bluuebunny/crossref_metadata_2025 huggingface dataset:
New Technologies in the Search for Dark Matter
Authors: Clark Kenneth , Piro Marie-Cécile , Giroux Guillaume , Savvidis George
Published: February 2025

The ongoing search for dark matter has prompted the exploration of parameter spaces with masses lighter than the traditional 10 GeV range. However, as experiments extend into this low-energy range, detectors encounter significant challenges related to background events. Innovative solutions have been proposed to tackle these hurdles, including developing new detector technologies and dedicated calibration methods. This paper delves into the nature of these emerging background sources and outlines current and future efforts to address these challenges.
Role of nitrogen cycling functional genes and their key influencing factors in eutrophic aquatic ecosystems
Authors: Wang Wenwen , Li Mengze , Chen Peng , Yuan Shengwu , Wang Kun , Wang Shuhang , Jiang Xia
Published: February 2025

The nitrogen cycle, essential for the transformation and circulation of nitrogen on earth, involves key processes such as nitrogen fixation, nitrification, denitrification, and ammonification. Microorganisms are crucial in these processes, influencing water quality through energy conversion and nitrogen cycling. Eutrophication, driven by human activities, increases exogenous nitrogen input, accelerating nitrogen cycling and loss, boosting greenhouse gas emissions, and impacting aquatic ecosystems. Functional genes in the nitrogen cycle, such as amoA (nitrification), nirS, nirK, nosZ (denitrification), and hzs and hzo (anaerobic ammonium oxidation), are indicators of nitrogen transformation in sediments. Environmental factors like temperature, dissolved oxygen (DO), organic matter content, nitrogen levels, pH, and salinity significantly influence these genes' expression and regulation. For example, temperature changes can affect nitrifying and denitrifying bacteria activities, DO levels impact microbial growth and metabolism, and higher organic matter content stimulates the expression of nitrogen cycle genes. Understanding how these environmental factors affect nitrogen cycling genes is crucial for addressing eutrophication in aquatic ecosystems. This review helps elucidate the adaptability and responses nitrogen-associated functional microorganisms and genes environmental changes, offering theoretical insights and practical guidance for sustainable ecosystem management.
Thermal fluctuation effects on shear viscosity to entropy ratio in five-dimensional Kerr–Newman black holes
Authors: Pourhassan Behnam , Sakallı İzzet , Brzo Aram Bahroz
Published: February 2025

We investigate how thermal fluctuations affect the properties of five-dimensional Kerr–Newman black holes, focusing particularly on the shear viscosity to entropy ratio. Our analysis incorporates logarithmic corrections to the Bekenstein–Hawking entropy and examines their impact on black hole thermodynamics. We explore three approaches to studying the shear viscosity-entropy ratio in the presence of thermal fluctuations: considering independent shear viscosity, thermally corrected shear viscosity, and an independent ratio assumption. Notably, we find that the lower bound of

$$\eta /S \ge 1/4\pi $$

η
/
S
≥
1
/
4
π

remains valid even with thermal fluctuations, though the specific behavior depends on the black hole mass and correction parameter. Our results suggest that thermal fluctuations generally decrease the ratio for massive black holes while maintaining the universal lower bound. This work extends our understanding of quantum corrections to black hole transport properties.
Cosmic ray sources and detectors
Authors: Mohanty Pravata Kumar
Published: February 2025

The origin of cosmic rays has remained an enduring mystery in astrophysics since their discovery by Victor Hess in 1912. However, extensive studies on the energy spectrum, mass composition, and angular distributions of cosmic rays have been conducted through both space-based and ground-based experiments. Although space-based experiments are limited to measuring cosmic rays with energies below about 10

$$^{14}$$

14

eV, their high precision has led to the discovery of several new features in the cosmic ray spectrum. However, ground-based detector arrays have provided valuable insights into cosmic rays at higher energies, offering new information on their spectrum, composition, and angular distributions. This paper reviews the major cosmic ray detectors currently in operation, both in space and on the ground, followed by an in-depth look at the GRAPES-3 experiment, which operates in Ooty, India, and focuses on the TeV-PeV energy range. Recent findings from GRAPES-3 on the cosmic ray spectrum and anisotropy are discussed, along with an update on the ongoing upgrade of the experiment.
A Method for Evaluating the Susceptibility of Hydrodynamic Landslides Based on Subcomponent Combination Prediction
Authors: Wang LiYing , Wang LinFeng
Published: February 2025

In order to accurately predict the probability of hydrodynamic landslide susceptibility, realize the effective assessment of landslide susceptibility, provide scientific reference basis and risk management tools for disaster management departments, and improve the disaster emergency response capability, a hydrodynamic landslide susceptibility evaluation method based on the combination of sub-indicators is proposed. The index system of hydrodynamic landslide susceptibility evaluation is constructed, and the geographic conditions, hydrological conditions and human activities in the index system are taken as sub-indicators, and the sub-indicator prediction model is established to predict the probability value of hydrodynamic landslide susceptibility corresponding to each sub-indicator; the hierarchical analysis method and entropy method are used to obtain the subjective and objective weights of hydrodynamic landslide susceptibility evaluation indexes respectively, and then the two of them are combined to get the comprehensive weights of evaluation indexes; the predicted probability values of the obtained sub-prediction of hydrodynamic landslide susceptibility and the comprehensive weights of the evaluation indicators are combined together, and the quantitative hydrodynamic landslide susceptibility evaluation results are obtained by utilizing the linear weighting method. Experiments have proved that this method can accurately assess the susceptibility of hydrodynamic landslides, and the sensitivity of perception can reach more than 95%.
Innovative Application and Practice of Intelligent Robots Enabled by the Internet of Things (IoT) in Basketball Teaching
Authors: Tang Dailin
Published: February 2025

