TargetScan Research Articles

Comprehensive cataloging of miR-363 as a therapeutic & non-invasive biomarker of prostate cancer

Background & objectives Overcoming the challenge of early diagnosis of prostate cancer (PCa) by exploring molecular biomarkers is urgently needed. With this objective, this study was designed to explore the biomarker and therapeutic potential of miRNA (miR)-363-3p in PCa pathogenesis. Methods Total participants (n=188) were enrolled, and blood and tissue samples were collected from individuals categorized into the control group (n=55), benign prostate hyperplasia (BPH) group (n=60), PCa group (n=48), and castration-resistant PCa (CRPC) group (n=25). MiR expression profiling was carried out using quantitative polymerase chain reaction (qPCR), and biomarker analysis was conducted employing receiver operating characteristics (ROC) curve. The miR-363 target genes were predicted by in silico tools like Target Scan and starBasev 2.0 and its expression was validated by qPCR and association among them was established by using the STRING database. Results The results showed that the tumour-suppressive nature of miR-363-3p in both PCa tissues and serum were significantly higher than the control with a greater area under curve (AUC) was 0.969 (sensitivity: 85%; specificity 100%) and 0.988 (sensitivity: 97.5%; specificity: 87.5%), respectively. The targetome analysis of miR-363-3p revealed five target genes-NRAS, E2F3, PTEN, MDM2, and CCNE2 which were strongly associated with cell division and proliferation. The expression analysis of the target genes showed a significant tumour-suppression of PTEN gene and significant upregulation of oncogenic genes such as NRAS, E2F3, MDM2, and CCNE2. Interpretation & conclusions Collectively, the findings of this study suggest that miR-363-3p may be a potential biomarker in differentiating individuals with PCa and CRPC from healthy controls. The miR-363-3p triggers various oncogenic genes (MDM2, NRAS, E2F3, CCNE2) and tumour suppressor genes (PTEN) that are actively involved in PCa progression and development.

High expression of CNOT6L contributes to the negative development of type 2 diabetes.

Type 2 diabetes (T2D) is a chronic metabolic disorder characterized by reduced responsiveness of body cells to insulin, leading to elevated blood sugar levels. CNOT6L is involved in glucose metabolism, insulin secretion regulation, pancreatic beta-cell proliferation, and apoptosis. These functions may be closely related to the pathogenesis of T2D. However, the exact molecular mechanisms linking CNOT6L to T2D remain unclear. Therefore, this study aims to elucidate the role of CNOT6L in T2D. The T2D datasets GSE163980 and GSE26168 profiles were downloaded from the Gene Expression Omnibusdatabase generated by GPL20115 and GPL6883.The R package limma was used to screen differentially expressed genes (DEGs). A weighted gene co-expression network analysis was performed. Construction and analysis of the protein-protein interaction (PPI) network, functional enrichment analysis, gene set enrichment analysis, and comparative toxicogenomics database (CTD) analysis were performed. Target Scan was used to screen miRNAs that regulate central DEGs. The results were verified by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), western blotting (WB), and blood glucose measurements in mice. A total of 1951 DEGs were identified. GO and KEGG enrichment analysis revealed that differentially expressed genes were mainly enriched in the insulin signaling pathway, ECM-receptor interaction, and PPAR signaling pathway. Metascape analysis indicated enrichment primarily in the cAMP signaling pathway and enzyme-linked receptor protein signaling pathway. WGCNA analysis yielded 50 intersecting genes. PPI network construction and algorithm identification identified two core genes (CNOT6L and GRIN2B), among which CNOT6L gene was associated with multiple miRNAs. CTD analysis revealed associations of core genes with type 2 diabetes, diabetic complications, dyslipidemia, hyperglycemia, and inflammation. WB and RT-qPCR results showed that in different pathways, CNOT6L protein and mRNA levels were upregulated in type 2 diabetes. CNOT6L is highly expressed in type 2 diabetes mellitus, and can cause diabetes complications, inflammation and other physiological processes by regulating miRNA, PPAR and other related signaling pathways, with poor prognosis. CNOT6L can be used as a potential therapeutic target for type 2 diabetes.

Spectral Background Calibration of Scanning Habitable Environments with Raman and Luminescence for Organics and Chemicals (SHERLOC) Spectrometer Onboard the Perseverance Rover Enables Identification of a Ubiquitous Martian Spectral Component.

