bluuebunny/crossref_metadata_2025 huggingface dataset:
meso-Halogenated nitrosylcobalamins: Formation and aerobic stability
Authors: Dereven’kov Ilia A. , Osokin Vladimir S. , Ershov Nikita A. , Makarov Sergei V.
Published: March 2024

Here, we report the results of the investigation of reactions between brominated and chlorinated at the C10 position of corrin cyano- (CNCbl-Hal, Hal = Br, Cl) and aquacobalamins (H2OCbl-Hal) and Angeli’s salt (AS). The reactions involving H2OCbl-Hal and AS proceed via two parallel routes, i.e., reactions between H2OCbl-Hal or HOCbl-Hal and HNO generated upon decomposition of AS and between H2OCbl-Hal and AS, and generate corresponding nitrosylcobalamins (NOCbl-Hal). In contrast to the process involving unmodified CNCbl, the reactions between CNCbl-Hal and HNO can be used to produce NOCbl-Hal, which can be explained by the labilization of the axial Co(III)-N[Formula: see text] bond upon meso-halogenation facilitating the reaction between HNO and the lower axial position of CNCbl-Hal. We indicated that the process is initiated by the relatively slow formation of CNCbl-Hal species with dissociated 5,6-dimethylbenzimidazole nucleotide and further rapid binding of HNO by Co(III) ion. Produced NOCbl-Hal reacts with dioxygen significantly slower than unmodified NOCbl and can be recommended for further investigation of their biological and medicinal properties.
L-Fuzzy Rough Proximity Spaces and Their Relationship with L-Fuzzy Rough Grills
Authors: Tiwari Surabhi , Kumar Virendra
Published: March 2024

In this paper, we study the concept of nearness between [Formula: see text]-fuzzy rough sets and the concepts of [Formula: see text]-fuzzy rough grills and [Formula: see text]-fuzzy rough [Formula: see text]-clusters on a Čech [Formula: see text]-fuzzy rough proximity space [Formula: see text]. Also, we investigate the relationship among [Formula: see text]-fuzzy rough grills, [Formula: see text]-fuzzy rough [Formula: see text]-clan and [Formula: see text]-fuzzy rough [Formula: see text]-clusters. Moreover, we introduce the [Formula: see text]-fuzzy rough LO-proximity spaces and study some of its basic properties. An adequate number of examples support the study.
Investigation of some finite integrals incorporating incomplete Aleph functions
Authors: Kumar Sachin , Mishra Manvendra Narayan , Dubey Ravi Shanker , Sharma Rahul
Published: March 2024

In this paper, we introduce a new approach to the study of finite integral formulas associated with special functions, bringing out the incomplete Aleph functions and other well-known special functions. Our method is based on the finite integral operator applied to incomplete Aleph functions. This approach makes it easier to derive integral formulas that connect the incomplete Aleph functions using well-known special functions and algebraic expressions. As a result, we can group integral formulas and special functions into different classes according to similar features. Furthermore, we can discover new integral formulas with this framework that are useful for computational applications.
An Analysis of the Cluster-Detecting Property of the BCM Neuron
Authors: Udeigwe Lawrence
Published: March 2024

The BCM learning rule, named for Elie Bienenstock, Leon Cooper, and Paul Munro, was first proposed to measure the selectivity of neurons in the primary visual cortex and its dependency on neuronal inputs. We show that an artificial BCM neuron has the ability to detect clusters in a dataset. By exploring the qualitative behaviors of an underlying system of differential equations, we present a rigorous mathematical analysis of this cluster-detecting property. While the focus of this work is not to develop a robust state-of-the-art clustering method, we also analyze and discuss the performance of a resulting preliminary clustering algorithm.
A Context-enhanced Adaptive Graph Network for Time-sensitive Question Answering
Authors: Li Jitong , Wu Shaojuan , Zhang Xiaowang , Feng Zhiyong
Published: March 2024

Time-sensitive question answering is to answer questions limited to certain timestamps based on the given long document, which mixes abundant temporal events with an explicit or implicit timestamp. While existing models make great progress in answering time-sensitive questions, their performance degrades dramatically when a long distance separates the correct answer from the timestamp mentioned in the question. In this paper, we propose a Context-enhanced Adaptive Graph network (CoAG) to capture long-distance dependencies between sentences within the extracted question-related episodes. Specifically, we propose a time-aware episode extraction module that obtains question-related context based on timestamps in the question and document. As the involvement of episodes confuses sentences with adjacent timestamps, an adaptive message passing mechanism is designed to capture and transfer inter-sentence differences. In addition, we present a hybrid text encoder to highlight question-related context built on global information. Experimental results show that CoAG significantly improves compared to state-of-the-art models on five benchmarks. Moreover, our model has a noticeable advantage in solving long-distance time-sensitive questions, improving the EM scores by 2.03% to 6.04% on TimeQA-Hard.
Hierarchical Convolutional Neural Network with Knowledge Complementation for Long-Tailed Classification
Authors: Zhao Hong , Li Zhengyu , He Wenwei , Zhao Yan
Published: March 2024