This paper aims to explore the application effect of intelligent robots enabled by the Internet of Things (IoT) in basketball teaching, to improve the quality and efficiency of basketball teaching. By choosing the basketball courses of two schools as practical cases, the paper deploys an intelligent robot teaching system and carries out a three-month teaching experiment. A variety of evaluation methods such as questionnaire survey, skill test, learning performance comparison and teacher feedback are used to evaluate the effect of an intelligent robot teaching system. The level of students’ basketball skills is improved, and their satisfaction with the intelligent robot teaching system is also high. Teachers believe that the system reduces the teaching burden and improves teaching efficiency. Therefore, this paper believes that intelligent robots enabled by IoT have significant value in basketball teaching, which can improve students’ skill levels and improve teaching efficiency.
Detection of Fan Blade Defects Based on Improved YOLOv8n
Authors: Wang Xu , Xu Wenfeng , Cui Qingyu , Yin Xiaolong , Hao Litong , Zhang Hao , Wang Hainan
Published: February 2025

As the core component of the wind turbine, blades are susceptible to deterioration caused by natural environmental factors. This can manifest as erosion, fissures, and gel coat detachment, which collectively impair the efficiency of wind power generation and the safe operation of the turbine. In order to address the issue of low detection accuracy for wind turbine blade defects in complex environments, an enhanced YOLOv8n wind turbine blade defect detection algorithm has been proposed. First, the Large Separable Kernel Attention (LSKA) attention mechanism is introduced into the Spatial Pyramid Pooling Fast (SPPF) module of the backbone network, thereby enhancing the network’s attention and improving the model’s feature extraction capability. Second, the neck employs a weighted bidirectional feature pyramid (Bi-FPN) structure and integrates a P2 small target detection layer, thus enhancing the model’s multi-scale feature fusion capability and improving its small target detection accuracy. Finally, the loss function of the original model is optimized using WIoU, thereby improving the model’s defect detection accuracy. The results of the defect detection experiments on wind turbine blade images demonstrate that the accuracy of the proposed method has been enhanced by 7.9%, the mAP 50 has been improved by 2.6%, and the number of parameters has been reduced by 23%.
Comprehensive Evaluation Method of Development Mode Transformation Level of Resource-Based Cities Based on Multi-Source Image Data
Authors: Zhu Yuanjiao
Published: February 2025

In order to improve the comprehensive evaluation ability of the transformation level of resource-based city development models, this paper proposes a comprehensive evaluation method based on multi-source image data. This method reconstructs a geographic vector field dataset based on the comprehensive evaluation features of resource-based urban development model transformation level through geographic vector field grid partitioning, block feature fusion analysis, urban edge area planning, and grid area overlay analysis. At the same time, by combining multi-source remote sensing data detection, global directional change information recombination, remote sensing image detection technology, and regional information recombination technology, this method can accurately identify the key features of the transformation of resource-based urban development models. The test results show that this method has high accuracy and comprehensiveness, solving the problems of traditional evaluation methods such as single data sources and limited evaluation dimensions. It provides strong support for the sustainable development of resource-based cities and is expected to play an important role in urban planning, policy formulation, and urban development monitoring in the future.
Luminescence properties of CdSe/PVA nanocomposite
Authors: Upadhyay Priyanka , Kuraria R. K. , Kuraria Shashi R.
Published: February 2025

An environmentally safe chemical process was used to create the CdSe nanocomposite films in a polyvinyl alcohol (PVA) matrix. X-ray diffraction was used to analyze these composites, revealing the cubic zinc blend structure of CdSe/PVA nanocomposite with crystal sizes as small as a few nm. These composite films have had their photoluminescence (PL) properties examined. At 420[Formula: see text]nm, the PL peak of CdSe/PVA was detected. The CdSe/PVA nanocomposite’s mechanoluminescence properties were investigated. It is found that the impact velocity determines the ML intensities. When ML is produced impulsively by the impact of a moving piston on the nanocomposite, two peaks in ML intensity arise with time, and it is found that the mechanoluminescence intensity is dependent on the magnitude of impact velocity. ML peak intensities of the first and second peaks ([Formula: see text] and [Formula: see text] increase with increasing impact velocity. However, the time corresponding to the first and second maxima ([Formula: see text] and [Formula: see text] shifts towards shorter time values with greater impact velocity. We examined the voltage-brightness and voltage-current characteristics curves for our electroluminescence investigations of CdSe/PVA nanocomposite. At higher frequencies, EL with greater brightness at low threshold voltage has been seen.
On the Limit Cycles Bifurcating from the Periodic Orbits of a Hamiltonian System
Authors: Anacona Gerardo H. , Llibre Jaume , Freitas Bruno
Published: February 2025

This paper concerns the weak 16th Hilbert problem and considers the Hamiltonian center [Formula: see text], [Formula: see text], and we perturb it by all polynomials of degree [Formula: see text] for [Formula: see text]. We prove that the maximum number of limit cycles that can bifurcate from the periodic orbits of this center for [Formula: see text], under the mentioned perturbations and using the averaging theory of first order, is [Formula: see text], respectively.
An Image Encryption Algorithm Based on HNN with Memristor
Authors: Liu Yian , Hu Hao , Gao Ya , Hu Shaogang , Yu Qi , Chen Tupei , Liu Yang
Published: February 2025

