bluuebunny/crossref_metadata_2025 huggingface dataset:
A Multi-Label Classification with an Adversarial-Based Denoising Autoencoder for Medical Image Annotation
Authors: Chai Yidong , Liu Hongyan , Xu Jie , Samtani Sagar , Jiang Yuanchun , Liu Haoxin
Published: September 2022

Medical image annotation aims to automatically describe the content of medical images. It helps doctors to understand the content of medical images and make better informed decisions like diagnoses. Existing methods mainly follow the approach for natural images and fail to emphasize the object abnormalities, which is the essence of medical images annotation. In light of this, we propose to transform the medical image annotation to a multi-label classification problem, where object abnormalities are focused directly. However, extant multi-label classification studies rely on arduous feature engineering, or do not solve label correlation issues well in medical images. To solve these problems, we propose a novel deep learning model where a frequent pattern mining component and an adversarial-based denoising autoencoder component are introduced. Extensive experiments are conducted on a real retinal image dataset to evaluate the performance of the proposed model. Results indicate that the proposed model significantly outperforms image captioning baselines and multi-label classification baselines.
Nonsurgical Orthodontic Treatment of a Severe Open Bite Case Using Miniscrews with Modified Multiloop Edgewise Arch Wire Technique
Authors: Hatrom Abdulkarim A. , Kanwal Bushra , Hamooda Fatima , Alzahrani Hashim A.
Published: September 2022

Skeletal open bite is one of the most challenging malocclusions to treat and maintain due to the difficulty and instability of correction. Although a combination of orthodontic treatment and orthognathic surgery may be the ideal approach in most cases, the complications, risks, and costs of surgery have sparked an interest in alternative treatment options that use temporary anchorage devices to achieve orthognathic-like effects. Adult patients can be treated without the need for special compliance using temporary anchorage devices such as miniscrews. This case report demonstrates a goal-oriented strategy for nonsurgical treatment of a complex skeletal open bite malocclusion in an adult patient using miniscrews and a modified multiloop edgewise arch wire (MEAW) technique, with the results evaluated clinically and cephalometrically.
Afatinib Targeted Therapy Affects the Immune Function and Serum Levels of EGFR and Gastrin-Releasing Peptide Precursor (pro-GRP) in Patients with Non-Small-Cell Lung Cancer (NSCLC)
Authors: Cao Wei , Ma Jun , Jiang Xuan , Gao Guangyi
Published: September 2022

Objective. This study is aimed at investigating the clinical intervention effect of afatinib targeted therapy in patients with non-small-cell lung cancer. Methods. The research object was a retrospective analysis of 86 patients with non-small-cell lung cancer who were admitted to our hospital from 1st January 2019 to 31st December 2021. The patients were divided into two groups. The patients in the two groups received conventional chemotherapy intervention, and the patients in group B received afatinib targeted therapy intervention on the basis of the treatment in group A. The clinical intervention effect, immune function, serum EGFR level, serum pro-GRP level, and incidence of adverse reactions were compared between the two groups of patients. Results. After afatinib targeted therapy intervention, the total intervention effective rate of patients in treatment group B was significantly higher than that in patients in treatment group A. Compared with the treatment group A, the CD3+, CD4+, CD8+, and CD4+/CD8+ of the treatment group were significantly upregulated. After the intervention, the serum EGFR levels of patients in treatment groups A and B were significantly decreased, and the serum EGFR levels in patients in treatment group B were significantly lower than those in patients in treatment group A. The serum pro-GRP level in group B patients was significantly decreased. The overall incidence of adverse reactions in treatment group B was significantly lower than that in treatment group A. Conclusion. Afatinib targeted therapy has a significant clinical intervention effect on patients with non-small-cell lung cancer, which not only helps to improve the immune function of patients but also effectively improves the serum EGFR and pro-GRP levels of patients.
Excessive Defense of Antitakeover Provisions and Enterprise Value Based on PSM-DID Quasi-Natural Experimental Method
Authors: Yao Zhang , Xuemeng Guo , Jia Jia
Published: September 2022

Under the background of frequent hostile takeover events in China and the autonomy of the articles of association, the articles of association of listed companies even set up antitakeover provisions with an excessive defense that unreasonably restricts the rights of shareholders. Based on the role orientation of innovative supervision, the Small and Medium-sized Investors Service Center (hereinafter referred to as “Investment and Service Center”) proposes to modify the relevant noncompliance provisions in the form of a shareholder recommendation letter. As the basic requirements of the articles of association and the relevant laws in corporate governance are increasingly inconsistent, the legal review of the articles of association is particularly important. This article determines the identification standard of excessive defense of antitakeover provisions by manually collecting the shareholder suggestion letter of the investment service center, the announcement on amendment of association articles of listed companies, and relevant laws and regulations. On this basis, the PSM-DID quasi-natural experimental method is used to empirically test the impact of excessive defense of antitakeover provisions on enterprise value. The study finds that the excessive defense of antitakeover provisions significantly inhibited the enterprise value. In this article, taking a-share listed companies in Shanghai and Shenzhen stock markets in 2007–2018 as an example, with the construction of antiacquisition clause excessive defense index (ATP end), relying on the “quasi-natural experiment” method of PSM-DID, we discuss the impact of excessive defense antiacquisition clause on the value of the enterprise and its impact path analysis. Further study found that its restriction mechanism was reflected in the significant increase of the first type of agency cost and the second type of agency cost. At the same time, the correlation between management compensation and performance and the correlation between management and professional performance are further reduced. Excessive defense against antiacquisition terms can reduce the corporate value, and the impact path of this effect is achieved by increasing type 1 agent costs and type 2 agent costs. It shows that the controlling shareholders are more likely to collude with the management, which aggravates the agency problem. The research conclusion of this article provides legal and economic empirical evidence for listed companies to reasonably set up antitakeover provisions and is also of great significance to innovative regulatory methods such as investment service centers. Excessive defense against antitakeover terms will enhance the dual-ditch effect and ultimately damage corporate value.
Intelligent Recognition System of Sports Athletes’ Wrong Actions Based on AI+IoT
Authors: Zhang Jie , Du Xiru , Bi Ran
Published: September 2022

