bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2688: Bombesin receptors regulate transactivation of HER4
Authors: Moody Terry W. , Ramos-Alvarez Irene , Mantey Samuel A. , Jensen Robert T. 
Published: June 2022

Abstract
Bombesin (BB) is an autocrine growth factor for non-small cell lung cancer (NSCLC) cells. BB stimulates whereas the BB2 receptor (R) antagonist PD176252 inhibits NSCLC growth (Moody et al., Eur. J. Pharmacol; 2003; 474:21). The BB2R regulates the transactivation of the EGFR, HER2 and HER3 (Lee et al., BBAMCR 2020; 1867: 118625). HER4 increases the proliferation of NSCLC cells (Mota et al., Oncotarget 2017; 8: 89284). It is unknown if BB2R stimulation increases tyrosine phosphorylation of HER4 and this was investigated using NSCLC cells. BB or neuregulin (NRG1) addition to NSCLC cells increased the phosphorylation of Tyr1284-HER4. By RT-PCR and western blot, mRNA and protein for BB2R, HER4 and NRG1 was present in NCI-H522 and NCI-H661 NSCLC cells. The addition of BB to NCI-H522 or NCI-H661 cells increased phosphorylation of HER4, ERK and AKT which was blocked by PD176252. By immunoprecipitation, HER4 heterodimerized with HER2 and the EGFR after addition of BB to NSCLC cells. PP2 and N-acetylcysteine impaired the ability of BB to increase P-Tyr1284-HER4 in NSCLC cells, which indicates that SRC and reactive oxygen species are essential. Ibrutinib, a tyrosine kinase inhibitor (Rauf et al., Oncogene 2018; 37: 2237), impaired the ability of BB2Rs to regulate HER4 transactivation. Ibrutinib or PD176252 inhibited the growth of NSCLC cells whereas BB or NRG1 increased NSCLC colony formation. The results indicate that peptide receptors for BB regulate HER4 transactivation and the proliferation of NSCLC cells.
Citation Format: Terry W. Moody, Irene Ramos-Alvarez, Samuel A. Mantey, Robert T. Jensen. Bombesin receptors regulate transactivation of HER4 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2688.
Abstract 2716: Cancer study of germline carriers of Inborn Errors of Metabolism (IEMs) related genes
Authors: Kim Seokhyeon , Song Seulki , Koh Youngil , Yoon Sung-Soo 
Published: June 2022

Abstract
Inborn Errors of Metabolism (IEMs) are a hereditary disorder in which a gene fails to operate correctly as a result of a mutation in a metabolic-related gene passed down from the parents. Majority of IEMs are inherited in autosomal recessive manner. Recent research has been undertaken on mutations in numerous metabolic genes, including IEMs and cancer development. This study used Next Generation Sequencing (NGS) data to confirm the link between metabolic-related genes and cancer development in heterozygous carriers. The PCAWG (Pan-Cancer Analysis of Whole Genomes) cancer patient cohort and the 1000G (1000 Genomes) healthy cohort were used in this work, with the TCGA (The Cancer Genome Atlas) cancer patient cohorts being used for validation analysis. A regression analysis was used to determine the statistical significance of the cancer patient group versus the normal cohort. We analyzed RNA-seq data by separating heterozygous carrier and non-carrier groups of IEM related genes. The proportion of IEM gene heterozygote carriers was higher in cancer patients than in the normal population. We chose genes showing similar patterns of occurrence with statistical significance across several cancer patient cohorts. Accordingly, the most important gene, phenylalanine hydroxylase (PAH), has been established, and the carrier frequency was consistently high in lung squamous cell carcinoma (LUSC). RNA analysis results revealed that the PAH heterozygous carrier group was enriched with various immune response pathways suggesting inflammation related carcinogenesis in PAH related LUSC. Our finding suggest that heterozygote carrier status of IEM genes may lead to generation of aberrant metabolites and the breakdown of metabolic networks, which eventually leads to cancer.
Citation Format: Seokhyeon Kim, Seulki Song, Youngil Koh, Sung-Soo Yoon. Cancer study of germline carriers of Inborn Errors of Metabolism (IEMs) related genes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2716.
Abstract 2748: High-speed and high-precision FL-based cell count and viability assays using the Cellaca™ MX high-throughput cell counter
Authors: Bell Jordan , Huang Yongyang , Yung Sun , Qazi Henry , Hernandez Charles , Lin Bo , Qiu Jean , Chan Leo 
Published: June 2022