Existing methods based on transfer learning leverage auxiliary information to help tail generalization and improve the performance of the tail classes. However, they cannot fully exploit the relationships between auxiliary information and tail classes and bring irrelevant knowledge to the tail classes. To solve this problem, we propose a hierarchical CNN with knowledge complementation, which regards hierarchical relationships as auxiliary information and transfers relevant knowledge to tail classes. First, we integrate semantics and clustering relationships as hierarchical knowledge into the CNN to guide feature learning. Then, we design a complementary strategy to jointly exploit the two types of knowledge, where semantic knowledge acts as a prior dependence and clustering knowledge reduces the negative information caused by excessive semantic dependence (i.e., semantic gaps). In this way, the CNN facilitates the utilization of the two complementary hierarchical relationships and transfers useful knowledge to tail data to improve long-tailed classification accuracy. Experimental results on public benchmarks show that the proposed model outperforms existing methods. In particular, our model improves accuracy by 3.46% compared with the second-best method on the long-tailed tieredImageNet dataset.
Space–Time Clustering and Climatic Risk Factors for Lumpy Skin Disease of Cattle in Uttar Pradesh, India, 2022
Authors: Agrawal Isha , Sharma Barkha , Varga Csaba
Published: March 2024

Lumpy skin disease (LSD), a transboundary infectious disease, negatively impacts cattle health and production. The first LSD outbreaks were reported in India in 2019, and since then, LSD spread to over 15 states, including Uttar Pradesh. This study evaluated LSD cases reported by veterinarians in Uttar Pradesh, India, during 2022. Using scan statistics, Poisson models that accounted for the background cattle population were constructed to identify spatial, temporal, and space–time clusters. A negative binomial regression model was built to assess the impact of temperature and humidity on the incidence rate (IR) of LSD. A total of 112,226 cases across 33 districts were reported in 2022. A purely temporal cluster with higher‐than‐expected LSD rates was identified between August and October 2022. Several purely spatial clusters were identified in the western part of the state. A primary space–time cluster was detected in west Utter Pradesh between August and October 2022 that overlapped with the spatial clusters. The secondary cluster occurred between September and October 2022 in the eastern part of the state. A rise in humidity (incidence rate ratio (IRR) = 1.39; 95% CI: 1.30–1.49) and temperature (IRR = 1.16; 95% CI: 1.06–1.27) increased the IR of LSD, suggesting a seasonality of the outbreaks. The results of this study can aid animal health authorities in developing effective LSD prevention, surveillance, and control strategies among cattle in India.
Semidirect Resin Composite Veneers in a Patient with Bruxism
Authors: Floriani Franciele , Coelho Natália Fiuza , de Azevedo Linhares Ludimila , Stolf Sheila Cristina , Lopes Guilherme Carpena
Published: March 2024

This case report details the esthetic rehabilitation of a 32‐year‐old male patient suffering from sleep bruxism, primarily manifesting as a fracture and significant loss of tooth structure in the anterior maxillary central incisors. To address these concerns, the patient underwent a restorative treatment involving the application of semidirect resin composite veneers on the maxillary incisors and direct resin composite restoration on the incisal regions of the maxillary canines. This approach not only restored the functional integrity of the teeth but also significantly enhanced the patient’s esthetic appearance.
In Vitro Gastrointestinal Digestion of Polyonatum cyrtonema Polysaccharides and Impact on Intestinal Flora
Authors: Gao Ya , Xiao Ying , Yu Ling , Tang Qingjiu , Ye Lin , Wang Jinyan , Wang Yipeng , Zhang Fangfang , Zhou Jianjin
Published: March 2024

Polyonatum cyrtonema polysaccharides (PCPs) have been illustrated to have pharmacological and biological activity. However, the impact of PCPs on gut microorganisms and their metabolic pathways has been scarcely addressed. In order to investigate the antidigestion potential of PCPs, an in vitro simulation of digestion was developed. Moreover, an in vitro fermentation model was used to explore the influence of PCPs on the microbial community. The findings revealed that the concentration of the remaining carbohydrate and reducing end of the polysaccharide chain grew slightly, while the molecular weight was unaltered after being digested. Notably, the gut microbiota degraded the PCPs during the subsequent fermentation, resulting in the production of short-chain fatty acids and a significant decrease in the pH level (p<0.05). Compared with the control group, the result of 16S rRNA sequencing revealed that PCPs could affect the diversity of the community and the composition of the gut microbes significantly (p<0.05), having similar regulatory effects to inulin on the structure of the microbial community. The PCPs group notably promoted the relative abundance of acetic-acid bacteria, such as Bifidobacterium. In addition, PICRUSt2 analysis demonstrated that PCPs mainly enhanced phenotypic KEGG pathways such as amino acid metabolism and carbohydrate metabolism. Overall, PCPs may have prebiotic properties that support human health by controlling the make-up and metabolic activity of intestinal flora.
Abstract 1007: Breast cancer related lymphedema patient characteristics and treatment patterns in a Hispanic majority population
Authors: Shaker Sarah A. , Rahman Lauren P. , Canola Marcela Mazo
Published: March 2024

Background: Studies have shown an increased risk of Breast Cancer Related Lymphedema (BCRL) in Hispanic populations, however there are limited studies examining factors specific to BCRL in this minority population. This retrospective study aims to characterize and analyze treatment and demographic related factors associated with BCRL in the Hispanic majority population of San Antonio.