This paper proposes an innovative approach to designing image encryption hardware by leveraging the Negative-resistance Memristor-based Hopfield Neural Network (NMHNN) model. In this method, the conventional Hopfield Neural Network (HNN) is modified by substituting one of its synaptic weights with a negative-resistance memristor model. This modification demonstrates enhanced complex dynamics while maintaining a simplified structure. The NMHNN generates a chaotic sequence, which serves as a key for image encryption and decryption through a confusion–diffusion process. As a result, this approach significantly improves the efficiency of both image encryption and decryption processes. To validate the practical feasibility of this memristor-based neural network for encryption, a 3-neuron HNN circuit with one [Formula: see text] memristor is constructed and tested. The hardware experiment demonstrates strong resilience against statistical analysis and entropy attacks. Notably, the operational efficiency of the proposed method is demonstrated to be 23 times greater than that of the Advanced Encryption Standard (AES), highlighting its substantial effectiveness and potential for hardware implementation in the field of image encryption.
Saddle–Node Bifurcation Leading to Survival Mosquitoes
Authors: Franco-Pérez L. , Núñez-López M.
Published: February 2025

A saddle–node bifurcation is demonstrated in a model of mosquito population growth. This bifurcation helps us understand the dynamics of two stages in the model: the aquatic stage (which includes the egg, larva, and pupa) and the nonaquatic stage (the mature mosquito). We calculated the basic offspring number using the next-generation matrix method and displayed it as the value of the saddle–node bifurcation. This means that the stable extinction state becomes unstable, while a survival state emerges as dominant in the dynamics, along with another coexistence state as a transition to surveillance. Numerical computations verify how factors such as limited logistic growth, the Allee effect, and the mortality density rate modify the mosquito dynamics.
The Existence of Traveling Fronts of a Generalized Reaction–Diffusion Equation
Authors: Zhang Panpan , Wu Kuilin
Published: February 2025

In this paper, we study the traveling wave solutions of a generalized reaction–diffusion system based on the classical Fisher-type system. Through qualitative analysis and blow-up techniques, we prove the existence of traveling fronts of the system. Moreover, we detect the limit points (i.e. [Formula: see text]-limit points or [Formula: see text]-limit points) of all traveling wave solutions by analyzing the global topological phase portraits of the equivalent system.
Multiparameter Bifurcation and Stability Analysis in Resource Allocation Problems of Swarm Robotic Systems
Authors: Zhang Xin , Sun Yi
Published: February 2025

Resource allocation is a pivotal concern in swarm robotic systems. This study delves into the chaotic dynamics within the yield game process of resource-providing robots in such systems, analyzing system bifurcation and stability across multiple parameters. We propose a market-based model for swarm robotic systems, employing game theory to formulate a yield game model among resource-providing robots. Through numerical analysis and simulation, we identify stable regions of system parameters and present bifurcation diagrams under varied parameters. Our findings indicate that increasing cost parameters or decreasing decision parameters stabilizes the yield and utility of resource-providing robots, while the opposite destabilizes them. Moreover, we examine the intricate influence of cost parameters on system stability by comparing linear and exponential resource cost functions. Results reveal that exponential cost functions lead to heightened system chaos and parameter sensitivity. These insights offer crucial theoretical groundwork and practical directions for the development and deployment of swarm robot systems.
Antitumor Effect of Apilactobacillus kosoi 10H by Inducing Immunostimulatory Macrophages
Authors: Takahashi Fuka , Chiou Tai-Ying , Tsukamoto Minako , Moriyama Satsuki , Takahashi Tomoya , Tanaka Sachi
Published: February 2025

Apilactobacillus kosoi (A. kosoi) 10H is a fructophilic lactic acid bacterium found in vegetable sugar fermentation liquid. It has been shown to possess high intestinal immunostimulatory activity. In this study, we investigated the potential for A. kosoi 10H heat‐killed cells and their components to convert the phenotype of immunosuppressive M2 macrophages to immunostimulatory M1 macrophages. We further investigated the ability of induction of apoptosis of cancer cell lines by macrophage culture medium in the presence of A. kosoi 10H heat‐killed cells. After induction of macrophage differentiation by phorbol 12‐myristate 13‐acetate in a THP‐1 human monocyte–derived cell line, immunosuppressive M2 macrophages were induced by interleukin‐6 (IL‐6). When A. kosoi 10H heat‐killed cells were brought into contact with IL‐6‐induced M2 macrophages, the expression of CD163 and CD209, which are M2 macrophage markers, markedly decreased, and the expression of CD80, human leukocyte antigen‐DR isotype (HLA‐DR), and tumor necrosis factor‐α (TNF‐α), markers of M1 macrophages, markedly increased. A similar effect was observed with a water‐soluble extract and a hydrophobic liquid extract from A. kosoi 10H heat‐killed cells. In addition, when M2 macrophage culture medium that had been conditioned with A. kosoi 10H heat‐killed cells was added to the MCF‐7 breast cancer cell line, we found that apoptosis was induced. TNF‐α in the medium conditioned with macrophages cultured in the presence of A. kosoi 10H heat‐killed cells was found to be partially responsible for MCF‐7 cell death. This study presents basic data on the potential to use lactic acid bacteria to improve the cancer microenvironment.
Prevalence of Obstructive Sleep Apnea Among Lebanese Patients With Chronic Kidney Disease: Its Repercussion on Disease Trajectory and Its Effect on Patients’ Quality of Life
Authors: Yakdan Salim , Rahhal Nazih , Al Chaar Soltan , Alhaddad Juliano , Al Akoum Monifa , Chahine Yaacoub , Najem Robert , Chahine Mirna N.
Published: February 2025

Background and Objectives: Chronic kidney disease (CKD) remains a public health threat and a major cause of morbidity and mortality worldwide. A bidirectional relationship is found between sleep disorders and CKD worldwide. However, to our knowledge, this study is the first to assess the prevalence of obstructive sleep apnea (OSA) and to evaluate its impact on the progression of other comorbidities among Lebanese patients with CKD.