Abstract
With multiple FDA-approved cell therapy products available and others in pre-clinical and clinical trials, now, more than ever, it is critical to provide fast and accurate measurements of cell concentration and viability. For certain clinical products, cell concentration is synonymous with dosage, and proper cell viability is crucial to avoidance of potential harmful side effects. The complex nature of patient-derived samples makes legacy cell analysis methods such as the trypan blue exclusion assay difficult or impossible to perform. Nonspecific objects such as RBCs and cellular debris can significantly increase cell counting variation. Meanwhile, the need for cell-based products that are custom-tailored to each patient has resulted in the analysis of smaller and more numerous batches, leading to cell counting bottlenecks. With the challenges of messy samples and counting bottlenecks in mind, we investigate the use of the Cellaca™ MX, a cell counter that can increase throughput and employ fluorescence-based cell counting methods. The high-throughput cell counter can utilize 5 fluorescence imaging channels for cell count and viability analysis in less than 3 min for 24 samples. In addition, fluorescent cell-based assays such as apoptosis (Annexin V or Caspase 3/7), cell cycle, reactive oxygen species measurement can be performed to better characterize the target cell samples. In this work, we demonstrate the capabilities of Cellaca™ MX for high-throughput fluorescence-based cell counting. Comparison among multiple instruments using Jurkat cells and fluorescent beads reveals high consistency between replicate counts, between Cellaca consumable plates, and between instruments. We also compare cell counts made on the Cellaca™ MX with manual counts performed using a hemocytometer, as well as counts performed using other fluorescence-based automated cell counting methods. In addition, we illustrate the use of the ISO 20391-II cell counting standard to evaluate and compare the performance of cell counting methods. Finally, we include real-world primary T cell linearity results over a 4-log range of concentration. The results of the experiments confirm the suitability of the Cellaca™ MX for general fluorescence-based assays applicable to cell therapy-related workflows.
Citation Format: Jordan Bell, Yongyang Huang, Sun Yung, Henry Qazi, Charles Hernandez, Bo Lin, Jean Qiu, Leo Chan. High-speed and high-precision FL-based cell count and viability assays using the Cellaca™ MX high-throughput cell counter [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2748.
Abstract 2763: Adaptation of therapeutic NK cell line to tumor microenvironment culture conditions increases potency and reduces cellular exhaustion
Authors: Lu Ann , Li Yunmin , Wong Albert , Lim James 
Published: June 2022

Abstract
NK-92 is a well-characterized human NK cell line that has demonstrated promising anti-cancer activities in clinical trials. However, the immunosuppressive solid tumor microenvironment (TME) can inhibit NK-92 function thereby limiting its cytolytic properties against tumor cells. To address this limitation, we serially passaged and maintained NK-92 cell lines under tumor microenvironment culture conditions (2.0 PSI + 1% O2) for a minimum of 5+ weeks. This adaptation period was used to precondition and expand NK cells with lower oxygen dependency, enriching for subset of cells that have metabolically adapted to pressurized and hypoxic culture conditions. NK cells maintained and expanded under TME culture conditions gradually reduced their cell-doubling times to reach parity with parental lines maintained under ambient oxygen (21% O2) culture conditions. This adaptation period coincided with improved tumor killing properties for TME-Adapted NK cells (TANK) against the non-small cell lung cancer cell line, A549. Enhanced cytolytic activity of TANK cells was two-fold greater than their non-TME adapted counterparts, with sustained potency observed in both ambient and TME screening conditions. Furthermore, serial exhaustion studies where TANK cells were repeatedly stimulated with tumor cells show sustained cytolytic activity, in contrast to non-TME adapted NK92 counterparts. TANK cells also exhibit higher levels of granzyme B and perforin as assessed by flow cytometry, while exhibiting distinct gene expression signatures as profiled with RNAseq. This initial study supports the hypothesis that immune cells can both react and adapt to different microenvironments. TME-adaptation strategies during cell expansion can enhance potency and efficacy of cell therapies designed to work in a solid tumor microenvironment.
Citation Format: Ann Lu, Yunmin Li, Albert Wong, James Lim. Adaptation of therapeutic NK cell line to tumor microenvironment culture conditions increases potency and reduces cellular exhaustion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2763.
Abstract 2767: Low baseline serum IFNg and TNFa are predictive of response to bexmarilimab
Authors: Koivunen Jussi , Jalkanen Juho , Björkman Mari , Mandelin Jami , Bono Petri 
Published: June 2022