Methods: This is a retrospective, single institution study of patients with BCRL between 2017-2020 at Mays Cancer Center in San Antonio, Texas, a city with a majority minority Hispanic population.Results: 110 subjects with BCRL were identified with a median age of 62 years old. 52.7% were Hispanic while 47.3% were non-Hispanic. The median BMI was 31 with over 75% of subjects with BCRL categorized with BMIs of overweight or obese. 89.1% received some form of chemotherapy, whether neoadjuvant or adjuvant, and 80.9% were exposed to some form of taxane. The majority underwent more invasive procedures and surgeries: 69.1% had axillary lymph node dissection and 71.2% underwent mastectomy. The median number of lymph nodes removed were 12. 62.7% of BCRL patients were exposed to nodal radiation. Regarding BCRL severity, 29.4% were classified as grade 1 while 11% classified as grade 3. Of note, there was a statistically significant correlation between more severe lymphedema (grade 2 and 3) and use of chemotherapy (P-value = 0.04.)Conclusion: The demographic composition of patients with BCRL closely mirrors the ethnically diverse population of San Antonio, with a large Hispanic predominance. Within this cohort of patients of BCRL, a large portion demonstrated factors of elevated BMI, more invasive surgical interventions, and exposure to chemotherapy, specifically taxane. The examination of these factors in a Hispanic-centric urban setting such as San Antonio offers crucial insight into a patient population at heightened risk for developing BCRL.

Citation Format: Sarah A. Shaker, Lauren P. Rahman, Marcela Mazo Canola. Breast cancer related lymphedema patient characteristics and treatment patterns in a Hispanic majority population [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1007.
Abstract 105: Understanding the role of β2-adrenergic receptor signaling on type II innate lymphoid cells in cancer
Authors: Choi Jee Eun , MacDonald Cameron R. , Daneshmandi Saeed , Wang Jianmin , Roberts Nathan T. , James Caitlin M. , McCarthy Philip L. , Mohammadpour Hemn , Repasky Elizabeth A.
Published: March 2024

Increased norepinephrine (NE) due to chronic stress can activate adrenergic receptors, specifically β2-adrenergic receptors (β2AR), to produce an immunosuppressive tumor microenvironment (TME). Previous work by our lab demonstrated that tumor burden can be reduced by blocking β2AR signaling. Immune cells express β2AR and we have shown that the increased β2AR signaling leads to increased T cell exhaustion and increased inhibitory activity of myeloid-derived suppressor cells (MDSCs). In general, the resulting immunosuppression leads to a modest but significant reduction in tumor growth. However, the lack of a dramatic reduction in tumor growth reveals the complex nature of adrenergic signaling dependent regulation of the cancer immune response. Recent literature revealed that innate lymphoid cells (ILCs), especially type II ILCs (ILC2s), express high levels of β2AR. Therefore, we evaluated whether β2AR regulates ILC2s within the TME and how that affects tumor prognosis. We generated a genetic conditional knockout murine model where β2AR expression in ILC2s was deleted by crossing IL5cre expressing mice to β2ARflox/flox mice (IL5creβ2ARf/f). AT3 breast cancer cell line was injected into the 4th mammary fat pad and tumor volume was measured every 2-3 days. Tumor infiltrating ILC2s (CD45+Lineage−CD90.2+CD127+ST2+) were analyzed by flow cytometry at D36 post-tumor implantation. Notably, IL5creβ2ARf/f mice had increased AT3 breast cancer tumor growth compared to control mice. Flow cytometric analysis of the tumor revealed an increased frequency of GATA3+ ILC2s within the TME. These GATA3+ ILC2s lacking β2AR expressed higher levels of the IL33 receptor, ST2, and a higher percentage of IL13+ cells compared to WT GATA3+ ILC2 cells. We also observe a trend toward increased MDSC population within the TME of mice lacking β2AR in ILC2s. These results reveal an anti-tumor function of β2AR signaling through the suppression of ILC2 activity within the TME that warrants further investigation. This work has been in part supported by NIH Grants F30 CA284763 (to J.C.), K99 HL155792 (to H.M.), and RO1 CA205246 (to E.R.)

Citation Format: Jee Eun Choi, Cameron R. MacDonald, Saeed Daneshmandi, Jianmin Wang, Nathan T. Roberts, Caitlin M. James, Philip L. McCarthy, Hemn Mohammadpour, Elizabeth A. Repasky. Understanding the role of β2-adrenergic receptor signaling on type II innate lymphoid cells in cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 105.
Abstract 1059: Biomarkers analysis on samples from patients in EVT801 clinical trial: Patient characterization and immunomonitoring
Authors: Davenne Lise , Fitzgerald Michael , Ancey Pierre-Benoit , Delpuech Oona , Poussereau-Pomié Céline , Esquerre Michael , Paillasse Michael R. , Mandron Marie , Rochaix Philippe , Ayyoub Maha , Scarlata Clara Maria , Caux Christine , Caux Christophe , Cassier Philippe , Gomez-Roca Carlos , Delord Jean-Pierre , Friend John , Fons Pierre
Published: March 2024

Introduction: EVT801 is a highly selective small molecule inhibitor of VEGFR3 that acts by inducing tumor (lymph)-angiogenesis normalization in and around the tumor. EVT801 has shown compelling activity in a wide range of cancer models showing a decrease of tumor hypoxia and an increase of CD8pos T-cells infiltration into the tumor micro-environment. EVT801 was well-tolerated in preclinical toxicology studies. A phase I study is underway focusing primarily on understanding the safety, tolerability, and pharmacokinetics of EVT801 across a range of doses (NCT05114668).

