Tumor Parenchyma Research Articles

Metformin exerted tumoricidal effects on colon cancer tumoroids via the regulation of autophagy pathway

BackgroundDespite the existence of promising outcomes from standard 2D culture systems, these data are not completely akin to in vivo tumor parenchyma. Therefore, the development and fabrication of various 3D culture systems can in part mimic intricate cell-to-cell interaction within the real tumor mass. Here, we aimed to evaluate the tumoricidal impacts of metformin (MTF) on colorectal cancer (CRC) tumoroids in an in vitro system via the modulation of autophagy.MethodsCRC tumoroids were developed using human umbilical vein endothelial cells (HUVECs), adenocarcinoma HT29 cells, and fibroblasts (HFFF2) in a ratio of 1: 2: 1 and 2.5% methylcellulose. Tumoroids were exposed to different concentrations of MTF, ranging from 20 to 1000 mM, for 72 h. The survival rate was detected using an LDH release assay. The expression and protein levels of autophagy-related factors were measured using PCR array and western blotting, respectively. Using H & E, and immunofluorescence staining (Ki-67), the integrity and proliferation rate of CRC tumoroids were examined.ResultsThe current protocol yielded typical compact tumoroids with a dark central region. Despite slight changes in released LDH contents, no statistically significant differences were achieved in terms of cell toxicity in MTF-exposed groups compared to the control tumoroids, indicating the insufficiency of MTF in the induction of tumor cell death (p > 0.05). Western blotting indicated that the LC3II/I ratio was reduced in tumoroids exposed to 120 mM MTF (p < 0.05). These data coincided with the reduction of intracellular p62 content in MTF 120 mM-treated tumoroids compared to MTF 40 mM and control groups (p < 0.05). PCR array analysis confirmed the up-regulation, and down-regulation of several genes related to various signaling transduction pathways associated with autophagy machinery and shared effectors between autophagy and apoptosis in 40 and 120 mM MTF groups compared to the non-treated control group (p < 0.05). These changes were more prominent in tumoroids incubated with 120 mM MTF. Histological examination confirmed the loosening integrity of tumoroids in MTF-treated groups, especially 120 mM MTF, with the increase in cell death via the induction of apoptosis (chromatin marginalization) and necrotic (pyknotic nuclei) changes. In the 120 mM MTF group, spindle-shaped cells with the remnants of a fibrillar matrix were detected. Data indicated the reduction of proliferating Ki-67+ cells within the tumoroids by increasing the MTF concentration from 40 to 120 mM.ConclusionsDifferent shared autophagy/apoptosis genes were modulated in CRC tumoroids after MTF treatment coinciding with both typical necrotic and apoptotic cells within the tumoroid structure. MTF can inhibit the integrity and proliferation of CRC tumoroids in dose-dependent manner.

Deep Learning Enabled Scoring of Pancreatic Neuroendocrine Tumors Based on Cancer Infiltration Patterns

Pancreatic neuroendocrine tumors (PanNETs) are a heterogeneous group of neoplasms that include tumors with different histomorphologic characteristics that can be correlated to sub-categories with different prognoses. In addition to the WHO grading scheme based on tumor proliferative activity, a new parameter based on the scoring of infiltration patterns at the interface of tumor and non-neoplastic parenchyma (tumor-NNP interface) has recently been proposed for PanNET categorization. Despite the known correlations, these categorizations can still be problematic due to the need for human judgment, which may involve intra- and inter-observer variability. Although there is a great need for automated systems working on quantitative metrics to reduce observer variability, there are no such systems for PanNET categorization. Addressing this gap, this study presents a computational pipeline that uses deep learning models to automatically categorize PanNETs for the first time. This pipeline proposes to quantitatively characterize PanNETs by constructing entity-graphs on the cells, and to learn the PanNET categorization using a graph neural network (GNN) trained on these graphs. Different than the previous studies, the proposed model integrates pathology domain knowledge into the GNN construction and training for the purpose of a deeper utilization of the tumor microenvironment and its architectural changes for PanNET categorization. We tested our model on 105 HE stained whole slide images of PanNET tissues. The experiments revealed that this domain knowledge integrated pipeline led to a 76.70% test set F1-score, resulting in significant improvements over its counterparts.