How to effectively identify errors in athletes’ movements, and then achieve the goal of correcting or eliminating them, is a problem that the sports world is currently trying to solve. Sports field is a field where sensor-based human movement recognition can play a role. Obtaining human body data through sensors can provide data basis for research on the characteristics of human movement, improvement of sports level, and technical movement analysis of sports. The main research direction of this paper is the AI+IoT joint recognition of human movements. The target human movements involved include standing, walking, running, shooting, jump shot, jumping, dribbling, walking dribbling, and running dribbling. Computer vision, as a branch of the artificial intelligence research field, uses computers to simulate human visual cognitive ability, excavate useful information from digitized images or videos without manual intervention, and strive to achieve human understanding of visual signals, and realize the ability to understand visual signals from a low level. Conversion of data input to high-level knowledge output. The system scoring module is the core functional module of the system. It extracts the angle feature of the trainer’s skeleton data, finds the corresponding frame to correspond, and prompts the trainer’s current standard degree of movement in the form of score. In terms of judgment accuracy, the accuracy of all subjects after watching the complete video is higher than the accuracy of the time blocking point, with extremely significant differences,

r

 = -10.80, df =52,

p
<
0.001

. This research improves the universality of target detection and extraction in the real natural environment of the machine vision system and improves the complexity and robustness of human action recognition.
Analysis of the Exploration of Security and Privacy for Healthcare Management Using Artificial Intelligence: Saudi Hospitals
Authors: Almalawi Abdulmohsen , Khan Asif Irshad , Alsolami Fawaz , Abushark Yoosef B. , Alfakeeh Ahmed S. , Mekuriyaw Walelign Dinku
Published: September 2022

A large component of the Health Information Systems now comprises numerous independent apps created in the past that need to be merged to provide a more uniform service. In addition to affecting the Intelligent Health Board Functionality and dependability, the quality of these additional apps may also have an impact. A critical characteristic of the SHS's management and upkeep is the SHS's reliance on the real benefits provided to it. In speaking, an HMIS (Healthcare Management Information System) is a computer-based device that benefits medical practitioners to perform their duties more efficiently by coordinating all of their data. Even though these systems are widely used by most of the world, there is a significant need to comprehend these technologies and indeed the potential they provide. Healthcare data warehouses in Saudi Arabia have evolved through time, and this research examines how key service improvements in Saudi present varied viewpoints on how premium initiative help may be attained in health as well as how this could be done. When it comes to understanding how different types of medical professionals interact with healthcare systems throughout history, researchers developed stages of the maturity model.
A Proposed Design Method for Sparse Array Antenna by Using the Spacing Coefficient Algorithm
Authors: Sandi Efri , Diamah Aodah , Yuliatmojo Pitoyo , Maruddani Baso
Published: September 2022

One of the major challenges in developing various practical communication systems is reducing device complexity and development costs. In this study, a linear sparse array antenna design problem is addressed and a new approach for density taper element spacing by using the spacing coefficient algorithm is proposed. This method is a mathematical approach to obtain the distance between array elements by developing a spacing coefficient

d

n
 

for each element to achieve radiation performances with a minimum number of antenna elements. The simulation and measurement results show a significant improvement in array performance compared to other sparse array design methods, such as the CDS sparse array method in our previous work.
Acetylation of Atp5f1c Mediates Cardiomyocyte Senescence via Metabolic Dysfunction in Radiation-Induced Heart Damage
Authors: Zeng Zhimin , Xu Peng , He Yanqing , Yi Yali , Liu Zhicheng , Cai Jing , Huang Long , Liu Anwen
Published: September 2022