The Perseverance rover landed at Jezero crater, Mars, on 18 February 2021, with a payload of scientific instruments to examine Mars' past habitability, look for signs of past life, and process samples for future return to Earth. The instrument payload includes the Scanning Habitable Environments with Raman and Luminescence for Organics and Chemicals (SHERLOC) deep ultraviolet Raman and fluorescence imaging spectrometer designed to detect, characterize, and map the presence of organics and minerals on the Martian surface. Operation and engineering constraints sometimes result in the acquisition of spectra with features near the detection limit. It is therefore important to separate instrumental (background) spectral components and spectral components inherent to Martian surface materials. For SHERLOC, the instrumental background is assessed by collecting spectra in the stowed-arm configuration where the instrument is pointed at the Martian nighttime sky with no surface sample present in its optical path. These measurements reveal weak Raman and fluorescence background spectral signatures as well as charged-coupled device pixels prone to erroneous intensity spikes separate from cosmic rays. We quantitatively describe these features and provide a subtraction procedure to remove the spectral background from surface spectra. By identifying and accounting for the SHERLOC Raman background features within the median Raman spectra of Martian target scans, we find that the undefined silicate spectral feature interpreted to be either amorphous silicate or plagioclase feldspar is ubiquitously found in every Mars target Raman scan collected through Sol 751.

Quantification of persulfidation on specific proteins: are we nearly there yet?

Hydrogen sulfide (H2S) played a pivotal role in the early evolution of life on Earth before the predominance of atmospheric oxygen. The legacy of a persistent role for H2S in life's processes recently emerged through its discovery in modern biochemistry as an endogenous cellular signalling modulator involved in numerous biological processes. One major mechanism through which H2S signals is protein cysteine persulfidation, an oxidative post-translational modification. In recent years, chemoproteomic technologies have been developed to allow the global scanning of protein persulfidation targets in mammalian cells and tissues, providing a powerful tool to elucidate the broader impact of altered H2S in organismal physiological health and human disease states. While hundreds of proteins were confirmed to be persulfidated by global persulfidome methodologies, the targeting of specific proteins of interest and the investigation of further mechanistic studies are still underdeveloped due to a lack of stringent specificity of the methods and the inherent instability of persulfides. This review provides an overview of the processes of endogenous H2S production, oxidation, and signalling and highlights the application and limitations of current persulfidation labelling approaches for investigation of this important evolutionarily conserved biological switch for protein function.

The value analysis of high-resolution thin-layer CT in the identification of early lung adenocarcinoma: An observation study.

The aim of this study was to explore the clinical value of high-resolution thin-layer computed tomography (CT) for the identification of early lung adenocarcinoma. Ninety patients with early lung adenocarcinoma who were diagnosed and treated in our hospital were selected as study subjects and divided into noninvasive (NIG, n = 51) and invasive (IG, n = 39) groups according to their pathological findings. Both groups underwent high-resolution target scanning. Differences in lesion size, density, and distribution between the 2 groups were compared. Intergroup differences in the CT signs were examined. A receiver-operating characteristic curve was established to calculate the diagnostic efficacy of high-resolution, thin-layer CT for early lung adenocarcinoma infiltration. The maximum diameter and density of the tumors were significantly higher in the IG than in the NIG (P < .05). The proportions of CT signs of lobulation, spicule, and vessel convergence were higher in the IG patients compared to the NIG (P < .05). High-resolution thin-layer CT for the diagnosis of lung adenocarcinoma infiltration had an AUC of 0.6702 (P < .05), a diagnostic sensitivity of 64.10%, and a diagnostic specificity of 60.78%. High-resolution thin-layer CT had certain differential diagnostic efficacy for early lung adenocarcinoma, which clearly presents various CT signs of early lung adenocarcinoma lesions.

Effects of Continuous Prenatal Low Dose Rate Irradiation on Neurobehavior, Hippocampal Cellularity, Messenger RNA and MicroRNA Expression on B6C3F1 Mice.