Methods: The first stage of the phase I trial aims to determine the maximum tolerated dose (MTD) and recommended Phase II dose (RP2D) for EVT801. Clinical samples from enrolled patients have been collected to explore preliminary signals of the biological activity of the drug and characterize tumor phenotypes. Patient characterization includes histology and mRNA signature in archival biopsies. Target engagement and pharmacodynamics effects are investigated by immunomonitoring as well as assessment of a defined set of proteins as markers of inflammation and angiogenesis as identified in preclinical in vivo models.

Conclusion: Our first biomarkers analyses on a subpopulation of patients with High Grade Serous Ovarian Cancer (HGS-OC) enrolled in the first stage of the study showed that levels of VEGFR3 expression had a negative correlation with CD8 positive T-cells infiltration in the tumor microenvironment and a tendency to positive correlation with a PD1 mAb resistance signature. VEGFR3 expression also tends to be correlated with higher levels of hypoxia (CAIX labelling). Our working hypothesis is that patients with hypoxic HGS-OC tumors with high VEGFR3 expression may benefit from EVT801 treatment; this will need to be reinforced by inclusion of additional patients in dedicated PD biomarkers cohorts during stage 2 of the clinical trial.

Citation Format: Lise Davenne, Michael Fitzgerald, Pierre-Benoit Ancey, Oona Delpuech, Céline Poussereau-Pomié, Michael Esquerre, Michael R. Paillasse, Marie Mandron, Philippe Rochaix, Maha Ayyoub, Clara Maria Scarlata, Christine Caux, Christophe Caux, Philippe Cassier, Carlos Gomez-Roca, Jean-Pierre Delord, John Friend, Pierre Fons. Biomarkers analysis on samples from patients in EVT801 clinical trial: Patient characterization and immunomonitoring [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1059.
Abstract 1067: Label free Raman spectroscopy of extracellular vesicles for early stage liquid biopsy cancer detection
Authors: Carney Randy P. , Navas-Moreno Maria , Birkeland Andrew
Published: March 2024

Aim: To improve patient outcomes, there remains a critical need to develop faster, less invasive platforms capable of identifying biomarkers. Multi-omics approaches, while promising, are high in cost and complexity, low throughput, slow, and require large sample volumes, thus are impractical for many stages of clinical care. Raman spectroscopy (RS) addresses many of these needs: it requires little to no sample preparation, is non-destructive, does not need exogenous dyes or labelling agents, and can be performed directly in aqueous solutions. Surface enhanced Raman spectroscopy (SERS) offers further improvements in terms of sensitivity but increases complexity. In this study we carried out comprehensive RS and SERS measurements on a >100-person cohort of blood and saliva from head and neck cancer (HNC) patients and benign controls. Analyses were carried out on whole biofluids, in addition to extracellular vesicles (EVs) isolated from the biofluids to investigate the added value. EVs are emerging biomarkers of interest found in high numbers in circulating biofluids with great promise to increase the specificity of diagnostic platforms.

Results: We analyzed a robust clinical dataset of HNC patient saliva and plasma to build a diagnostic model. We found distinct Raman and SERS signatures of metabolites in all cancer samples uniquely present compared to non-cancer controls. Raman spectral measurements fused together for EVs and whole biofluids across saliva and plasma delivered accuracies of >95% using a novel machine learning module based on convolutional neural networks. Our platform features an automated method to isolate EVs amenable to rapid clinical use.

Conclusions: The results of this study indicate an exciting step in validating Raman as a robust diagnostic tool revealing that complementary chemical information spread out across biofluids and biomarkers of interest can strongly improve simple, quick Raman based diagnostics of whole biofluids and EVs.

Citation Format: Randy P. Carney, Maria Navas-Moreno, Andrew Birkeland. Label free Raman spectroscopy of extracellular vesicles for early stage liquid biopsy cancer detection [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1067.
Abstract 1085: Exploiting bioenergetic vulnerabilities in chemotherapy resistant osteosarcoma
Authors: Nguyen Valerie U. , Graves Laurie , Colmenares Veronica , Macabangon Abol , Syal Casey , Fisher-Wellman Kelsey , Eward William C. , Kim So Young , Somarelli Jason A.
Published: March 2024

BACKGROUND: Osteosarcoma (OS) is the most common bone cancer in children and young adults. Standard treatment involves chemotherapy and surgical resection, yet nearly 4 in 10 patients experience disease relapse, and almost all of these patients are resistant to additional chemotherapy. Therefore, there is an urgent need to understand the biology of chemo-resistant OS and identify new therapeutic strategies to treat this therapy-resistant disease state.

METHODS: Using a CRISPR-Cas9 based lineage tracing system, RNA sequencing, and Seahorse XF assays in chemo-naïve, -resistant, and -released human 143b and patient-derived 173x OS cell lines, we modeled the evolution of chemoresistance and release from the selective pressure of chemotherapy in osteosarcoma. The phenotype of each cell state was assessed through in vitro and ex vivo cell proliferation assays, a comparison of cell viability upon re-challenge to standard-of-care chemotherapy, and comparison of cell viability in response to novel therapies targeting alterations in cell respiration.

RESULTS: Chemo-resistant OS cells demonstrate a decrease in expression of proliferation-associated gene pathways and shift their resources toward expression and maintenance of multi-drug resistance ABC transporters. Chemo-released cells maintain elevated expression of ABC transporters while re-activating proliferation. Across the spectrum of chemotherapy exposure, OS cells demonstrate changes in respiratory capacity, with unique bioenergetic phenotypes observed in chemo-naïve, -resistant, and -released cells.