Exploratory profiling of metabolites in cerebrospinal fluid using a commercially available targeted LC-MS based metabolomics kit to discriminate leptomeningeal metastasis

BackgroundLeptomeningeal metastasis (LM) is a devastating complication of cancer that is difficult to treat. Thus, early diagnosis is essential for LM patients. However, cerebrospinal fluid (CSF) cytology has low sensitivity, and imaging approaches are ineffective. We explored targeted CSF metabolic profiling to discriminate among LM and other conditions affecting the central nervous system (CNS).MethodsWe quantitatively measured amino acids, biogenic amines, hexoses, acylcarnitines (AC), cholesteryl esters (CE), glycerides, phosphatidylcholines (PC), lysophosphatidylcholines (LPC), sphingomyelins (SM), and ceramides (Cer) in 117 CSF samples from various groups of healthy controls (HC, n = 10), patients with LM (LM, n = 47), parenchymal brain tumor (PBT, n = 45), and inflammatory disease (ID, n = 13) with internal standards using the Absolute IDQ- p400® targeted mass spectrometry kit. Metabolites detected in > 90% of samples or showing a difference in proportional level between groups ≥ 75% were used in logistic regression models when there was no single metabolite with AUC = 1 for the groups of comparison.ResultsPC and SM had higher levels in LM than in PBT or HC, whereas LPC had lower level in PBT than the other groups. Glycerides and Cer levels were higher in PBT and LM than in HC. Long-chain AC level in PBT was lower than in LM or HC. A regression model including Ala, PC (42:7), PC (30:3), PC (37:0), and Tyr achieved complete discrimination (AUC = 1.0) between LM and HC. In comparison of PBT and HC, twenty-six individual metabolites allowed complete discrimination between two groups, and between ID and HC fourty-six individual lipid metabolites allowed complete discrimination. Twenty-one individual metabolites (18 ACs and 3 PCs) allowed complete discrimination between LM and PBT.ConclusionsUsing a commercial targeted liquid chromatography-mass spectrometry (LC-MS) metabolomics kit, we were able to differentiate LM from HC and PBT. Most of the discriminative metabolites among different diseases were lipid metabolites, for which their CNS distribution and quantification in different cell types are largely unknown, whereas amino acids, biogenic amines, and hexoses failed to show significant differences. Future validation studies with larger, controlled cohorts should be performed, and hopefully, the kit may expand its metabolite coverage for unique cancer cell glucose metabolism.

3D synergistic tumor-liver analysis further improves the efficacy prediction in hepatocellular carcinoma: a multi-center study

BackgroundBesides tumorous information, synergistic liver parenchyma assessments may provide additional insights into the prognosis of hepatocellular carcinoma (HCC). This study aimed to investigate whether 3D synergistic tumor-liver analysis could improve the prediction accuracy for HCC prognosis.MethodsA total of 422 HCC patients from six centers were included. Datasets were divided into training and external validation datasets. Besides tumor, we also performed automatic 3D assessment of liver parenchyma by extracting morphological and high-dimensional data, respectively. Subsequently, we constructed a tumor model, a tumor-liver model, a clinical model and an integrated model combining information from clinical factors, tumor and liver parenchyma. Their discrimination and calibration were compared to determine the optimal model. Subgroup analysis was conducted to test the robustness, and survival analysis was conducted to identify high- and low-risk populations.ResultsThe tumor-liver model was superior to the tumor model in terms of both discrimination (training dataset: 0.747 vs. 0.722; validation dataset: 0.719 vs. 0.683) and calibration. Moreover, the integrated model was superior to the clinical model and tumor-liver model, particularly in discrimination (training dataset: 0.765 vs. 0.695 vs. 0.747; validation dataset: 0.739 vs. 0.628 vs. 0.719). The AUC of the integrated model was not influenced by AFP level, BCLC stage, Child–Pugh grade, and treatment style in training (6 months p value: 0.245–0.452; 12 months p value: 0.357–0.845) and validation (6 months p value: 0.294–0.638; 12 months p value: 0.365–0.937) datasets. With a risk score of 1.06, high- and low-risk populations demonstrated significant difference for progression-free survival (p < 0.001 in both datasets).ConclusionsCombined with clinical factors, 3D synergistic tumor-liver assessment improved the efficacy prediction of HCC.

Optimal virtual monoenergy for the detection of pancreatic adenocarcinoma during the pancreatic parenchymal phase on photon counting CT.

