Heterogeneous Infiltration Research Articles

Characterization of tumor-infiltrating lymphocytes and their spatial distribution in triple-negative breast cancer

BackgroundThe tumor immune microenvironment, particularly tumor-infiltrating lymphocytes (TILs), plays a critical role in disease progression and treatment response in triple-negative breast cancers (TNBCs). This study was aimed to characterize the composition of TILs and investigate their clinicopathological and prognostic significance with a special focus on the spatial distribution of TILs in TNBCs.MethodsWe analyzed TNBC samples through PanCancer Immune Profiling using NanoString nCounter assays to identify immune-related genes that are expressed differentially in relation to TIL levels and evaluated protein expression of selected markers through immunohistochemical staining on tissue microarrays. For a comprehensive assessment of the expression of cytotoxic T lymphocyte (CTL) and natural killer (NK) cell markers, a CTL-NK score was devised based on CD8+, CD56+, CD57+, GNLY+, and GZMB+ TIL levels.ResultsGene expression analysis revealed significant upregulation of CTL and NK cell-associated genes including GNLY, KLRC2, and GZMB in TIL-high TNBCs. Immunohistochemical validation confirmed that TNBCs with higher TILs had a greater amount of CD56+, CD57+, GNLY+, and GZMB+ TILs not only in absolute number but also in proportion relative to CD4+ or CD8+ TILs. High TIL and its subset (CD4+, CD8+, CD56+, CD57+, GNLY+, and GZMB+ TIL) infiltration correlated with favorable clinicopathological features of tumor. In survival analysis, high CTL-NK score was found to be an independent prognostic factor for better disease-free survival (DFS) of the patients. Furthermore, uniformly high TIL infiltration was linked to better DFS, whereas cases with heterogeneous TIL infiltration showed no difference in survival compared to those with uniformly low TIL infiltration.ConclusionOur study showed that CTL and NK cell-associated gene expression and protein levels differ significantly according to TIL levels and that CTL-NK score and distribution of TILs within tumors have a prognostic value. These findings emphasize the importance of CTLs and NK cells as well as the spatial uniformity of TIL infiltration in clinical outcome of TNBC patients, providing valuable insights for refining prognostic assessments and guiding immunotherapeutic strategies.

Impact of textural layers in soils on irrigation infiltration processes in an oasis farmland, northwestern China

AbstractThe Hexi Corridor Oasis in China is an important agricultural area supported by extensive irrigation. The sandy soil in the area exhibits textural layering, which has marked effects on its properties. Textural layered soils play an important role in distributing and regulating water regimes in irrigated fields. However, little is known about the textural layering‐dependent variability in soil water infiltration characteristics following irrigation and the potential effects on soil hydrology and water regimes in these soils. The objective of this study was to determine water infiltration processes and their influencing factors for six soil profile types in an oasis farmland. Six in situ constant‐head infiltration measurements (with three replications in each of six sites) were conducted with a single‐ring infiltrometer in different soil texture configurations, and Hydrus‐2D model was applied to evaluate soil water distribution. The results showed that the wetting front depth for all treatments was in the following order: ST‐1 > ST‐5 > ST‐6 > ST‐3 > ST‐2 > ST‐4; the stable infiltration rate ranged from 1.2 to 3.3 cm/min during the investigation period in all treatments. The presence of a finer‐textured soil layer decreased water infiltration rates through the profiles, and the thickness and burial depth of textured layers had obvious effects on infiltration, compared with homogeneous sand profiles. Presence of a medium‐textured layer affected the infiltration characteristics and soil water spatial variability; driven by increased field capacity and the wilting point, hydraulic conductivity was correlated with soil variables that were related to soil structure, such as wilting point and field capacity, and less so with variables that were related to soil texture, such as the clay fractions (p < 0.001). These results suggested that textural layering altered the soil structure and caused heterogeneous infiltration of water, with implications for the distribution of irrigation water and prevention of deep percolation in this oasis farmland. The result of this study offers a new analysis of ponded single‐ring water infiltrometer data that the soil water infiltration is also influenced by the type of textural layered than just the irrigation system, which supports crop irrigation management in study area and other regions with similar soil regimes.

Histologic, Phenotypic, and Molecular Characterization of Feline Hodgkin-Like Lymphoma With Classical Reed-Sternberg Cells.