CONCLUSIONS: Chemo-resistant and chemo-released OS cells demonstrate unique bioenergetic patterns that correlate with gene expression signatures and growth phenotypes. Evolution imposed by the selective pressure of chemotherapy may render OS cells vulnerable to therapies that disrupt cellular energetics and exploit multi-drug resistant ABC transporters.

Citation Format: Valerie U. Nguyen, Laurie Graves, Veronica Colmenares, Abol Macabangon, Casey Syal, Kelsey Fisher-Wellman, William C. Eward, So Young Kim, Jason A. Somarelli. Exploiting bioenergetic vulnerabilities in chemotherapy resistant osteosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1085.
Abstract 1103: Molecular signature for treatment-responsive and recurrent Wilms tumors
Authors: Jiang Jieyun , Allison Derek B. , Burikhanov Ravshan , Ganguly Saptadwipa , Goettl Ryan A. , Liu Jinpeng , Wang Chi , Saltzman Amanda F. , Tucker Thomas , Spielmann Peter H. , Durbin Eric B. , D’Orazio John A. , Rangnekar Vivek M.
Published: March 2024

Wilms tumor is the most common solid renal malignancy in children. To better understand the biology of Wilms tumors and develop drugs especially against tumors that relapse, we sought to identify mutations in the exome, differential gene expression patterns, and tumor suppressor expression status of Wilms tumors in Kentucky. The Kentucky Cancer Registry (KCR) Virtual Tissue Repository identified 29 cases of Wilms tumor diagnosed between 2015 and 2021 at hospitals across the Commonwealth of Kentucky. The Markey Cancer Center Biospecimen Procurement and Translational Pathology Facility prepared tumor microarrays (TMAs) using Wilms tumor and matching normal tissue cores for immunohistochemistry (IHC) analysis. RNA-Seq analysis performed at the Markey Cancer Center Oncogenomics Facility identified a gene signature that was further validated by IHC and reverse-transcription quantitative-PCR (RT-qPCR). Outcome data provided by KCR indicated that increased expression of an anti-apoptotic protein in the tumor counter-intuitively provided better prognosis and outcome to treatment, indicating that the treatments either directly or indirectly targeted this protein. Loss of function of the tumor suppressor proteins in Par-4/PAWR, WT1 and p53 did not explain the higher expression of the cell survival protein. Our multidisciplinary team approach involves basic science/translational researchers, population scientists, biomedical informaticists, and clinicians and has resulted in well-characterized, Wilms tumor tissue microarrays. The next phase of the study is generating patient derived xenografts (PDXs) for a better understanding of Wilms tumor biology in children.

Citation Format: Jieyun Jiang, Derek B. Allison, Ravshan Burikhanov, Saptadwipa Ganguly, Ryan A. Goettl, Jinpeng Liu, Chi Wang, Amanda F. Saltzman, Thomas Tucker, Peter H. Spielmann, Eric B. Durbin, John A. D’Orazio, Vivek M. Rangnekar. Molecular signature for treatment-responsive and recurrent Wilms tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1103.
Abstract 1122: Inhibition of HDAC6 enhanced radiation therapy induced antitumor response in melanoma
Authors: Li Xintang , Suresh Manasa , Coulibaly Hawa , Quiceno-Torres David , Gajendran Nithya , Tan Karen , Noonepalle Satish , Grindrod Scott , Dritschilo Anatoly , Villagra Alejandro
Published: March 2024

Patients with advanced melanoma are often treated with radiation therapy (RT) in combination with immunotherapies. Immunogenic tumor cell death induced by radiation is known to activate innate and adaptive immunity. Macrophage activity in the irradiated tumor microenvironment (TME) has been implicated to play a critical role in post-RT innate immune responses. However, tumor-associated macrophages (TAMs) due to their phenotypic plasticity convert from initial antitumor M1 phenotype to protumor M2 macrophages resulting in relapse and metastasis of any surviving tumor cells. Therefore, strategies regulating tumor macrophages post-radiation can enhance the therapeutic effectiveness of radiation therapy. Here, we report targeting HDAC6 function with a novel selective inhibitor (SP-2-225) as a potential therapeutic candidate for combination with RT. When administered intraperitoneally at a dose of 25mg/kg, SP-2-225 resulted in significant tumor suppression in the immunocompetent SM1 murine melanoma model. Phenotyping of tumor immune infiltrate indicated a profound decrease in M2 macrophages resulting in an M1/M2 macrophage ratio and an increase in CD8 effector memory T-cells. Further, in-vitro co-culture studies using trans-well assays indicated that irradiated tumor cells attract M2 macrophages, and treatment with SP-2-225 abrogated the M2 macrophage migration but did not alter M1 macrophage migration. In addition, 24 hours post-RT, intratumor administration of SP-2-225 treated M1 macrophages as adoptive cell therapy resulted in diminished tumor growth. The combination of conditioned M1 macrophages with an irradiated tumor microenvironment resulted in an increased M1/M2 ratio and infiltration of effector memory and central memory CD8 T-cells underscoring the influence of transplanted macrophages on the local antitumor immunity. Therefore, our findings demonstrate the ability of HDAC6 inhibitors to regulate macrophage function and phenotype as a cell therapy for use in combination with radiation therapy.

