bluuebunny/crossref_metadata_2025 huggingface dataset:
Fingerprinting dissolved organic compounds: a potential tool for identifying the surface infiltration environments of meteoric groundwaters
Authors: Stillings M. , Lunn R. J. , Shipton Z. K. , Lord R. A. , Thompson S. , Knapp M.
Published: March 2024

Current methods for tracing decades-old groundwaters rely on isotope geochemistry to determine groundwater age and altitude at the point of infiltration. Temporal and spatial variability in atmospheric conditions, and water–rock interactions, can make the interpretation of isotopes uncertain. Here, we propose a new method of groundwater tracing based on the fingerprinting of natural dissolved organics. We present our initial findings from the Grimsel Test Site in Switzerland, located within a fractured granite. Using 2D gas chromatography, we derive detailed organic fingerprints from surface soils at several locations and show that different soils produce distinctly different dissolved organic signatures. We then compare the soils with groundwater and lake water using a non-targeted approach employing principal component analysis and hierarchical cluster analysis. Our analysis finds three statistically significant clusters. Most groundwaters are clustered with the lake-water samples but two are clustered with soil from the highest altitude surface sampling location. We hypothesize that for samples to form a significant cluster, they must have been derived from a common environment, with a unique combination of organic compounds. For groundwaters to cluster with soil samples or lake water, we theorize there must be a hydraulic connection between the type of infiltration environment and the groundwater sampling locations within each cluster. Our research demonstrates that organic molecules derived from the surface environment can be used to discriminate near-surface environment(s) through which meteoric groundwater has infiltrated. Organic fingerprinting could prove a powerful tool for improved understanding of groundwater flow systems, particularly when combined with other complementary techniques.

Supplementary material
:
Compound alignment data set and supplementary tables are available at
https://doi.org/10.6084/m9.figshare.c.7129987

Thematic collection:
This article is part of the Sustainable geological disposal and containment of radioactive waste collection available at:
https://www.lyellcollection.org/topic/collections/radioactive
Multiwalled-Carbon-Nanotube-Modified Li2ZnTi3O8 as Anode with Improved Cycling Stability and Rate Capability for Lithium-Ion Batteries
Authors: Yıldız Süleyman , Şahan Halil , Şanlı Ferhat , Patat Şaban
Published: March 2024

Due to its better electronic conductivity and larger lithium intercalation capacity, a multiwalled-carbon-nanotube (MWCNT)-modified Li2ZnTi3O8 (LZTO) has significant potential for use as an anode material for Li-ion batteries. The rapid and affordable ball-mill-aided solid state approach is used to synthesize LZTO with MWCNT-modified Li2ZnTi3O8 (LZTO@MWCNTs). Electrochemical performance tests of the anode material were carried out using cyclic voltammetry (CV), galvanostatic charge-discharge measurement, and electrochemical impedance spectroscopy (EIS). The CV experiments reveal that the LZTO@MWCNT electrode has a lower anodic and cathodic peak potential difference (0.184 V) than the LZTO electrode (0.211 V) and LZTO@MWCNT electrode has better electrochemical reversibility than that of pristine electrode after five cycles utilizing the same procedure. LZTO@ MWCNTs anode material has a higher charge-discharge capacity than LZTO anode material at 0.1–5 C rates. After 100 cycles, the initial discharge capacities of LZTO@MWCNT electrode are 227 and 142 mAh g−1 at the 1C–5C rate, respectively, whereas the initial discharge capacities of LZTO electrode are only144 and 28 mAh g−1 in the same condition. The charge transfer and SEI resistance of the LZTO anode are 19.31 and 62.74 ohms, respectively, after 30 cycles at 1C, according to the EIS data. Additionally, the values for the LZTO@MWCNTs anode are 7.93 and 12.06 ohms for charge transfer and SEI resistance, respectively.
Regulation of renal aquaporin water channels in acute pyelonephritis
Authors: Ernstsen Christina V. , Ranieri Marianna , Login Frédéric H. , Mahmoud Isra K. , Therkildsen Jacob R. , Valenti Giovanna , Praetorius Helle , Nørregaard Rikke , Nejsum Lene N.
Published: March 2024

Acute pyelonephritis (APN) is associated with reduced renal concentration capacity and increased aquaporin-2 (AQP2) excretion. Uropathogenic Escherichia coli (UPEC) mediates changes in the subcellular localization of AQP2 and we show that in vitro, these changes could be elicited by two pathogen-associated molecular patterns (PAMPs), namely, lipopolysaccharide and peptidoglycan. UPEC infection was unaltered by AQP2 expression and the other renal AQPs (AQP1, AQP3, and AQP4) were unaltered in APN.
Serum Ghrelin and Leptin Concentrations in Patients with Major Depressive Disorder before and after Supplementation with Vitamin D3
Authors: AlGhamdi Shareefa , Alsulami Nabilah , Khoja Sawsan , Alsufiani Hadeil , Tayeb Haythum O. , Alshaibi Huda , Tarazi Frank I.
Published: March 2024