Hodgkin-like lymphoma (HLL) is a rare neoplasm in cats that shares characteristics with the human disease. The hallmark of HLL is the presence of Reed-Sternberg (RS) cells expressing CD30 and CD20. This study aimed to elucidate the clinicopathologic features, immunophenotype and clonality patterns of feline HLL. A comprehensive retrospective review of clinicopathologic and molecular data of nodal lymphomas over a 6-year period was conducted in MyLav laboratory. Twenty-four cases were identified. All cats presented with submandibular or retropharyngeal lymphadenopathy. Histopathologic examination revealed a multifocal to diffuse proliferation of medium-to-large lymphoid cells with low mitotic activity, interspersed RS cells, and a heterogeneous inflammatory infiltrate comprising T-cells, plasma cells and neutrophils. In addition, extensive necrosis was a consistent finding. Immunohistochemistry showed a variable membranous CD20 and nuclear PAX5 expression in neoplastic cells, while RS cells displayed only mild to moderate CD20 positivity and were negative to PAX5. In 21/24 cases (87.5%), RS cells were diffusely CD30-positive. PARR analysis demonstrated clonal B-cell expansion in 60% of cases, with the remaining 40% exhibiting polyclonality. For the 10 cats with available follow-up, the prognosis was generally favourable, with only two cats succumbing to progressive disease. In conclusion, diagnosing feline HLL is challenging. The expression of CD30 and CD20 by RS cells should be considered a hallmark of the disease, but only after excluding differential diagnoses such as anaplastic B-cell lymphoma and granulomatous lymphadenopathy.

Integrative single-cell analysis of longitudinal t(8;21) AML reveals heterogeneous immune cell infiltration and prognostic signatures.

Immunotherapies targeting T cells in solid cancers are revolutionizing clinical treatment. Novel immunotherapies have had extremely limited benefit for acute myeloid leukemia (AML). Here, we characterized the immune microenvironment of t(8;21) AML patients to determine how immune cell infiltration status influenced prognosis. Through multi-omics studies of primary and longitudinal t(8;21) AML samples, we characterized the heterogeneous immune cell infiltration in the tumor microenvironment and their immune checkpoint gene expression. Further external cohorts were also included in this research. CD8+ T cells were enriched and HAVCR2 and TIGIT were upregulated in the CD34+CD117dim%-High group; these features are known to be associated with immune exhaustion. Data integration analysis of single-cell dynamics revealed that a subset of T cells (cluster_2) (highly expressing GZMB, NKG7, PRF1 and GNLY) evolved and expanded markedly in the drug-resistant stage after relapse. External cohort analysis confirmed that the cluster_2 T-cell signature could be utilized to stratify patients by overall survival outcome. In conclusion, we discovered a distinct T-cell signature by scRNA-seq that was correlated with disease progression and drug resistance. Our research provides a novel system for classifying patients based on their immune microenvironment.

CCDC58 is a potential biomarker for diagnosis, prognosis, immunity, and genomic heterogeneity in pan-cancer

Coiled-coil domain-containing 58 (CCDC58) is a member of the CCDC protein family. Similar to other members, CCDC58 exhibits potential tumorigenic roles in a variety of malignancies. However, there is no systematic and comprehensive pan-cancer analysis to investigate the diagnosis, prognosis, immune infiltration, and other related functions of CCDC58. We used several online websites and databases, such as TCGA, GTEx, UALCAN, HPA, CancerSEA, BioGRID, GEPIA 2.0, TIMER 2.0, and TISIDB, to extract CCDC58 expression data and clinical data of patients in pan-cancer. Then, the relationship between CCDC58 expression and diagnosis, prognosis, genetic alterations, DNA methylation, genomic heterogeneity, and immune infiltration level were determined. In addition, the biological function of CCDC58 in liver hepatocellular carcinoma (LIHC) was investigated. Pan-cancer analysis results showed that CCDC58 was differentially expressed in most tumors and showed excellent performance in diagnosis and prediction of prognosis. The expression of CCDC58 was highly correlated with genetic alterations, DNA methylation, and genomic heterogeneity in some tumors. In addition, the correlation analysis of CCDC58 with the level of immune infiltration and immune checkpoint marker genes indicated that CCDC58 might affect the composition of the tumor immune microenvironment. Enrichment analysis showed that CCDC58-related genes were mainly linked to mitosis, chromosome, and cell cycle. Finally, biological function experiments demonstrated that CCDC58 plays an important role in tumor cell proliferation and migration. CCDC58 was first identified as a pan-cancer biomarker. It may be used as a potential therapeutic target to improve the prognosis of patients in the future.

Bioinformatic validation and machine learning-based exploration of purine metabolism-related gene signatures in the context of immunotherapeutic strategies for nonspecific orbital inflammation.