Materials and Methods: The study is an observational cross‐sectional study, carried out between September and November 2021. Lebanese patients with any stage of CKD were included. Patients’ characteristics were collected via electronic health record and baseline questionnaires. We screened for obstructive sleep apnea using the STOP‐Bang questionnaire.

Results: We included 168 patients. The prevalence of OSA among our patients was 47.6%. The prevalence of OSA is higher in males compared with females (81.2% vs. 18.8%, p = 0.002). Obesity was more prevalent in patients with OSA compared with patients without OSA (42.5% vs. 19.3%, p = 0.002). Among the 168 patients, 69.6% had hypertension, with a significantly higher prevalence among those with OSA compared with those without OSA (81.2% vs. 59.1%, p = 0.003). Patients with OSA reported significantly lower scores compared with those without OSA in several domains of physical and emotional health, including physical functioning (54.06 vs. 66.88, p = 0.002), role limitations due to physical health (42.19 vs. 63.07, p = 0.001), role limitations due to emotional problems (49.17 vs. 69.32, p = 0.004), pain (61.31 vs. 70.45, p = 0.019), and physical component score (52.53 vs. 69.53, p = 0.002). All the abovementioned parameters were also examined in two subpopulations: patients with CKD and ESRD. Similarly, some comorbidities and a lower physical QOL score were observed more in patients with OSA in these two subpopulations.

Conclusion: Patients with OSA in our study have higher probability of being male, obese, and hypertensive as well as poorer QOL compared with their counterparts without OSA. Implementing more effective screening and treatment of OSA in CKD patients is necessary.
Intratumoral Fusobacterium nucleatum Recruits Tumor-Associated Neutrophils to Promote Gastric Cancer Progression and Immune Evasion
Authors: Zhang Tianhao , Li Ying , Zhai Ertao , Zhao Risheng , Qian Yan , Huang Zhixin , Liu Yinan , Zhao Zeyu , Xu Xiang , Liu Jianqiu , Li Zikang , Liang Zhi , Wei Ran , Ye Linying , Ma Jinping , Wu Qingping , Chen Jianhui , Cai Shirong
Published: February 2025

Intratumoral microbiota can impact the development and progression of many types of cancer, including gastric cancer (GC). A better understanding of the precise mechanisms by which microbiota support GC could lead to improved therapeutic approaches. Here, we investigated the effect of intratumoral microbiota on the tumor immune microenvironment (TIME) during GC malignant progression. Analysis of human GC tissues with 16S rRNA amplicon sequencing revealed that Fusobacterium nucleatum (F. nucleatum) was significantly enriched in GC tissues with lymph node metastasis and correlated with a poor prognosis. F. nucleatum infection spontaneously induced chronic gastritis and promoted gastric mucosa dysplasia in mice. Furthermore, GC cells infected with F. nucleatum showed accelerated growth in immunocompetent mice compared to immunodeficient mice. Single-cell RNA sequencing uncovered that F. nucleatum recruited tumor-associated neutrophils (TANs) to reshape the tumor immune microenvironment. Mechanistically, F. nucleatum invaded GC cells and activated IL-17/NF-κB/RelB signaling, inducing TAN recruitment. F. nucleatum also stimulated TAN differentiation into the pro-tumoral subtype and subsequent promotion of PD-L1 expression, further facilitating GC immune evasion while also enhancing the efficacy of anti-PD-L1 antibody therapy. Together, this data uncovers mechanisms by which F. nucleatum affects GC immune evasion and immunotherapy efficacy, providing insights for developing effective treatment strategies.
Germline pathogenic DROSHA variants are linked to pineoblastoma and Wilms tumor predisposition
Authors: Fiorica Peter N. , Golmard Lisa , Kim Jung , Bao Riyue , Lin Frank Y. , Roy Angshumoy , Pribnow Allison , Perrino Melissa R. , Masliah-Planchon Julien , Michalak-Provost Sophie , Wong Jennifer , Filser Mathilde , Stoppa-Lyonnet Dominique , Bourdeaut Franck , Brahimi Afane , Ingster Olivier , Saulnier Sholler Giselle , Jackson Sarah A. , Sasaki Mark M. , Fowler Trent , Ng Anita , Corbett Ryan J. , Kaufman Rebecca S. , Haley Jeremy S. , Carey David J. , Huang Kuan-lin , Diskin Sharon J. , Rokita Jo Lynne , Al-Kateb Hussam , McGee Rose B. , Schiffman Joshua D. , Chen Kenneth S. , Stewart Douglas R. , Williams Parsons D. , Plon Sharon E. , Schultz Kris Ann P. , Onel Kenan
Published: February 2025

