bluuebunny/crossref_metadata_2025 huggingface dataset:
Design of Edge Computing Online Classroom Based on College English Teaching
Authors: Zhen Changhui , Hu Kun
Published: June 2022

This study expects to solve the network delay in college English online teaching, prevent disrupting the normal teaching order of college English classroom caused by network problems, increase college students’ interest in learning English, and improve the time spent by college students in English learning. First, the study investigates and analyzes the current needs and situation of college English online learning and puts forward the improvement standards for college English online classroom. Second, the study applies edge computing (EC) technology to improve online teaching system and designs an online learning system for college specific English based on EC. Finally, the improvement effect is tested, and it is concluded that the effect of using EC to improve the online classroom system for college English is better than that of the unmodified online teaching system. The results reveal that the online classroom system for college English based on EC designed can effectively reduce the network delay of the traditional teaching system, ensure a high reliability, and increase the time for students to learn English, so it is helpful to improve the teaching quality. The results provide a new direction for applying emerging computer technology in online teaching in colleges and universities, a guarantee for improving college students’ English level, and set an example for the relevant research in the future.
Heme Oxygenase 1/Peroxisome Proliferator-Activated Receptor Gamma Pathway Protects Intimal Hyperplasia and Mitigates Arteriovenous Fistula Dysfunction by Regulating Oxidative Stress and Inflammatory Response
Authors: Xie Tingfei , Xu Yunpeng , Ji Lecai , Sui Xiaolu , Zhang Aisha , Zhang Yanzi , Chen Jihong
Published: June 2022

Purpose. An arteriovenous fistula (AVF) is the preferred vascular access mode for maintenance hemodialysis, and access stenosis and thrombosis are the primary causes of AVF dysfunction. This study is aimed at exploring the molecular mechanisms underlying AVF development and the roles of the heme oxygenase 1/peroxisome proliferator-activated receptor gamma (HO-1/PPAR-γ) pathway in AVF. Method. AVF model mice were established, and the vascular tissues from the arteriovenous anastomosis site were sent for mRNA sequencing. Differentially expressed mRNAs (DEmRNAs) were screened and subjected to functional analysis. Thereafter, the mice with HO-1 knockdown and coprotoporphyrin IX chloride (COPP) pretreatment were used to investigate the roles of the HO-1/PPAR-γ pathway in AVF. Results. By sequencing, 2514 DEmRNAs, including 1323 upregulated and 1191 downregulated genes, were identified. These DEmRNAs were significantly enriched in the PPAR signaling pathway, AMPK signaling pathway, glucagon signaling pathway, IL-17 signaling pathway, and Toll-like receptor signaling pathway. High expression of HO-1 and PPAR-γ reduced endothelial damage and intimal hyperplasia during AVF maturation. After AVF was established, the levels of transforming growth factor-β (TGF-β), interleukin-1β (IL-1β), interleukin-18 (IL-18), and reactive oxygen species (ROS) were significantly increased (

P
<
0.05

), and HO-1 normal expression and COPP pretreatment evidently decreased their levels in AVF (

P
<
0.05

). Additionally, AVF significantly upregulated HO-1 and PPAR-γ and downregulated MMP9, and COPP pretreatment and HO-1 normal expression further upregulated and downregulated their expression. Conclusion. The HO-1/PPAR-γ pathway may suppress intimal hyperplasia induced by AVF and protect the intima of blood vessels by regulating MMP9 and ROS, thus mitigating AVF dysfunction.
Research on the Multimodal Digital Teaching Quality Data Evaluation Model Based on Fuzzy BP Neural Network
Authors: Feng Wenyan , Feng Fan
Published: June 2022

We propose in this paper a fuzzy BP neural network model and DDAE-SVR deep neural network model to analyze multimodal digital teaching, establish a multimodal digital teaching quality data evaluation model based on a fuzzy BP neural network, and optimize the initial weights and thresholds of BP neural network by using adaptive variation genetic algorithm. Since the BP neural network is highly dependent on the initial weights and points, the improved genetic algorithm is used to optimize the initial weights and thresholds of the BP neural network, reduce the time for the BP neural network to find the importance and points that satisfy the training termination conditions, and improve the prediction accuracy and convergence speed of the neural network on the teaching quality evaluation results. The entropy value method, a data-based objectivity evaluation method, is introduced as the guidance mechanism of the BP neural network. The a priori guidance sample is obtained by the entropy method. Then, the adaptive variational genetic algorithm is used to optimize the BP neural network model to learn the a priori sample knowledge and establish the evaluation model, which reduces the subjectivity of the BP neural network learning sample. To better reflect and compare the effects of the two neural network evaluation models, BP and GA-BP, the sample data were continued to be input into the original GA and BSA to obtain the evaluation results and errors; then, the evaluation results of the two evaluation models, BP and GA-BP, were compared with the evaluation results of the two algorithms, GA and BSA. It was found that the GA-BP neural network evaluation model has higher accuracy and can be used for multimodal digital teaching quality evaluation, providing a more feasible solution.
Water Absorption Behavior of Teff (Eragrostis tef) Straw Fiber-Reinforced Epoxy Composite: RSM-Based Statistical Modeling and Kinetic Analysis
Authors: Prabhu S. Venkatesa , Wilson Vincent Herald , Anand K. T. , Jose S. , Sivamani S. , Gomadurai Chinnasamy , Kifetew Melkamu
Published: June 2022