Aim. To determine serum concentrations of leptin and ghrelin in patients with major depressive disorder (MDD) before and after vitamin D3 supplementation. Methods. A total of 72 participants were recruited in this study (40 MDD patients and 32 healthy controls). MDD was diagnosed by using Beck’s Depression Inventory (BDI) scale. Blood samples were collected from all participants at the beginning of the study to determine baseline serum 25(OH)D3, leptin, and ghrelin concentrations. Patients were then treated weekly with vitamin D3 (50,000 IU) for 3 months, and blood samples were collected again by the end of the study. Results. At baseline, serum leptin concentrations were significantly higher in MDD patients than in healthy controls. In contrast, serum ghrelin concentrations were significantly lower compared to those in healthy controls. After supplementation with vitamin D3 for three months, MDD patients showed improvements characterized by a decrease in their BDI’s scores and an increase in their serum vitamin D and ghrelin concentrations. No effects of vitamin D3 supplementation were seen on serum leptin concentration. Conclusions. The antidepressant effects of vitamin D3 supplementation could be mediated by ghrelin but not leptin.
Rheological and Biological Properties of Adhesive Skin Secretions from Eupsophus vertebralis (Anura: Alsodidae)
Authors: Contreras Felipe A. , Sepúlveda Daniela P. , Amaral André Capaldo , Nuñez José J. , Trovatti Eliane , Suárez-Villota Elkin Y.
Published: March 2024

Skin secretions from Patagonian ground frogs, Eupsophus vertebralis, have previously been reported as a potent proteinaceous adhesive with potential biomedical applications. Here, we conducted a rheological analysis indicating the mechanical robustness of these secretions, with a storage modulus ranging from 1 to 10 Pa. In addition, antimicrobial and cytotoxicity assays were performed, revealing no antimicrobial activity against both the Gram-positive and Gram-negative bacteria. The cytotoxicity results were intriguing, as three samples showed no harm, and one exhibited a severe cytotoxic effect on the human cell line MG63. These properties, as indicated by these preliminary results, reinforce their potential for practical applications in the industrial and medical sectors.
First Molecular Detection of SARS-CoV-2 in Sewage and Wastewater in Ghana
Authors: Duker Ewurabena Oduma , Obodai Evangeline , Addo Seth Offei , Kwasah Lorreta , Mensah Edna Serwah , Gberbi Emmanuel , Anane Abraham , Attiku Keren O. , Boakye Jessica , Agbotse Gayheart Deladem , Dickson Angelina Evelyn , Quarcoo Joseph Ahia , Darko Patience Akosua , Larbi Yaw Awuku , Ntim Nana Afia Asante , Dzudzor Bartholomew , Odoom John Kofi
Published: March 2024

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is shed in the stool of infected individuals and can be detected in sewage and wastewater contaminated with infected stool. This study is aimed at detecting the virus and its potential survival in sewage and wastewater in Ghana. The cross-sectional study included samples from 16 validated environmental surveillance sites in 7 regions of Ghana. A total of 354 samples composed of wastewater (280) and sewage (74) were collected from November 2020 to November 2022. Overall, 17% of the samples were positive for SARS-CoV-2 by real-time PCR, with 6% in sewage and 11% in wastewater. The highest number of positive samples was collected from the Greater Accra Region (7.3%) with the least recorded in the Bono East Region (0.6%). Further characterization of the positive samples using the next-generation sequencing (NGS) approach yielded two variants: Alpha (B.1.1.7) and Delta (AY.36). Attempts to isolate SARS-CoV-2 in the Vero cell line were not successful probably due to the low viral load concentrations (Ct values>35) or prolonged exposure to high temperatures rendering the virus noninfectious. Our findings suggest that SARS-CoV-2 RNA in sewage and wastewater may not be infectious, but the prevalence shows that the virus persists in the communities within Ghana.
Abstract 1028: Assessment of the diagnostic accuracy of Oncuria-Detect® for detection of upper tract urothelial carcinoma
Authors: Sakatani Toru , Tanaka Sunao , Fujita Kazutoshi , Linehan Jennifer , Murakami Kaoru , Lee Riko , Pagano Ian , Rosser Charles J. , Furuya Hideki
Published: March 2024

Introduction and Objective: Due to insufficient accuracy, urine-based assays currently have a limited role in the management of patients with upper tract urothelial carcinoma (UTUC). The application of a robust urine-based multiplex assay, Oncuria-Detect®, to aid in the detection of UTUC has the potential to address this deficiency and to assist with accurate, non-invasive diagnosis.

Methods: To evaluate the performance of Oncuria-Detect®, a multiplex immunoassay capable of querying a voided urine sample for 10 protein biomarkers associated with urothelial carcinoma (A1AT, APOE, ANG, CA9, IL8, MMP9, MMP10, PAI1, SDC1 and VEGFA). Its application to UTUC was evaluated in a multi-institutional cohort of 35 prospectively collected subjects presenting for evaluation of upper tract mass along with 46 prospective collected matched controls (i.e., non-tumor bearing). The ability of the test to identify patients harboring UTUC was assessed. UTCU status was confirmed by endoscopy and tissue biopsy or definitive surgery. Diagnostic performance was assessed using ROC curves.

Results: Oncuria-Detect® provided an AUC of 0.886 (95% CI: 0.807-0.964) with an overall sensitivity of 91.4%, specificity of 73.9%, NPV 91.9% and PPV 72.7%. Sensitivity values of the diagnostic panel for low-grade UTUC, high-grade UTUC, non-invasive UTUC and invasive UTUC were 86.7%, 90.9%, 88.9% and 94.1%, respectively. Urinary cytology or selective ureteral washing/cytology was associated with an overall sensitivity of 54.2%, specificity of 100%, NPV 63.3% and PPV 100%. Sensitivity values of cytology for low-grade UTUC, high-grade UTUC, non-invasive UTUC and invasive UTUC were 100%, 100%, 100%, and 77.8%, respectively.