Objective. Ionizing radiation (IR) causes cardiac senescence, which eventually manifests as radiation-induced heart damage (RIHD). This study is aimed at exploring the mechanisms underlying IR-induced senescence using acetylation proteomics. Methods. Irradiated mouse hearts and H9C2 cells were harvested for senescence detection. Acetylation proteomics was used to investigate alterations in lysine acetylation. Atp5f1c acetylation after IR was verified using coimmunoprecipitation (Co-IP). Atp5f1c lysine 55 site acetylation (Atp5f1c K55-Ac) point mutation plasmids were used to evaluate the influence of Atp5f1c K55-Ac on energy metabolism and cellular senescence. Deacetylation inhibitors, plasmids, and siRNA transfection were used to determine the mechanism of Atp5f1c K55-Ac regulation. Results. The mice showed cardiomyocyte and cardiac aging phenotypes after IR. We identified 90 lysine acetylation sites from 70 protein alterations in the heart in response to IR. Hyperacetylated proteins are primarily involved in energy metabolism. Among them, Atp5f1c was hyperacetylated, as confirmed by Co-IP. Atp5f1c K55-Ac decreased ATP enzyme activity and synthesis. Atp5f1c K55 acetylation induced cardiomyocyte senescence, and Sirt4 and Sirt5 regulated Atp5f1c K55 deacetylation. Conclusion. Our findings reveal a mechanism of RIHD through which Atp5f1c K55-Ac leads to cardiac aging and Sirt4 or Sirt5 modulates Atp5f1c acetylation. Therefore, the regulation of Atp5f1c K55-Ac might be a potential target for the treatment of RIHD.
Convolution and Coefficient Estimates for (p,q)-Convex Harmonic Functions Associated with Subordination
Authors: Bayram Hasan , Yalçın Sibel
Published: September 2022

We preface and examine classes of (p, q)-convex harmonic locally univalent functions associated with subordination. We acquired a coefficient characterization of (p, q)-convex harmonic univalent functions. We give necessary and sufficient convolution terms for the functions we will introduce.
Adaptive Control Strategy of Multiemergency Power Supply Network System Connected to New Energy
Authors: Zhang Wei , Zhong Ming , A Minfu , Sun Yumei , Wang Yang
Published: September 2022

In order to improve the adaptive control effect of the multiemergency power supply networking system connected to new energy (NE), this paper studies the adaptive control strategy (ACS) of the multiemergency power supply networking system connected to NE by combining intelligent algorithms and determines the appropriate installation node of the distributed power supply according to the voltage stability index. Moreover, this paper proposes a method to optimize the distribution network feeder reorganization and distributed power configuration problems based on the fireworks algorithm to reduce the network loss and improve the voltage distribution. In addition, distribution network reorganization and distributed power configuration optimization can more effectively minimize network losses and improve voltage distribution. Through the experimental research, it can be seen that the ACS of the multiemergency power supply network system connected to NE proposed in this paper has the established effect.
An In-Place Simplification on Mixed Boolean-Arithmetic Expressions
Authors: Liu Binbin , Zheng Qilong , Li Jing , Xu Dongpeng
Published: September 2022

Mixed Boolean-arithmetic (MBA) expression, which involves both bitwise operations (e.g., NOT, AND, and OR) and arithmetic operations (e.g.,

+

,

−

, and

∗

), is a software obfuscation scheme. On the other side, multiple methods have been proposed to simplify MBA expressions. Among them, table-based solutions are the most powerful simplification research. However, a fundamental limitation of the table-based solutions is that the space complexity of the transformation table drastically explodes with the number of variables in the MBA expression. In this study, we propose a novel method to simplify MBA expressions without any precomputed requirements. First, a bitwise expression can be transformed into a unified form, and we provide a mathematical proof to guarantee the correctness of this transformation. Then, the arithmetic reduction is smoothly performed to further simplify the expression and produce a concise result. We implement the proposed scheme as an open-source tool, named MBA-Flatten, and evaluate it on two comprehensive benchmarks. The evaluation results show that MBA-Flatten is a general and effective MBA simplification method. Furthermore, MBA-Flatten can assist malware analysis and boost SMT solvers’ performance on solving MBA equations.
Analyzing the Coupling Degree of Coordinated Development between Ecological Environment and Regional Economy in Underdeveloped Areas
Authors: Zhang Shiwen , Sentosa Bin Anwar Malik Ilham , Zhong Yibo
Published: September 2022

Accelerating economic development necessitates the rapid consumption of more ecological environmental resources. Excessive consumption of ecological environmental resources will seriously damage the overall coordination between the ecological environment and the local economy. We examine the objective issues of regional economic development and environmental protection in the three eastern provinces. Local governments should prioritize the degree of interdependence between regional economic development and the ecological environment. A healthy development strategy includes managing and improving the regional environment while also ensuring the orderly and stable development of the local economy. The grey correlation analysis model is based on the coupling relationship analysis and uses the underdeveloped Northeast Economic Zone as the study goal. So, this study proposes that the regional economic circle of three northeastern provinces be the main topic of a quantitative analysis of the coupling relationship and influencing factors of the coordinated development of the ecological environment and regional economy from 2013 to 2020. Inferred from the perspective of spatial coupling relationships is the calculation of the coupling degree model based on the grey correlation degree. Specifically, Harbin (0.661), Changchun (0.650), Jixi (0.615), and Liaoyuan (0.629) fall into the antagonistic type and low-level coupling type categories. The degree of coupling between Northeast China’s economic development and the natural environment significantly decreased from 2015 to 2018 from the perspective of the time series coupling relationship. The changing trend of coupling degree makes it abundantly clear that the problems with the ecological environment’s constraints in the underdeveloped areas of the three northeastern provinces have not been fundamentally resolved. If no action is taken rapidly, the ecological environment will continue to deteriorate because environmental problems do not improve significantly in a short time.
The Effects of Dietary Thyme Oil (Thymus vulgaris) Essential Oils for Common Carp (Cyprinus carpio): Growth Performance, Digestive Enzyme Activity, Antioxidant Defense, Tissue and Mucus Immune Parameters, and Resistance against Aeromonas hydrophila
Authors: Ghafarifarsani Hamed , Hoseinifar Seyed Hossein , Sheikhlar Atefeh , Raissy Mehdi , Chaharmahali Fatemeh Heidarinezhad , Maneepitaksanti Worawit , Faheem Mehwish , Van Doan Hien
Published: September 2022