Abstract
Introduction: Bexmarilimab is a novel macrophage re-programming agent targeting CLEVER-1 on tumor associated macrophages. CLEVER-1 is a highly immunosuppressive macrophage scavenger receptor. Inhibition of CLEVER-1 has been shown remove T cell exhaustion, induce clonal expansion of cytotoxic CD8 cells, and improve their infiltration into the tumor. Recently in a first-in-human Phase I/II study named MATINS (NCT03733990) bexmarilimab was reported to have a survival benefit in 30% of patients with advanced gastric cancer, cutaneous melanoma and cholangiocarcinoma as a single agent (ESMO 2021).
Methods & Aims: As a part of the mode of action of bexmarilimab, it is known to induce TNFa and IFNg responses, especially when IFNg levels are low. Using ROC analysis we aimed to test the performance of baseline cytokine serum IFNg and TNFa levels to identify patients that experienced a survival benefit from bexmarilimab compared to patients that did not benefit from bexmarilimab.
Results: 30 patients, 10 per each tumor type, were treated with bexmarilimab at 1mg/kg Q3W. Mean age 59 years (SD, 9 years), 73% male, with an average of 3 previous lines of therapy, median survival not yet reached. At baseline, patients that saw a clinical benefit (CBR) from bexmarilimab had average serum IFNg levels of 5.11 pg/ml (SD, +/- 4.99 pg/ml) compared to 13.07 pg/ml (SD, +/- 13.26) in patients that did not respond to the treatment. Similarly serum TNFa levels 0.96 pg/ml (SD, +/- 0.74 pg/ml) in patients that benefited from bexmarilimab compared to 2.37 pg/ml (SD, +/- 1.43 pg/ml) in patients that did not respond to the treatment. Log transformed values were used for ROC analyses. Used together IFNg and TNFa performed well to identify patients that are responsive to bexmarilimab (AUC 0.83, P &lt; 0.0001).
Conclusion: Bexmarilimab works well in patients with low IFNg and TNFa levels, which is opposite to what is usually seen with checkpoint inhibitors and other T cell activating agents.
Citation Format: Jussi Koivunen, Juho Jalkanen, Mari Björkman, Jami Mandelin, Petri Bono. Low baseline serum IFNg and TNFa are predictive of response to bexmarilimab [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2767.
Abstract 2769: Advance understanding of the tumor microenvironment with multiplex immunofluorescence
Authors: Patel Bhavika , Allen Stephanie , Boldt Brittney , Mammadova Najiba , Haggerty Agnes , Mehra Navi 
Published: June 2022