Purpose: DROSHA, DGCR8, and DICER1 regulate microRNA biogenesis and are commonly mutated in cancer. Whereas DGCR8 and DICER1 germline pathogenic variants (GPVs) cause autosomal dominant tumor predisposition, no association between DROSHA GPVs and clinical phenotypes has been reported. Experimental Design: After obtaining informed consent, sequencing was performed on germline and tumor samples from all patients. The occurrence of germline DROSHA GPVs was investigated in large pediatric and adult cancer datasets. The population prevalence of DROSHA GPVs was investigated in the UK Biobank and Geisinger DiscovEHR cohorts. Results: We describe nine children from eight families with heterozygous DROSHA GPVs and a diagnosis of pineoblastoma (n=8) or Wilms tumor (WT, n=1). A somatic second hit in DROSHA was detected in all eight tumors analyzed. All pineoblastoma tumors analyzed were classified as miRNA processing altered-1 (PB-miRNA1) subtype. We estimate the population prevalence of germline DROSHA loss-of-function variants to be 1:3,875-1:4,843, but find no evidence for increased adult cancer risk. Conclusions: This is the first report of DROSHA-related tumor predisposition. As pineoblastoma and WT are also associated with DICER1 GPVs, our results suggest the tissues-of-origin for these tumors are uniquely tolerant of general microRNA loss. The PB-miRNA1 pineoblastoma subtype is associated with older age of diagnosis and better outcome than other subtypes, suggesting DROSHA GPV status may have important clinical and prognostic significance. We suggest genetic testing for DROSHA GPVs be considered for patients with pineoblastoma, WT, or other DICER1/DGCR8-related conditions and propose surveillance recommendations through research studies for individuals with DROSHA GPVs.
Abstract A002: cloneTCR: A high-throughput TCR assembly platform for highly multiplexed screening
Authors: Hamberger Moritz , Neuhoff Marie-Therese , Platten Michael , Green Edward Wilhelm
Published: February 2025

Manufacturing T cell receptors (TCRs) de novo has previously been considered costly and time-consuming, limiting the widespread application of TCR transgenic T cell immunotherapies. Here, we introduce a fully automated, cost-effective, and high-throughput TCR cloning platform capable of assembling and quality controlling thousands of TCRs per week. Our pipeline leverages the widely adopted AIRR (VDJ sequence) format as input, generating scripts automatically for streamlined operation of our lab automation. The platform is highly modular and adaptable, accommodating both human and murine V(D)J sequences alongside a broad spectrum of modular constant regions. These constant regions are available in germline-encoded or engineered versions, all designed for orthogonal compatibility. Additionally, the system supports multiple vector formats, enabling diverse delivery solutions such as lentiviral vectors, transposons, direct in vitro transcription, and recombination-based vectors targeting loci like AAVS1 and TRAC. A key innovation of our platform is the integration of Oxford Nanopore-based bulk long-read sequencing for quality control, enabling precise assessment of library uniformity and cloning fidelity. Compared to existing approaches, our method achieves superior library uniformity (frequency 2.5x around the median) and high sequence accuracy of 95%. With its unparalleled capacity to clone and quality-check 2,000 TCRs a week, our platform sets a new benchmark for high-throughput TCR production and screening. It enables large-scale, multiplexed exploration of entire tumor-infiltrating lymphocyte (TIL) repertoires, paving the way for transformative advances in T cell immunotherapy.

Citation Format: Moritz Hamberger, Marie-Therese Neuhoff, Michael Platten, Edward Wilhelm Green. cloneTCR: A high-throughput TCR assembly platform for highly multiplexed screening [abstract]. In: Proceedings of the AACR IO Conference: Discovery and Innovation in Cancer Immunology: Revolutionizing Treatment through Immunotherapy; 2025 Feb 23-26; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Immunol Res 2025;13(2 Suppl):Abstract nr A002.
Abstract A003: Investigating the effect of lipid metabolism on cDC1 dendritic cell cross-presentation in tumor microenvironment
Authors: Kar Uddipan , Balan Sreekumar , Bhardwaj Nina
Published: February 2025

Dendritic cells, or primary antigen-presenting cells, are critical immune sentinels essential in bridging the body’s adaptive and innate immune responses. Among DC subsets, conventional dendritic cell subset 1 (cDC1) stands out for its unique ability to cross-present antigens, which is crucial for initiating effective cytotoxic T-cell responses against tumors. Unlike normal cells, abnormal lipid accumulation (leading to lipid droplet formation) is observed in several cancers, which is considered one of the major reasons for DC dysfunction. However, very few reports are available on the impact of lipid metabolism on dendritic cells' antigen cross-presentation ability. To achieve this, in-vitro generated cDC1s were cocultured with heat-killed dying tumor cells and subsequently subjected to bulk RNA sequencing. Transcriptomic profiling was performed on cDC1s in the presence or absence of dead cells. Our analysis revealed distinct transcriptome profiles highlighting differential gene expression patterns linked to lipid metabolism, cholesterol regulation, and lipid droplet formation. Furthermore, we carried out various in-vitro metabolic functional assays, such as lipid staining assay, and observed increased lipid drop formation on antigen uptake. We validated this observation using confocal microscopy. In the future, we will reprogram lipid metabolism (using pathway inhibitors and gene knockdowns) and test dendritic cell-based antigen cross-presentation. Our findings suggest a crucial role for lipid metabolism in modulating the cross-presentation machinery of cDC1s within the tumor microenvironment. Understanding the mechanisms underlying this interplay between lipid metabolism and. DC cross-presentation may offer novel insights into the design of immunotherapeutic strategies aimed at harnessing DC-mediated immune responses against cancer.