This study evaluated the effects of dietary supplementation of thyme (Thymus vulgaris) essential oil (TVO) on growth performance, digestive enzymes, biochemical parameters, hematological indices, liver enzymes, and pathogen resistance in common carp (Cyprinus carpio). Triplicate groups of fish (15.36 ± 0.10 g) were fed daily with diets supplemented with TVO at 0, 0.5, 1, and 2 percent for 60 days then challenged with Aeromonas hydrophila. The results determined that supplementation of thyme resulted in significantly higher final body weights (FBW) and lower feed conversion ratios (FCR). Furthermore, no mortality was observed in the thyme-supplemented treatments. Regression analysis showed that fish growth parameters were polynomially related to dietary TVO levels. The optimum dietary TVO level, based upon the varied growth parameters, was 1.344 to 1.436%. Digestive enzymes activity, including amylase and protease, significantly increased in fish fed the supplemented diets. The thyme-supplemented diets also significantly increased the biochemical parameters, including total protein, albumin, and acid phosphatase (ACP), compared to the control group. We also observed significant increases in hematological indices, including red blood cells (RBC), white blood cells (WBC), hematocrit (Hct), and hemoglobin (Hb) in common carp fed diets containing thyme oil (

P
<
0.05

). Liver enzymes activity including alanine aminotransferase (ALT), alkaline phosphatase (ALP), and aspartate aminotransferase (AST) was also reduced (

P
<
0.05

). Immune parameters, including total protein and total immunoglobulin (total Ig) levels, alternative complement pathway hemolytic (ACH50), lysozyme, protease, and ALP in the skin mucus, and lysozyme, total Ig, and ACH50 in the intestine were higher (

P
<
0.05

) in TVO-supplemented fish. Catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR), and glutathione peroxidase (GPx) in the liver were also elevated (

P
<
0.05

) in TVO administered groups. Lastly, thyme-supplementation resulted in higher survival rates after the A. hydrophila challenge compared to the control (

P
<
0.05

). In conclusion, dietary inclusion of thyme oil (1 and 2%) effectively improved fish growth, immune systems, and resistance to A. hydrophila.
Modeling and Simulation of Restorative Indoor Environment Based on Neural Network
Authors: Ding Xianyao , Feng Haocheng , Shan Weilun
Published: September 2022

In the busy modern society, after experiencing fast-paced work, what people most desire is to have a relatively relaxed and comfortable indoor rest scene. At the same time, with the improvement of material living standards, people have higher and higher requirements for the office and indoor rest environment, and the indoor temperature environment is very important to create a sense of comfort. A good indoor somatosensory temperature environment can not only convey the degree of somatosensory comfort but also relax people’s bodies, in which the air conditioning control system can restore the indoor temperature environment. Based on the abovementioned problems, this paper uses the neural network modeling method, based on the indoor comfort index SET

∗

value and other factors that can affect the environmental temperature, aiming at the human body temperature comfort, constructs a BP neural network model with indoor environmental parameters as the input index, and uses the improved particle swarm optimization algorithm to optimize the model, so as to realize the real-time control of SET

∗

value, and then analyzes the relationship between indoor environmental thermal factors and SET

∗

value, The model is optimized through simulation experiments to improve the optimization degree of the model. The practice shows that the restorative indoor environment model based on improved BP neural network constructed in this paper can optimize the room parameters well, so as to improve indoor comfort and give people a warm accommodation environment.
A Comprehensive Evaluation Method for the Effectiveness of Public Health‐Oriented Music Performance Art Based on Blockchain Technology
Authors: Shang Yiran
Published: September 2022

Music can promote the development of physical and mental health, and its therapeutic utility has begun to receive widespread attention from scholars, who have systematically studied the therapeutic methods, processes, and utility of music with the theme of music therapy, and the topic has been extended from music therapy at the individual level to the public health domain. Therefore, it is worthwhile to further explore how the effectiveness of music as a performing arts activity in the public health domain is evaluated. However, most of the current studies focus on the evaluation of the effects of music on individual physical and mental health, and few involve the evaluation of music performing arts activities in the public health domain, which greatly hinders the potential of music performing arts applications in the public health domain. Therefore, this study proposes a dynamic and comprehensive evaluation scheme based on the cross‐chain technology in blockchain and establishes a cross‐chain‐based information exchange model for the feasibility of information exchange between music performing arts and public health. The research findings can not only provide theoretical guidance for the formulation of public health policies but also provide technical support for the comprehensive evaluation of the effectiveness of music performing arts activities.
Support Personalized Weighted Local Differential Privacy Skyline Query
Authors: Zhang Guopeng , Chen Xuebin , Jia Yuanli , Zhai Ran
Published: September 2022