Nonspecific orbital inflammation (NSOI) represents a perplexing and persistent proliferative inflammatory disorder of idiopathic nature, characterized by a heterogeneous lymphoid infiltration within the orbital region. This condition, marked by the aberrant metabolic activities of its cellular constituents, starkly contrasts with the metabolic equilibrium found in healthy cells. Among the myriad pathways integral to cellular metabolism, purine metabolism emerges as a critical player, providing the building blocks for nucleic acid synthesis, such as DNA and RNA. Despite its significance, the contribution of Purine Metabolism Genes (PMGs) to the pathophysiological landscape of NSOI remains a mystery, highlighting a critical gap in our understanding of the disease's molecular underpinnings. To bridge this knowledge gap, our study embarked on an exploratory journey to identify and validate PMGs implicated in NSOI, employing a comprehensive bioinformatics strategy. By intersecting differential gene expression analyses with a curated list of 92 known PMGs, we aimed to pinpoint those with potential roles in NSOI. Advanced methodologies, including Gene Set Enrichment Analysis (GSEA) and Gene Set Variation Analysis (GSVA), facilitated a deep dive into the biological functions and pathways associated with these PMGs. Further refinement through Lasso regression and Support Vector Machine-Recursive Feature Elimination (SVM-RFE) enabled the identification of key hub genes and the evaluation of their diagnostic prowess for NSOI. Additionally, the relationship between these hub PMGs and relevant clinical parameters was thoroughly investigated. To corroborate our findings, we analyzed expression data from datasets GSE58331 and GSE105149, focusing on the seven PMGs identified as potentially crucial to NSOI pathology. Our investigation unveiled seven PMGs (ENTPD1, POLR2K, NPR2, PDE6D, PDE6H, PDE4B, and ALLC) as intimately connected to NSOI. Functional analyses shed light on their involvement in processes such as peroxisome targeting sequence binding, seminiferous tubule development, and ciliary transition zone organization. Importantly, the diagnostic capabilities of these PMGs demonstrated promising efficacy in distinguishing NSOI from non-affected states. Through rigorous bioinformatics analyses, this study unveils seven PMGs as novel biomarker candidates for NSOI, elucidating their potential roles in the disease's pathogenesis. These discoveries not only enhance our understanding of NSOI at the molecular level but also pave the way for innovative approaches to monitor and study its progression, offering a beacon of hope for individuals afflicted by this enigmatic condition.

Morphological characteristics of in-stent restenosis with different degrees of area stenosis: an optical coherence tomography study.

The morphological characteristics of in-stent restenosis (ISR) in relation to varying degrees of area stenosis have not been comprehensively examined. This study aimed to explore the tissue characteristics of patients experiencing ISR with different degrees of area stenosis through the utilization of optical coherence tomography (OCT). In total, 230 patients with ISR who underwent OCT were divided into the following three groups: area stenosis (AS) < 70% (n = 26); 70-80% (n = 119) and AS ≥ 80% (n = 85). Among the 230 patients, the clinical presentation as stable angina was 61.5% in AS < 70%, followed by 47.2% in 70% < AS ≤ 80%, and 31.8% in AS ≥ 80% (P = 0.010). The OCT findings showed that heterogeneous neointima, ISNA, LRP, neointima rupture, TCFA-like pattern, macrophage infiltration, red and white thrombus was more common with AS increased. Ordinal logistic regression analysis showed that higher AS was associated with previous dyslipidemia (odds ratio [OR], 4.754; 95% confidence interval [CI], 1.419-15.927, P = 0.011), neointimal rupture (OR: 3.640; 95% CI, 1.169-11.325, P = 0.026), red thrombus (OR: 4.482; 95% CI, 1.269-15.816, P = 0.020) and white thrombus (OR: 5.259; 95% CI, 1.660-16.659, P = 0.005). Patients with higher degrees of area stenosis in the context of ISR exhibited a greater number of discernible morphological characteristics as identified through OCT analysis. Furthermore, previous dyslipidemia, neointimal rupture, white thrombus and red thrombus were highly associated with and the progression of ISR lesions.

Revolutionizing cancer treatment: the power of bi- and tri-specific T-cell engagers in oncolytic virotherapy.