As the pancreas is a low contrast visibility organ, pancreatic ductal adenocarcinoma detection is challenging due to subtle attenuation differences between tumor and pancreatic parenchyma. Photon counting CT (PCCT) has superior iodine contrast-to-noise ratio than conventional CT and also affords the creation of low keV virtual monoenergetic images, both of which increase adenocarcinoma conspicuity. The purpose therefore was to identify the optimal virtual monoenergy for visualizing PDAC during the pancreatic parenchymal phase of enhancement at PCCT using both quantitative and qualitative analyses. Consecutive patients with pancreatic parenchymal phase PCCT source data were retrospectively identified by PACS search. For the quantitative analysis, region of interest (ROI) measurements were drawn in the pancreatic head, body, tail, pancreatic adenocarcinoma (if present), and psoas muscles on 40-120keV virtual monoenergetic images in 10keV increments. Based on the quantitative analysis results and vendor recommendations, four virtual monoenergies(40keV, 55keV, 70keV, and 85keV) were selected for additional qualitative analysis. Three radiologists blinded to four virtual monoenergies assessed overall image quality, image noise, pancreatic enhancement, and pancreatic mass conspicuity on 5-point Likert scales. 54 patients (28/54 male, mean[SD] age: 62 [13] years) were included. Quantitatively, 40keV had the highest pancreatic parenchymal CNR and attenuation difference between the adenocarcinoma and parenchyma, but also the highest noise (HUsd). Qualitatively, 70keV had the best overall image quality (Mean [SE]: 3.7[0.1]) and lower noise than 40 and 55keV (3.6[0.08] vs. 1.8[0.07] and 2.7[0.05], respectively, p < .001). 40keV had the greatest pancreatic enhancement (mean[SE] 4.6[0.11]). Adenocarcinoma conspicuity ratings were greatest at 40keV and 55keV, and not significantly different from each other (mean[SE] 4.4[0.13] and 4.3[0.14], respectively, Tukey adj-p =.20). 55keV had greater overall image quality and lower noise than 40keV (mean[SE] 3.4[0.08] vs. 2.5[0.08], Tukey adj-p < .001 and 2.7[0.05] vs. 1.8[0.07], Tukey adj-p < .001 respectively). 55keV pancreatic parenchymal phase virtual monoenergetic images afford optimal pancreatic assessment at PCCT for the visualization of pancreatic adenocarcinoma. Routinely viewing 55keV virtual monoenergetic images at PCCT may improve PDAC detection.

Agathisflavone Inhibits Viability and Modulates the Expression of miR-125b, miR-155, IL-6, and Arginase in Glioblastoma Cells and Microglia/Macrophage Activation.

Glioblastomas (GBM) are malignant tumours with poor prognosis. Treatment involves chemotherapy and/or radiotherapy; however, there is currently no standard treatment for recurrence, and prognosis remains unfavourable. Inflammatory mediators and microRNAs (miRNAs) influence the aggressiveness of GBM, being involved in the communication with the cells of the tumour parenchyma, including microglia/macrophages, and maintaining an immunosuppressive microenvironment. Hence, the modulation of miRNAs and inflammatory factors may improve GBM treatments. In this study, we investigated the effects of agathisflavone, a biflavonoid purified from Cenostigma pyramidale (Tul.), on the growth and migration of GBM cells, on the expression of inflammatory cytokines and microRNAs, as well on the response of microglia. Agathisflavone (5-30 μM) induced a dose- and time-dependent reduction in the viability of both human GL-15 and rat C6 cells, as determined by the MTT test, and reduced cell migration, as determined by cell scratch assay. RT-qPCR analysis revealed that agathisflavone (5 μM) down-regulated the expression of miR-125b and miR-155 in the secretome derived from GL-15 cells, which was associated with upregulation of the mRNA expression of IL-6 and arginase-1 immunoregulatory factors. Exposure of human microglia/macrophage to the secretome from GL-15 GMB cells modulated proliferation and morphology, effects that were modulated by agathisflavone treatment. These results demonstrate the effect of flavonoids on the growth of GBM cells, which impacts cells in the microenvironment and can be considered for preclinical studies for adjuvant treatments.

Deciphering functional tumor-immune crosstalk through highly multiplexed imaging and deep visual proteomics.

Deciphering the intricate tumor-immune interactions within the microenvironment is crucial for advancing cancer immunotherapy. Here, we introduce mipDVP, an advanced approach integrating highly multiplexed imaging, single-cell laser microdissection, and sensitive mass spectrometry to spatially profile the proteomes of distinct cell populations in a human colorectal and tonsil cancer with high sensitivity. In a colorectal tumor-a representative cold tumor-we uncovered spatial compartmentalization of an immunosuppressive macrophage barrier that potentially impedes Tcell infiltration. Spatial proteomic analysis revealed distinct functional states of Tcells in different tumor compartments. In a tonsil cancer sample-a hot tumor-we identified significant proteomic heterogeneity among cells influenced by proximity to cytotoxic Tcell subtypes. Tcells in the tumor parenchyma exhibit metabolic adaptations to hypoxic regions. Our spatially resolved, highly multiplexed strategy deciphers the complex cellular interplay within the tumor microenvironment, offering valuable insights for identifying immunotherapy targets and predictive signatures.