Recently, reinforced polymeric composites prepared from natural fibers have received a significant interest among the researchers because of its appreciable sustainability, environmentally friendly, and low cost. However, one particular issue, that is, hydrophilic property, still needs to be addressed for its successful applications. Since the hydrophilic tendency of natural fibers is extremely undesirable, it leads to the quick degradation of fiber-based polymer composites. Hence, the fiber property, hydrophilic nature, is influenced by the presence of noncrystalline and voids part of these fibers that significantly influences the polymer matrix adhesion. Hence, it is very important to understand the water absorption behavior of reinforced fiber composites. In this study, a crop residual material specific to Ethiopia, teff straw (Eragrostis tef), was used as fiber material. The fiber was treated with 1% NaOH followed by 1% CH2=CHCOOH at room temperature for improving the bonding strength between the fiber and polymer, which leads to suppress the water absorption. The investigation on mathematical model for water absorption property at different fiber loadings (4%, 8%, 12%, 16, and 20%) was carried out, and the analysis on the kinetic behavior of water absorption was also investigated. In addition, the response surface-based statistical modeling which correlates water absorption, fiber loading, and time has been analyzed.
Effect Analysis of Clinical Pathway Nursing Combined with Humanized Nursing in Cerebral Infarction
Authors: Xiao Lili , Xiao Tingting , Yu Fang
Published: June 2022

Objective. To make the application effect of clinical pathway nursing combined with humanized nursing clear in cerebral infarction. Methods. Hundred patients who suffered from cerebral infarction had been hospitalized in our hospital and they become our study objects. By means of the random number table, they were separated into 50 cases both in the control group and in the observation group. The control group accepted the routine nursing, and the observation group accepted the clinical pathway nursing mixed with humanized nursing. The followings were compared including the hospitalization expenses and length of stay, National Institutes of Health Stroke Scale (NIHSS), Barthel index rating scale (BI), Fugl-Meyer Assessment (FMA), and the incidence of complications and recurrence rate. A comparison was made between the two groups about satisfaction with nursing. Results. In contrast with the control group, the treatment expenses and length of stay in the observation group were considerably reduced (

P
<
0.05

). In contrast, before nursing, the NIHSS marks of the two groups decreased considerably after nursing, and the scores of BI and FMA increased considerably, the NIHSS scores of the observation group were considerably lower than those of the control group, and the scores of BI and FMA were considerably higher than those of the control group (

P
<
0.05

). In contrast with the control group (44.00%, 12.00%), the incidence of complications (22.00%) and recurrence rate (2.00%) in the observation group were considerably lower (

P
<
0.05

). In contrast with the control group (72.00%), the satisfaction of nursing in the observation group (96.00%) was considerably higher (

P
<
0.05

). Conclusion. The application effect of clinical pathway nursing combined with humanized nursing in cerebral infarction is significant. It can not only effectively reduce hospitalization expenses and hospitalization time, improve neurological function, ability of daily living, and limb motor function but also reduce complications and recurrence and improve patients’ satisfaction with nursing, which has a high clinical reference value.
Analysis of English Education Classroom Student Interaction and Performance Based on Virtual Reality
Authors: Li Meng , Madina Zamira
Published: June 2022

In order to improve the interaction of students in English teaching and improve the effect of English teaching, this paper combines virtual reality algorithms to construct an interactive virtual space for English teaching. Furthermore, this article advances the virtual reality method from a physics standpoint and uses the Fourier transform to superimpose the cross-correlation function in the frequency domain. At the same time, this study uses the Fourier transform to convert the overlaid result into a cross-correlation function in the time domain, eliminating the noise of virtual reality algorithms, improving immersion in the virtual environment of English instruction, and encouraging student participation. In addition, on the basis of virtual reality analysis, this paper constructs an English virtual reality education platform. Finally, after the platform is built, the effect is verified through experimental teaching methods. The experimental teaching shows that the English education classroom student interaction and performance analysis system based on virtual reality proposed in this paper has certain advantages over traditional algorithms.
Efficacy of Rosuvastatin Combined with rt-PA Intravenous Thrombolytic Therapy for Elderly Acute Ischemic Stroke Patients
Authors: Zhu Jianzhong , Wang Shan , Chen Zhenqiu , Cheng Qiqi
Published: June 2022