Bi- or tri-specific T cell engagers (BiTE or TriTE) are recombinant bispecific proteins designed to stimulate T-cell immunity directly, bypassing antigen presentation by antigen-presenting cells (APCs). However, these molecules suffer from limitations such as short biological half-life and poor residence time in the tumor microenvironment (TME). Fortunately, these challenges can be overcome when combined with OVs. Various strategies have been developed, such as encoding secretory BiTEs within OV vectors, resulting in improved targeting and activation of T cells, secretion of key cytokines, and bystander killing of tumor cells. Additionally, oncolytic viruses armed with BiTEs have shown promising outcomes in enhancing major histocompatibility complex I antigen (MHC-I) presentation, T-cell proliferation, activation, and cytotoxicity against tumor cells. These combined approaches address tumor heterogeneity, drug delivery, and T-cell infiltration, offering a comprehensive and effective solution. This review article aims to provide a comprehensive overview of Bi- or TriTEs and OVs as promising therapeutic approaches in the field of cancer treatment. We summarize the cutting-edge advancements in oncolytic virotherapy immune-related genetic engineering, focusing on the innovative combination of BiTE or TriTE with OVs.

Genomic discordances and heterogeneous mutational burden, PD-L1 expression and immune infiltrates of non-small cell lung cancer metastasis

AimsTo investigate the genomic discordances and heterogeneous mutational burden, PD-L1 expression and immune cell (IC) infiltrates of non-small cell lung cancer (NSCLC) metastasis.MethodsSurgical samples from 41 cases of NSCLC with...

Lymphocyte-Infiltrated Periportal Region Detection With Structurally-Refined Deep Portal Segmentation and Heterogeneous Infiltration Features.

Goal: The early diagnosis and treatment of hepatitis is essential to reduce hepatitis-related liver function deterioration and mortality. One component of the widely-used Ishak grading system for the grading of periportal interface hepatitis is based on the percentage of portal borders infiltrated by lymphocytes. Thus, the accurate detection of lymphocyte-infiltrated periportal regions is critical in the diagnosis of hepatitis. However, the infiltrating lymphocytes usually result in the formation of ambiguous and highly-irregular portal boundaries, and thus identifying the infiltrated portal boundary regions precisely using automated methods is challenging. This study aims to develop a deep-learning-based automatic detection framework to assist diagnosis. Methods: The present study proposes a framework consisting of a Structurally-REfined Deep Portal Segmentation module and an Infiltrated Periportal Region Detection module based on heterogeneous infiltration features to accurately identify the infiltrated periportal regions in liver Whole Slide Images. Results: The proposed method achieves 0.725 in F1-score of lymphocyte-infiltrated periportal region detection. Moreover, the statistics of the ratio of the detected infiltrated portal boundary have high correlation to the Ishak grade (Spearman's correlations more than 0.87 with p-values less than 0.001) and medium correlation to the liver function index aspartate aminotransferase and alanine aminotransferase (Spearman's correlations more than 0.63 and 0.57 with p-values less than 0.001). Conclusions: The study shows the statistics of the ratio of infiltrated portal boundary have correlation to the Ishak grade and liver function index. The proposed framework provides pathologists with a useful and reliable tool for hepatitis diagnosis.

CAR T cell therapy: Current limitations and advancements in treating solid tumors

An emerging technology that tackles cancer via immunotherapy known as CAR T cell therapy has been gaining great popularity after its success in treating leukemia and other forms of blood cancer. However, major advancement to treat other forms of cancer have not been attained as this treatment is still facing major obstacles in addressing solid tumors due to tumor heterogeneity, hostile microenvironment, on-target/off-tumor toxicities, antigen specificity, and difficult infiltration. Moreover, this treatment has also been associated with threatening side-effects like mass dieoff of antibody producing B cells, CRS, and ICANS. Thus, because of simplified manipulation yet lack of knowledge of the physiology behind the mechanisms, significant progress has not yet been achieved-though on its way. Scientists explored multiple ways of addressing these major challenges by researching on feasible techniques, antigens, and mechanisms that can increase T cell reproduction, decrease toxicity, overcome and infiltrate into the hostile microenvironment, and target multiple antigens via TAA. It is a technology that is not yet fully developed; therefore, physicians still opt to target cancer via other forms of immunotherapies, adjuvant and neoadjuvant therapies including chemotherapy, drug consumption, and surgery. As clinical results for CAR T cell therapy have not presented prosperous results in targeting solid tumors, the future for research on solid tumors is still unclear. Scientists might either opt for other methods like microwave ablation, the use of TT fields, different combinations of current therapies to maximize efficiency, and ADCs.