Conclusions: Urinary levels of a biomarker panel enabled the accurate discrimination of UTUC and controls non-tumor bearing individuals. The multiplex Oncuria-Detect® test can achieve the efficient and accurate detection of UTUC in a non-invasive patient setting.

Citation Format: Toru Sakatani, Sunao Tanaka, Kazutoshi Fujita, Jennifer Linehan, Kaoru Murakami, Riko Lee, Ian Pagano, Charles J. Rosser, Hideki Furuya. Assessment of the diagnostic accuracy of Oncuria-Detect® for detection of upper tract urothelial carcinoma [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 1028.
Abstract 1036: Development of a comprehensive RNA-based NGS panel for detecting tyrosine kinase fusion genes associated with primary hypereosinophilia
Authors: Lee Bongyong , Chern Andrew , Zhang Aiguo , Lu Chuanyi M. , Sha Michael Y.
Published: March 2024

Hypereosinophilia (HE) is a medical condition diagnosed when the count of eosinophils in the blood is greater than 1.5 × 109 per liter. Primary HE is a clonal stem cell disorder manifesting as myeloid and/or lymphoid malignancies and is typically driven by aberrant fusion genes involving a tyrosine kinase (TK) gene (e.g., PDGFRA, PDGFRB, FGFR1, JAK2, FLT3, ABL1). The precise identification of the underlying molecular genetic abnormalities is crucial for timely diagnosing and managing primary HE. There has been no HE-specific assay that can concomitantly detect all known or novel TK fusion genes. We've developed an RNA-based panel for detecting TK fusion genes in HE. This panel, developed using 5' RACE technology, allows for the identification of fusion transcripts in total RNA extracted from whole blood sample. It covers 48 known TK fusion genes and also has the capability of detecting novel gene fusions. We balanced and optimized the panel using an in-house fusion RNA mix containing 11 fusion transcripts. We additionally validated the panel using samples from five patients with hematopoietic neoplasms and two fusion-positive cell lines to ensure its accuracy. The Limit of Detection (LoD) was established by analyzing the in-house RNA mix and normal whole blood RNA. It was found that the minimum RNA input required to detect all 11 fusion transcripts was 3540 copies in the presence of a background of 10 ng normal whole blood RNA. The panel successfully identified two fusion transcripts using 10 ng of total RNA obtained from cancer cell lines. The panel was then used to test five patient RNA samples extracted from whole blood. We successfully confirmed the presence of BCR-ABL1 fusion in 3 patient samples and ETV6-PDGFRB fusion in 1 patient sample, respectively. In summary, we developed HE-specific NGS panel for detecting all known and novel TK fusion genes and demonstrated its analytical accuracy and clinical effectiveness. The next phase of our study is to further validate the tests on a large patient cohort.

Citation Format: Bongyong Lee, Andrew Chern, Aiguo Zhang, Chuanyi M. Lu, Michael Y. Sha. Development of a comprehensive RNA-based NGS panel for detecting tyrosine kinase fusion genes associated with primary hypereosinophilia [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 1036.
Abstract 1114: Evaluating strategies to harness metabolic vulnerabilities in cervical
Authors: Schwarz Dr Julie K. , Gabriel Dr Nishanth , Banik Aunannya
Published: March 2024

Tumor cells frequently alter their metabolism to generate biomolecular intermediates for tumor maintenance and proliferation. Previous reports from Dr. Julie Schwarz’s laboratory have shown that cervical cancer cells with altered AKT/mTOR pathways show increased reliance on glucose and exogenous glutamine. The laboratory has introduced PIK3CA E545K mutation and PTEN null mutation in cervical cancer cell line SiHa. Further, to evaluate the role of glutamine metabolism in these genotypes, these cells were cultured in sequentially lower levels of glutamine until these cells were able to proliferate without exogenous glutamine supplementation. These isogenic long-term glutamine-deprived (QD) cells allow us to tease out the role of these specific mutations in cellular metabolism. In our study, long-term glutamine deprivation inhibits the proliferation of rapidly proliferating PI3K-AKT-altered cell lines. Even though glutamine is required for GSH synthesis, which quenches reactive oxygen species, QD cells were refractory to cisplatin and irradiation. Since PI3K-AKT altered were dependent on exogenous glutamine, it was expected that QD cells would have lower GLS levels to inhibit glutaminolysis. However, GLS levels in WT and QD were similar. Altogether, these data indicate that in PI3K-AKT altered cervical tumor glutamine antagonists can reduce tumor proliferation. Further glutamine reduction does not alter GLS levels and hence GLS can be targeted as well to further improve treatment outcomes. To evaluate the alternate metabolic dependencies in cervical cancer tumors, both cellular as well as secreted metabolites were evaluated. To identify alternative pathways that are activated in these glutamine-deprived cell lines, we cultured these cells in uniformly labeled 13C glucose. Mass spectrometric studies also evaluated the protein levels of various metabolic intermediates in glutamine-deprived cell lines compared to control cells. To confirm our observations, we used kit-based assays to evaluate metabolites in the cells. To further confirm the change in the metabolite pathway we evaluated changes in transcriptional changes of regulating proteins/enzymes. This pre-clinical study reinforced efforts that are anticipated to reveal new metabolic vulnerabilities of cancer cells that can be targeted by therapeutic interventions. This study also helped us evaluate druggable metabolic targets that can be targeted for effective tumor eradication along with conventional treatment. This understanding will help advance the pre-clinical trials persistent to the cure of cervical cancer.

