bluuebunny/crossref_metadata_2025 huggingface dataset:
Low-Intensity Pulsed Ultrasound Modulates RhoA/ROCK Signaling of Rat Mandibular Bone Marrow Mesenchymal Stem Cells to Rescue Their Damaged Cytoskeletal Organization and Cell Biological Function Induced by Radiation
Authors: Zhang Rong , