment method; however, for advanced stages, a multimodal treatment approach is utilized, resulting in a relatively favorable prognosis. Immunotherapy represents a promising and effective treatment modality for patients with middle to advanced stage disease exhibiting high PD-L1 expression levels.

Engineering ADSCs by manipulating YAP for lymphedema treatment in a mouse tail model.

Secondary lymphedema is a chronic disease associated with deformity of limbs and dysfunction; however, conventional therapies are not curative. Adipose-derived stem cells (ADSCs) based therapy is a promising way, but a single transplantation of ADSCs has limited efficacy. In this study, ADSCs were engineered in vitro and then transplanted into the site of lymphedema. Yes-associated protein (YAP), a crucial regulator of Hippo pathway, plays an important role in regulating stem cell functions. We examined the YAP expression in a mouse tail lymphedema model, and found that transplanted ADSCs exhibited high expression level of YAP and a large number of YAP positive cells existed in lymphedema environment. In vitro, the downregulation of YAP in ADSCs resulted in higher expression levels of genes related to lymphangiogenesis such as Lyve-1, VEGFR-3 and Prox-1. In vivo, YAP-engineered ADSCs generated abundant VEGFR-3-positive lymphatic vessels and significantly improved subcutaneous fibrosis. These results indicated that the transplantation of pre-engineered ADSCs by manipulating YAP is a promising strategy for lymphatic reconstruction.

Exploring SLC30A6 as a potential prognostic, immunomodulatory, and therapeutic biomarker in various cancers with a focus on pancreatic cancer: a pan-cancer analysis

BackgroundOur study investigates the multifaceted role of the gene SLC30A6 across 33 distinct cancer types. Understanding the function and impact of SLC30A6 in cancer biology is crucial, as previous studies have hinted at its aberrant expression and potential involvement in tumor progression. This research aims to elucidate the expression patterns, methylation variations, mutational signatures, and survival implications of SLC30A6, along with its engagement with the immune system in various cancers and specially in pancreatic cancer.MethodsWe conducted a comprehensive analysis of SLC30A6 using data from 33 different cancer types, and all data were analyzed in silico. The study involved examining expression levels, performing correlation analyses with clinical outcomes, assessing methylation variations, and identifying mutational signatures. Functional enrichment analyses were carried out to understand the gene's involvement in biological pathways. Additionally, we evaluated the relationship between SLC30A6 expression and immune cell infiltration levels to uncover its role in the tumor microenvironment.ResultsSLC30A6 was found to be significantly up-regulated in the majority of the 33 cancer types analyzed. High expression levels of SLC30A6 were consistently correlated with poor survival outcomes, indicating its potential role in cancer prognosis. Functional enrichment analyses revealed that SLC30A6 is involved in key pathways related to tumor progression, including cell proliferation and apoptosis. Moreover, SLC30A6 showed significant associations with diverse immune pathways, suggesting its involvement in immune regulation. Notably, our analysis demonstrated a significant correlation between SLC30A6 expression and the infiltration levels of key immune cells, highlighting its dual immunosuppressive and immunostimulatory roles in a pan-cancer context. Moreover, SLC30A6 is significantly overexpression in pancreatic cancer and have shown different clinical associations as above.ConclusionOur study provides comprehensive insights into the complex interplay between SLC30A6 and cancer development. These findings position SLC30A6 as a promising prognostic biomarker and therapeutic target across diverse cancer types, highlighting its importance in future cancer research and treatment strategies.

Identification of key ferroptosis‑related biomarkers in Kawasaki disease by clinical and experimental validation.

Kawasaki disease (KD) is an acute febrile rash that is primarily characterized by systemic vasculitis and is the leading cause of childhood-acquired heart disease. At present, a KD diagnosis is solely dependent on clinical symptoms and effective diagnostic markers are unavailable. Ferroptosis, a novel form of programmed cell death, contributes to the pathophysiology of infectious diseases. The present study aimed to identify key ferroptosis-related genes (FRGs) involved in the pathological process of KD and thus potential diagnostic biomarkers for this disease. For this purpose, differentially expressed-FRGs (DE-FRGs) between patients with KD and healthy controls were screened. The least absolute shrinkage and selection operator (LASSO) algorithm and a logistic regression model combined with receiver operating characteristic analysis were then used to identify and assess ferroptosis-related markers. Additionally, immune cell infiltration landscapes in the KD and control groups were evaluated using CIBERSORT. Moreover, the predictive value of the identified markers was validated in the clinical samples as well as vascular endothelial cells. A total of 10 DE-FRGs were screened from the KD and control samples. These 10 DE-FRGs were then applied to the LASSO model and 6 key ferroptosis-related markers were obtained. The subsequent Gene Set Variation Analysis results suggested that high expression levels of these markers were closely associated with innate immune activation and metabolism, while low expression was mainly linked to adaptive immune-related pathways. In addition to validating each gene in the training and validation sets, the diagnostic potential of these markers was assessed utilizing KD samples obtained from Shenzhen Baoan Women's and Children's Hospital. As a result, MAPK14, SLC2A3 and PGD were selected as potential diagnostic markers for KD. Additionally, changes in the expression of marker genes during inflammatory activation of vascular endothelial cells were measured by reverse transcription-quantitative PCR. The results of the present study will help to understand the role of FRGs in the pathogenesis of KD. Moreover, the identified FRGs may serve as diagnostic biomarkers, providing new strategies for KD prediction and treatment.

Toxicity of Flonicamid to Diaphorina citri (Hemiptera: Liviidae) and Its Identification and Expression of Kir Channel Genes.

Flonicamid is a selective insecticide effective against piercing-sucking insects. Although its molecular target has been identified in other species, the specific effects and detailed mechanism of action in Diaphorina citri Kuwayama remain poorly understood. In this study, we determined that the LC50 of flonicamid for D. citri adults was 16.6 mg AI L-1 after 4 days of exposure. To explore the relevant mechanisms, the treatments with acetone and with 20 mg AI L-1 flonicamid for 96 h were collected as samples for RNA-Seq. The analysis of the transcriptomes revealed 345 differentially expressed genes (DEGs) in D. citri adults subjected to different treatments. Among these DEGs, we focused on the inward-rectifying potassium (Kir) channel genes, which have been extensively studied as potential targets of flonicamid. Three Kir subunit genes (Dckir1, Dckir2, Dckir3) in D. citri were successfully cloned and identified. Furthermore, the expression profiles of these DcKirs were investigated using RT-qPCR and showed that their expression significantly increased after D. citri eclosion to adulthood, particularly for DcKir3. The DcKirs were predominantly expressed in gut tissues, with DcKir1 and DcKir2 exhibiting high expression levels in the hindgut and midgut, respectively, while DcKir3 showing high expression in the midgut and Malpighian tubules. This study provides insights into the potential roles of Kir subunits in D. citri and enhances our understanding of the physiological effects of flonicamid in this pest.

Inhibition of NPC1 suppresses cell proliferation and β-catenin signaling activation of liver cancer.

Niemann-Pick Type C1 (NPC1) plays a significant role in the development of liver diseases and liver cancer. Our objective was to investigate the involvement of NPC1 in regulating liver cancer development. We observed that high levels of NPC1 expression in tumor tissues from patients with liver cancer were associated with a poor prognosis. Through in vitro experiments, we found that inhibiting NPC1 expression reduced the proliferation, invasion, and migration of liver cancer cells, while also inducing apoptosis in these cells. Additionally, the inhibition of NPC1 led to decreased activation of Wnt/β-catenin signaling. In vivo studies further supported our findings by demonstrating that the suppression of liver cancer cell growth was effectively achieved through the inhibition of NPC1. Overall, our results strongly indicate that the inhibition of NPC1 suppresses liver cancer cell proliferation. Targeting NPC1 is a promising potential therapeutic strategy for liver cancer.