Trans-Vessel Wall Cell Transplantation, Engraftment, and Tumor Access in the VX2 Rabbit Model

The trans-vessel wall device (TW-device) is a new endovascular tool for precise and safe delivery of various payloads (cells, viral, modified RNA, chemotherapy, growth factors) in oncology and regenerative medicine. The twofold aim of this study was to assess cell engraftment and tumor growth using the TW-device for endovascular transplantation and to evaluate its ability to directly access solid tumors. We used the VX2 model in the rabbit kidney to compare percutaneously implanted fresh VX2 cells with TW-device injections of cryopreserved VX2 cells. We demonstrated the feasibility of endovascular transplantation (n = 7) of tumor cells, achieving a 57.1% engraftment rate despite cryopreservation, comparable with 70% for percutaneous delivery of fresh cells (n = 10). Re-access using the TW-device was 100% successful (n = 11) with super-selective intratumoral contrast administration without complications. In conclusion, endovascular transplantation of VX2 cells using the TW-device resulted in proliferating cell grafts in the rabbit kidney establishing functional proof that cells indeed survive handling, preparation, and device passage. We also show the TW-device is able to access solid tumor parenchyma allowing precise intraparenchymal administration.This proof-of-concept study open up possibilities for repeated direct parenchymal injections via the endovascular route in any hard to reach organ.

Multiomics reveals tumor microenvironment remodeling in locally advanced gastric and gastroesophageal junction cancer following neoadjuvant immunotherapy and chemotherapy

BackgroundPerioperative chemotherapy is the standard of care for patients with locally advanced gastric and gastroesophageal junction cancer. Recent evidence demonstrated the addition of programmed cell death protein 1 (PD-1) inhibitors...

Assessing Therapeutic Nanoparticle Accumulation in Tumors Using Nanobubble-Based Contrast-Enhanced Ultrasound Imaging.

This study explores the challenges associated with nanoparticle-based drug delivery to the tumor parenchyma, focusing on the widely utilized enhanced permeability and retention effect (EPR). While EPR has been a key strategy, its inconsistent clinical success lacks clear mechanistic understanding and is hindered by limited tools for studying relevant phenomena. This work introduces an approach that employs multiparametric dynamic contrast-enhanced ultrasound (CEUS) with a nanoscale contrast agent for noninvasive, real-time examination of tumor microenvironment characteristics. We demonstrate that CEUS imaging can: (1) evaluate tumor microenvironment features, (2) be used to help predict the distribution of doxorubicin-loaded liposomes in the tumor parenchyma, and (3) be used to predict nanotherapeutic efficacy. CEUS using nanobubbles (NBs) was carried out in two tumor types of high (LS174T) and low (U87) vascular permeability. LS174T tumors consistently showed significantly different time intensity curve (TIC) parameters, including area under the rising curve (AUCR, 2.7×) and time to peak intensity (TTP, 1.9×) compared to U87 tumors. Crucially, a recently developed decorrelation time (DT) parameter specific to NB CEUS dynamics successfully predicted the distribution of doxorubicin-loaded liposomes within the tumor parenchyma (r = 0.86 ± 0.13). AUCR, TTP, and DT were used to correlate imaging findings to nanotherapeutic response with 100% accuracy in SKOV-3 tumors. These findings suggest that NB-CEUS parameters can effectively discern tumor vascular permeability, serving as a biomarker for identifying tumor characteristics and predicting the responsiveness to nanoparticle-based therapies. The observed differences between LS174T and U87 tumors and the accurate prediction of nanotherapeutic efficacy in SKOV-3 tumors indicate the potential utility of this method in predicting treatment efficacy and evaluating EPR in diseases characterized by pathologically permeable vasculature. Ultimately, this research contributes valuable insights into refining drug delivery strategies and assessing the broader applicability of EPR-based approaches.

Abstract B011: Sensory neurons regulate intracellular and intercellular mitochondrial dynamics in breast cancer cells

Abstract Crosstalk between cancer and the nervous system regulates tumor progression. These interactions occur at both the local (i.e., within the tumor parenchyma) and systemic (e.g., endocrine signaling) levels. More recent work has determined that cancer cells not only interact with their local environment via secreted molecules, but in some cases, via acquiring proteins or organelles from other cells within the tumor microenvironment (TME). Here, we demonstrate that breast cancer cells acquire mitochondria from neurons. Breast cancer cells with higher metastatic potential acquire more mitochondria in NGF dependent manner compared to the cell line with lower metastatic potential. These results suggest that in addition to their known roles in electrochemical and paracrine signaling, tumor-innervating nerves can also enhance cancer cell fitness via transfer of mitochondria. We are now characterizing the mechanisms underlying this transfer, determining the fate of transferred mitochondria in recipient cells, and defining how transferred mitochondria drive cancer development, progression, and metastasis. Further, sensory neurons at the tumor microenvironment dictate the intracellular mitochondria dynamics affecting morphology and function. The change in mitochondria dynamics enhances cancer cell survival and metastasis, we are uncovering how sensory neuron induced mitochondria dynamics drives tumor progression. Citation Format: Ankit Tiwari, Yue Wu, Jeremy C. Borniger. Sensory neurons regulate intracellular and intercellular mitochondrial dynamics in breast cancer cells [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr B011.