Epidemiological, experimental, and ecological data have indicated the controversial effect of in utero chronic low dose rate (<6 mGy/h) with accumulative low (≤100 mGy) or high (>100 mGy) dose radiation exposure. Our main goal of this study was to examine if different low dose rates of chronic pre- and/or post-natal radiation exposure with accumulative high doses could induce hippocampal cellular, mRNA, and miRNA changes leading to neuropsychiatric disorders. The comprehensive mouse phenotypic traits, organ weight, pathological, and blood mRNA and miRNA changes were also studied. Using different approaches including SmithKline, Harwell, Imperial College, Royal Hospital, Phenotype Assessment (SHIRPA), neurobehavioral tests, pathological examination, immunohistochemistry, mRNA and miRNA sequencing, and real-time quantitative polymerase chain reaction (qRT-PCR) validation, we found that in prenatally irradiated (100 mGy/d for 18 days with an accumulative dose of 1.8 Gy) 1-year-old mice, no cellular changes, including immature neurons in the subgranular zone, mature neurons and glial cells in the hilus of the dentate gyrus and development of cognitive impairment, neuropsychiatric disorders, occurred. However, a significant reduction in body weight and mass index (BMI) was indicated by the SHIRPA test. A reduced exploratory behavior was shown by an open field test. Organ weights showed significant reductions in the testes, kidneys, heart, liver and epididymides with no abnormal pathology. mRNA and miRNA sequencing and qRT-PCR validation revealed the upregulation of Rubcnl and Abhd14b, and downregulation of Hspa1b, P4ha1, and Banp genes in both the hippocampus and blood of mice prenatally irradiated with 100 mGy/d. Meanwhile, downregulation of miR-448-3p and miR1298-5p in the hippocampus, miR-320-3p, miR-423-5p, miR-486b-5p, miR-486b-3p, miR-423-3p, miR-652-3p, miR-324-3p, miR-181b-5p, miR-let-7b, and miR-6904-5p in the blood was induced. The target scan revealed that Rubcnl is one of the miR-181b-5p targets in the blood. We, therefore, concluded that prenatal chronic irradiation with a low dose rate of 100 mGy/d and accumulative dose of 1.8 Gy or below might not induce significant adverse health effects on the offspring. Further study of different low dose rate radiation exposures with accumulative high doses may provide threshold doses for authorities or regulators to set new radiation safety guidelines to replace those extrapolated from acute high dose/dose rate irradiation to reduce unnecessary emergency evacuation or spending once a nuclear accident or leakage occurs.

Quantitative parameters of HRCT target scan to predict the risk of lung adenocarcinoma based on the detection of lung ground-glass nodules.

To explore the value of high-resolution computed tomography (HRCT) in the differential diagnosis of benign and malignant ground-glass nodules (GGNs), and to provide a theoretical basis for the clinical application of HRCT. A total of 208 patients with GGN who had been clinically confirmed by surgical pathology and clinical confirmation were collected, and HRCT target scanning technology was used to scan and collect general information of patients, and observe the distribution of GGN, GGN size, GGN cross-sectional area, diameter, transverse diameter, solid composition, relationship with bronchi, and relationship with blood vessels and other indicators. Multivariate regression analysis and risk factor prediction are performed. The differences were statistically significant in multivariate regression analysis, such as nodule location, maximum diameter, maximum cross-sectional area, GGN status, nodule boundary and relationship with blood vessels (P < 0.05). The results of ROC curve showed that the AUC value of nodule site and nodule boundary was greater than 0.5, and the nodule boundary AUC value was 0.676, which was more sensitive to predict whether GGN deteriorated to lung adenocarcinoma (LUAD). Nodule site and nodule boundary are effective risk predictors for LUAD in patients with GGN, and nodule boundary is the most valuable independent predictor.

Construction of cirRNA-miRNA-mRNA network and MAPK1 protein signaling pathway in patients with valvular disease affected by artificial heart valve replacement surgery