EXTH-73. IN VIVO EFFECTS OF EPCAM-DIRECTED CAR T CELLS IN COMBINATION WITH PD-1 CHECKPOINT BLOCKADE IN LUNG CANCER BRAIN METASTASIS

Abstract BACKGROUND Metastatic lung cancer is incurable once it spreads to the brain. Chimeric antigen receptor (CAR) T cells show promise in hematologic malignancies but face challenges in solid tumors, particularly in brain tumors, due to a highly immunosuppressive tumor microenvironment (TME), tumor antigen (TA) heterogeneity and impeded infiltration and persistence at the tumor site. To test the efficacy of CAR T cells and to find strategies for potentially enhancing its efficacy, CAR T cells were applied with and without checkpoint-blockade to treat lung cancer brain metastasis. METHODS We used an immunocompetent syngeneic orthotopic cerebral metastasis model combined with a chronic cranial window and repetitive intracerebral two-photon laser scanning microscopy (TPLSM). This allowed us to observe fluorescent tumor cells and CAR T cells in vivo at a single cell level over time. We injected red fluorescent EpCAM-transduced Lewis Lung carcinoma cells (EpCAM-LL/2tdT) into the brain tissue, followed by injection of EpCAM-directed or mock CAR T cells. Animals received anti-PD-1 and the respective isotype control via intraperitoneal injections. RESULTS EpCAM-directed CAR T cells were generated and showed substantial anti-tumor effects in vitro. Local injection into EpCAM-LL/2 tumor-bearing mice led to intratumoral accumulation of CAR T cells, resulting in reduced tumor growth compared to undirected CAR T cells translating into long-term survival in a fraction of animals. However, intratumoral CAR T cell numbers decrease over time pointing towards insufficient persistence. Interestingly, anti-PD-1 treatment did not enhance intratumoral CAR T accumulation, persistence and anti-tumor efficacy. CONCLUSION Our findings demonstrate the great potential of cell-based treatment approaches for the treatment of lung cancer brain metastases and underlines the necessity of further investigations to enhance infiltration and persistence in solid tumors.

NIMG-31. AXONS INTERSECTING TUMOR ENHANCEMENT (AXITE) RADIOMICS: A NOVEL PREDICTOR FOR OVERALL SURVIVAL IN GLIOBLASTOMA

Abstract AIM The diffuse infiltration of glioblastoma (GBM) along the intricate network of white matter (WM) tracts poses a major challenge. Our aim is to investigate the potential of axons intersecting tumor enhancement (AXITE)-derived radiomic features in predicting one-year overall survival (OS), in comparison to conventional tumor-based radiomics. METHODS This study included data from the publicly available UCSF-PDGM-v2 (n=97). T1+C images and their corresponding tumor mask were rigidly registered to a common brain template (MNI152) using ANTs. WM tracts within the tumor-defined region of interest were generated using the HCP1065-1mm tractography atlas in DSI Studio Software, which mapped the AXITEs. Radiomic features were then extracted using PyRadiomics from subsequently masked AXITE maps as well as contrast-enhancing tumor masks for a comparative analysis. Four machine learning models, including a Random Forest, Support Vector Machine (SVM), Decision Tree, and Logistic Regression, were employed to predict OS using an 80/20 train/test split. RESULTS Among all models, SVM was the most effective in predicting OS. AXITE radiomics demonstrated remarkable precision (0.73 for OS &amp;lt; 1 yr and 0.78 for OS &amp;gt;1yr), accuracy (0.75), F1-Score (0.75), recall (0.75), and AUC of 0.84 in ROC analysis outclassing the performance of traditional tumor radiomics which had an accuracy (0.60), precision (0.66), F1-Score (0.56) and AUC of 0.60. CONCLUSIONS AXITE Radiomics holds promising potential in capturing tumoral heterogeneity and white matter tract infiltration of GBM, demonstrating superior prediction over traditional tumor radiomics. Our approach presents a more accurate representation of tumor invasiveness and tumor resistance associated with OS more accurately and promises a potential tool for improving patient prognosis and patient management.

Next Generation of Brain Cancer Nanomedicines to Overcome the Blood–Brain Barrier (BBB): Insights on Transcytosis, Perivascular Tumor Growth, and BBB Models

AbstractBrain cancers, particularly malignant gliomas such as glioblastoma, are highly invasive and characterized by elevated complexity, heterogeneity, and high infiltration ability. Therefore, they pose a significant challenge to conventional treatments due to the limited drug permeability of the blood–brain barrier (BBB), the involvement of numerous acquired and intrinsic drug resistance mechanisms in metastatic brain tumors, and the high sensitivity of surrounding healthy tissues. Despite recent advances in diagnosis and treatment, their prognosis remains poor, with their median overall survival rarely exceeding 12 months. To overcome these limitations, different nanomedicine‐based therapeutic approaches have recently been proposed, aiming to provide more effective and safer drug delivery for targeting brain cancers. However, most reported nanomedicines to date have failed to meet the high expectations in the clinic. This fact can be attributed to limited understanding of brain tumor biology and lack of knowledge about bio‐nanoparticle interactions, among other factors. This review discusses recent progress in brain cancer nanomedicines, with a particular focus in understanding intracellular sorting mechanisms, perivascular tumor growth, and the design of advanced BBB models. It also highlights how an improved understanding of brain tumor biology can pave the way for designing safer and more effective nanomedicines for brain cancer treatment.