Abstract A025: RNA-Based Therapeutics to target the p53 pathway in high-grade serous ovarian cancer: TAp63-Regulated Oncogenic LncRNAs (TROLLs) as novel therapeutic targets in cancer

Abstract High-grade serous ovarian cancer (HGSOC) is the most common and malignant type of ovarian cancer with a high frequency of p53 mutation and hyperactivation of the AKT pathway. Even though roughly 85% of patients achieve remission after initial surgical debulking and chemotherapy, four out of five HGSOC patients will experience disease recurrence, resulting in a 5-year survival rate of 43%. We have identified and characterized two TAp63 Regulated Oncogenic Long non-coding RNAs (TROLL-2 and TROLL-3) whose transcription is induced by mutant p53 and high expression level correlate with a more aggressive ovarian cancer grade and phosphorylation of AKT. In-depth analysis demonstrated that TROLL-2 and TROLL-3 directly activate AKT in tumor cells, thus linking p53 alterations to AKT activity. The use of AKT inhibitors in the clinic is limited because of frequent adverse effects, likely due to the constitutive expression of AKT in all cells. TROLL-2 and TROLL-3 expression is limited to tumor cells and provides a more specific target for decreasing AKT activation, potentially minimizing off-target effects. We have optimized reduction of TROLL-2 and TROLL-3 expression using antisense oligonucleotides (ASOs) in an ex vivo perfusion platform for 3D culture of microtumors from primary patient derived tumors. Samples show maximum reduction of proliferation (EdU) and phosphorylation of AKT when TROLL-2 and TROLL-3 ASOs are used concurrently. We also performed phospho-proteomics and single cell RNAseq to compare individual ASO-mediated knockdown of each lncRNA to combined targeted ASO treatments. We have developed primary patient derived xenograft models using ovarian tumors orthotopically implanted into NSG mice generating a relevant preclinical model to determine the efficacy of ASO treatment. To reduce the rapid degradation and clearance of ASOs in circulation, ASOs were encapsulated into lipid nanoparticles and metal-organic frameworks (MOFs) and we are currently testing these in vivo. Since AKT activation is associated with increased staging and resistance to platinum and targeted therapies in ovarian cancer patients, our data provides preclinical rationale for the implementation of ASOs and the use of new delivery methods as a relevant translational therapy and novel method to exploit the specificity of TROLL-2 and TROLL-3 expression in HGSOC. Citation Format: Ashley Lui, Marco Napoli, Jaden R Baldwin, Christina L Carr, Angie Rivera, Duy Nguyen, W. Gregory Sawyer, Michael R Dunne, Erin George, Omar K Farha, Michelle Teplensky, Elsa R Flores. RNA-Based Therapeutics to target the p53 pathway in high-grade serous ovarian cancer: TAp63-Regulated Oncogenic LncRNAs (TROLLs) as novel therapeutic targets in cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: RNAs as Drivers, Targets, and Therapeutics in Cancer; 2024 Nov 14-17; Bellevue, Washington. Philadelphia (PA): AACR; Mol Cancer Ther 2024;23(11_Suppl):Abstract nr A025.

Linalool and 1,8-Cineole as Constitutive Disease-Resistant Factors of Norway Spruce Against Necrotrophic Pathogen Heterobasidion Parviporum.