Abstract
Immunotherapy and precision medicine are rapidly developing approaches to cancer therapy. Biomarkers that detect the tumor and tumor microenvironment allow for the development of strategies that accelerate the advancement of treatments to enhance a patient’s immune system. Akoya’s MOTiF™ PD-1/PD-L1 Panel is a validated, multiplex immunoassay enabling detection of the 6 most clinically relevant immuno-oncology and spatial biomarkers: PD-1, PD-L1, FoxP3, CD8, CD68, and PanCK. The MOTiF™ PD-1/PD-L1 Panel was used to analyze the tumor microenvironment and specifically assess immune phenotypes of different types of cancers: non-small cell lung cancer (NSCLC), colon adenocarcinoma, head and neck squamous cell carcinoma (HNSCC), and breast cancer. We demonstrate the utility of Akoya’s MOTiF™ PD-1/PD-L1 panel kit in studying the cellular diversity of various cancers while retaining spatial context. Stained slides were analyzed using the InForm® and PhenoptrReports image analysis platforms to identify and better understand spatial relationships between a variety of simple and complex cell phenotypes. The MOTiF™ PD-1/PD-L1 panel kit provides reproducibility across different tissue types. These data provide insight into the innate and adaptive immune environment for targeted design of new immunotherapies and have implications for improving the treatment paradigm across many tumor types.
Citation Format: Bhavika Patel, Stephanie Allen, Brittney Boldt, Najiba Mammadova, Agnes Haggerty, Navi Mehra. Advance understanding of the tumor microenvironment with multiplex immunofluorescence [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2769.
Abstract 2815: Exploring the role of telomerase in senescence and exhaustion in CAR T cell immunotherapy
Authors: Murty Tara , Ramello Maria C. , Sotillo Elena , Chen Lu , Artandi Steven A. , Mackall Crystal L. 
Published: June 2022

Abstract
Chimeric antigen receptor (CAR) T cell therapy has revolutionized oncology through engineered targeting of antigens on previously untreatable cancers. However, less than half of patients on CAR T cell therapy experience long-term disease control, with better outcomes observed in pediatric compared to adult populations. Senescent T cells have been shown to manifest defective killing abilities and the development of negative regulatory functions, with evidence suggesting that senescence may play a role in decreasing CAR T cell efficacy and persistence. Moreover, levels of telomerase have been shown to control the lifespan of human T cells, with increased levels delaying senescence. It has been shown that T cell exhaustion limits CAR T cell efficacy in the context of solid tumors, where CAR T cells have yet to demonstrate sustained responses. Using a model of CAR T cell exhaustion developed in our lab, and CAR T cells manufactured from younger and older donors, we have developed a comprehensive method to profile T cell functionality, phenotype, and proliferation. We are able to characterize features of senescence via cell surface markers, intracellular activity, telomere length, and telomerase activity as well as features of exhaustion. Using CRISPR/Cas9, we have inactivated TERT in primary human T cells and CAR T cells and interrogated the effects of hTERT knockout as well as of hTERT overexpression on key markers of T cell identity, exhaustion, senescence, and cytotoxic activity. Understanding telomerase in the context of CAR T cell therapy will provide mechanistic insights into senescence and exhaustion cellular programs and has the potential to inform increasingly effective CAR T cell cancer treatments.
Citation Format: Tara Murty, Maria C. Ramello, Elena Sotillo, Lu Chen, Steven A. Artandi, Crystal L. Mackall. Exploring the role of telomerase in senescence and exhaustion in CAR T cell immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2815.
Abstract 2818: Targeting solid tumors with GDT002, a first-in-class γδTCR-based T cell therapy
Authors: Drent Esther , Bisso Andrea , Baardman Sjoerd , Verweij Dagmar , Salcedo Estefania , Coomans Chris , Braem Steven , van de Weg Sander , Melief Sara , Norell Haakan , van Loenen Marleen , Gobessi Stefania , Throsby Mark 
Published: June 2022