Citation Format: Uddipan Kar, Sreekumar Balan, Nina Bhardwaj. Investigating the effect of lipid metabolism on cDC1 dendritic cell cross-presentation in tumor microenvironment [abstract]. In: Proceedings of the AACR IO Conference: Discovery and Innovation in Cancer Immunology: Revolutionizing Treatment through Immunotherapy; 2025 Feb 23-26; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Immunol Res 2025;13(2 Suppl):Abstract nr A003.
Abstract A007: A meta-analysis of cancer neoantigens curated from the literature
Authors: Kosaloglu-Yalcin Zeynep , Carter Hannah , Nielsen Morten , Sette Alessandro , Peters Bjoern
Published: February 2025

The Cancer Epitope Database and Analysis Resource (CEDAR) aggregates cancer epitope data from peer-reviewed studies, providing a structured repository of epitope-specific T cell, antibody, and MHC ligand assays. Neoantigens, arising from somatic mutations in cancer cells, represent a class of tumor-specific epitopes with potential for precision immunotherapy and as biomarkers for patient prognosis and therapeutic response. Our study harnesses CEDAR’s curated dataset to conduct a meta-analysis on neoantigens across various cancers, identifying factors contributing to neoantigen immunogenicity and their prevalence among patient populations.

Citation Format: Zeynep Kosaloglu-Yalcin, Hannah Carter, Morten Nielsen, Alessandro Sette, Bjoern Peters. A meta-analysis of cancer neoantigens curated from the literature [abstract]. In: Proceedings of the AACR IO Conference: Discovery and Innovation in Cancer Immunology: Revolutionizing Treatment through Immunotherapy; 2025 Feb 23-26; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Immunol Res 2025;13(2 Suppl):Abstract nr A007.
Abstract A012: Targeting macrophage galactose binding lectin -tumor associated MUC1 immune axis in cancer
Authors: Cudic Mare , Ayyalasomayajula Ramya , Boneva Ivet , Beckwith Donella M. , André Sabine , Kaltner Herbert
Published: February 2025

Carbohydrate-binding proteins, lectins, are the main binding partners of tumor-associated carbohydrate antigens (TACAs) on the cell surface. Interactions between the tumor-associated carbohydrate antigens of Mucin 1 (TA MUC1) and the carbohydrate-binding proteins, lectins, often lead to the creation of a pro-tumor microenvironment favoring tumor initiation, progression, metastasis, and immune evasion. Macrophage galactose binding lectin (MGL) is a C-type lectin receptor found on antigen-presenting cells that facilitates the uptake of carbohydrate antigens for antigen presentation, modulating the immune response homeostasis, autoimmunity, and cancer. MGL has been identified as a receptor of Thomsen-nouvelle (Tn, CD175) and sialylated Tn (sialyl-Tn, CD175s) antigens. The interaction between Tn and sialyl-Tn antigens of tumor-associated MUC1 and hMGL likely plays a critical role in establishing the immunological tolerance to tumor cells. Yet, MGL targeting is an attractive approach to enhance cancer vaccine efficacy. The ability of hMGL to regulate immune response by engaging tumor-associated MUC1 to either augment the anti-tumorigenic effects or minimize the pro-tumorigenic effects is poorly understood. Thus, a thorough understanding of their binding interaction is essential for it to be exploited for cancer vaccine strategies. The synthesis of MUC1 glycopeptide models carrying a single or multiple Tn and/or sialyl-Tn antigen(s) is described. A novel approach for the sialyl-Tn threonine building block suitable for the solid phase peptide synthesis was developed. The thermodynamic profile of the binding interaction between the human MGL and MUC1 glycopeptide models was analyzed using isothermal titration calorimetry. All studied interactions, regardless of the glycan’s site of attachment or density, exhibited enthalpy-driven thermodynamics. The thermodynamic analysis pointed out the critical role of entropy in driving the affinity enhancement of multivalent interactions. Further studies into the nature of the hMGL ligands and their role in shaping the immune responses are warranted.

Citation Format: Mare Cudic, Ramya Ayyalasomayajula, Ivet Boneva, Donella M. Beckwith, Sabine André, Herbert Kaltner. Targeting macrophage galactose binding lectin-tumor associated MUC1 immune axis in cancer [abstract]. In: Proceedings of the AACR IO Conference: Discovery and Innovation in Cancer Immunology: Revolutionizing Treatment through Immunotherapy; 2025 Feb 23-26; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Immunol Res 2025;13(2 Suppl):Abstract nr A012.
Abstract A016: Targeting axonal guidance dependencies in glioblastoma with ROBO1 CAR T cells
Authors: Singh Sheila K , Chokshi Chirayu R , Shaikh Vaseem , Brakel Benjamin , Rossotti Martin , Lazo John , Venugopal Chitra , Kislinger Thomas , Henry Kevin , Moffat Jason
Published: February 2025

Resistance to genotoxic therapies and tumor recurrence are hallmarks of glioblastoma (GBM), an aggressive brain tumor. In this study, we investigated functional drivers of post-treatment recurrent GBM through integrative genomic analyses, genome-wide genetic perturbation screens in patient-derived GBM models and independent lines of validation. Specific genetic dependencies were found consistent across recurrent tumor models, accompanied by increased mutational burden and differential transcript and protein expression compared to its primary GBM predecessor. Our observations suggest a multi-layered genetic response to drive tumor recurrence and implicate PTP4A2 (protein tyrosine phosphatase 4A2) as a modulator of self-renewal, proliferation and tumorigenicity in recurrent GBM. Genetic perturbation or small-molecule inhibition of PTP4A2 acts through a dephosphorylation axis with roundabout guidance receptor 1 (ROBO1) and its downstream molecular players, exploiting a functional dependency on ROBO signaling. Because a pan-PTP4A inhibitor was limited by poor penetrance across the blood–brain barrier in vivo, we engineered a second-generation chimeric antigen receptor (CAR) T cell therapy against ROBO1, a cell surface receptor enriched across recurrent GBM specimens. A single dose of ROBO1-targeted CAR T cells doubled median survival in cell-line-derived xenograft (CDX) models of recurrent GBM. Moreover, in CDX models of adult lung-to-brain metastases and pediatric relapsed medulloblastoma, ROBO1 CAR T cells eradicated tumors in 50–100% of mice. Our study identifies a promising multi-targetable PTP4A–ROBO1 signaling axis that drives tumorigenicity in recurrent GBM, with potential in other malignant brain tumors.