The potential privacy risks in certain situations are of concern because of the frequent sharing of data during skyline queries, leading to leakage of users’ private information. The most common privacy-preserving technique is to anonymize data by removing or changing certain information, for which an attack with specific background knowledge would render the privacy protection ineffective. To overcome these difficulties, this study proposes a personalized weighted local differential privacy method (PWLDP) to protect data privacy during skyline querying. Compared with existing studies of skyline queries under privacy protection, the degree of privacy protection can be quantitatively analyzed, and the processing of data privacy lies with the user, who quantitatively perturbs the processing according to the sensitivity of the weights of different attributes to avoid substantial information loss. The performance of the proposed PWLDP is verified by comparing PWLDP and LDP on different datasets, the average privacy leakage reduction of 62.22% and 51.67% is obtained for experiments conducted on different datasets relative to the iDP-SC algorithm, and the experimental results demonstrate the efficiency and advantages of the proposed method.
The Usage of Designing the Urban Sculpture Scene Based on Edge Computing
Authors: Zhu Junru
Published: September 2022

To not only achieve the goal of urban cultural construction but also save the cost of urban sculpture space design, EC (edge computing) is combined with urban sculpture space design and planning first. Then it briefly discusses the service category, system architecture, advantages, and characteristics of urban sculpture, as well as the key points and difficulties of its construction, and the layered architecture of EC for urban sculpture spaces is proposed. Secondly, the cloud edge combination technology is adopted, and the urban sculpture is used as a specific function of the edge system node to conduct an in-depth analysis to build an urban sculpture safety supervision system architecture platform. Finally, the actual energy required for implementation is predicted and evaluated, the specific monitoring system coverage is set up, and some equations are made for calculating the energy consumption of the monitored machines according to the number of devices and route planning required by the urban sculpture safety supervision system. An optimization algorithm for energy consumption is proposed based on reinforcement learning and compared with the three control groups. The results show that when the seven monitoring devices cover detection points less than 800, the required energy consumption increases linearly. When the detection devices cover more than 800 detection points, the required energy consumption is stable and varies from 10000 to 12000; that is, when the number of monitoring devices is 7, the optimal number of monitoring points is about 800. When the number of detection points is fixed, increasing the number of monitoring devices in a small range can reduce the total energy consumption. The optimization algorithm based on the reinforcement learning proposal can obtain an approximate optimal solution. The research results show that the combination of edge computing and urban sculpture can expand the function of urban sculpture and make it serve people better.
The MicroRNA-106a/20b Strongly Enhances the Antitumour Immune Responses of Dendritic Cells Pulsed with Glioma Stem Cells by Targeting STAT3
Authors: Zhou Hui , Sun Chengmei , Li Cong , Hua Shiting , Li Feng , Li Ruichun , Cai Dongpeng , Zou Yuxi , Cai Yingqian , Jiang Xiaodan
Published: September 2022

Background. Evaluate the effect of the miRNA-106a/20b on the efficacy of DCs pulsed with GSCs in activating GSC-specific T cell responses. Methods. We cultured GSCs and prepared GSC antigen lysates by apoptosis. Then, immature DCs were pulsed with GSC antigen lysates in vitro. STAT3 levels in DCs were assessed by Western blotting, and the expression of CD80, CD86, and MHC-II was tested by fluorescence-activated cell sorting. The production and secretion of the cytokines IL-6, IL-12, TNF-α, and IL-10 in DCs induced by GSCs were determined by enzyme-linked immunosorbent assay. Finally, the cytotoxic functions of T cells stimulated by GSC-DC fusion cells transfected with a miR-106a/20b mimic in vitro and the antitumour activity in vivo were detected. Results. We found that the levels of miR-106a/20b were downregulated, but the expression of STAT3 was significantly upregulated. Simultaneously, the inhibition of STAT3 in the fusion cells by STAT3-specific siRNA caused significant upregulation of the expression of CD80, CD86, and MHC-II, and the secretion of the cytokines IL-6 and IL-12 was substantially increased, IL-10 was markedly decreased. These findings revealed that STAT3 is an important regulator of DC maturation. Furthermore, the interactional binding sites between the 3

′

-untranslated region (3

′

-UTR) of STAT3 mRNA and miR-106a/20b were predicted by bioinformatics and verified by a dual-luciferase assay. Moreover, the reduction in STAT3 levels in GSC-DCs enhanced the generation of CD8+ T cells and reduced the generation of Foxp3+ regulatory T cells. Meanwhile, the secretion of the T cell cytokine IFN-γ was significantly increased. Further research showed that DCs after miR-106a/20b-mimics transfection could promote the inhibition of GSC proliferation by T cells in vitro and suppress tumour growth in vivo. Conclusions. This study indicted that the miR-106a/20b activation could be one of the important molecular mechanisms leading to enhance antitumour immune responses of GSC-mediated DCs, which downregulated the expression of STAT3 to alleviate its the inhibitory effect.
Abstract A012: Targeting the dysregulation of transcription and splicing in Ewing sarcoma
Authors: Lawrence Liesl A. , Miller Henry , Bishop Alexander
Published: September 2022