Background. Acute ischemic stroke (AIS) is a fatal and disabling disease. Given the unsatisfactory results by current treatment strategies, optimizing the treatment of AIS is still an urgent problem to be solved. Objective. To determine the therapeutic efficacy of rosuvastatin (ROS) combined with thrombolytic therapy using recombinant tissue plasminogen activator (rt-PA) on senile AIS patients and analyze its effects on serum inflammatory responses and neurological function. Methods. A retrospective study was conducted on 150 senile AIS patients who visited the Longmen County People’s Hospital between January 2019 and June 2021. Of them, 100 cases treated by ROS combined with rt-PA intravenous thrombolytic therapy (ivTT) were set as the observation group and the rest 50 cases receiving rt-PA alone were included in the control group. Intergroup comparisons were conducted with respect to the following parameters: neurological function (National Institutes of Health Stroke Scale, NIHSS; Scandinavian Stroke Scale, SSS), serum neuron-specific enolase (NSE) and high-sensitivity C-reactive protein (hs-CRP), therapeutic efficacy, incidence of adverse reactions, and patient satisfaction. Results. The observation group had lower NIHSS and SSS scores and serum NSE and hs-CRP than the control group. In addition, the observation group was found with a higher overall response rate, higher patient satisfaction, and fewer adverse reactions. Conclusion. ROS combined with rt-PA ivTT can better enhance the therapeutic efficacy of elderly patients with AIS, improve their neurological function, and reduce serum inflammatory responses.
The Internet‐Based Business Model and Corporate Risk‐Taking: An Empirical Study from the Information Empowerment Perspective
Authors: Li Lei , Yang Shuili
Published: June 2022

Reasonable risk‐taking acts as a solid foreground for sustaining corporate growth. Having companies trading on Chinese stock exchanges between 2010 and 2019 sampled, this paper explored how an Internet‐based business model would affect corporate risk‐taking from the perspective of information empowerment. Through the mediation effect model and quantitative text analysis, the following findings were obtained here. First, a network‐powered business model could significantly enhance the risk tolerance of companies. Second, mechanism testing showed that such a novel model would help reduce the asymmetry of corporate information and thus enhance corporate risk‐taking capacity. Third, an analysis on heterogeneity revealed that businesses that enjoy a freer market and fewer financing constraints could better feel the positive impact of an Internet‐based business model on corporate risk‐taking. Fourth, an examination of economic consequences showed that risk‐taking under Internet‐based business models allowed enterprises to create sustainable value. Overall, the present work confirmed the positive impact of an Internet‐based business model on corporate risk‐taking from the information empowerment angle, and it is expected to provide a theoretical basis for enterprises to optimize their investment decision‐making strategies and increase their risk‐taking willingness and capacity.
Abstract 10: Development of single cell spheroid as a tool to capture functional heterogeneity in head and neck cancer
Authors: Pandey Jyoti , Shyanti Ritis , Malhotra Sanjay V. , Chaturvedi Rupesh , Singh Rana P.
Published: June 2022

Introduction: Head and neck cancer squamous cell carcinomas (HNSCC) are diverse and complex with inter-tumor and intra-tumor heterogeneity and discrepancies in therapy response regardless of the clinical stage. Tumors are thought to be heterogeneous at the single-cell level involving cancer stem cells. Therefore, cancer diagnosis and treatment are difficult because single cells might have intrinsic functional and genetic heterogeneity. Single cell transcriptome is used for mapping the genetic heterogeneity of a tumor. Currently, no model is available to provide a genotype to phenotype correlations for the heterogeneity, a crucial step for successful treat of a cancer. Here, our focus is to develop a single-cell clonal spheroid model system as a valuable tool for studying functional heterogeneity, including growth, proliferation, angiogenesis, metastasis, and genetic heterogeneity.

Material and methods: HNSCC cell lines namely, UMSCC-22B, FaDu and A-253, and patients derived primary cells were cultured in 96-well ultra-low attachment to develop spheroids. Spheroids were cultured from 2, 4 and 128 cells per well with media, growth factors and supplements. Confocal microscopy was done using cocktail dyes including Hoechst, Calcein-AM and Ethidium bromide to check the functionality of spheroids.

Results: We have established a method for the development of single-cell derived clonal spheroids. The frequency of spheroids formation from 2, 4 and 128 UMSCC-22B cells per well were higher than FaDu and A-253 cell lines. The growth measurement showed clones of single cell derived spheroids with hyperproliferation as well as with low proliferative activity. UMSCC-22B and A-253 hypoproliferative spheroids had significant higher migration capabilities compared to spheroids with hyperproliferative activity. Confocal microscopy of single-cell derived clonally propagated spheroids revealed a different zone viz proliferating, quiescent and necrotic zone in intact spheroids. This condition mimics in vivo conditions and suitable tool for drug penetration studies.

Conclusion: Together, our results suggest that single cell-derived clonal spheroid from different wells would represent the genetic and functional heterogeneity of the cell population. Such a spheroid model can reveal heterogeneity inside a tumor formed by the proliferation of a single-cell and heterogeneity between tumors resulting from a different single cell and thus has great potential for drug discovery and tumor biology research.

Citation Format: Jyoti Pandey, Ritis Shyanti, Sanjay V. Malhotra, Rupesh Chaturvedi, Rana P. Singh. Development of single cell spheroid as a tool to capture functional heterogeneity in head and neck cancer [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 10.
Abstract 1101: CTNNB1 mutation can mediate resistance to EGFR, ALK and KRAS targeted therapies
Authors: Majumder Anurima , Meyer Benjamin S. , Hicks J Kevin , Boyle Theresa A. , Haura Eric B.
Published: June 2022