Citation Format: Dr Julie K. Schwarz, Dr Nishanth Gabriel, Aunannya Banik. Evaluating strategies to harness metabolic vulnerabilities in cervical [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 1114.
Abstract 1247: PPARsing epigenetic memory in intestinal stem cells: High-fat diet and oncogenic susceptibility
Authors: Saiz Dominic R. , Hartley-McDerrmott Thomas , Barrera-Millan Yesenia , Castro-Ochoa Karla Fabiola , Shukla Abhigyan , Torel Matthew , Mana Miyeko D.
Published: March 2024

Diet has a profound impact on health and susceptibility to cancer. Adaptation to a Western diet is strongly associated with increased colorectal cancer (CRC) and obesity prevalence. This study demonstrates the epigenetic changes in intestinal stem cells (ISCs) induced by a pro-obesity high-fat Western diet (HFD) and the heightened risk of oncogenic transformation. A HFD induces a phenotype characterized by increased ISC frequency, proliferation, regenerative capacity, and ability to initiate adenomas. We hypothesized that a HFD generates lasting epigenetic alterations in ISCs thereby enhancing oncogenic susceptibility. We evaluated the chromatin landscape of intestinal stem cells (ISCs) and the reversibility of the epigenetic changes. Mice carrying a fluorescent reporter of the ISC marker Lgr5, were subjected to a control diet, HFD, or HFD with a return to control. We isolated ISCs by flow cytometry and used an assay for transposase-accessible chromatin sequencing (ATAC-seq) to identify differentially accessible genomic regions correlated with the ISC phenotype. We found that a HFD induces distinct changes in the chromatin landscape that persist after removal from the HFD. These accessible regions are associated with genes involved in lipid metabolism and are enriched with PPAR (Peroxisome Proliferator-Activated Receptor) binding motifs. We demonstrate that these lipid metabolism regulators, PPAR-delta and PPAR-alpha, are necessary for the majority of significant accessible regions. We further show that HFD-induced accessibility significantly correlates with early chromatin alterations seen by loss of tumor suppressor APC. Our data supports a mechanistic pathway for diet-induced epigenetic reprogramming due to PPAR activity and suggests that the HFD imposes a preneoplastic stem cell chromatin state that stokes the risk of oncogenic change.

Citation Format: Dominic R. Saiz, Thomas Hartley-McDerrmott, Yesenia Barrera-Millan, Karla Fabiola Castro-Ochoa, Abhigyan Shukla, Matthew Torel, Miyeko D. Mana. PPARsing epigenetic memory in intestinal stem cells: High-fat diet and oncogenic susceptibility [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 1247.
Abstract 1267: Projected impact of liquid biopsy screening strategies with high sensitivity in focused populations and high specificity in broad populations
Authors: Bach Peter B. , Trivedi Niti U. , Cotton Lindsey
Published: March 2024

Purpose: Two 'liquid biopsy' cancer screening approaches exist: Focused and Broad. The Focused approach targets specific cancers and populations lacking adequate screening, emphasizing high sensitivity. The Broad approach aims to detect various cancers in the population, prioritizing high specificity to minimize false-positives but potentially compromising sensitivity. The public health impact and testing burden of these approaches remain unexplored.

Methods: Projected cancer mortality reduction and annual test volumes are based on published data for liquid biopsy tests. Focused screening is modeled for patients not undergoing guideline-recommended screening for lung, liver, and colorectal cancers. Broad screening is modeled for individuals aged 50-80, assuming the detection of all cancer types. Averted deaths are calculated by combining published mortality reduction estimates or assumed at 20% if not available. Reductions are adjusted for reported test sensitivities. Liquid biopsy cannot prevent deaths from cancers recommended for screening if the person has already undergone the recommended test.

Results: Focused and broad screening strategies annually prevented around 20,000 cancer deaths. Focused screening necessitated testing only one-fifth of the people compared to broad screening (30MM vs. 150MM). The projected reduction in mortality for focused testing relied entirely on randomized studies, while the equivalent statistic for the broad strategy was 53%.

Conclusions: Equivalent reductions in cancer deaths can be achieved with focused screening at high sensitivity and broad screening at high specificity, with only one-fifth of the testing burden. The influence of false positives in focused testing and false reassurance in broad testing may impact these findings, which could also be influenced by testing and follow-up costs.

Table 1 Annual cancer deaths averted (thousands) Focused cancer testing Broad cancer testing Lung 10-12 4-8 Liver 4-5 1 Colorectal 3-4 1 Other screened cancers* Not modeled 1-2 Unscreened cancers Not modeled 6-10 Total 17-20 14-22 Annual test volume (millions) 30 150 *Other screened cancers includes prostate, breast, cervical. Range reflects the application of reported Stage I or overall sensitivity, respectively.