The goals of this study are to investigate how EWS-FLI1 disrupts regulation of transcription and splicing in Ewing sarcoma, and to evaluate this dysregulation as a potential therapeutic target. Ewing Sarcoma is an aggressive pediatric bone and soft tissue cancer driven primarily by the EWS-FLI1 fusion oncogene. EWS-FLI1 acts as an oncogenic transcription factor and also interferes with normal regulation of transcription and transcription-associated RNA processing. Chemotherapy has substantially improved overall survival, but the prognosis is still very poor for those with metastatic or recurrent disease, and toxicity is a major concern in pediatric patients. Consequently, effective, less toxic, targeted treatment strategies are much needed. We performed a genome-wide RNAi screen in the Ewing sarcoma cell line TC32 and found that one of the top pathways required for cell survival was RNA processing/splicing. Previous studies have reported increased amounts of alternative splicing events in Ewing sarcoma. RNA processing, including splicing, is functionally coupled with transcription. We recently reported that Ewing sarcoma cells have high levels of phosphorylated RNA polymerase II and increased transcription activity, accumulate R-loops, and fail to regulate transcription appropriately in response to DNA damage. We proposed that EWS-FLI1 driven hyper-activation of transcription causes a targetable dependence on RNA splicing in Ewing sarcoma. Ewing sarcoma cells showed increased sensitivity to depletion of key splicing factors compared to mesenchymal stem cell controls; this was partially rescued by depleting EWS-FLI1 or by overexpressing RNASEH1, which degrades R-loops. Ewing sarcoma cells were extremely sensitive to splicing inhibition, which showed synergy with multiple chemotherapeutic agents. Splicing inhibition in Ewing sarcoma cells caused cell cycle arrest, induced apoptosis, and altered global gene expression, alternative splicing, and R-loop profiles as seen by RNA-seq and DRIP-seq. This work provides novel insight into transcription regulation and its dysregulation by EWS-FLI1. In addition, our results point to RNA splicing as a potential new therapeutic target in Ewing sarcoma, which has the potential to benefit all Ewing sarcoma patients, especially those with chemo-resistant disease.

Citation Format: Liesl A. Lawrence, Henry Miller, Alexander Bishop. Targeting the dysregulation of transcription and splicing in Ewing sarcoma [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr A012.
Abstract A026: PROX1 transcription factor regulates rhabdomyosarcoma growth, myogenic properties and therapeutic targets
Authors: Gizaw Nebeyu Y. , Kallio Pauliina , Gucciardo Erika , Haglund Caj , Bohling Tom , Lehti Kaisa , Sampo Mika , Alitalo Kari , Kivela Riikka
Published: September 2022

Rhabdomyosarcoma (RMS) is an aggressive pediatric soft tissue cancer with features of skeletal muscle. Because of poor survival of RMS patients and severe long-term side effects of RMS therapies, novel RMS therapies are urgently needed. Here we show that the PROX1 transcription factor is highly expressed in RMS tumors regardless of their cell type of origin. We demonstrate that PROX1 is needed for RMS cell clonogenicity, growth and tumor formation. PROX1 gene silencing repressed several myogenic and tumorigenic transcripts and altered the RD (ERMS) transcriptome to resemble that of benign mesenchymal stem cells. Importantly, we found that fibroblast growth factor receptors (FGFR) mediated the growth effects of PROX1 in RMS. Because of receptor cross-compensation, inhibition of either alone did not mimic the effects of PROX1 silencing, whereas a pan-FGFR inhibitor ablated RMS cell proliferation and decreased tumor xenograft growth. Our findings uncover the critical role of PROX1 in RMS and offer new insights to the mechanisms that regulate RMS development and growth. As FGFR inhibitors have already been tested in unrelated clinical trials, our findings provide a new promising option for RMS treatment.

Citation Format: Nebeyu Y. Gizaw, Pauliina Kallio, Erika Gucciardo, Caj Haglund, Tom Bohling, Kaisa Lehti, Mika Sampo, Kari Alitalo, Riikka Kivela. PROX1 transcription factor regulates rhabdomyosarcoma growth, myogenic properties and therapeutic targets [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr A026.
Abstract A037: TT-00420, a novel kinase inhibitor potentially benefiting c-Myc amplified/overexpressed osteosarcoma
Authors: Hua Yingqi , Qiang Xiaoyan , Tian Kai , Jiang Yafei , Wu Frank , Peng Peng
Published: September 2022