The progress of modern medical technology has made artificial heart valve replacement an effective means to treat valvular disease, but the impact of cardiac function on patients after surgery is still a key issue. The purpose of this study was to construct the cirRNA-miRNA-mRNA network after artificial heart valve replacement in valvular disease patients, and to explore the regulatory mechanism related to MAPK1 protein, so as to reveal its potential role in affecting cardiac function. We downloaded cyclic cRNA expression profiles from the GEO database. Use the limma package to identify dec. WGCNA is used to identify key modules of circular rna. The target miRNAs of circular rna and the corresponding target genes of miRNAs were screened by ring intertome and target scan database. GO and KEGG analysis using the DAVID database. The genes associated with iron sag disease were derived from FerrDb database. The overlapping genes were obtained by Wien analysis. Next, the CircrNa-mirNa-mrna network was constructed based on the circRNA-miRNA pair and miRNA-mRNA pair and their cyclic landscape software. This study revealed the changes in the structure and expression of MAPK1 protein in the cirRNA-miRNA-mRNA network after artificial heart valve replacement in valvular disease patients, suggesting the potential role of MAPK1 protein in regulating cardiac function, and laying a foundation for further revealing its mechanism and clinical application.

Autonomous Trajectory Planning for Ultrasound-Guided Real-Time Tracking of Suspicious Breast Tumor Targets

Ultrasound image guidance could display the movement of soft tissue in real time, which provides an important basis for the operation path selection of tumor for examinations and interventions such as precise localization and puncture biopsy. However, factors such as tissue deformation, patient motion, and instrument contact pose great challenges for real-time smooth localization tracking of target tissues and maintaining a suitable acoustic window. Not only does it reduce the accuracy of ultrasound examination, but also may prolong the examination time. In this paper, a real-time autonomous ultrasound robot trajectory planning framework is proposed to achieve global scanning of breast tissue and real-time local tracking of suspicious tumor targets. In addition, an ultrasound suspicious tumor target next moment motion attitude and position estimation algorithm and an acceleration-continuous online trajectory generation (ACOTG) algorithm are proposed. Experiments on breast phantom showed that the fluctuation difference of both global scanning and tracking scanning force was less than 5 N <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$\pm$</tex-math> </inline-formula> 15%, and ACOTG takes less than 1 microsecond to calculate the next moment state. The position, velocity, and acceleration of the tracking path did not change abruptly when the unknown target changed. The ultrasound image of the suspected tumor target could always be smoothly maintained in a suitable acoustic window, while the errors between the center of the suspected tumor target image and the center of the acoustic window in the horizontal direction and the depth direction were less than <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$\pm$</tex-math> </inline-formula> 1 mm and 30 mm <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$\pm$</tex-math> </inline-formula> 10%, respectively. <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">Note to Practitioners</i> —The motivation of this study was to solve the clinical problem of difficulty in keeping the suspected breast tumor target stably in the center of the field of view under the influence of tissue deformation, patient motion and instrument contact in ultrasound guidance. We proposed the idea of global conventional scanning and local online tracking to achieve the autonomous scanning, detecting and tracking of suspicious tumor targets within the global scope of the breast. ACOTG algorithm is proposed and implemented, which can make the ultrasound image of suspicious tumor target can be displayed stably in the center of the acoustic window. With the help of this system, the accuracy of ultrasound examination can be improved and the time of examination can be reduced. The proposed system can also be applied to ultrasound scanning of other parts of the body.

Mechanism of BRD4 Inhibitor-Mediated c-MYC Expression and Regulation of AR Expression to Inhibit Prostate Cancer

Prostate cancer is a major health challenge, probably because of regulatory role of hormones that has not been clearly studied. Our study explored BRD4’s role in the development of prostate cancer. BRD4 expression was detected in tumor tissues by RT-PCR method. Transwell detected cell function after different BRD4 expressions, while Target scan detected the relationship between c-MYC and BRD4, including protein expression between the two luciferase activity and Western-blot. Western-blot detected the protein expression after addition of c-MYC, and further verified the effect of c-MYC expression on AR. We proved that, BRD4 was highly expressed in tumor tissues, and inhibiting BRD4 expression significantly inhibited tumor cell invasion and proliferation. BRD4 targets negative feedback to regulate the expression of c-MYC. Further results showed that, addition of BRD4 activated c-MYC signal transduction and inhibited prostate cancer development. Our study reveals that, BRD4 regulates AR to inhibit prostate cancer by regulating c-MYC. BRD4 is involved in AR-mediated regulation of PCa cells. In addition, inhibiting the expression of BRD4 can inhibit pathological progression of prostate cancer cells.

MiR-181d-5p ameliorates hypercholesterolemia by targeting PCSK9.