Citation Format: Peter B. Bach, Niti U. Trivedi, Lindsey Cotton. Projected impact of liquid biopsy screening strategies with high sensitivity in focused populations and high specificity in broad populations [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 1267.
Abstract 1318: Targeting multiple myeloma with BCMA-CAR NK cells expressing a GPRC5D-NKG2D bispecific antibody
Authors: Tian Lei , Chen Victoria , Barr Tasha , Li Zhiyao , Chen Yuqing , Zhang Jiaying , Caligiuri Michael A. , Yu Jianhua
Published: March 2024

Recently developed therapeutics for multiple myeloma (MM) include targeted therapy, immunomodulatory drugs, and immunotherapy, the latter of which consists of B-cell maturation antigen (BCMA)-CAR cells, bispecific antibodies (BsAbs) to engage T cells. However, challenges including relapse and toxicity still exist. The orphan G protein-coupled receptor, class C group 5 member D (GPRC5D), emerged as a novel target for MM therapy. Therefore, following our established system [Cancer Immunol Res PMID: 29769244; Leukemia, PMID:24067492], we generated a BsAb by fusing anti-GPCR5D single-chain variable fragment (scFv) and an anti-NKG2D scFv (GPRC5D-NKG2D BsAb) in a human IgG4 platform and expressed it from BCMA-CAR and soluble IL15 natural killer (NK) cells (s15.BsAb.BCMA-CAR NK) to address the current known limitations. Our data showed that s15.BsAb.BCMA-CAR NK cells secreted GPRC5D-NKG2D BsAb both in vitro and in vivo. To evaluate the function of GPRC5D- NKG2D BsAb expressed from NK cells in vivo, we treated MM tumor-bearing mice with the purified antibody. The data showed that GPRC5D-NKG2D BsAb demonstrated a significantly better survival effect than saline (P≤0.01). We tested s15.BsAb.BCMA-CAR NK cells for cytotoxicity against a homogenous tumor cell mode (BCMA + GPRC5D + ) and a heterogeneous MM tumor cell model, including 50% BCMA + GPRC5D + , 25% BCMA ─ GPRC5D + , and 25% BCMA + GPRC5D ─ cells. Real-time cell analysis (RTCA) data showed s15.BsAb.BCMA-CAR NK cells killed tumor cells in vitro better than both BCMA-CAR NK cells and empty vector-transduced NK cells in an antigen-dependent manner. To test the efficacy of s15.BsAb.BCMA-CAR NK cells in vivo, we established a heterogeneous MM tumor cell mouse model, by mixing BCMA + GPRC5D + OPM2 cells and BCMA + GPRC5D ─ OPM2 cells at the ratio of 2:1. MM mice were treated with “off-the-shelf” different construct NK cells. The results showed that s15.BsAb.BCMA-CAR NK cells significantly prolonged survival compared to other NK cells(s15.BsAb.BCMA-CAR vs. BCMA-CAR P = 0.0012). Our data demonstrated that this all-in-one, multifaceted modality provides 2 ready-to-use biologics simultaneously, CAR NK cells and BsAb, for MM treatment.

Citation Format: Lei Tian, Victoria Chen, Tasha Barr, Zhiyao Li, Yuqing Chen, Jiaying Zhang, Michael A. Caligiuri, Jianhua Yu. Targeting multiple myeloma with BCMA-CAR NK cells expressing a GPRC5D-NKG2D bispecific antibody [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 1318.
Abstract 1515: Tertiary lymphoid structures restrain cancer evolution by supporting stem-like CD8+ T cells for sustained anti-tumor immunity
Authors: Wang Pengxiang , Zhou Chi , Wei Zhiting , Wang Yue , Gao Yuli , Meng Fangliangzi , Zhou Jian , Fan Jia , Liu Qi , Sun Yunfan
Published: March 2024

Currently, our understanding of the mechanism underlying the immune response on extending the survival of patients with aggressive tumors remains limited. In this study, we generated multi-regional genomic and transcriptional data coupled with digital pathology from 28 long-term survivors (LTSs) and 34 stage-matched short-term survivors, with intrahepatic cholangiocarcinoma (iCCA). Our investigation unveiled that mature tertiary lymphoid structures (TLSs) within the tumors of LTSs exerted significant immune pressure on the tumor, leading to the restriction in generating clonal mutations and increased subclonal diversity. Employing single-cell profiling, we identified significant enrichment of IgM+ naïve B cells and TCF7+ stem-like CD8+ T cells in TLS-positive tumors. Spatial profiling further highlighted that TCF7+ stem-like CD8+ T cells preferentially localized in TLS, facilitating reduced vulnerability to unfavorable environmental factors. Functionally, TCF7+ stem-like CD8+ T cells displayed strong capacity to differentiate into effector subsets while maintaining a diverse T-cell receptor repertoire. Mechanistically, we identified that naïve B cells promote activation and differentiation of stem-like CD8+ T cells within the TLS niche through IgM-FcμR axis, which primed these T cells for enhanced anti-tumor immunity. In summary, our findings underscore that TLS plays a pivotal role in orchestrating enduring and effective anti-tumor immunity, which yield strong immune stress on cancer evolution and ultimately contributing to better prognosis of patients.

Citation Format: Pengxiang Wang, Chi Zhou, Zhiting Wei, Yue Wang, Yuli Gao, Fangliangzi Meng, Jian Zhou, Jia Fan, Qi Liu, Yunfan Sun. Tertiary lymphoid structures restrain cancer evolution by supporting stem-like CD8+ T cells for sustained anti-tumor immunity [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 1515.
Abstract 1533: Characterizing tumor-immune surveillance in Li-Fraumeni syndrome
Authors: Giovino Camilla , Fischer Nicholas , Ong Noel , Kissoondoyal Ashby , Kafri Ran , Malkin David
Published: March 2024