HEV-associated dendritic cells are observed in metastatic tumor-draining lymph nodes of cutaneous melanoma patients with longer distant metastasis-free survival after adjuvant immunotherapy.

Tissue biomarkers that aid in identifying cutaneous melanoma (CM) patients who will benefit from adjuvant immunotherapy are of crucial interest. Metastatic tumor-draining lymph nodes (mTDLN) are the first encounter site between the metastatic CM cells and an organized immune structure. Therefore, their study may reveal mechanisms that could influence patients´ outcomes. Twenty-nine stage-III CM patients enrolled in clinical trials to study the vaccine VACCIMEL were included in this retrospective study. After radical mTDLN dissection, patients were treated with VACCIMEL (n=22) or IFNα-2b (n=6), unless rapid progression (n=1). Distant Metastasis-Free Survival (DMFS) was selected as an end-point. Two cohorts of patients were selected: one with a good outcome (GO) (n=17; median DMFS 130.0 months), and another with a bad outcome (BO) (n=12; median DMFS 8.5 months). We analyzed by immunohistochemistry and immunofluorescence the expression of relevant biomarkers to tumor-cell biology and immune cells and structures in mTDLN, both in the tumor and peritumoral areas. In BO patients, highly replicating Ki-67+ tumor cells, low tumor HLA-I expression and abundant FoxP3+ lymphocytes were found (p=0.037; p=0.056 and p=0.021). In GO patients, the most favorable biomarkers for prolonged DMFS were the abundance of peri- and intra-tumoral CD11c+ cells (p=0.0002 and p=0.001), peri-tumoral DC-LAMP+ dendritic cells (DCs) (p=0.001), and PNAd+ High Endothelial Venules (HEVs) (p=0.004). Most strikingly, we describe in GO patients a peculiar, heterogeneous structure that we named FAPS (Favoring Antigen-Presenting Structure), a triad composed of DC, HEV and CD62L+ naïve lymphocytes, whose postulated role would be to favor tumor antigen (Ag) priming of incoming naïve lymphocytes. We also found in GO patients a preferential tumor infiltration of CD8+ and CD20+ lymphocytes (p=0.004 and p=0.027), as well as peritumoral CD20+ aggregates, with no CD21+ follicular dendritic cells detected (p=0.023). Heterogeneous infiltration with CD64+CD68-CD163-, CD64+CD68+CD163- and CD64+CD68+CD163+ macrophages were observed in both cohorts. The analysis of mTDLN in GO and BO patients revealed marked differences. This work highlights the importance of analyzing resected mTDLN from CM patients and suggests a correlation between tumor and immune characteristics that may be associated with a spontaneous or vaccine-induced long DMFS. These results should be confirmed in prospective studies.

Spatial distribution and functional relevance of FGFR1 and FGFR2 expression for glioblastoma tumor invasion

Glioblastoma is the most lethal brain cancer in adults. These incurable tumors are characterized by profound heterogeneity, therapy resistance, and diffuse infiltration. These traits have been linked to cancer stem cells, which are important for glioblastoma tumor progression and recurrence. The fibroblast growth factor receptor 1 (FGFR1) signaling pathway is a known regulator of therapy resistance and cancer stemness in glioblastoma. FGFR1 expression shows intertumoral heterogeneity and higher FGFR1 expression is associated with a significantly poorer survival in glioblastoma patients. The role of FGFR1 in tumor invasion has been studied in many cancers, but whether and how FGFR1 mediates glioblastoma invasion remains to be determined. Here, we investigated the distribution and functional relevance of FGFR1 and FGFR2 in human glioblastoma xenograft models. We found FGFR1, but not FGFR2, expressed in invasive glioblastoma cells. Loss of FGFR1, but not FGFR2, significantly reduced cell migration in vitro and tumor invasion in human glioblastoma xenografts. Comparative analysis of RNA-sequencing data of FGFR1 and FGFR2 knockdown glioblastoma cells revealed a FGFR1-specific gene regulatory network associated with tumor invasion. Our study reveals new gene candidates linked to FGFR1-mediated glioblastoma invasion.