Hypercholesterolemia is an independent risk factor for cardiovascular disease and lowering circulating levels of low-density lipoprotein cholesterol (LDL-C) can prevent and reduce cardiovascular events. MicroRNA-181d (miR-181d) can reduce the levels of triglycerides and cholesterol esters in cells. However, it is not known whether miR-181d-5p can lower levels of circulating LDL-C. Here, we generated two animal models of hypercholesterolemia to analyze the potential relationship between miR-181d-5p and LDL-C. In hypercholesterolemia model mice, adeno-associated virus (AAV)-mediated liver-directed overexpression of miR-181d-5p decreased the serum levels of cholesterol and LDL-C and the levels of cholesterol and triglyceride in the liver compared with control mice. Target Scan 8.0 indicated Proprotein convertase subtilisin/kexin type 9 (PCSK9) to be a possible target gene of miR-181d-5p, which was confirmed by in vitro experiments. miR-181d-5p could directly interact with both the PCSK9 3'-UTR and promoter to inhibit PCSK9 translation and transcription. Furthermore, Dil-LDL uptake assays in PCSK9 knockdown Huh7 cells demonstrated that miR-181d-5p promotion of LDL-C absorption was dependent on PCSK9. Collectively, our findings show that miR-181d-5p targets the PCSK9 3'-UTR to inhibit PCSK9 expression and to reduce serum LDL-C. miR-181d-5p is therefore a new therapeutic target for the development of anti-hypercholesterolemia drugs.

Experimental validation of a PNS-optimized whole-body gradient coil.

Peripheral nerve stimulation (PNS) limits the usability of state-of-the-art whole-body and head-only MRI gradient coils. We used detailed electromagnetic and neurodynamic modeling to set an explicit PNS constraint during the design of a whole-body gradient coil and constructed it to compare the predicted and experimentally measured PNS thresholds to those of a matched design without PNS constraints. We designed, constructed, and tested two actively shielded whole-body Y-axis gradient coil winding patterns: YG1 is a conventional symmetric design without PNS-optimization, whereas YG2's design used an additional constraint on the allowable PNS threshold in the head-imaging landmark, yielding an asymmetric winding pattern. We measured PNS thresholds in 18 healthy subjects at five landmark positions (head, cardiac, abdominal, pelvic, and knee). The PNS-optimized design YG2 achieved 46% higher average experimental thresholds for a head-imaging landmark than YG1 while incurring a 15% inductance penalty. For cardiac, pelvic, and knee imaging landmarks, the PNS thresholds increased between +22% and +35%. For abdominal imaging, PNS thresholds did not change significantly between YG1 and YG2 (-3.6%). The agreement between predicted and experimental PNS thresholds was within 11.4% normalized root mean square error for both coils and all landmarks. The PNS model also produced plausible predictions of the stimulation sites when compared to the sites of perception reported by the subjects. The PNS-optimization improved the PNS thresholds for the target scan landmark as well as most other studied landmarks, potentially yielding a significant improvement in image encoding performance that can be safely used in humans.

Optimizing Ion Optical Design for Laser Ablation Source in Mass Spectrometry

The study focuses on optimizing a Laser Ablation Source (LAS) for use in mass spectrometry, particularly aiming to enhance transmission of ions and and determination of isotopic compositions in diverse matrices at TRIUMF’s Ion Trap for Atomic and Nuclear Science (TITAN). Critical variables affecting mass spectrometer resolution, such as ion kinetic energy distribution and ion transport are optimized through LAS. The paper explores the importance of consistent initial conditions, ion transport efficiency, and ion capture for accurate and reproducible measurements. Using SIMION software, we optimized ion optical design to tackle challenges like collimating ions travelling off-axis and mass filtering. A significant part of the study is dedicated to ion optic design to enable low abundance measurement of ions of interest (e.g., 10 ions of interest out of 1014 total ions). Techniques like high voltage switching and beam shaping were explored, and the final design aims for high transport efficiency and a 4 mm effective target scan area with maximum transport efficiency. The integration of LAS with a Multiple-Reflection Time-Of-Flight Mass Spectrometer (MR-TOF-MS) is expected to yield an analytical tool with high spatial resolution and transport efficiency for isotope abundance measurement.

Diffusion MRI harmonization via personalized template mapping.