Li-Fraumeni Syndrome (LFS) is a cancer predisposition syndrome associated with germline TP53 mutations and increased lifetime risk of developing multiple cancers. p53, the protein encoded by TP53, plays a role in regulating various aspects of immune cell biology. Accordingly, inactivation of p53 in tumor cells shapes interactions with the immune system and influences the anti-tumor immune response. Mutation or deletion of p53 in immune cells also favors tumor development and progression. However, to date, a comprehensive study of immune cell populations and the anti-tumor immune response has not been undertaken in the context of LFS. We hypothesized that germline TP53 mutation carriers possess dysfunctional immune cells and/or immune responses, supporting LFS-associated tumor progression. Our preliminary data indicate a profound defect in tumor-immune surveillance in an LFS murine model. We observed that MC38 cancer cells develop significantly faster into tumors when injected into LFS mice (harboring a gain-of-function mutation Trp53R172H/WT) compared to wildtype (WT) control mice. Importantly, we identified significant alterations in immune cell compositions and phenotypes within the tumor microenvironment as well as immune organs using flow cytometry. We are currently using ex vivo proliferation assays and cytotoxicity assays to investigate the impact of mutant p53 on the tumor-killing ability of immune effector cells. Altogether, an improved understanding of anti-tumor immune dysfunction in the context of germline TP53 mutation may reveal the potential for innovative therapies to target different facets of the immune system to intercept cancer progression in LFS patients.

Citation Format: Camilla Giovino, Nicholas Fischer, Noel Ong, Ashby Kissoondoyal, Ran Kafri, David Malkin. Characterizing tumor-immune surveillance in Li-Fraumeni syndrome [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 1533.
Abstract 1565: USP13 influences metastatic features in fallopian tube-originated high-grade serous carcinoma
Authors: Mok Boram , Kwon Juntae , Zhang Jinmin , Allsup Samuel , Choi Hyeongjwa , Han Cecil
Published: March 2024

High-grade serous ovarian carcinoma (HGSOC), originating from the fallopian tube, stands as a formidable challenging gynecologic malignancy with the highest mortality rate worldwide. Ubiquitin-specific peptidase 13 (USP13) has garnered attention due to the highly frequent USP13 gene copy amplification in human ovarian cancer. However, its precise pathological role in ovarian cancer remained elusive. To uncover the underlying role of USP13 in HGSOC, we developed a novel genetically engineered mouse model (GEMM) of HGSOC. Ovgp1-iCreERT2;Trp53flox/flox;Ptenflox/flox mouse (OPT) was crossed with Rosa26−LSL-Usp13 knockin mouse model (U), generating Ovgp1-iCreERT2;Trp53flox/flox;Ptenflox/flox ; Rosa26−LSL-Usp13 mouse model (OPTU). Usp13 overexpression with loss of p53 and Pten was induced specifically in fallopian tube secretory epithelial cells in OPTU mice. OPTU mice demonstrated the development of STIC and early HGSOC in the fallopian tube as well as invasive metastatic HGSOC. While OPT tumors largely retained mucinous and HGSOC subtypes, displaying limited metastatic behavior, OPTU tumors exhibited more complex ovarian tumor subtypes, including mucinous, HGSOC, and Mixed Mesodermal Mullerian Tumor (MMMT) histology. Furthermore, OPTU tumors showed invasive and metastatic phenotypes, characterized by an increased incidence of ovary invasion, metastasis to distant organs, and ascites development. Importantly, OPTU-driven HGSOC tumors closely resemble human HGSOC histology features, making the OPTU model a valuable tool for exploring fallopian tube-originated ovarian cancer. Primary ovarian cancer cells established from ascites of the OPTU mouse reproduced metastatic HGSOC tumorigenesis in the syngeneic mouse study. Stable USP13 knockdown in OPTU cells exhibited reduced cell proliferation and metastatic abilities in 2D and 3D cultures. In summary, this study underscores the pivotal role of USP13 in facilitating metastasis in fallopian tube-originated HGOSC. Our findings may provide a promising avenue for future therapeutic strategies aimed at mitigating the aggressive nature of USP13-amplified fallopian tube-originated HGOSC.

Citation Format: Boram Mok, Juntae Kwon, Jinmin Zhang, Samuel Allsup, Hyeongjwa Choi, Cecil Han. USP13 influences metastatic features in fallopian tube-originated high-grade serous carcinoma [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 1565.
Abstract 163: Role of macrophages in the development of ovarian cancer stem-like cells during chemotherapy
Authors: Cruz Luisjesus S. , Stevenson Denay , Robinson Mikella , Mathew Steffy , Amador Isabella , Jordan Gregory , House Carrie D.
Published: March 2024

Ovarian cancer is a poorly understood disease with 75% death rate when identified after metastasis. Drug resistance and tumor recurrence are likely due to cancer stem-like cells (CSCs) that evade chemotherapy and induce relapse with phenotypes that are distinct from bulk tumor cells. It remains unclear how CSCs facilitate relapse and what role the tumor microenvironment (TME) plays in this process. Our preliminary data show that a secreted cytokine, tumor necrosis factor-like weak inducer of apoptosis (TWEAK), and its receptor Fn14 are overexpressed in ovarian tumors and increase after chemotherapy. TWEAK is a strong inducer of stem cell features and enhances survival of CSCs during chemotherapy exposure. The source of TWEAK has not been clarified, although clinical data from human ovarian tumors show that TWEAK mRNA was primarily observed in a subset of infiltrating immune cells known as tumor associated macrophages (TAMs). When we evaluated TWEAK expression in cells found in the ovarian TME, including PBMC and THP-1-derived macrophages, IMR-90 activated fibroblasts, and ovarian cancer cell lines, we found that PBMC macrophages were the primary source of TWEAK production. Since cytotoxic chemotherapy can enrich for different TAM populations, we propose that TWEAK producing TAMs support CSC survival and expansion and contribute to the high rate of relapse in ovarian cancer patients. We further found that treatment with a small molecule inhibitor of TAM polarization leads to decreased TWEAK production, fewer CSCs, and prolonged remission in mouse models of ovarian cancer relapse as compared to vehicle. Currently we are testing whether knockout of Fn14 in ovarian cancer cells co-cultured with TAMs reduces stem cell features dependent on TWEAK-Fn14 signaling. Identification of immune populations responsible for TWEAK production could lead to new therapeutic strategies for patients with high rates of relapse.