Citation Format: Xintang Li, Manasa Suresh, Hawa Coulibaly, David Quiceno-Torres, Nithya Gajendran, Karen Tan, Satish Noonepalle, Scott Grindrod, Anatoly Dritschilo, Alejandro Villagra. Inhibition of HDAC6 enhanced radiation therapy induced antitumor response in melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1122.
Abstract 1128: Association between gut microbial change and acute gastrointestinal toxicity in patients with prostate cancer undergoing definitive radiation therapy
Authors: Jang Bum-Sup , Lee Dong Soo
Published: March 2024

Purpose: This prospective cohort study investigated the association between gut microbial changes and acute gastrointestinal toxicities in prostate cancer patients undergoing definitive radiation therapy (RT).

Methods: Seventy-nine fecal samples were analyzed. Stool samples were collected at the following timepoints: pre-RT (prRT), 2 weeks after the start of RT (RT-2w), 5 weeks after the start of RT (RT-5w), 1 month after completion of RT (poRT-1m), and 3 months after completion of RT (poRT-3m). We computed the microbial community polarization index (MCPI) as an indicator of RT-induced dysbiosis.

Results: Patients experiencing toxicity had lower alpha diversity, especially at RT-2w (P=0.037) and RT-5w (P=0.003). Compared to patients without toxicity, the MCPI in those experiencing toxicities was significantly elevated (P=0.019). In terms of predicted metabolic pathways, we found linearly decreasing pathways including carbon fixation pathways in prokaryotes (P=0.035) and the bacterial secretion system (P=0.005), in patients who experienced toxicities.

Conclusions: We showed RT-induced dysbiosis among patients who experienced toxicities. Reduced diversity and elevated RT-related MCPI could be helpfully used for developing individualized RT approaches.

Citation Format: Bum-Sup Jang, Dong Soo Lee. Association between gut microbial change and acute gastrointestinal toxicity in patients with prostate cancer undergoing definitive radiation therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1128.
Abstract 117: Improved PBMC humanized mouse models for immuno-oncology drugs evaluation
Authors: Xie Bin , Hou Xin , Yang Pengfei , Zhou Yanbing , Wen Danyi
Published: March 2024

It has been demonstrated that anti-PD(L)1 therapy could significantly improve survival outcomes for patients with metastatic solid cancers, such as melanoma, non-small cell lung cancer and MSI-H colorectal cancer. However, more than half of these patients (NCT02563002, NCT02142738, NCT02563002) do not respond to immune checkpoint blockade (ICB) therapy, and 90% of colorectal cancer patients have MSS/pMMR characteristics and almost no response at all, which drives scientists to design novel drug candidates urgently and integrate tumor and tumor microenvironment (TME) signals systemically to rejuvenate exhausted immune system and overcome ICB resistance. LIDE has developed both traditional PBMC humanized mice model, and human immune cells and cancer co-inoculated model for IO drugs functional evaluation in the last 5-years by using unique PBMC donor selection and standard cancer biobank (PDX patient-derived xenografts, CDX cell derived xenograft) establishment. Cancer biopsies with MDM2 amplification or DNMT3A alternation have been shown resistant to PD1-Ab therapy but sensitive to chemical combo-therapy, and pMMR cancer PDX models seem responsible well to EGFR targeted, CD39-CD73-Adenosine targeted therapy. Human IL15 transgenic mice are used as one useful tool to investigate in vivo function of NK cells, NK engagers/modulators as well as drugs with ADCC (Antibody dependent cell-mediated cytotoxicity) effect. The activated immune cells and cancer co-transfer model, can significantly delay the graft-versus-host-reaction for another month compared with traditional PBMC humanized model, and archives maximal therapeutic effect of the testing drugs by pre-activating T cells with specific tumor antigens. Dendritic cells or macrophages could be successfully introduced into this co-inoculated system to work together with T cells, mimic human tumor tissue complexity, to help determine the overall effect induced by integrated signals. It seems CD40 agonist or CD47 antagonist could well synergies with anti-PD(L)1 to control melanoma and triple negative breast cancer development, respectively. This optimized PBMC humanized cancer-bearing mice models have been tested for various cancer targets, e.g. PD(L)1, Tim3, Lag3, Tigit, PVRIG, TGFβ1, CD73/CD39, CD47/Sirpa, DLL3/CD3, Claudin18.2/4-1BB, B7-H3, GITR, to better understand the interaction between cancer cells and immune cells.

Citation Format: Bin Xie, Xin Hou, Pengfei Yang, Yanbing Zhou, Danyi Wen. Improved PBMC humanized mouse models for immuno-oncology drugs evaluation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 117.
Abstract 1171: Estimating sensitivity of exploratory cellular biomarkers in oncology clinical trials
Authors: Xue Chengsen , Swenson Christina D. , Mc Closkey Thomas W.
Published: March 2024

Background: In the prolonged and complex process of assessing immunotherapeutic oncology drug candidates, flow cytometry plays an essential role. In oncology studies, biomarker measurement requires robust and reliable flow cytometric assays. A crucial element of these assays is to understand the lower limits of measurable surface biomarkers.

Methods: We incorporated into the development and validation of flow cytometry assays the measurement of the lower limit of quantitation (LLOQ). The LLOQ is the lowest number of a reportable parameter in a blood or bone marrow study sample that can be quantitatively determined with acceptable precision and accuracy. Two steps are necessary to determine the LLOQ. First in these assessments a measurable range of lower limits of a particular cell type or receptor is estimated. That is, in validating an assay LLOQ, there are three options to select cells expressing target surface biomarkers of interest: using cultured or bioengineered cells, using samples of known high expression of a biomarker, or using pre-titered positive samples. Then decreasing concentrations of the target cell or receptor may be used to precisely determine the LLOQ of a specific assay reportable parameter.