One fundamental challenge in diffusion magnetic resonance imaging (dMRI) harmonization is to disentangle the contributions of scanner-related effects from the variable brain anatomy for the observed imaging signals. Conventional harmonization methods rely on establishing an atlas space to resolve anatomical variability and generate a unified inter-site mapping function. However, this approach is limited in accounting for the misalignment of neuroanatomy that still widely persists even after registration, especially in regions close to cortical boundaries. To overcome this challenge, we propose a personalized framework in this paper to more effectively address the confounding from the misalignment of neuroanatomy in dMRI harmonization. Instead of using a common template representing site-effects for all subjects, the main novelty of our method is the adaptive computation of personalized templates for both source and target scanning sites to estimate the inter-site mapping function. We integrate our method with the rotation invariant spherical harmonics (RISH) features to achieve the harmonization of dMRI signals between sites. In our experiments, the proposed approach is applied to harmonize the dMRI data acquired from two scanning platforms: Siemens Prisma and GE MR750 from the Adolescent Brain Cognitive Development dataset and compared with a state-of-the-art method based on RISH features. Our results indicate that the proposed harmonization framework achieves superior performance not only in reducing inter-site variations due to scanner differences but also in preserving sex-related biological variability in original cohorts. Moreover, we assess the impact of harmonization on the estimation of fiber orientation distributions and show the robustness of the personalized harmonization procedure in preserving the fiber orientation of original dMRI signals.

Abstract 470: AR-induced miRNA124-3p.2 suppresses the metastasis-promoting factor CCL2

Abstract Objective: The standard treatment for advanced prostate cancer is androgen deprivation therapy that blocks androgen receptor (AR) signaling, but this therapy becomes ineffective within a few years, resulting in castration-resistant prostate cancer. We have previously shown that blockade of AR signaling induces CCL2 and activates cancer cells through an autocrine mechanism, but why AR blockade induces CCL2 secretion has not been elucidated. Micro RNAs (miRs) are endogenous non-coding RNAs that regulate the translation of various proteins. In this study, we investigated whether miRs are involved in CCL2 induction. Subjects and Methods: We used miRNA PCR Plate Array and Target Scan (https://www.targetscan.org) to search for miRs that are controlled by AR and bind to and repress the CCL2 gene. We knocked down the AR of human prostate cancer cell lines LNCaP and C4-2B with siRNA (siAR) and examined the expression of target miRs and CCL2. siAR-transfected LNCaP and C4-2B were transfected with target miR mimic and CCL2 expression was examined. siAR-transfected LNCaP, C4-2B, and C4-2B were transfected with target miR mimic and CCL2 expression was examined. siAR-transfected LNCaP and C4-2B were transfected with target miR mimic and CCL2 expression was examined. The target miR mimic was introduced into them, and the changes in migration ability were evaluated. Results: miR-124-3p.2 was extracted as a candidate from miRNA PCR Plate Array and Target Scan. miR-124-3p.2 expression was upregulated in both LNCaP and C4-2B when DHT was added and downregulated when CCL2 was added. In siAR LNCaP and C4-2B, the expression of CCL2 was decreased when miR-124-3p.2 mimic was introduced. Furthermore, in siAR-transfected LNCaP, C4-2B, the introduction of miR-124-3p.2 mimic suppressed the migration. Conclusion: AR suppresses CCL2 via miR124-3p.2. miR124-3p.2-mediated regulation of CCL2 may be a new therapeutic target for advanced prostate cancer. Citation Format: Shuhei Aoyama, Kouji Izumi, Atsushi Mizokami. AR-induced miRNA124-3p.2 suppresses the metastasis-promoting factor CCL2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 470.