Citation Format: Luisjesus S. Cruz, Denay Stevenson, Mikella Robinson, Steffy Mathew, Isabella Amador, Gregory Jordan, Carrie D. House. Role of macrophages in the development of ovarian cancer stem-like cells during chemotherapy [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 163.
Abstract 1662: Evaluation of TEAD inhibitor, VT03989 treatment on growth of aggressive meningioma preclinical models
Authors: Desai Siddhi , Shivanna Anil Thaghalli , Stavnichuk Mariya , Hayes Elizabeth , Patel Sonam , McNamara James , DeSantis Anita , Sanai Nader , Tien An-Chi , Mehta Shwetal
Published: March 2024

Background: Aggressive meningiomas (WHO grade II/III) are incurable brain tumors and despite surgery and radiation therapy, tumors recur frequently without approved systemic treatment options. Hence, there is an unmet need for targeted inhibition therapy and biomarkers that predict vulnerability to targeted therapies in aggressive meningioma patients. Hippo signaling pathway has been shown to be dysregulated in meningioma, resulting in activation of its target TEAD as a transcriptional factor. We hypothesize that targeted inhibition of TEAD in recurrent high-grade meningioma with NF2 loss will lead to increased inhibition of tumor growth. Here, we investigated the effect of VT3989, a potent TEAD inhibitor, in meningioma cell lines with NF2 wild-type or NF2 loss genotype.

Methods: Two meningioma cell-lines: IOMM-Lee (NF2 wild-type) and MG309, a grade III meningioma patient-derived cell line with NF2 loss were treated with various doses of VT3989 and cell viability was assessed. Relative expression of direct targets of TEAD including CTGF and CYR61 was determined with quantitative PCR (qPCR) to ensure target engagement. Global transcriptomic changes were examined via RNAseq analysis on RNA isolated from MG309 cells treated with VT03989 compared to vehicle treated cells.

Results: VT3989 treatment significantly decreased viability of both IOMM-Lee and MG309 cells. Canonical downstream targets of TEAD such as CTGF, CYR61, BIRC5, BCL-2 were downregulated in VT3989 treated cells compared to control. Furthermore, VT03989 treatment resulted in downregulation in cell cycle regulation and DNA damage response (DDR) pathway genes. Ongoing work is focused on combining VT3989 with agents targeting cell cycle and DDR pathways.

Conclusion: VT3989 treatment led to downregulation of TEAD direct targets CTGF and CYR61 and inhibited cell viability of both IOMM-Lee cell and MG309 meningioma lines. RNA-seq analysis identified cell cycle and DDR pathway genes to be downregulated upon TEAD inhibition.

Citation Format: Siddhi Desai, Anil Thaghalli Shivanna, Mariya Stavnichuk, Elizabeth Hayes, Sonam Patel, James McNamara, Anita DeSantis, Nader Sanai, An-Chi Tien, Shwetal Mehta. Evaluation of TEAD inhibitor, VT03989 treatment on growth of aggressive meningioma preclinical models [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 1662.
Abstract 1681: Characterizing signaling and localization of Daple-FLT3 gene fusion in leukemia cells
Authors: Acquazzino Michael , Ear Jason
Published: March 2024

Receptor tyrosine kinases (RTKs) are cell surface receptors that regulate cell survival, cell proliferation, and cell migration upon ligand binding. Genetic mutations leading to hyperactivation of RTKs are often seen in cancer and allow signaling pathways to be constitutively active. In some hematological malignanices, the coiled-coil domain (CCD) of Daple, a cell signaling scaffold protein, has been found in gene fusions with the RTKs FLT3 and PDGFRB, and to the intracellular kinase JAK2. It is hypothesized that this gene fusion product causes the tyrosine kinase domain of FLT3 to become activated and mislocalizes in the cell, leading to rampant cell signaling of either the MAPK, AKT, and STAT5 signal transduction cascades. Using fluorescently tagged Daple-FLT3, it was shown that the gene fusion product is subcellularly localizes to the peri-centrosome space across all leukemia cell lines tested. Ectopic expression demonstrated activation of the kinase domain and subsequent increases in phosphorylation of STAT5α and AKT, but no significant increase in the phosphorylation of MAPK was observed. Further investigation is needed to identify direct substrates of the Daple-FLT3 gene fusion and transcriptional changes. Identifying the direct kinase substrates and insights gained through understanding the gene fusion localization, affects on cell signaling, and changes in gene expression, may unveil novel therapeutic strategies into targeting this protein in cancer.