Osteosarcoma (OS) is the most common bone tumor in pediatric patients, whose effective therapies are of great unmet medical needs, particularly for those of high-risk features. c-Myc, a well-known oncogene, is commonly amplified/overexpressed in OS patients, contributes to dismal prognosis and associates with lower 5-year overall survival rate, which is also confirmed by our retrospective analysis for the correlation between c-Myc gene amplification/overexpression and patient prognosis in our patient cohort. TT-00420, a clinical-stage small molecule targeting Aurora A/B, VEGFRs, FGFRs, JAKs and CSF1R, has demonstrated down-regulation on c-Myc expression in TNBC studies. Moreover, as reported, Aurora kinases and c-Myc regulate each other back and forth, triggering maintenance of the malignant state. In this study, we examined the efficacy of TT-00420 in c-Myc-amplified/overexpressed OS. In pre-clinical clonogenic assay, TT-00420 significantly inhibited the colony formation in c-Myc-overexpressed OS cell lines in a dose-dependent manner. In MNNG/HOS cell line, we demonstrated that TT-00420 dramatically suppressed Aurora kinase pathway and blunted c-Myc protein expression level, indicating TT-00420 might down-regulate c-Myc by Aurora kinases inhibition. In pre-clinical in-vivo efficacy studies, 143B cells were injected into the medullary cavity of the tibia to make the orthotopic OS xenograft model. With 15 mg/kg/day treatment of TT-00420, tumor growth was obviously inhibited as compared to vehicle group. Immunohistochemistry (IHC) analyses in tumors revealed that TT-00420-treated tumors exhibited increased apoptosis with upregulated expression of cleaved caspase 3 (CC3) and H2AX, and significantly decreased proliferations with raised expression of Ki67. Moreover, in a c-Myc-overexpressed OS patient-derived xenograft (PDX) model from a male pediatric patient, 15 mg/kg/day TT-00420 dramatically blunted the tumor growth with a TGI of 87.6%. In China, an investigator-initiated trial is currently ongoing to evaluate the safety and tolerability of TT-00420 in OS patients with c-Myc overexpression (ChiCTR2000036618). Taken together, TT-00420 depicted robust anti-tumor activity in c-Myc overexpressed OS both in vitro and in vivo with downregulation of c-Myc expression, indicating its potential as a novel monotherapy for OS patients with c-Myc overexpression. Future clinical trial result is of great expectation.

Citation Format: Yingqi Hua, Xiaoyan Qiang, Kai Tian, Yafei Jiang, Frank Wu, Peng Peng. TT-00420, a novel kinase inhibitor potentially benefiting c-Myc amplified/overexpressed osteosarcoma [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr A037.
Abstract A039: IGFBP5 is induced by tumor-suppressive naïve stromal cells and regulates GAS2-associated apoptosis in rhabdomyosarcoma
Authors: Ehnman Monika , Katkhada Karim , Meng Liu Zhen , Zhang Yue , Zhao Binbin , Tong Shanlin , Kallai Aditi , Östman Arne , Tobin Nick
Published: September 2022

Stromal cells can have either suppressive or stimulatory effects on tumor progression. However, this diverse collection of cells is still poorly characterized in soft-tissue sarcomas. Rhabdomyosarcoma (RMS) is the most common soft-tissue sarcoma in childhood with molecular features of skeletal muscle differentiation. In this study, we explored tumor growth-regulatory signaling between differentially educated non-malignant mesenchymal stromal cells and malignant tumor cells in RMS. Xenograft experiments in mice were designed to provide functional evidence for alterations in tumor behavior linked to tumor-associated stromal cells. Molecular characterization was performed on the Clariom D platform, by gene set enrichment analysis, cell culture validation of deregulated genes, and finally, by clinicopathological validation in a discovery cohort (N=101) and a validation cohort (N=147). The results revealed transient tumor growth-suppressive paracrine effects of naïve orthotopic stromal cells derived from skeletal muscle. These effects were easily abolished if the stromal cells were pre-exposed to RMS cells, either short-term in vitro, or long-term in hindlimb muscle in vivo. Explorative analyses of RMS cells exposed to naïve stromal cells identified insulin-like growth factor binding protein 5 (IGFBP5) as the top upregulated gene, with an ability to induce Caspase 3/7 activation and growth arrest. The gene coding for connective tissue growth factor (CTGF) was identified as the major prosurvival factor being downregulated. Analysis of patient cohorts consistently demonstrated an association between IGFBP5, growth arrest specific protein 2 (GAS2), lower disease stage and favorable survival. Altogether, our study identifies a novel anti-tumor growth mechanism with conditionally deregulated genes linked to apoptosis and differentiation in RMS. The results provide support for continued studies on tumor growth-regulatory mechanisms induced by tumor-surrounding stromal cell subsets at primary and metastatic sites in sarcoma.

Citation Format: Monika Ehnman, Karim Katkhada, Liu Zhen Meng, Yue Zhang, Binbin Zhao, Shanlin Tong, Aditi Kallai, Arne Östman, Nick Tobin. IGFBP5 is induced by tumor-suppressive naïve stromal cells and regulates GAS2-associated apoptosis in rhabdomyosarcoma [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr A039.
Abstract B009: Establishing new cell lines from undifferentiated pleomorphic sarcoma for sarcoma research
Authors: You Hye Jin , Lee Eun-Young , Rayhan Abu , Kang Hyun Guy , Kim June Hyuk , Park Jongwoong
Published: September 2022

Undifferentiated pleomorphic sarcoma (UPS), also known as malignant fibrous histiocytoma (MFH), is a high-grade soft-tissue sarcoma (STS). Despite of importance, there are very few models for study. Here, we have established and characterized UPS-derived cell lines. Cells were obtained UPS tissues by mincing followed by extracting or dissociating using enzymes and cultured by regular culture environment. Cells were maintained and immortal for months without artificial treatment and tumorigenic in vivo study. The tissues from in vivo study and tissues from patients were compared if it is representative for UPS by immunohistochemistry. Transcriptome from tissues and cell lines were compared. Several genes were identified as specific in tumor tissues and cell lines. Fusion genes were also identified. This study showed that new UPS cell lines might be a good resource for UPS study to get new insights.