Resistance to targeted therapy remains an ongoing and elusive challenge in lung cancer. β-Catenin (CTNNB1) mutations have been reported in lung cancer patients, often co-occurring with known oncogenic drivers such as rearranged ALK, and mutant EGFR and KRAS. Oncogenic CTNNB1 mutations result in aberrant accumulation of the encoded β-catenin protein and may cause resistance to targeted therapy. However, different CTNNB1 mutations may activate β-catenin differentially and thereby have varying phenotypic effects. A CTNNB1 T41A mutation was identified in donated lung cancer tissue from a patient with a clinical history of an EGFR exon 20 insertion. To investigate the functional significance of this mutation, CTNNB1 T41A was transduced into an EGFR-mutated cell line (HCC827) and one cell line harboring ALK translocation (STE1). Response to EGFR and ALK targeted therapy was then compared in the GFP-transduced control cell lines versus the CTNNB1 T41A transduced cell lines to assess for effects of this CTNNB1 mutation on targeted therapy. Both HCC827 and STE1 cells harboring the CTNNB1 T41A mutation were more resistant to EGFR and ALK targeted therapy than the control cell lines. Since, different CTNNB1 mutations may exert different phenotypic effects, the CTNNB1 mutation spectrum was analyzed in lung cancer patients at the Moffitt Cancer Center. Mutations in 19 unique residues of CTNNB1 (D32, S33, G34, S37, T41, S45, A215, C419, S29, G757, K335, N287, Q72, R151, R710, S352, S681, V62 and W383) were detected in 50 lung cancer patients. Of these, 23 had some type of EGFR abnormality that co-existed with the CTNNB1 mutation while others had ROS1, RET, BRAF, NTRK3, ALK or KRAS mutations. The most frequent Exon 3 hotspot CTNNB1 mutations (at residues D32, S33, G34, S37, and S45) were chosen for further analysis wherein these mutations were transduced into HCC827, STE1 and two KRAS mutant cell lines (H358 and LU65) to assess their ability to drive resistance to EGFR, ALK and KRAS targeted therapies in ongoing studies. Our data shows that CTNNB1 mutations can cause resistance to EGFR, ALK and KRAS inhibitor therapy to different extents. Hence, further investigation of the mechanism of action of these beta-catenin mutations and the ability of beta-catenin/other inhibitors to reverse resistance is needed. This data also suggests that CTNNB1 may be an important co-occurring alteration in the context of development of resistance to targeted therapy and should be included in comprehensive clinical genetic testing panels.

Citation Format: Anurima Majumder, Benjamin S. Meyer, J Kevin Hicks, Theresa A. Boyle, Eric B. Haura. CTNNB1 mutation can mediate resistance to EGFR, ALK and KRAS targeted therapies [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 1101.
Abstract 1330: Role of ENPP1 mediated extracellular cGAMP hydrolysis in cancer metastasis and immune evasion
Authors: Kwon John Young Ho , Li Jun , Duran Mercedes , Dhanota Ninjit , Bakhoum Samuel
Published: June 2022

Chromosome instability (CIN) promotes metastasis through sustained tumor-cell autonomous response to cytosolic DNA. Chromosome mis-segregation engenders the formation of micronuclei, which upon rupture chronically activates the cGAS-STING DNA sensing pathway. Tumor cGAS recognizes cytosolic DNA to generate cGAMP, a potent immune-stimulatory molecule that is readily exported into the extracellular space to enforce a type 1 IFN-mediated anti-tumor response in a host STING dependent manner. However, it is unclear how highly aggressive, unstable tumors have evolved to co-opt this chronic inflammatory signaling to drive tumorigenic behaviors and immune evasion. Here, we show that ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) may drive tumor metastasis through degradation of extracellular cGAMP. Using RNA sequencing of isogenic tumors, we uncovered a link between ENPP1 and CIN. We leveraged syngeneic mouse models to demonstrate that genetic perturbation of tumor ENPP1 and overexpression of a catalytically inactive ENPP1 mutant suppress metastasis. Furthermore, loss of ENPP1 increases tumor immune infiltration, reduces extracellular adenosine, and enhances response to immune checkpoint blockade in a manner dependent on tumor cGAS and host STING. In line with these findings, ENPP1 expression correlates with immune suppression in human cancers. These results suggest that ENPP1-mediated hydrolysis of cGAMP may facilitate metastasis through evasion of immune surveillance, perhaps in part through cGAMP breakdown into immune-suppressive adenosine, in which the contributions of tumor and host adenosine can be further explored. Taken together, our study demonstrates how ENPP1-mediated hydrolysis of cGAMP can transform an otherwise inflammatory pathway into an immune-suppressive mechanism to promote tumor progression and metastasis.

Citation Format: John Young Ho Kwon, Jun Li, Mercedes Duran, Ninjit Dhanota, Samuel Bakhoum. Role of ENPP1 mediated extracellular cGAMP hydrolysis in cancer metastasis and immune evasion [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 1330.
Abstract 1402: Monitoring the concentration of cGAMP using homogenous bioluminescent HTS-formatted assay
Authors: Goueli Said A. , Hsiao kevin
Published: June 2022