Abstract 5583: Helicobacter pylori induces the wee1, promoting immune suppressive in human gastric adenocarcinoma

Abstract Background: Gastric cancer (GC) is the fourth most common cause of cancer-related death worldwide. GC has a dismal 5-year survival rate of about 32%. Helicobacter pylori (H. pylori) infection is the strongest risk factor for gastric carcinogenesis. 75-90% of gastric cancer patients exhibit positive H. pylori serology. WEE1 is a nuclear kinase that regulates cell cycle progression by inhibiting cyclin-dependent kinases. Methods &amp; Results: Analysis of non-cancer normal stomach tissue (n=360) and human GC samples (n=1221) identified that WEE1 mRNA was significantly overexpressed in GC. Using immunohistochemistry (IHC) staining in 41 normal and 44 GC human samples, the data indicated that WEE1 protein was significantly overexpressed in GC. Western blot and immunofluorescence (IF) staining confirmed that H. pylori infection induces WEE1 overexpression in GC cells with an unexpected cytosolic localization of WEE1 in GC cells. Tff1 knockout GC mouse model (Tff1-/-) was infected with mouse-adapted H. pylori strain PMSS1. Western blot and immunohistochemistry data demonstrated that H. pylori infection induced WEE1 expression in Tff1 -/- HGD/cancer mouse gastric tissues compared to control Tff1 -/- mouse stomach. To study how H. pylori infection induces WEE1 in GC, analysis from miRTAR, Target Scan, and miRDB demonstrated that WEE1 is predicted to be targeted by miR-497-5p. Using qRT-PCR analysis, we assessed 30 normal and 30 GC human tissue samples, revealing a pronounced downregulation of miR-497-5p in GC specimens. Western blot analysis confirmed that the restoration of miR-497-5p in GC cells markedly reduced WEE1 protein expression. Interestingly, H. pylori infection further decreased miR-497-5p levels in GC cells, upregulating WEE1 protein expression. Since the miR-497-5P host gene promoter region contains CpG islands, we confirmed that treatment with 5 Azacytidine (demethylating agent) or DNMT1 knockdown induced expression of miR-497-5p with decreased WEE1 level in GC cells. Furthermore, our preliminary results indicate that aberrant WEE1 overexpression in GC demonstrates a significant positive correlation with immune-suppression gene signatures, underscoring its predictive value for unfavorable patient survival outcomes in GC. Using the TFF1 knockout mouse GC model, we found that WEE1 inhibition notably decreases the number and activation of CD4+ and CD8+ T cells in neoplastic gastric tissues. These findings also indicate a crucial relationship between WEE1 and the immunosuppressive microenvironment in GC, establishing WEE1 as a promising therapeutic target to enhance therapeutic efficacy and overcome immunotherapy resistance in GC. Conclusions: Our findings demonstrated a novel mechanism in which H. pylori activates the WEE1 in GC, promoting an immune suppressive microenvironment. WEE1 inhibition with immunotherapy could be a promising therapeutic method for GC patients. Citation Format: Md Obaidul Islam, Krishnapriya Thangaretnam, Heng Lu, Dunfa Peng, Nadeem Sidiq Bhat, Mohammed Soutto, Wael El-Rifai, Zheng Chen. Helicobacter pylori induces the wee1, promoting immune suppressive in human gastric adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5583.

The critical role and functional mechanism of microRNA-146a in doxorubicin-induced apoptosis in breast cancer cells

Background Breast cancer among women is the most frequently diagnosed cancer and the leading cause of death worldwide. There many advances in diagnosing and treating this disease, early diagnosis and treatment are still a significant challenge in the early stages. In recent years, microRNAs have attracted much attention in cancer diagnosis and treatment. However, the role of miR-146a in breast cancer is still controversial. We aimed to investigate the roles of miR-146a in apoptosis in breast cancer cells. Methods A microarray dataset from the GEO database was selected, and using the GEO2R tool, the gene expression profile of this dataset was extracted. Then, the target scan database was used to explore the miR-146a target genes. The link between the signaling pathways was collected. We used miR-146a mimic, which was transfected to the MCF-7 cells to investigate the miR-146a roles in the apoptosis. The expression levels of miR-146a and BAX, BCL-2, and p-21(most essential genes in the apoptosis) were quantified by qPCR and western blot analysis. Results Our findings indicated that doxorubicin induces miR-146a expression. In addition, overexpression of miR-146a affected MCF-7 cell viability, induced apoptosis, and led to reduced expression levels of BCL-2 and P-21, as well as increased BAX expression levels. Conclusion Considering the role of doxorubicin in inducing apoptosis and increasing the expression of miR-146a, it can be suggested that this miR is involved in inducing apoptosis in BC cells. In addition, miR-146a can be considered a therapeutic candidate.