Citation Format: Hye Jin You, Eun-Young Lee, Abu Rayhan, Hyun Guy Kang, June Hyuk Kim, Jongwoong Park. Establishing new cell lines from undifferentiated pleomorphic sarcoma for sarcoma research [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr B009.
Abstract B011: Two epigenetically distinct cellular states in osteosarcoma are regulated by a cluster-specific set of pioneer transcription factors
Authors: Fuentes Eunice Lopez , Clugston Andrew , Sayles Leanne C. , Sweet-Cordero E. Alejandro
Published: September 2022

The Osteosarcoma (OS) genome is characterized by aneuploidy and dramatic structural rearrangements often leading to copy number gain and loss. There is significant heterogeneity between patients. Additionally, there is less known regarding the epigenetic heterogeneity within osteosarcoma and how this may impact critical phenotypes such as therapy response and metastasis. To characterize the osteosarcoma epigenome, we integrated ATAC-seq and RNA-seq across a unique set of patient samples, PDXs and PDX-derived cell lines obtained across the disease continuum and including biopsies, resections, and metastases. Assay for Transposase Accessible Chromatin with high-throughput sequencing (ATAC-seq) analysis revealed the presence of two distinct epigenetic cell states. Differential chromatin accessibility between the clusters showed ~3000 unique open regions per cluster. Using GREAT to associate the differential genomic regions to genes, the enriched pathways of Cluster 1 (C1) are related to mesenchymal cell proliferation, skeletal system morphogenesis, osteoblast differentiation, and Wnt signaling pathway, whereas Cluster 2 (C2) enriched pathways are related to cell motility and migration, extracellular matrix organization and regulation of protein serine/threonine kinase activity. Accessible regions in the chromatin were enriched for a unique set of transcription factor binding sites. C1 accessible regions are enriched with transcription factor binding motifs (TFBM) related to development including RUNX2/3, MEOX2, HOXA2/5, VAX1/2, MEF, VSX and DLX; whereas members of AP1 complex including FOSL1/2, c-FOS, JUN are TFBM enriched in open regions in C2. This is correlated with high protein expression of these transcription factors in a cluster-specific manner and differential binding to chromatin. As expected, gene expression showed a positive correlation with accessible regions in promoters. RUNX2 plays a major role in chromatin remodeling in C1, modifying the chromatin accessibility of 5300 regions, most located in <1kb from the promoters. In conclusion, we uncovered two epigenetically cell states regulated by a cluster-specific set of transcription factors which are able to remodel the chromatin.

Citation Format: Eunice Lopez Fuentes, Andrew Clugston, Leanne C. Sayles, E. Alejandro Sweet-Cordero. Two epigenetically distinct cellular states in osteosarcoma are regulated by a cluster-specific set of pioneer transcription factors [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr B011.
Abstract B017: Tumor-secreted collagen VI weakens endothelium and promotes metastasis
Authors: Liu Ying , Fuller Ashley M. , Murazzi Ileana , Devine Ann , Skuli Nicolas
Published: September 2022

Metastasis is the leading cause of cancer-related deaths. However, the mechanisms behind the metastatic cascade remain poorly understood. Tumor’s metastatic potential is strongly influenced by microenvironmental cues such as low oxygen (hypoxia). Our published work reported that lung metastases in sarcoma are associated with increased primary tumor expression of the hypoxia-inducible collagen-modifying lysyl hydroxylase, Plod2; in multiple subtypes of sarcoma, excessive collagen lysyl hydroxylation results in secretion of immature collagen aggregates able to physically associate with tumor cells and promote both intravasation and extravasation. In the current study, we observed that Plod2 is colocalized with collagen VI and is important for its extracellular network structure, suggesting that collagen VI is a putative substrate of Plod2. Using an impedance sensing assay and a zebrafish intravital microinjection model, we demonstrated that tumoral Plod2 and collagen VI weaken the endothelial barrier and promote extravasation. With a tail vein injection mouse model, we determined that collagen VI is essential for lung metastasis. Clinically, we detected high levels of collagen VI in metastatic sarcoma in surgically resected patient lungs. Furthermore, by analyzing patient data in The Cancer Genome Atlas (TCGA), we found a strong correlation between the expression of both PLOD2 and collagen VI with disease outcomes. Together, our study identifies a novel mechanism of sarcoma lung metastasis, opening up opportunities for therapeutic intervention.

Citation Format: Ying Liu, Ashley M. Fuller, Ileana Murazzi, Ann Devine, Nicolas Skuli. Tumor-secreted collagen VI weakens endothelium and promotes metastasis [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr B017.