Abstract
Broad application of cell therapies like CAR-T have been hampered by a lack of tumor-specific targets. Gadeta leverages the natural HLA-independent tumor recognition capabilities of γδTCRs combined with the proliferative capacity and robust tumor killing of αβT cells to develop tumor-specific cell therapies. γδT cells expressing Vγ9Vδ2 TCRs are the most common subtype found in peripheral blood. They sense the presence of phosphoantigens (pAgs) upregulated in malignant cells due to a dysregulated mevalonate pathway. The Vγ9Vδ2 TCR expressed by GDT002 was isolated from a PBMC-derived γδT cell of a healthy donor and was selected for its broad and strong tumor reactivity. GDT002 is currently being evaluated in a multicenter first-in-human phase 1/2 study for the treatment of Multiple Myeloma (NCT04688853). Here we describe studies carried out to identify potential solid tumor indications which could be effectively targeted by GDT002. Tumor reactivity was tested against cell lines from different cancer types in the presence of pamidronate, a clinically approved N-bisphosphonate that boosts pAgs levels. Significant GDT002 reactivity, in terms of cytotoxicity and cytokine release, was observed against most tumor targets. In a more complex ex vivo 3D co-culture system, GDT002 reactivity was also observed towards a broad panel of patient-derived tumor organoids. In contrast, there was no or very limited GDT002 reactivity against primary healthy cells. From our broad screening of solid tumor types, we identified ovarian cancer as a promising indication for GDT002. GDT002 displayed significant reactivity towards 4/5 ovarian cell lines and 9/10 patient-derived ovarian cancer organoids in the presence of pamidronate. To study GDT002 in vivo, we investigated the biodistribution of i.v. administered pamidronate. In tumor-bearing mice, intra-tumor pamidronate concentrations were detected during the 7-day observation period, allowing targeting by GDT002. Taken together, our results demonstrate that GDT002 has a potent anti-tumor activity across a broad spectrum of solid and liquid tumor types. We have identified ovarian cancer as a potential indication to explore the clinical effect of GDT002 against solid tumors.
Citation Format: Esther Drent, Andrea Bisso, Sjoerd Baardman, Dagmar Verweij, Estefania Salcedo, Chris Coomans, Steven Braem, Sander van de Weg, Sara Melief, Haakan Norell, Marleen van Loenen, Stefania Gobessi, Mark Throsby. Targeting solid tumors with GDT002, a first-in-class γδTCR-based T cell therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2818.
Abstract 2822: Enhanced effector activity of mediator CDK8 kinase module deficient CAR-T Cells
Authors: Freitas Katherine A. , Belk Julia A. , Sotillo Elena , Daniel Bence , Sandor Katalin , Klysz Dorota , Duong Vandon T. , Xu Peng , Malipatlolla Meena , Weber Evan W. , Majzner Robbie G. , Chang Howard Y. , Satpathy Ansuman T. , Mackall Crystal 
Published: June 2022

Abstract
Adoptive T cell immune therapies mediate impressive clinical benefit in a fraction of patients, but anti-tumor effects are often limited by inadequate T cell potency. To identify genes that limit T cell effector function, we conducted genome-wide CRISPR knock-out screens in human primary CAR-T cells. The top hits were components of the CDK8 kinase module of the Mediator complex, an evolutionarily conserved regulator of gene transcription. CDK8 kinase module deficient CAR-T cells manifest increased expansion, cytokine production, metabolic fitness, effector function, anti-tumor activity and reduced terminal effector differentiation. CDK8 kinase module deficient CAR-T cells showed widespread but selective increases in chromatin accessibility, MED1 chromatin occupancy, and H3K27 acetylation most notably involving transcription factors that play a critical role in T cell fate, including several STAT and AP1 family members. The most pronounced enhancement was observed for STAT5 which manifested as increased sensitivity to IL-2 in CDK8 kinase module deficient CAR-T cells. These results link Mediator induced transcriptional coactivation with T cell effector programming and identify the CDK8 kinase module as a target for enhancing the potency of anti-tumor T cell responses.
Citation Format: Katherine A. Freitas, Julia A. Belk, Elena Sotillo, Bence Daniel, Katalin Sandor, Dorota Klysz, Vandon T. Duong, Peng Xu, Meena Malipatlolla, Evan W. Weber, Robbie G. Majzner, Howard Y. Chang, Ansuman T. Satpathy, Crystal Mackall. Enhanced effector activity of mediator CDK8 kinase module deficient CAR-T Cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2822.