Hand-eye calibration of line structured-light sensor by scanning and reconstruction of a free-placed standard cylindrical target

This paper proposed a hand-eye calibration method for a line structured-light (LSL) scanner mounted on industrial robot. Unlike widely used spherical targets, the target used in our work is a cylinder, which is inexpensive and easy to be manufactured in larger size to cover a wider calibration space. In order to address the profile self-occlusion and drastically shape changing of conic section during calibration, an unconstrained dimensionality reduction method is proposed to fit the scanned profiles. Based on fitted ellipse centers under various robot poses, the hand-eye calibration model is developed based on the difference between cylinder radius and the distance from the profile point to the cylinder axis. Simulation and actual experiments have shown that the proposed ellipse fitting method is more accurate and robust, and the calibration method achieves higher reconstruction accuracy under the same robot positioning error, need less robot operations to obtain more accurate hand-eye parameters.

Diagnostic significance and potential function of miR-320d in schizophrenia.

Schizophrenia is a chronic brain disorder and needs objective diagnostic biomarkers. MicroRNAs are highly expressed in the nervous system. The study investigated the expression and clinical values of serum miR-320d in schizophrenia patients. In addition, the underlying mechanism was preliminarily examined via bioinformatic analysis. Serum samples were collected from 57 patients with first-episode schizophrenia and 62 healthy controls. The cognitive function of patients was assessed via Measurement and Treatment Research to Improve Cognition in Schizophrenia Consensus Cognitive Battery (MCCB) consisting of seven domains. Serum miR-320d levels were tested via qRT-PCR. The miRNA target predictions were obtained from Target Scan, and annotated through Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Based on the GSE167630 dataset, downregulated serum miR-320d in schizophrenia was identified, which was determined in the serum of schizophrenia patients. Serum miR-320d presented a conspicuous relationship with MCCB score in both the control group and the schizophrenia group. After adjusting for age, sex, BMI, and education, serum miR-320d was still independently related to the occurrence of schizophrenia. It can identify schizophrenia cases from healthy ones with an AUC of 0.931. The Go enrichment analysis indicated that the target genes were mainly enriched in homophilic cell adhesion and cell-cell adhesion via plasma-membrane adhesion molecules, and GTPase activity and guanosine diphosphate (GDP) binding. Rap1 signaling pathway was enriched via KEGG analysis. Serum miR-320d can be taken as a candidate marker for the diagnosis of schizophrenia. Its regulatory role in neuronal cell adhesion and Rap1 signaling pathway might be the potential underlying mechanism of miR-320d in schizophrenia.

Circ_0000877 accelerates proliferation and immune escape of non-small cell lung cancer cells by regulating microRNA-637/E2F2 axis.

Recently, circular RNA (circRNA) has become a vital targeted therapy gene for non-small-cell lung cancer (NSCLC) cells. CircRNA_0000877 (Circ_0000877) has been researched in diffuse large B-cell lymphoma (DLBCL). However, whether circ_0000877 regulated NSCLC cell progression is still poorly investigated. The research attempted to investigate the influence of circ_0000877 in NSCLC. Circ_0000877 levels in NSCLC tissues and cell lines were determined applying RT-qPCR. Cell functions were evaluated by CCK-8, EdU, flow cytometry, ELISA, and western blot. Gene interactions were predicted by Cirular RNA interactome database and Target Scan website and certified by dual-luciferase reporter, RIP, and RNA pull-down assays. Finally, mice experimental model was established to explore the effects of circ_0000877 on tumor growth in vivo. The elevated trend of circ_0000877 expression was discovered in NSCLC tissues compared to para-carcinoma tissues. The clinicopathological data uncovered that up-regulated circ_0000877 was linked to tumor size, differentiation, and TNM stages of NSCLC patients. Knockdown of circ_0000877 inhibited the proliferation, triggered apoptosis, and prohibited immune escape in NSCLC cells. It was certified that miR-637 was directly interacted with circ_0000877 and targeted by E2F2. Overexpressed E2F2 strongly overturned the functions of circ_0000877 knockdown in NSCLC cells. Mice experimental data demonstrated that circ_0000877 knockdown suppressed tumor growth in vivo. The research demonstrated that circ_0000877 exhibited the promotive effect on NSCLC cells proliferation and immune escape by regulating miR-637/E2F2 axis.