Genomic instability engages the cGAS (cytosolic DNA sensor cyclic AMP-GMP synthase)-STING (stimulator of interferon genes) signaling (cGAS-cGAMP-STI NG) pathway on both sides of cancer either by being instrumental for antitumor immunity and blocking malignancy or by promoting inflammation-driven carcinogenesis and metastasis. The cGAS-STING axis also plays an essential role in initiating host immune defense against microbial invasion and STING-deficient mice are more susceptible to some viral infections. Credible evidence suggests that the cGAS-cGAMP-STI NG pathway makes fundamental contributions to at least three major cancer therapies, radiation therapy, chemotherapy, immunotherapy, and thus represents a promising pharmaceutical target. cGAS is well-positioned to act as a DNA sensor that triggers innate immune responses through production of the second messenger cyclic GMP-AMP (cGAMP) upon binding to DNA and initiating host immune responses against pathogens and cancer. However, inappropriate activation of STING signaling causes severe and often fatal autoimmune or autoinflammatory diseases. Hence, STING is an attractive drug target for the treatment of STING-driven autoimmune and inflammatory disorders. Therefore, there is a need to identify lead compounds that effectively inhibit human STING for further drug development. An attempt to search for such molecules have been described recently with the report on a STING-specific inhibitor that showed high efficiency, specificity, and safety, paving the way for therapeutically manipulating STING-mediated clinical diseases. Towards this goal, we have developed a bioluminescent assay to monitor the concentration of cGAMP in intracellular as well as extracellular compartments. The assay relies on the use of ectonucleotide pyrophosphatase phosphodiesterase 1 (ENPP1) to release AMP which can be converted to ATP via a coupled enzyme reaction. The ATP generated can be quantified using luciferase/luciferin detection reagents. The assay is homogenous, HTS formatted and can be completed in less than one hour. The assay is sensitive and can detect as low as 10 nM cGAMP. We will show data on the use of agents that stimulate GAS activity such as dsDNA, and the use of reporter assays to detect cytokine release upon cGAS-STING activation.

Citation Format: Said A. Goueli, kevin Hsiao. Monitoring the concentration of cGAMP using homogenous bioluminescent HTS-formatted assay [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 1402.
Abstract 1640: Enhanced development of functional human innate immune cells in a novel FLT3nullNSG mouse strain expressing human FLT3L
Authors: Yao Li-Chin , Vaidya Shantashri , Kaur Pali , Keck James G. , Shultz Leonard D. , Greiner Dale L. , Brehm MIchael A.
Published: June 2022

Humanized mice are being applied widely to study human immune system homeostasis, function, and as a testing platform for cancer immunotherapies. A major limitation for many humanized mouse models is the lack of functional and mature human innate immune cells, which are critical for effective human immune system-tumor interactions. Previous studies have demonstrated that delivery of human FLT3L into immunodeficient mice that lack mouse FLT3, promotes the development of human innate immune cell subsets following engraftment with human hematopoietic stem cells (HSC). Here we describe a NOD-scid IL2rgnull (NSG) mouse that lacks the expression of mouse FLT3 and expresses human FLT3L transgenically (NSG-mFLT3nullTg (HuFLT3L) or NSG-FLT3L. In these studies, NSG-FLT3L and NSG mice were engrafted with human umbilical cord blood (UCB) CD34+ HSC and compared for human immune system development and function. HSC-engrafted NSG-FLT3L and NSG mice show similar levels of total human CD45+ cells in blood over the course of 18 weeks post-engraftment. However, HSC-engrafted NSG-FLT3L mice show significantly higher levels of human CD141+ and CD1c+ DC subsets, CD123+ pDC, CD14+ monocytes, CD56+ NK cells and CD3+ T cells in the blood as compared to NSG mice. CD34-FLT3L mice also show increased levels of human immune cell infiltration into the gut mucosa. CD56+ NK cells and CD3+ T cells from HSC-engrafted NSG-FLT3L mice express granzyme A and granzyme B, indicating cytotoxic activity. Following treatment of HSC-engrafted NSG-FLT3L mice with LPS, heightened levels of human cytokines were detected in serum samples, confirming innate immune system function. The growth kinetics of tumor cells from a triple negative breast cancer cell line MDA-MB-231 and a lung PDX model LG1306 in HSC-engrafted NSG-FLT3L mice are work in progress and will be compared with HSG-engrafted NSG mice. Overall these results demonstrate that the NSG-FLT3L model supports enhanced development of functional, human innate immune cells and is a novel tool to study human immuno-oncology.

Citation Format: Li-Chin Yao, Shantashri Vaidya, Pali Kaur, James G. Keck, Leonard D. Shultz, Dale L. Greiner, MIchael A. Brehm. Enhanced development of functional human innate immune cells in a novel FLT3nullNSG mouse strain expressing human FLT3L [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 1640.
Abstract 168: DDR2 regulates angiogenesis via HSP-90 modulation in ovarian cancer
Authors: Akinjiyan Favour A. , Dave Ritu M. , Longmore Gregory , Fuh Katherine C.
Published: June 2022