Norway spruce is an important coniferous species in boreal forests. Root and stem rot diseases caused by the necrotrophic pathogen Heterobasidion parviporum threaten the wood production of Norway spruce which necessitates the search for durable control and management strategies. Breeding for resistant traits is considered a viable long-term strategy. However, identification of potential resistant traits and markers remains a major challenge. In this study, short-term disease resistance screening was conducted using 218 Norway spruce clones from 17 families. Disease resistance was evaluated based on the size of necrosis lesion length following infection with the pathogen. A subset of needles/branches from clones with small (partial resistant) or large (susceptible) lesions were used for terpene analysis and transcriptomic profiling. The results revealed that the content of monoterpene linalool and 1,8-cineole and their respective encoded genes were significantly more abundant and highly expressed in the partial resistant group. Furthermore, linalool and 1,8-cineole were demonstrated to have inhibitory effect on the growth of the pathogen H. parviporum, with morphological distortion of the hyphae. RNAseq analysis revealed that transcript of pathogen genes involved in the regulation of carbohydrate metabolism and stress responses were significantly decreased in presence of the terpenes. The results suggest the relevance of monoterpenes together with jasmonic acid precursor and some genes involved in phenylpropanoid biosynthesis, as constitutive tolerance factors for Norway spruce tolerance against necrotrophic pathogen. The high level of necrosis related cell death gene expression might be factors critical for host susceptibility and disease development.

Semi-supervised spatial-temporal calibration and semantic refinement network for video polyp segmentation

Automated video polyp segmentation (VPS) was of vitality for the early prevention and diagnosis of colorectal cancer (CRC). However, existing deep learning-based automatic polyp segmentation methods mainly focused on independent static images and struggled to perform well due to neglecting spatial–temporal relationships among successive video frames, while requiring massive frame-by-frame annotations. To better alleviate these challenges, we proposed a novel semi-supervised spatial–temporal calibration and semantic refinement network (STCSR-Net) dedicated to VPS, which simultaneously considered both the inter-frame temporal consistency in video clips and intra-frame semantic-spatial information. It was composed of a segmentation pathway and a propagation pathway by use of a co-training scheme for supervising the predictions on un-annotated images in a semi-supervised learning fashion. Specifically, we proposed an adaptive sequence calibration (ASC) block in segmentation pathway and a dynamic transmission calibration (DTC) block in propagation pathway to fully take advantage of valuable temporal cues and maintain the prediction temporally consistent among consecutive frames. Meanwhile, in these two branches, we introduced residual block (RB) to suppress irrelevant noisy information and highlight rich local boundary details of polyp lesions, while constructed multi-scale context extraction (MCE) module to enhance multi-scale high-level semantic feature expression. On that basis, we designed progressive adaptive context fusion (PACF) module to gradually aggregate multi-level features under the guidance of reinforced high-level semantic information for eliminating semantic gaps among them and promoting the discrimination capacity of features for targeting polyp objects. Through synergistic combination of RB, MCE and PACF modules, semantic-spatial correlations on polyp lesions within each frame could be established. Coupled with the context-free loss, our model merged feature representations of neighboring frames to diminish the dependency on varying contexts within consecutive frames and strengthen its robustness. Extensive experiments substantiated that our model with 100% annotation ratio achieved state-of-the-art performance on challenging datasets. Even trained under 50% annotation ratio, our model exceled significantly existing state-of-the-art image-based and video-based polyp segmentation models on the newly-built local TRPolyp dataset by at least 1.3% and 0.9% enhancements in both mDice and mIoU, whilst exhibited comparable performance to top rivals attained through using fully supervised approach on publicly available CVC-612, CVC-300 and ASU-Mayo-Clinic benchmarks. Notably, our model showcased exceptionally well in videos containing complex scenarios like motion blur and occlusion. Beyond that, it also harvested approximately 0.794 mDice and 0.707 mIoU at an inference of 0.036 s per frame in endoscopist-machine competition, which outperformed junior and senior endoscopists as well as almost matched with those of expert ones. The strong capability of the proposed STCSR-Net held promise in improving quality of VPS, accentuating model’s adaptability and potential in real-world clinical scenarios.

Granzyme mRNA-miRNA interaction and its implication to functional impact.