Citation Format: Sheila K Singh, Chirayu R Chokshi, Vaseem Shaikh, Benjamin Brakel, Martin Rossotti, John Lazo, Chitra Venugopal, Thomas Kislinger, Kevin Henry, Jason Moffat. Targeting axonal guidance dependencies in glioblastoma with ROBO1 CAR T cells [abstract]. In: Proceedings of the AACR IO Conference: Discovery and Innovation in Cancer Immunology: Revolutionizing Treatment through Immunotherapy; 2025 Feb 23-26; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Immunol Res 2025;13(2 Suppl):Abstract nr A016.
Abstract A021: Spatiotemporal Regulation of Chimeric Antigen Receptor T-cells Using Focused Ultrasound Therapy to Enable Use of High-Affinity Binders
Authors: Trope Edward , Su Yun-Yun , Nakata Rie , Golyska Magda , Richman Sarah
Published: February 2025

Pediatric High Grade Glioma (HGG) is an aggressive cancer with a 5-year survival rate of just 10-20% (Fangusaro, 2012). While previous studies using chimeric antigen receptor (CAR) T-cells for brain malignancies demonstrated tolerability (Majzner et al., 2022; Vitanza et al., 2021; Vitanza et al., 2023), the low survival rates demand more efficacious therapies (Majzner et al., 2022). In this study, we perform in vitro modeling of spatiotemporally regulated high-affinity CARs targeting the GD2 antigen to enable use of highly potent CAR variants while mitigating on-target, off-tumor toxicity. By placing CAR affinity variants under control of a heat shock promoter, thermal energy delivered to the tumor tissue by focused ultrasound (FUS) induces CAR expression (Wu et al., 2021) at an optimal temperature of 43oC. We first determine FUS-controlled CAR induction and decay kinetics in vitro for selected signaling domain architectures as well as affinity variants. Following codon optimization, differences in basal expression at 37oC from the heat shock promoter were noted for the three signaling domain architectures, with 28z and BBz displaying the highest and lowest levels of aberrant expression, respectively. Next, we compare expression of CAR constructs with and without 3 cycles of heat shock at 43oC in CAR-T cells. Then, using live cell imaging, we analyze the kinetics of expression and decay of the CAR following heat shock. Finally, we perform effector function assays of the CAR-T cells to compare the cytotoxicity and persistence for each construct. We identify the CAR architecture harboring the CD28 transmembrane domain and the 4-1BB cytoplasmic domain as the optimal balance between basal and induced expression. We plan to move this architecture forward to in vivo studies to assess the ability to maintain robust tumor lysis and minimal on-target, off-tumor toxicity.

Citation Format: Edward Trope, Yun-Yun Su, Rie Nakata, Magda Golyska, Sarah Richman. Spatiotemporal Regulation of Chimeric Antigen Receptor T-cells Using Focused Ultrasound Therapy to Enable Use of High-Affinity Binders [abstract]. In: Proceedings of the AACR IO Conference: Discovery and Innovation in Cancer Immunology: Revolutionizing Treatment through Immunotherapy; 2025 Feb 23-26; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Immunol Res 2025;13(2 Suppl):Abstract nr A021.
Abstract A024: CD3ζ ITAM diversity determines chimeric antigen receptor force and function
Authors: Echelibe Hilda , Majumdar Shubhabrata , Kolawole Elizabeth , Liu Baoyu , Bettini Maria , Evavold Brian , Bettini Matt
Published: February 2025

The Chimeric antigen receptor (CAR) T cell therapy is a powerful treatment for hematologic malignancies; however, clinical responses vary depending on CAR T cell function and persistence. The optimization of CAR design has gained considerable interest with a new focus on calibrating CAR intracellular signals via the immunoreceptor tyrosine-based activation motifs (ITAMs) to increase the fitness of CAR T cells. Evidence suggests that unique ITAM motifs at positions 1, 2, or 3 (ITAMs-A, B, and C respectively) of the CD3ζ signaling domain may have non-redundant roles in signal strength, T cell functionality, persistence, and memory formation. However, the individual contributions of each CD3ζ ITAM in transmitting force and activating mechanosensitive signals are yet unexplored. We have found that human CARs expressing specific ITAM sequences (ITAM-A, ITAM-B, or ITAM-C only) generate differing amounts of force and bond lifetimes after interacting with their target antigen, CD19. Therefore, a CAR may act as a mechanosensor similar to a TCR, discriminating the quality of the interaction with antigen as cellular-derived forces are applied to the bond. The higher transmission of force directly correlates with better anti-tumor efficacy in vitro and in vivo. Hence, understanding the mechanotransduction in CARs has important implications in “tuning” the activation of a CAR T cell without changing specificity.