Results: By acquiring at least ten negative control sample results, the limit of the blank (LOB) and limit of detection (LOD) of a biomarker is determined. LOB is calculated as mean +1.645 SD, and LOD as mean +3 SD. The negative control samples either lack the fluorescent monoclonal antibodies against the markers of interest or lack the target cell population. Different reportable parameters, detected by one or more biomarkers have varying LLOQ. The LLOQ for a reportable parameter is a precise measurement of a parameter above or equal to the LOD. We found that typical LLOQ values are from 4 to 145 positive cells, with a frequency of 0.01 to 0.1% among a parent population. For example, in the case of tetramer assays, flow cytometry can reliably detect as few as 25 CD8+ antigen-specific cells among three million lymphocytes.

Conclusions: LLOQ determinations for exploratory flow cytometry biomarkers are required for clinical research in cancer immunotherapy and cancer vaccine development. This valuable information allows for interpretation of data at the low end of a measurable range.

Citation Format: Chengsen Xue, Christina D. Swenson, Thomas W. Mc Closkey. Estimating sensitivity of exploratory cellular biomarkers in oncology clinical trials [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1171.
Abstract 1215: DIRAS3 inhibits oncogenic RAS signaling and RAS-dependent cell growth driven by prevalent KRAS hot spot mutations
Authors: Bildik Gamze , Liu Junchen , Mao Weiqun , Yang Hailing , Hancock John F. , Bast Robert C. , Lu Zhen
Published: March 2024

RAS mutations are found in 30% of human cancers and are characterized by hotspot mutations at codons G12, G13, and Q61. This results in constitutively activated RAS proteins. Despite recent advances in drug design and the emergence of covalent inhibitors, effectively targeting mutant RAS remains challenging. Our group previously identified the tumor suppressor protein DIRAS3, which directly binds Ras and inhibits its function by disrupting KRAS dimers/nanoclusters and blocking effector activation. DIRAS3 expression reduced cancer cell growth in pancreatic and ovarian cancers. To determine whether DIRAS3 plays a definitive role in inhibiting mutant KRAS, we comprehensively characterized DIRAS3's RAS-inhibitory effects using RASless mouse embryo fibroblast (MEF) cells as a model system lacking endogenous RAS isoforms. We ectopically expressed various KRAS mutations in these cells and then examined DIRAS3's effect on each KRAS mutant clone. We analyzed downstream RAS signaling, colony formation, cell viability, and toxicity. Our results showed that DIRAS3 significantly inhibited RAS-dependent colony formation in KRAS mutant (G12C, G12D, G12V, G13D, G12R, Q61R, Q61L) but not wild-type KRAS clones. However, when we added epidermal growth factor (EGF) to the medium, DIRAS3 expression inhibited wild-type KRAS activity, decreasing colony formation. This suggests that DIRAS3 selectively affects the active, GTP-bound form of KRAS and downstream signaling. Additionally, DIRAS3 expression decreased Erk1/2 phosphorylation in MEF cells with mutant KRAS, but not in the cells with a BRAFV600E mutation. This indicates that DIRAS3 specifically inhibits the KRAS-mediated MEK/ERK signaling, cell proliferation, and cytotoxicity. Moreover, to determine the critical region for DIRAS3's RAS-inhibitory function, we compared wild-type DIRAS3 and deletions (ΔNTE, ΔCTE, ΔNCTE). Our results showed both termini are required to effectively suppress clonogenic growth, emphasizing the requirement for DIRAS3 to anchor to the plasma membrane and interact with the RAS dimer interface to inhibit RAS-dependent tumor growth. In conclusion, our findings underscore the potent and specific inhibitory effects of DIRAS3 on RAS-driven oncogenesis, positioning it as a promising pan-RAS inhibitor for RAS-mutant cancers.

Citation Format: Gamze Bildik, Junchen Liu, Weiqun Mao, Hailing Yang, John F. Hancock, Robert C. Bast, Zhen Lu. DIRAS3 inhibits oncogenic RAS signaling and RAS-dependent cell growth driven by prevalent KRAS hot spot mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1215.
Abstract 1319: In vivo programming of natural killer cells and T cells using mRNA delivered cytotoxic chimeric antigen receptors
Authors: Diwanji Neha , Getts Daniel , Wang Yuxiao
Published: March 2024

Introduction: Current generation of adoptive cell therapy relies on ex vivo engineering of immune cells with tumor-targeting chimeric antigen receptors. In addition to manufacturing challenges, ex vivo engineered cells such as CAR-NK and CAR-T show limited tumor infiltration and persistency within tumor microenvironment. A method leveraging mRNA-based delivery for in-vivo engineering of these cells could address these issues but has not been established. Conventional mRNA/LNP when infused systemically result in significant accumulation in liver tissue and the CAR has no immune cell specificity in their expression or function.

Method: Many immune receptors are heterodimeric complexes composed of a ligand binding subunit (e.g. CD16 and TCR) and a signaling adaptor containing Immunoreceptor Tyrosine-based Activation Motif (ITAM) (e.g. FcRγ and CD3ζ). Notably, the cell surface expression of the ligand binding subunit often depends on the expression of the ITAM adaptor. Based on this we fused HER2-targeting scFv to multiple activating receptors whose expression is restricted to NK cells and T cells, and evaluated their expression and anti-tumor function.