Ovarian cancer is often diagnosed in later stages and patients have low five-year survival. There is a need to develop better therapies for this patient population. Solid tumors like ovarian cancer respond to hypoxia by inducing the sprouting of new blood cells. This process is called angiogenesis. Angiogenesis leads to formation of disordered and immature vessels that precludes efficient drug delivery. Prior work has shown that overexpressing DDR2 in endothelial cells leads to increased angiogenic activity, measured by tube formation and sprouting. We show that DDR2 expression in ovarian cancer tumor cells modulates angiogenic activity in endothelial cells. Human umbilical vein endothelial cells (HUVECs) cultured in conditioned media from DDR2-expressing tumor cells (ES2 shSCRM) have 20% increase in tube length and 18% increase in sprout network area than HUVECs cultured in conditioned media from DDR2-depleted tumor cells (ES2 shDDR2) (P=0.032). This difference in HUVEC angiogenesis is mediated by vascular endothelial growth factor (VEGF). We observed a 16% decrease in VEGF levels in conditioned media from ES2 shDDR2 cells compared to media from ES2 shSCRM cells (P​​<0.0001). DDR2 WT mice bearing ID8 Trp53-/- BRCA2-/- tumors have 53% more CD31+ microvessels than DDR2 KO mice (P​​<0.0001). We also identified that DDR2’s role in angiogenesis is mediated by the Hsp70/90 chaperone machinery. In summary, we have identified that DDR2 regulates angiogenesis in ovarian cancer.

Citation Format: Favour A. Akinjiyan, Ritu M. Dave, Gregory Longmore, Katherine C. Fuh. DDR2 regulates angiogenesis via HSP-90 modulation in ovarian cancer [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 168.
Abstract 2116: VPS34 inhibitor SB02024 activates cGAS-STING signaling and sensitizes tumors to STING agonist
Authors: Yu Yasmin , Noman Muhammad Z. , Parpal Santiago , Kleinendorst Simone C. , Saether Kristine B. , Alexeyenko Andrey , Viklund Jenny , Andersson Martin , Martinsson Jessica , Tamm Katja P. , De Milito Angelo , Janji Bassam
Published: June 2022

Novel approaches to reduce tumor immunosuppression and improve responses to anti-cancer immunotherapies based on immune-checkpoint inhibitors are needed. Emerging evidence demonstrates that autophagy inhibition enhances anti-tumor immunity by tumor cell-intrinsic and extrinsic mechanisms. Recently, we reported that pharmacological inhibition of VPS34 (PIK3C3), a lipid kinase regulating autophagy initiation, decreases tumor growth and improve the efficacy of anti-PD-1/PD-L1 therapy in melanoma and colorectal cancer mouse models. This effect was dependent on increasing T and NK cell infiltration as well as the expression of CCL5 and CXCL10 chemokines in the tumor microenvironment. Here, we explored the signaling mechanisms underlying the chemokine release following treatment with VPS34 inhibitor SB02024. We found that both pharmacological and RNAi-mediated inhibition of VPS34 activated a cGAS-STING-dependent type I interferon response in renal cell carcinoma (RCC) and melanoma cell lines. Furthermore, combination treatment of VPS34 inhibitor SB02024 with STING agonist ADU-S100 or cGAMP increased the mRNA expression and release of proinflammatory cytokines in human and murine RCC and melanoma cancer cell lines. Oral administration of SB02024 in combination with intratumoral injections of ADU-S100 significantly decreased tumor growth and weight and improved mice survival of B16-F10 tumor-bearing mice. Taken together, our data demonstrates that targeting of VPS34 results in a cGAS/STING-mediated increase of pro-inflammatory cytokine secretion and synergizes with a STING agonist. We believe that systemic VPS34 inhibition using SB02024 would be of major interest in combination or as an alternative to STING agonists to improve anti-tumor immune responses.

Citation Format: Yasmin Yu, Muhammad Z. Noman, Santiago Parpal, Simone C. Kleinendorst, Kristine B. Saether, Andrey Alexeyenko, Jenny Viklund, Martin Andersson, Jessica Martinsson, Katja P. Tamm, Angelo De Milito, Bassam Janji. VPS34 inhibitor SB02024 activates cGAS-STING signaling and sensitizes tumors to STING agonist [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 2116.
Abstract 2166: Ibuprofen induces ferroptosis to potentiate antitumor immunity
Authors: Talty Ronan , Ferreira Michelle , Krykbaeva Irina , Bosenberg Marcus
Published: June 2022

Ferroptosis is an iron-dependent regulated cell death pathway that has recently emerged as a promising target for cancer therapy. PTGS2 has been identified as a marker of ferroptosis due to its upregulation in cells after treatment with ferroptosis inducers. Nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit the protein product of PTGS2 and have been shown to enhance antitumor immunity. We hypothesized that NSAIDs induce ferroptosis and that this mechanism is essential for their effect on antitumor immunity. To test this, we first determined that ibuprofen and other NSAIDs induce cell death in YUMMER-1.7 melanoma and MC38 colorectal cancer cells in vitro. Cell death occurred progressively over 24 hours and could be blocked by multiple ferroptosis inhibitors. Treatment with NSAIDs also increased intracellular lipid peroxides and decreased levels of reduced glutathione and ferrous iron. This indicated that ibuprofen and other NSAIDs induce ferroptosis in vitro, so we next evaluated whether ibuprofen also induces ferroptosis in vivo. Analysis of ibuprofen-treated YUMMER-1.7 or MC38 tumors showed increased levels of lipid peroxides and lipid peroxide intermediates, 4-hydroxynonenal and malondialdehyde, compared to untreated tumors. Treatment of tumor-bearing mice with ibuprofen alone resulted in approximately 60% tumor clearance and survival, whereas a combination of multiple ferroptosis inhibitors with ibuprofen completely abrogated this effect. Since it has been demonstrated by that CD8+ T cells can release IFN-γ to induce ferroptosis in tumor cells, we next tested the dependence of the antitumor effect of ibuprofen on CD4+ T cells, CD8+ T cells, and IFN-γ using depleting antibodies. Depletion of any of these was sufficient to prevent the antitumor effect of ibuprofen. Given the need to identify pathways that enhance responses to existing immunotherapies, we determined whether treatment with ibuprofen synergizes with PD-1 checkpoint blockade. The combination of α-PD-1 with ibuprofen resulted in a 100% tumor clearance and survival rate, whereas α-PD-1 or ibuprofen alone resulted in 40% and 60% tumor clearance and survival rates, respectively. Furthermore, treatment with ibuprofen also sensitized α-PD-1-resistant tumors to PD-1 in two distinct models of α-PD-1 resistance. Overall, these findings establish ibuprofen and other NSAIDs as inducers of ferroptosis and underscore the potential for ferroptosis inducers to be used in combination with existing immunotherapy regimens.