Misdiagnosis in breast imaging: a statement paper from European Society Breast Imaging (EUSOBI)-Part 2: Main causes of errors in breast imaging and recommendations from European Society of Breast Imaging to limit misdiagnosis.

Breast cancer is one of the leading causes of negligence claims in radiology. The objective of this document is to describe the specific main causes of errors in breast imaging and provide European Society of Breast Imaging (EUSOBI) recommendations to try to minimize these. Technical failures represent 17% of all mammographic diagnostic negligence claims. Mammography quality control protocol and dedicated training for technologists and radiologists are essential. Lack of consideration of the clinical context is a second critical issue, as a clinical abnormality is found in 80% of malpractice claims. EUSOBI emphasizes the importance of communication and clinical examination before the diagnostic investigation. Detection errors or misapplications of the lexicon or Breast Imaging Reporting Data System (BI-RADS) score account for 5% of malpractice claims and should be reduced by limiting radiologists' distraction or fatigue, and being aware of satisfaction of search errors and the importance of a personal systematic review. Errors related to pathological concordance and MDT review can be limited by the use of markers after biopsy and the use of standardized reports, which can aid communication with other specialities. Finally, errors related to tumor or patient factors should be discussed, considering the use of contrast-enhanced mammography and magnetic resonance imaging. Several factors are responsible for misdiagnosis in breast cancer, including errors in the practice of the technician and/or radiologist (technical failures, lack of consideration of the clinical context, incorrect application of the BI-RADS score, false reassurances), lack of communication with other specialists or with the patient, and the type of tumor and breast parenchyma. Question What factors most contribute to and what implications stem from misdiagnosis in breast imaging? Findings Ongoing training and education for radiologists and other healthcare providers, as well as interdisciplinary collaboration and communication is paramount. Clinical relevance Misdiagnosis in breast imaging can have significant implications for patients, healthcare providers, and the entire healthcare system.

EXTH-48. ADENO-ASSOCIATED VIRUS TARGETING OF THE TUMOR MICROENVIRONMENT TO IMPROVE LYMPHOCYTE RECRUITMENT IN GLIOBLASTOMA

Abstract BACKGROUND Glioblastoma (GBM) is notoriously devoid of lymphocytes driven in part by a paucity of lymphocyte trafficking factors necessary to prompt their recruitment, infiltration, and activation. We have developed a novel recombinant adeno-associated virus (AAV) gene therapy strategy that enables focal and stable reconstitution of the GBM tumor microenvironment (TME) with C-X-C motif ligand 9 (CXCL9), a powerful call-and-receive chemokine for cytotoxic T lymphocytes (CTLs). By increasing lymphocyte recruitment in GBM, we HYPOTHESIZE that these tumors will be more responsive to immunotherapy. METHODS 3D fluorescence microscopy was used to characterize the distribution, tropism, and durability of AAV6 transgene expression in preclinical GBM. Flow cytometry and 3D fluorescence microscopy was used to quantify T cell infiltration and visualize their biodistribution in preclinical GBM treated with AAV-CXCL9 in combination with anti-PD-1 immune checkpoint blockade (ICB). Single-cell RNA sequencing was used to characterize intratumor lymphocyte subsets following treatment. Survival studies with and without lymphocyte depletion were done to evaluate immunogenic responsiveness of therapy. RESULTS Focal delivery of CXCL9 with AAV6 results in stable transduction of peri-tumoral astrocytes. When combined with anti-PD-1 ICB, lymphocyte infiltration was improved, with cells found disseminated across the tumor stroma and within the tumor parenchyma. CXCR3, the chemokine receptor for CXCL9, was expressed across lymphocyte subsets and NKT cells, validating treatment selectivity of AAV6-CXCL9 for lymphocytes. Combination treatment considerably improved overall survival in two distinct GBM models, with durable responses dependent on CD8 T-lymphocyte infiltration. CONCLUSIONS By manipulating local chemokine directional guidance, AAV-CXCL9 increases tumor infiltration by CD8-postive cytotoxic lymphocytes, sensitizing GBM to anti-PD-1 ICB. These effects are accompanied by immunologic signatures evocative of an inflamed and responsive TME. These findings support targeted AAV gene therapy as a promising adjuvant strategy for reconditioning GBM immunogenicity given its excellent safety profile, TME-tropism, modularity, and off-the-shelf capability.