Granzyme activity can affect the processing and stability of miRNAs within target cells. They also could induce changes in miRNA expression that impact apoptotic signaling. Granzyme-induced apoptosis might result in changes to the miRNA profile, which can further influence the apoptosis and inflammation processes. The aim of this study was to bioinformatically analyze which miRNAs and transcription factors bind to the CDS and UTR regions of the granzyme family to regulate gene expression in relation to granzyme evolution and their association with human cancer diseases. The expression patterns of granzyme genes were analyzed in various human tissues. MiRNAs binding to the CDS and UTR of the granzyme family were examined, and the transcription factors binding to these miRNAs binding sites were also analyzed. Cytoscape program was used to visualize and analyze the networks of interactions between granzyme mRNA and miRNAs. Additionally, the evolutionary patterns of the granzyme family in relation to miRNAs and transcription factors binding were investigated. Analysis of the expression patterns of the granzyme family in various human tissues shows that GZMA and GZMK are strongly expressed in lymph nodes. GZMB exhibits strong expression in the bone marrow, while GZMA is prominently expressed in the spleen. Twenty-two miRNAs bind to both GZMK and GZMB mRNA, while six miRNAs bind to both GZMK and GZMM mRNA. The only miRNA that binds to GZMK, GZMB, GZMM, and GZMA mRNA is hsa-miR-146a-5p. Transcription factors JUND, FOS, and JUN are distinctly interconnected with has-miR-5696 and GZMK. Association data between the granzyme family and cancers showed that various miRNAs were consistently implicated and exhibited either upregulation or downregulation. Although the granzyme family possesses distinct genetic information, it shows relatively high expression levels in the lymph node, spleen, and bone marrow. Many miRNAs specifically regulate granzyme gene expression, and various transcription factors are involved. Analyzing the granzyme genes-miRNAs-transcription factors-related network will provide crucial insights into the mechanisms of cancer development and suppression.

BIOM-09. CLINICAL EFFECT OF CD25 ON THE MTX-BASED CHEMOSENSITIVITY OF PRIMARY CENTRAL NERVUS SYSTEM LYMPHOMA

Abstract INTRODUCTION Primary central nervous system diffuse large B-cell lymphoma (PCNSL) is treated with chemotherapy, mainly with high-dose methotrexate (HD-MTX), but some cases are resistant to treatment and relapse in a short period. However, a biomarker that predicts the chemosensitivity and recurrence has not yet been identified. In this study, we used surface marker analysis of tumor tissue using flow cytometry to search for biomarkers related to chemosensitivity and early recurrence. METHODS From 2017 to 2023, 25 patients were diagnosed with PCNSL by biopsy in Kobe University Hospital, and flow cytometry of the biopsy tissues were performed. Cerebrospinal fluid (CSF) examination including sIL2R, β2MG, and IL-10 was performed in all cases before surgery, and treatment based on methotrexate was administered postoperatively. We retrospectively analyzed patient characteristics, flow cytometry data, treatment details, response rate, time to recurrence, and overall survival. RESULTS The mean age of the patients was 71 (range, 37-80) years. In univariate analysis, high expression level of CD25 (interleukin-2 receptor, IL-2R) in the tissue was associated with a lower response rate to MTX-based chemotherapy (Fisher’s exact test, p=0.0072) and had a tendency of shorter time to recurrence. However, there were no significant differences in progression-free survival and overall survival. There was no correlation between expression level of CD25 in tissue and sIL-2R in CSF. Expression level of sIL-2R in CSF was independent of CD25 expression in tissue. CONCLUSION There are some reports that CD25 can be a prognostic factor in follicular lymphoma, but no reports in PCNSL. Here, we suggested that high expression level of CD25 in PCNSL tissue was associated to chemoresistance and early recurrence. In cases of PCNSL with high CD25 expression level, additional whole-brain irradiation and transition to tirabrutinib be considered because there are possibilities of chemoresistance and early recurrence.