Citation Format: Ronan Talty, Michelle Ferreira, Irina Krykbaeva, Marcus Bosenberg. Ibuprofen induces ferroptosis to potentiate antitumor immunity [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 2166.
Abstract 245: Interleukin-4 receptor-targeted Abraxane inhibits tumor growth by enhancing drug delivery and reprogramming of M2-type macrophages into M1 phenotype
Authors: Murugan Poongkavithai Vadevoo Sri , Rangaswamy Gunassekaran Gowri , Lee Byungheon
Published: June 2022

Abraxane, an albumin-bound paclitaxel nanoparticle, is widely used in treating patients with cancer such as breast cancer. The present study aims at a targeted delivery of Abraxane to tumors by conjugating it with IL4RPep-1, an interleukin-4 receptor (IL4R)-binding peptide. IL4R is up-regulated in many tumor cells and M2-polarized macrophages. IL4R-targeted Abraxane (IL4RPep-1-Abx) showed more efficient binding and internalization into IL4R-high tumor cells and M2-type macrophages than untargeted Abraxane, which was inhibited in part by an inhibitor of macropinocytotsis and receptor-mediated endocytosis, respectively, while being completely inhibited by an IL4R-blocking antibody. In the same context, IL4RPep-1-Abx induced higher levels of cytotoxicity to IL4R-high tumor cells and reprogramming of M2-type macrophages into M1-like phenotype than untargeted Abraxane. In vivo fluorescence imaging showed that IL4RPep-1-Abx homed to tumor, with little accumulation in the liver and lung, more efficiently than untargeted Abraxane. In addition, IL4RPep-1-Abx inhibited tumor growth and metastasis to the lung in 4T1 or LLC tumor-bearing mice and reduced the number of tumor nodules in the lung of K-ras transgenic mice more efficiently than untargeted Abraxane. Liver function was maintained within the normal ranges after treatment with IL4RPep-1-Abx. In 4T1 and LLC tumor tissues, IL4RPep-1-Abx reduced the population of immune-suppressive cells such as M2-type macrophages and increased that of immune-stimulatory cells such as CD8+ T cells compared with untargeted Abraxane. These results suggest that IL4R-targeted Abraxane can enhance the therapeutic efficacy and anti-tumor immunity of Abraxane and is a promising strategy for targeted cancer therapy.

Citation Format: Poongkavithai Vadevoo Sri Murugan, Gunassekaran Gowri Rangaswamy, Byungheon Lee. Interleukin-4 receptor-targeted Abraxane inhibits tumor growth by enhancing drug delivery and reprogramming of M2-type macrophages into M1 phenotype [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 245.
Abstract 2501: Identification and characterization of key players in the Ewing Sarcoma metastatic process
Authors: Serra Sara Sanchez , Bosch Mariona Chicon , Velazquez Judith Besalú , Marcos Susana Maqueda , Monaco Paola , Viera Laura Santana , Alemany Roser Lopez , Tirado Oscar M.
Published: June 2022

Metastasis at diagnosis is the most adverse prognostic factor in Ewing Sarcoma (ES), drastically decreasing patients’ overall survival down to <30%1. However, mechanisms leading to metastasis in this disease remain poorly understood. To deepen our knowledge on the main players in ES metastasis, Microarray (Clariom™ D Array, Affymetrix) and phosphoarray (Phospho Explorer Antibody Arrays, Full Moon Biosystems) analyses were performed on primary tumours (n=5) and metastases (n=12) samples from an spontaneous metastatic ES mouse model2. After normalisation and bioinformatics analysis, a list of differentially expressed genes/proteins between the 2 sample groups was obtained. Candidates were validated by qPCR (TaqMan™ probes) and Western Blot, and further characterised in vitro through loss-of-function phenotypic assays (proliferation, clonogenic and migration) using siRNAs and/or inhibitors. Several genes and proteins of interest, including LOXHD1, confirmed to be overexpressed in metastases samples compared to primary tumours as identified by microarray/phosphoarray analyses and qPCR/Western Blot validation. Also, candidates subjected to transient loss-of-function phenotypic assays significantly decreased in all cases tumorigenic and/or metastatic capabilities of ES cells when suppressed. Additionally, LOXHD1 showed upregulation in sphere-growing ES cells, validating its role in ES hypoxia and metastasis as demonstrated previously3. Using stable knock-out models of the genes of interest, the exact mechanism and pathways in which these candidates play a role in ES metastasis will be further studied in vitro and in vivo. Also, their clinical relevance will be explored in several cohorts of patients by immunohistochemistry on Tissue Microarrays. [1] Grünewald, TGP et al. 2018, Nat Rev Dis Prim, 4(1), pp.1-22. [2] Lagares-Tena, L et al. 2016, Oncotarget, 7(35), pp.56889-903. [3] Deng, Q et al. 2021, bioRxiv 2021.02.22.432287.