Citation Format: Michael Acquazzino, Jason Ear. Characterizing signaling and localization of Daple-FLT3 gene fusion in leukemia cells [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 1681.
Abstract 17: Metabolic screening of T cells guides the enrichment of stem cell memory phenotype
Authors: Lauer Anthony , Karassina Natasha , Sylvester Kayla , Vidugiriene Jolanta
Published: March 2024

The efficacy of T cell-mediated immunotherapy is improved by the proliferation of T cells with a stem cell memory phenotype (Tscm). These cells are capable of long-term persistence, self-renewal, multipotency, and robust engagement with cancer cells. This study focuses on refining T cell activation protocols to bolster the presence of Tscm within the therapeutic arsenal against cancer. Through a series of cell health and metabolic assays, we traced the activation-induced metabolic reprogramming that results from Tscm enrichment. Ex vivo activation was characterized by a rapid 2.5-3 fold increase in ATP production and a 2 fold shift in intracellular reducing potential, paralleling the metabolic demands of Tscm in the cancer microenvironment. Progression of activation intensified the metabolic flux through catabolic and anabolic pathways, with significant fluctuations in metabolites such as glucose, lactate, glutamine, glutamate, branched chain amino acids, and pyruvate. These metabolic alterations were accompanied by a 2 fold increase in the prevalence of Tscm cells, and demonstrated a dependency on media composition. Our findings reveal that metabolic profiling during T cell activation provides early and actionable insights into the enrichment of Tscm, and may serve as a strategic guideline for the enhancement of cancer immunotherapies.

Citation Format: Anthony Lauer, Natasha Karassina, Kayla Sylvester, Jolanta Vidugiriene. Metabolic screening of T cells guides the enrichment of stem cell memory phenotype [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 17.
Abstract 1768: Investigating oncogene amplifications in brain metastasis of esophageal adenocarcinoma samples
Authors: Lawson Nora , Ye Lingqun , Zhao Bo , Futreal Andy , Akdemir Kadir
Published: March 2024

Esophageal Adenocarcinoma (EAC) exhibits a rising incidence and dismal survival rate in the US, primarily attributed to frequent distant metastases. Brain metastasis, though uncommon is an aggressive tumor type, remains poorly understood in EAC. To address this knowledge gap, we conducted a comprehensive analysis of 9 brain metastasis samples, profiling their whole-genome with matched normal and comparing them to the primary EAC tumors, other EAC metastasis types and other brain metastasis types, including the Hartwig Institute and PCAWG. Overall, our sample size comprised over 4,500 tumor whole-genome data and we utilized a uniform data analysis pipeline for robust comparisons between cohorts. Our analysis revealed that brain metastasis samples have significantly higher single-nucleotide variants, however, the structural variant burden in brain metastases closely resembled those observed in the primary tumor. Notably, the incidence of whole genome doubling is significantly higher in brain metastasis compared to other primary cancer types and metastases. This significant difference stems from the higher WGD incidence in primary EACs compared to other tumor types. Mutational signature analysis revealed no significant changes compared to the primary tumor, and we observed an enrichment of SBS 17a/b, previously associated with tumor progression in primary esophageal adenocarcinoma, along with a prevalent presence of the APOBEC mutagenesis signature in most samples. Notably, ERBB2 and TP53 emerged as prominent driver genes present in most brain metastasis samples. We also observed an upregulation of ERBB2 expression in brain metastasis samples. Interestingly, ERBB2 appeared to play a more significant role in brain metastasis progression compared to the primary tumor and other metastasis types, suggesting its potential as a therapeutic target. These findings advance our comprehension of brain metastasis in EAC, providing crucial insights into genomic alterations and mutational processes that could inform future therapeutic strategies and clinical management for this previously understudied metastatic type.

Citation Format: Nora Lawson, Lingqun Ye, Bo Zhao, Andy Futreal, Kadir Akdemir. Investigating oncogene amplifications in brain metastasis of esophageal adenocarcinoma samples [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 1768.
Abstract 1785: The role of BRCA1 on metabolic pathways in an in vivo system
Authors: Hembruff Stacey L. , Dekonenko Alexander , Thyfault John , Sardiu Mihaela , Washburn Michael , Jensen Roy A. , Harlan-Williams Lisa M.
Published: March 2024

The role of BRCA1 in cellular metabolism is not well characterized and what we do understand has been mostly demonstrated in vitro. Our studies aim to fully characterize the role of BRCA1 in metabolic pathways in a whole-body system. In vivo studies using C57BL/6 wild-type and transgenic humanized BRCA1 mice demonstrate the effect of human BRCA1 on the whole-body metabolic phenotype and start to elucidate the mechanism by which this occurs. We used Promethion metabolic chambers and glucose tolerance tests to measure a number of metabolic outputs of male and female mice that had either normal mouse Brca1 gene expression (wild-type/WT mice) or a knock-out mouse Brca1/knock-in human BRCA1 (humanized/HU mice). Humanized BRCA1 mice are more lean, hyperactive and demonstrate a sexual dimorphism in glucose tolerance when compared to wild-type mice on the same genetic background. To begin to elucidate the mechanisms behind the observed metabolic phenotype, we used a metabolic tissue, female mouse skeletal muscle, to perform mass spectrometry, SuperArray, and Western blot analysis. Proteomic samples were sent to the IDeA National Resource for Quantitative Proteomics at the University of Arkansas Medical Sciences for processing and analysis. Proteomic and genomic analysis showed changes in a number of metabolic pathways that may be implicated in the observed whole body metabolic phenotype. We can conclude that changing the expression levels of BRCA1 in an in vivo model altered the overall metabolic profile of C57BL/6 mice. This is the first in vivo evidence demonstrating the effects of BRCA1 expression in whole body metabolism.

Citation Format: Stacey L. Hembruff, Alexander Dekonenko, John Thyfault, Mihaela Sardiu, Michael Washburn, Roy A. Jensen, Lisa M. Harlan-Williams. The role of BRCA1 on metabolic pathways in an in vivo system [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 1785.