Results: We developed a series of Natural Cytotoxic Receptor (NCR)-based in vivo CAR constructs by fusing HER2-targeting scFv to the native extracellular and transmembrane sequences of the receptor NKp30, NKp44 and NKp46, These CARs engage signaling adaptor FcRγ or DAP12 expressed in NK cells, and exhibit potent tumoricidal activity against HER2+ ovarian cancer cells as well as mediate tumor-induced cytokine and chemokine response. Using Huh7 liver cell as a model for off-target expression in hepatocytes, we found that NKp44 and NKp46-based CAR requires their cognate signaling adaptors for stable cell surface expression. We also showed that similar strategy can be used to design receptors for selective expression on the surface of T cells utilizing CD3ε chain fused with tumor targeting scFv.

Conclusion: These results demonstrated the feasibility of engineering primary NK cells with CARs based on the NCR family of receptors NKp44 and NKp46, whose expression requires the immune-restricted signaling adaptors. These CAR-NK receptors do not contain intracellular signaling motifs but can engage and activate their signaling natural adaptors to initiate tumor killing and cytokine response. An mRNA/LNP based delivery of NKp44-based CAR for cancer treatment is currently under clinical development.

Citation Format: Neha Diwanji, Daniel Getts, Yuxiao Wang. In vivo programming of natural killer cells and T cells using mRNA delivered cytotoxic chimeric antigen receptors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1319.
Abstract 1336: Verification of the effect of human allogeneic iPS cell-derived gene-engineered NK cells (eNK cells HLCN061) on mesothelioma
Authors: Matsumoto Seiji , Fujita Yoshiko , Goto Kumiko , Sato Yuka , Nishigaki Fusako , Higashi Yasuyuki , Kimura Hironobu , Tamura Kouichi
Published: March 2024

Malignant pleural mesothelioma is a malignant tumor that is caused by exposure to asbestos. Because it is a rare cancer, there are few drugs that can be used to treat it. Healios is developing iPSC-derived NK cells (eNK cells) (HLCN061) engineered with NKG2D, DAP10, IL-15, CD16, CCL19, and CCR2B genes for the treatment of refractory solid tumors. This study evaluated the antitumor effect of eNK cells against human malignant pleural mesothelioma cell lines.(Method) The antitumor effect of eNK cells on human malignant pleural mesothelioma cell lines in vitro was evaluated using the Viability/Cytotoxicity Multiplex Assay Kit (WST-8/LDH).In vivo, immunodeficient mice were subcutaneously inoculated with human malignant pleural mesothelioma cell lines. After subcutaneous tumor formation, eNK cells were inoculated into or near the tumor, the size of the subcutaneous tumor was measured, and the tumor volume was compared with that of an untreated group. After the study was completed, the tumors were removed and the tumors in the untreated group and the eNK cell-administered group were analyzed using RNA-seq.(Results) The antitumor effect of eNK cells against human malignant pleural mesothelioma cell lines was confirmed in vitro. In a mouse subcutaneous tumor model, the eNK cell treatment group suppressed tumor growth compared to the untreated group (p≦0.05). RNA-seq showed increased expression of granulysin, IL2RB, cathepsin W, etc. in subcutaneous tumors treated with eNK cells.

Citation Format: Seiji Matsumoto, Yoshiko Fujita, Kumiko Goto, Yuka Sato, Fusako Nishigaki, Yasuyuki Higashi, Hironobu Kimura, Kouichi Tamura. Verification of the effect of human allogeneic iPS cell-derived gene-engineered NK cells (eNK cells HLCN061) on mesothelioma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1336.
Abstract 1337: Characterization of anti-AXL CAR-NK cell therapy for the treatment of melanoma
Authors: Hibler Winston , Merlino Glenn , Yu Yan Lin
Published: March 2024

AXL is a receptor tyrosine kinase (RTK) that is commonly expressed in metastatic melanoma and several other forms of cancer. Activation of AXL signaling is linked to tumor cell growth, invasion, metastatic transformation, drug resistance, and immune response regulation. However, treatment of metastatic melanoma continues to present clinical challenges due to its capacity to rapidly develop drug resistance, which manifests under the influence of AXL signaling. As such, AXL may be a viable antigen target for immunotherapies. Despite their successes in hematological malignancies, chimeric antigen receptor (CAR)-T cell therapies remain limited by their susceptibility to causing off-target toxicity. In contrast, natural killer (NK) cells are not associated with such risks, which makes them clinically appealing as an alternative to T cells. However, studies on the application of CAR-NK cells to treat solid tumors remain acutely scarce. To explore the clinical viability of CAR-NK cells for the treatment of metastatic melanoma, AXL-specific CAR-NK cells were generated by transfecting NK cells obtained from NK cell lines. Preliminary data from a cytotoxicity assay indicated that the cytotoxicity of CAR-NK92-MI cells against AXL-positive melanoma cell lines was significantly stronger (up to a 10-fold difference) than the corresponding parental cells. Further in vitro characterization of this CAR-NK cell line and other CAR-NK cell lines, as well as in vivo evaluation of their efficacy in a mouse xenograft model of melanoma metastasis, is underway.

Citation Format: Winston Hibler, Glenn Merlino, Yan Lin Yu. Characterization of anti-AXL CAR-NK cell therapy for the treatment of melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1337.