HERV-W envelope protein is present in microglial cells of the human glioma tumor microenvironment and differentially modulates neoplastic cell behavior

Gliomas are the most common parenchymal tumors of the central nervous system (CNS). With regard to their still unclear etiology, several recent studies have provided evidence of a new category of pathogenic elements called human endogenous retroviruses (HERVs) which seem to contribute to the evolution and progression of many neurological diseases such as amyotrophic lateral sclerosis (ALS), schizophrenia, chronic inflammatory polyneuropathy (CIDP) and, particularly, multiple sclerosis (MS). In these diseases, HERVs exert effects on cellular processes such as inflammation, proliferation, and migration. In previous studies, we demonstrated that in MS, the human endogenous retrovirus type-W envelope protein (HERV-W ENV) interferes with lesion repair through the activation of microglia (MG), the innate myeloid immune cells of the CNS. Here, we now show that HERV-W ENV is also present in the microglial cells (MG) of the tumor microenvironment (TME) in gliomas. It modulates the behavior of glioblastoma (GBM) cell lines in GBM/MG cocultures by altering their gene expression, secreted cytokines, morphology, proliferation, and migration properties and could thereby contribute to key tumor properties.

Simple Analysis Using Immunohistochemical Staining for Tumor-Infiltrating Lymphocytes in Brain Metastasis of Small Cell Lung Cancer

Background: We investigated the distribution of tumor-infiltrating lymphocytes (TILs) and the expression of programmed cell death-ligand 1 (PD-L1) in patients with brain metastasis (BM) from small cell lung cancer (SCLC). Methods: A retrospective analysis was performed on 12 surgical specimens of BMs from SCLC at our institute for 5 years. The Immunofluorescence-based Tissue Microenvironment Analysis Panel (MAP) was utilized for the detection of TILs, including CD3, CD8, PD-1, and PD-L1, in pathological archival specimens of BMs. The correlation between the overall survival (OS) and the above-mentioned markers was analyzed in the patients.Results: Positive rates of CD3+ TILs in the tumor parenchyma versus tumor stroma were 0.60±0.94% versus 1.76±2.72% (p=0.010), respectively; positive rates of CD8+ TILs in the tumor parenchyma versus tumor stroma were 0.80±0.78% versus 2.46±3.72% (p=0.016), respectively. There was no co-expression of CD8+ and PD-1+ TILs in the tumor parenchyma of 11 cases, and the infiltration density of co-expressed CD3+ and PD-1+ TILs was more than 10/mm2 in only 1 case. There was no co-expression of CD3+ and PD-1+ TIL in the stroma of 10 cases, and the infiltration density of CD8+ and PD-1+ TILs was more than 10/mm2 in 2 cases. Immunohistochemistry was used to detect the expression of PD-L1 in 12 cases of BMs, and 3 cases (25%) were positive. Survival analysis showed that patients with positive CD3+ TILs had significantly longer OS (p=0.040). Conclusions: The distribution of TILs in BM of SCLC is low and mainly distributed in the stroma, with the low expression of PD-L1 in the tumor tissues.

Inflammation-Associated Long Non-Coding RNAs (lncRNAs) in Chronic Viral Hepatitis- Associated Hepatocellular Carcinoma.

This study aimed to identify the expression profile and prognostic significance of inflammation-associated lncRNAs in chronic viral hepatitis (CVH) and CVH-associated hepatocellular carcinoma (CVH-HCC). In the first step, lncRNA expression analysis was performed by real-time polymerase chain reaction (RT-PCR) using an array panel of 84 inflammation-associated lncRNAs in 48 formalin-fixed paraffin-embedded (FFPE) tissue samples (12 CVH-HCC, 12 peritumoral cirrhotic parenchyma, 12 nontumoral cirrhotic CVH parenchyma, 12 normal liver samples). In the second step, 7 lncRNAs (DLEU2, HOTAIR, LINC00635, LINC00662, RP11-549J18.1, SNHG16 and XIST) were chosen for RT-PCR assay testing in 72 samples (24 CVH-HCC, 24 peritumoral cirrhotic parenchyma, 24 nontumoral cirrhotic CVH parenchyma samples). Fifty-six inflammation-associated lncRNAs were significantly up-regulated in the peritumoral cirrhotic parenchyma compared to the normal liver. Expression of 71 lncRNAs was significantly higher in peritumoral cirrhotic parenchyma compared to cirrhotic CVH parenchyma. DLEU2 and SNHG16 were up-regulated both in the tumor and peritumoral cirrhotic parenchyma compared to cirrhotic CVH parenchyma. Expression of LINC00662 was significantly higher in CVH-HCC than in cirrhotic CVH parenchyma. Expression of XIST was also increased in both tumor and peritumoral parenchyma samples, albeit without statistical significance. No significant association was found between lncRNA expressions and survival. Inflammation-associated lncRNAs DLEU2, SNHG16, LINC00662, and XIST are candidate diagnostic biomarkers in CVH-HCC. More evidence is needed to prove their utility as prognostic markers.