Citation Format: Sara Sanchez Serra, Mariona Chicon Bosch, Judith Besalú Velazquez, Susana Maqueda Marcos, Paola Monaco, Laura Santana Viera, Roser Lopez Alemany, Oscar M. Tirado. Identification and characterization of key players in the Ewing Sarcoma metastatic process [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 2501.
Abstract 2695: Transcriptomic profiling of human patients with acute myeloid leukemia reveals critical effects of growth hormone responsiveness on patient survival and mechanisms of tumor therapy resistance
Authors: Davis Emily , Basu Reetobrata , Kopchick John J.
Published: June 2022

Growth hormone (GH) has been reported to drive leukemia cell proliferation as early as 1977, while downstream effectors of the GH signaling pathway (i.e., insulin-like growth factor 1; IGF1) have more recently been widely implicated in the aggressive pathology and robust therapy resistance phenotypes of acute myeloid leukemia (AML). It is also known that patients with acromegaly (high levels of circulating GH and IGF1) develop AML at a significantly higher rate than the general population, while individuals with GH resistance remain completely resistant to all types of cancer. Despite all of the aforementioned evidence pointing to a critical role of GH in AML progression and prognosis, a detailed transcriptomic analysis of human patients is lacking. We aim to address this gap by thorough in silico studies aimed at revealing the nature and extent to which the GH axis can be suitable targeted in AML. Here, we present a summary of our analyses of clinical and transcriptomic data from 200 human patients with AML from The Cancer Genome Atlas database, using multiple bioinformatics software programs in the public domain. Our study revealed that GH receptor (GHR) is expressed more highly in AML than in normal myeloid cells, and a robust correlation exists between GH responsiveness and patient survival. A sex-specific profiling of the patient transcriptomic data identified critical pathways related to therapy resistance, extracellular matrix remodeling, metastasis, and immune evasion to be upregulated differentially in male and female AML patients with relatively elevated GHR expression compared to patients with relatively lower GHR expression. Furthermore, we identified microRNAs which are differentially expressed in AML patients with higher than median GHR expression. We validated these miRNAs to be effective indicators of disease progression and prognosis and can be valuable markers in liquid biopsy approaches.

Acknowledgement: This work was supported in part by the State of Ohio’s Eminent Scholar Program that includes a gift from Milton and Lawrence Goll, by the AMVETS, and Ohio University’s Edison Biotechnology Institute.

Citation Format: Emily Davis, Reetobrata Basu, John J. Kopchick. Transcriptomic profiling of human patients with acute myeloid leukemia reveals critical effects of growth hormone responsiveness on patient survival and mechanisms of tumor therapy resistance [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 2695.
Abstract 2699: Glucocorticoid receptor biology in lobular breast cancer
Authors: Frerich Candace , Durdana Ishrat , Hanker Ariella , Arteaga Carlos L. , Bennett Lynda , Conzen Suzanne
Published: June 2022

While considered uncommon, there are an estimated 39,000 new invasive lobular carcinoma (ILC) cases in the US yearly, making it the sixth most common among all tumor types. ILC is recognized for unique metastatic organotropism. Its propensity to metastasize to serosal surfaces lining the gut (peritoneum) and lung (pleura) make it particularly challenging to diagnose and consequently is associated with overall unfavorable prognosis. We hypothesize that GR activation will slow cell proliferation and reduce metastatic potential of ILC cells in vitro and in vivo. Using transcriptome data from 142 ILC tumors with long-term follow up clinical data from the METABRIC dataset we found a trend towards improved overall patient survival with increased GR (NR3C1) expression (and presumably activity). Preliminary in vitro data showed slowed cell proliferation in an ILC cell line (MDA-MB-134IV) treated with GR agonist dexamethasone relative to vehicle control. ILC cell lines (MDA-MB-134IV and SUM44PE) have dramatically different adhesion preference for extracellular components and we will study whether GR mediates the metastatic organotrophism of ILC cells. Preliminary transcriptome data lends insight into the proliferative and adhesive gene expression changes elicited by both ER and GR activation. Two GR+ ILC patient derived organoid lines have GR dependent proliferative and adhesive phenotypes. In future studies we will utilize an in vivo mouse model that recapitulates ILC progression from an in situ to invasive breast carcinoma and finally to metastatic colonization of distant organs to determine the role of GR activity in proliferation and metastasis to the serosal surfaces.

Citation Format: Candace Frerich, Ishrat Durdana, Ariella Hanker, Carlos L. Arteaga, Lynda Bennett, Suzanne Conzen. Glucocorticoid receptor biology in lobular breast cancer [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 2699.