Citation Format: Hilda Echelibe, Shubhabrata Majumdar, Elizabeth Kolawole, Baoyu Liu, Maria Bettini, Brian Evavold, Matt Bettini. CD3ζ ITAM diversity determines chimeric antigen receptor force and function [abstract]. In: Proceedings of the AACR IO Conference: Discovery and Innovation in Cancer Immunology: Revolutionizing Treatment through Immunotherapy; 2025 Feb 23-26; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Immunol Res 2025;13(2 Suppl):Abstract nr A024.
Abstract A030: A deeper understanding of tumor microenvironment modulation induced by IL13Ra2 CAR T cells in a patient with grade 3 ependymoma
Authors: Aftabizadeh Maryam , Aguilar Brenda , D'Apuzzo Massimo , Badie Behnam , Brown Christine E
Published: February 2025

Grade 3 ependymoma is classified as a fast-growing solid tumor that often occurs in the base of the brain with a median survival of less than two years. Despite aggressive standard treatments (surgery, radiation therapy and chemotherapy), grade 3 ependymoma remains uniformly fatal, especially if the tumor cannot be completely removed or if it spreads to other areas of the central nervous system (CNS). Achieving a responsive therapy with chimeric antigen receptor (CAR) T cell therapy has been a significant challenge due to evasive tumor microenvironment, specifically utilized by slid tumors, to resist therapy. Recent studies have highlighted the substantial role of glioma-associated stromal cells in solid tumors by depositing various collagen subunits, including COL1A1, within the extracellular matrix (ECM). This collagen deposition leads to increased ECM stiffness, modulating the tumor microenvironment, inducing immune suppression (e.g. SPP1 overexpression), and hindering T cell trafficking, ultimately worsening clinical outcomes and patient survival. To gain a better understanding of the role of the tumor microenvironment in CAR T persistence and efficacy, we designed and performed ultra-high plex immunofluorescent staining on tissue biopsies obtained from a patient with grade 3 ependymoma before and after treatment with IL13Ra2 CAR T cells. The patient has been received three CAR T infusions at weekly intervals. The tissues were stained for CD3, CD19 (CAR T cells), CD68, CD163, HLA-DR (macrophages), CD3, CD8 (endogenous T cells), SOX2 (stem cells), SPP1 (suppression) and COL1A1 (extracellular matrix). The presence of CAR T cells in tissue biopsies indicated persistence of IL13Ra2 CAR T cells for more than a year. Our preliminary analysis revealed an increase in the number of HLA-DR+ CD163+ macrophages within cellular neighborhoods associated with CAR T cells (around 50% of CD163+ express HLA-DR after treatment) as compared to baseline tumors, which suggests a dynamic interaction between myeloid subsets and CAR T cells within the tumor microenvironment. Ongoing experiments are currently investigating the presence of antigen presenting natural killer cells (CD56, HLA-DR), proliferative T cells (CD3, Ki67) and inflammatory macrophages and/or T cells (IFNg) to gain a better understanding of IL13Ra2 persistence in brain tumor patients.

Citation Format: Maryam Aftabizadeh, Brenda Aguilar, Massimo D'Apuzzo, Behnam Badie, Christine E Brown. A deeper understanding of tumor microenvironment modulation induced by IL13Ra2 CAR T cells in a patient with grade 3 ependymoma [abstract]. In: Proceedings of the AACR IO Conference: Discovery and Innovation in Cancer Immunology: Revolutionizing Treatment through Immunotherapy; 2025 Feb 23-26; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Immunol Res 2025;13(2 Suppl):Abstract nr A030.
Abstract A038: Inducing tumor associated tertiary lymphoid structures using cellular therapy
Authors: Amwas Nour , Alizadeh Darya , Brown Christine , Chiu Victor , Egelston Colt , Gumber Diana , Wang Hanmin , Williams John , Wang Leo David
Published: February 2025

Targeting the tumor microenvironment (TME) to be more immune permissive is a potential strategy for enhancing immunotherapies, such as chimeric antigen receptor T cell (CAR-T) therapy, providing a promising avenue for treating aggressive tumors such as glioblastoma multiforme (GBM). Tertiary lymphoid structures (TLS) are ectopic lymphoid aggregates that arise in response to chronic inflammation and mimic the structure and function of secondary lymphoid organs. The spontaneous presence of TLS in some solid tumors, including GBM, is associated with improved clinical outcome and responsiveness to immunotherapy. I hypothesize that membrane tethered anti-LTβR Sc-Fv (mTa) T cells can function as lymphoid tissue inducer cells (LTi) and interact with lymphoid tissue organizer cells (LTo) in the TME, that express lymphotoxin-β receptor (LTβR), to nucleate TLS. To this end, we screened commercially available murine LTβR agonist antibodies to induce NFκB signaling. When a lead candidate with robust NFkB activation was injected in situ into mice orthotopically injected with a murine glioma cell line, we found multiple TLS-like lymphoid aggregates. We therefore sequenced and cloned a membrane-tethered version of this antibody alongside a CAR construct to generate mTa-CAR-T cells. mTa-CAR T cells could activate LTβR in an NFκB-GFP reporter assay. We proceeded to intratumorally inject these cells into a syngeneic mouse GBM tumor model to evaluate TLS induction and assess the ability of mTa-CAR T cells to improve antitumor efficacy and survival. This study attempts to identify strategies to remodel the TME of GBM and nucleate TLS using cellular therapy to improve the response to CAR-T therapy and survival of GBM patients.

Citation Format: Nour Amwas, Darya Alizadeh, Christine Brown, Victor Chiu, Colt Egelston, Diana Gumber, Hanmin Wang, John Williams, Leo David Wang. Inducing tumor associated tertiary lymphoid structures using cellular therapy [abstract]. In: Proceedings of the AACR IO Conference: Discovery and Innovation in Cancer Immunology: Revolutionizing Treatment through Immunotherapy; 2025 Feb 23-26; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Immunol Res 2025;13(2 Suppl):Abstract nr A038.