Expression of CD68 and CD163 in malignant epithelial cells of oral squamous cell carcinoma: Phenotypic shift or mere cell fusion

Background / purposeThe progression of epithelial to mesenchymal tissue (EMT) is a highly intricate process that facilitates the transformation of cancer cells, allowing them to changeover their characteristic epithelial properties to mesenchymal attributes. This notable change empowers the cells with enhanced mobility and the ability to migrate to distant locations. Furthermore, it is imperative to adopt the idea of macrophage tumor cell fusion to achieve comprehensive considerate of this phenomenon. Our primary objective was to conduct a thorough investigation of macrophage-restricted antigens expression, specifically CD68 and CD163, in malignant epithelial cells of oral cavity squamous cell carcinoma (OSCC) to elucidate aforementioned perceptions. Materials and methodsCD68 and CD163 immunohistochemical expression were assessed in oral squamous cell carcinoma (OSCC), encompassing both the neoplastic cells and the tumor-associated macrophages (TAMs). ResultsBoth CD68 and CD163 antigens were revealed in OSCC malignant epithelial cells in a granular cell pattern, localized in membrane and cytoplasm of tumor cells respectively as well as in the infiltrating TAMs. ConclusionThe macrophage antigens were not limited to the infiltrating tumor-associated macrophages (TAMs), but were also observed in a substantial proportion of OSCC malignant epithelial cells within the tumor parenchyma. This particular expression pattern may represent a subset of tumor cells that have undergone an epithelial to a mesenchymal phenotypic transition. In addition, fusion of macrophages with tumor cells cannot be excluded; both might be associated with increased metastatic activity of OSCC.

Risk factors and drug resistance of adult community-onset urinary tract infections caused by Escherichia coli-producing extended-spectrum β-lactamase in the Chongqing region, China: a retrospective case-control study.

To evaluate the prevalence, resistance and risk factors of community-onset urinary tract infections (COUTIs) caused by extended-spectrum β-lactamase-producing Escherichia coli (ESBL-EC) for providing a basis for the selection of clinical therapeutic agents. A retrospective case-control study. The Affiliated Dazu Hospital of Chongqing Medical University (also known as The People's Hospital of Dazu Chongqing), a 1000-bed tertiary hospital in China. This study encompassed adult patients diagnosed with community-acquired urinary tract infections (UTIs) caused by E. coli between May 2017 and December 2022 with exclusion criteria including incomplete clinical data, disagreement to participate in the study, hospitalisation duration exceeding 48 hours prior to confirmation of diagnosis and prior history of urinary tract infection caused by E. coli. The risk factors for COUTIs caused by ESBL-EC were evaluated using a case-control design, defining patients who were diagnosed with UTIs and had an ESBL-positive urine culture as the case group and patients who were diagnosed with UTIs and had an ESBL-negative urine culture as the control group. Perform drug susceptibility testing and resistance analysis on isolated ESBL-EC. In total, 394 cases of COUTIs caused by E. coli were included; 192 cases were ESBL-positive with a detection rate of 48.7% (192/394). Parenchymal tumour, history of urolithiasis stone fragmentation, history of urological surgery, hospitalisation within 6 months, indwelling catheter outside the hospital and antibiotic use (mainly third-generation cephalosporins) were the factors significantly associated with COUTIs caused by ESBL-EC (p<0.05) through logistic regression for univariate analysis. Multivariate analysis revealed that a history of urolithiasis stone fragmentation (OR=2.450; 95% CI: 1.342 to 4.473; p=0.004), urological surgery (OR=3.102; 95% CI: 1.534 to 6.270; p=0.002), indwelling catheter outside hospital (OR=2.059; 95% CI: 1.025 to 4.133; p=0.042), hospitalisation within 6 months (OR=2.127; 95% CI: 1.207 to 3.748; p=0.009) and use of third-generation cephalosporins (OR=1.903; 95% CI: 1.069 to 3.389; p=0.029) were the independent risk factors for COUTIs caused by ESBL-EC. The results of the drug susceptibility testing revealed that ESBL-EC exhibited the highest resistance rates to ampicillin, ceftriaxone and cefixime, all at 100%. Mezlocillin followed with a resistance rate of 98.7%. On the other hand, ESBL-EC strains displayed the highest sensitivity to carbapenem antibiotics (imipenem, meropenem, ertapenem) and amikacin, all at 100%. Sensitivity rates were also high for cefotetan at 96.6%, piperacillin/tazobactam at 95.3% and nitrofurantoin at 87.9%. Our results revealed high ESBL-EC detection rates. COUTIs caused by ESBL-EC are more likely to occur in patients with parenchymal tumour, a history of urolithiasis stone fragmentation, a history of urological surgery, hospitalisation within 6 months, indwelling catheter outside the hospital and use of third-generation cephalosporins. These patients were highly resistant to penicillins, cephalosporins and quinolones.