MTHFR act as a potential cancer biomarker in immune checkpoints blockades, heterogeneity, tumor microenvironment and immune infiltration

PurposeTo evaluate the role and landscape of 5-10-Methylenetetrahydrofolate reductase (MTHFR) to immune infiltration, tumor microenvironment, heterogeneity, immune checkpoints blockades, prognostic significance across cancer types.MethodsData sets of genomic, transcriptomic and clinic features of MTHFR across > 60,000 patients and up to 44 cancer types were comprehensively analyzed using R software.ResultsExpression of MTHFR gene is significantly lower in 17 tumors and correlated with overall survival (OS), disease-specific survival (DSS), progression-free interval (PFI) in specific tumors. Gene alterations of MTHFR are observed significant differences across tumor types. Expression of MTHFR is negatively correlated with the stemness index (mDNAsi, mRNAsi, DMPsi, ENHsi, EREG-mDNAsi and EREG-mRNAsi) in the most cancers. MTHFR showed significantly correlated with 67 types of immune cell infiltration scores in 44 cancer types by XCELL algorithm. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis are conducted to show the core tumor mechanism and biological process. Correlations between MTHFR and biomarkers of heterogeneity (MSI, TMB, MATH, HRD, LOH, Neoantigen, ploidy and purity) are also significant in specific tumors. MTHFR is significantly positively correlated with biomarkers of immune related genes (CD19, CD274, CD80, CD86) and mismatched repair genes (MLH1, PMS2, MSH2, MSH6, EPCAM, MLH3, PMS1, EXO1) in most cancer types. Receiver Operating Characteristics (ROC) analyses show MTHFR could act as a potential biomarker in anti-PD-1 (nivolumab to melanoma) and anti-CTLA4 (ipilimumab to melanoma) group of ontreatment, in anti-PD-1 (pembrolizumab to melanoma) group of pretreatment. Two immunohistochemistry antibodies HPA076180 and HPA077255 are verified in 20 types of tumor and could be used to detect the expression of MTHFR efficiently in clinic.ConclusionsMTHFR could predict the response of immune checkpoints blockades, heterogeneity, tumor microenvironment and immune infiltration.

CD73/adenosine pathway-related gene signature for intratumor heterogeneity and immune infiltration in prognosis of lung adenocarcinoma.

e20524 Background: Adenosine is an immunosuppressive molecule that can inhibit the activity of immune cells, particularly T cells, and promote tumor growth. CD73, also known as ecto-5'-nucleotidase, is a cell-surface enzyme that plays a role in the adenosine pathway can catalyzes the conversion of AMP to adenosine. Establish and evaluate a prognosis model based on CD73/ Adenosine pathway will help to understanding of the biological mechanisms in lung adenocarcinoma (LUAD) and providing more accurate predictions for patients' treatment. Methods: The RNA sequencing (RNA-seq), whole exome sequencing (WES) and clinical data were obtained from LUAD cohorts of the Cancer Genome Atlas (TCGA). CD73/ Adenosine pathway-related genes were identified by screening for genes that were in both the CD73 STRING network (confidence = 0.9, 1st shell ≤50 interactors) and the KEGG purine metabolism pathway gene list(map00230). Least absolute shrinkage and selection operator (LASSO) regression was conducted to develop a CD73/ Adenosine pathway-related risk score (CD73RS). Time-dependent ROC curve and survival analysis were conducted in LUAD-TCGA cohort to verify the predictive performance of CD73RS. The MATH (Mutant-Allele Tumor Heterogeneity) ITH scores were calculated to evaluate intratumor heterogeneity (ITH) in LUAD-TCGA WES "maf" files using the R package "maftools" and the "math.score" function. TIMER 2.0 database(http://timer.cistrome.org/) was conducted to estimation of tumor microenvironment to further understand the molecular features of CD73RS. An independent cohort (GSE72094) was used to verify the prognostic effect of CD73RS. Results: A prognosis prediction model based on 21 CD73/ Adenosine pathway-related genes was established using LASSO regression. Kaplan-Meier survival analysis of the risk model from LUAD-TCGA cohort showed that the high-risk group had a significantly worse prognosis (HR = 3.298, P = 1.26e-08). The area under the curve (AUC) in time-dependent ROC curve at 1, 3 and 5 years were turn out to be 0.794, 0.722 and 0.686, respectively. The validation set also showed worse prognosis in high-risk group(HR = 1.473,p = 0.0403). In the high-risk group, there was a higher ITH score compared to the low-risk group (p = 0.0094), along with lower infiltration levels of B cells(p = 7.23e-15), CD4+ T cells(p = 0.003), macrophage cells(p = 0.016), and myeloid dendritic cells(p = 0.037). Conclusions: We established and validated a CD73/ Adenosine pathway risk model that exhibits a robust predictive performance for the outcome of patients with LUAD. The CD73/Adenosine pathway has been shown to play a role in both self-driven genomic changes, such as intra-tumor heterogeneity (ITH), as well as non-autonomous processes, such as immune microenvironment adaptation, which contribute to tumor plasticity and development.