ERBB2 is a potential diagnostic and prognostic biomarker in renal clear cell carcinoma

Renal clear cell carcinoma (ccRCC) is a common parenchymal tumor of the kidney, and the discovery of biomarkers for early and effective diagnosis of ccRCC can improve the early diagnosis of patients and thus improve long-term survival. Erb-b2 receptor tyrosine kinase 2 (ERBB2) mediates the processes of cell proliferation, differentiation, and apoptosis inhibition. The purpose of this study was to investigate the diagnostic and prognostic role of ERBB2 in ccRCC. We analyzed the expression levels of ERBB2 in various cancers from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. RNA-seq data were analyzed using R packages to identify differentially expressed genes between the high and low ERBB2 expression groups in the TCGA-KIRC dataset. Spearman correlation analysis was performed to determine the correlation between ERBB2 expression and immune cell infiltration, immune checkpoint expression, and PTEN expression. DNA methylation changes and genetic alterations in ERBB2 were assessed using the MethSurv and cBioPortal databases. Logistic regression analysis was performed to determine the correlation between ERBB2 expression and the clinicopathological characteristics of ccRCC patients. The diagnostic and prognostic value of ERBB2 was assessed using Kaplan‒Meier (K‒M) survival curves, diagnostic ROC curves, time-dependent ROC curves, nomogram models, and Cox regression models. The expression level of ERBB2 is lower in tumor tissues of ccRCC patients than in the corresponding control tissues. Differentially expressed genes associated with ERBB2 were significantly enriched in the pathways “BMP2WNT4FOXO1 pathway in primary endometrial stromal cell differentiation” and “AMAN pathway”. In ccRCC tissues, ERBB2 expression levels were associated with immune cell infiltration, immune checkpoints, and PTEN. The DNA methylation status of 10 CpG islands in the ERBB2 gene was associated with the prognosis of ccRCC. ERBB2 expression levels in ccRCC tissues were associated with race, sex, T stage, M stage, histological grade, and pathological stage. Cox regression analysis showed that ERBB2 was a potential independent predictor of overall survival (OS), disease-specific survival (DSS), and progression-free interval (PFI) in ccRCC patients. ROC curve analysis showed that the expression level of ERBB2 could accurately distinguish between ccRCC tissue and adjacent normal renal tissue. Our study showed that ERBB2 expression in ccRCC tissues can be of clinical importance as a potential diagnostic and prognostic biomarker.

Characterization of Classic Meningioma with Use of Conventional Magnetic Resonance and Diffusion Tensor Imaging

Introduction: The conventional magnetic resonance imaging (MRI) method is widely considered ‘with limited success’ in differentiating the meningioma types but may fail to localize the tumor occupation of white-matter fiber bundles accurately. Diffusion tensor imaging (DTI) is considered as an imaging modality that may elucidate the microstructure of brain tumors. We provide characteristics of meningioma using DTI-based-three-dimensional tracing of white matter to portray meningioma in a noninvasive approach and its structural contact to contiguous tumors and elucidate the influence of occupying lesions on white-matter fiber bundles. Case Presentation: A 28-year-old female presented with visuospatial disturbances and persistent headaches for 2 years. Conventional and advanced MRI studies were performed. Diffusion-weighted Images (DWI) and Apparent Diffusion Coefficient (ADC) values were measured in the lesion using routine MRI sequences. Advanced MRI using DTI was also performed. Conventional MRI outcomes showed tumor parenchyma, peritumoral edema, and compression on the circumnavigated brain tissue. There was hyperintense on DW trace image and isointense on ADC map. On T2-weighted image (T2WI) and Fluid-Attenuated Inversion Recovery (FLAIR) images, there was an increased signal intensity that demonstrated an extra-axial lesion, while T1-weighted imaging signals showed hypointensity. DTI fractional anisotropy (FA) marker is an unclear optic radiation in the concerned area, indicating the shift or destruction of the optic radiation. The mean FA values of solid-enhancing areas of meningioma were 0.28 ± 0.17. Mean ADC values (103 mm2/s) were 0.764 ± 0.172. Conclusions: Classic meningioma in this case has low intratumoral FA and high ADC. DTI displayed that intratumoral microscopic water motion is disorganized.