Heterogeneous baseline immune cell infiltration landscape as a predictor of pathological complete response in locally advanced esophageal squamous cell carcinoma (ESCC) following neoadjuvant chemotherapy and immunotherapy: Results from a single-arm, phase II clinical trial (SEEK-01).

4047 Background: Neoadjuvant immunotherapy and chemotherapy have shown promise in improving outcomes for patients with locally advanced ESCC, but the response to treatment can vary among patients. Therefore, there is a need to identify predictive biomarkers that can guide treatment decisions and improve patient outcomes. The infiltration of immune cells within the tumor microenvironment has been shown to be an important factor in cancer progression and response to treatment. In this study, we sought to explore the immune microenvironment prospectively related to treatment response of Neoadjuvant immunotherapy by scRNAseq. Methods: This was a single-arm phase II clinical trial, planned to enroll 20 locally advanced ESCC patients (cT2-4N0-1M0). After neoadjuvant chemotherapy (paclitaxel-albumin + carboplatin) combined with tislelizumab, patients received minimal invasive esophagectomy (MIE).Prior to neoadjuvant therapy, fresh tumor tissues were obtained via endoscopy and used for scRNAseq analysis, to evaluate the predictive capacity of the immune microenvironment. Pathological outcomes were assessed and divided into pCR, MPR defined as &lt;10% residual tumor excluding pCR, and non-MPR defined as &gt;10%. Results: Out of 18 enrolled patients, 15 completed neoadjuvant therapy and MIE surgery since March 2022. Of these 15 patients, 3 achieved pCR, 2 got MPR, and 10 were non-MPR, with a total of 51,208 single cells sequenced. Both groups showed similarly low expression of PD-L1 (5.7% vs. 2.3%). Comparing pCR to non-MPR patients, the former showed a significant increase in T lymphocytes (CD8+(17.1% vs.1.1%), Regulatory T(20.2% vs.4.0%)), plasma cell(22.0% vs.8.3%), and mature dendritic cell(1.5% vs.0.5%); a significant decrease in B cells(0.9% vs. 1.9%), neutrophils(1.7% vs.11.0%), and tumor cell, as well as a similar expression of macrophage(5.6% vs.6.6%) and monocyte, indicating a clear difference in the immune cell infiltration landscape between two groups. Furthermore, we identified a new subtype of plasma cell which was highly enriched in the non-MPR group, even though the pCR group showed a great increase in plasma cells in total. Conclusions: This study demonstrates the heterogeneity of baseline immune cell infiltration landscape in Locally Advanced ESCC, in which pCR and non-MPR patients exhibited entirely different profiles, and provide a potential predictor of response to neoadjuvant immunotherapy. Our findings suggest that baseline immune cell infiltration status, rather than pd-L1 expression, could accurately predict pCR or non-MPR before treatment, which could guide personalizing and minimize the need for trial-and-error approaches.

KIF22 in the Prognosis and Immune Biomarking of Pan-Cancer

KIF22, also known as kinesin-like DNA-binding protein (Kid), is a member of the Kinesin superfamily proteins (KIFs). Available evidence indicated that KIF22 was associated with cancer occurrence and development. However, the functions and underlying mechanisms of KIF22 in carcinogenesis and cancer progression remain largely unknown. In this study, we examined the expression profile and methylation status of KIF22 in different cancers, as well as its associations with prognosis, tumor stemness, genomic heterogeneity, immune evasion, immune infiltration, and therapeutic response in various tumor types. The results demonstrated that the expression level of KIF22 was higher in tumors than nontumor tissues and had strong relationships with prognosis, genomic heterogeneity, tumor stemness, neoantigen, ESTIMATE, and immune infiltration. KIF22 methylation status showed strong relationships with immunomodulators and chemokines. KIF22 had a significant relevance with drug susceptibility and could be a useful biomarker for forecasting survival probability and therapeutic reaction. Furthermore, KIF22 interaction and coexpression networks were mainly involved in cell division, cell cycle, DNA repair, and antigen processing and presentation. KIF22 could be used as a pan-cancer biomarker for clinical diagnosis, therapeutic schedule, prognosis, and cancer monitoring.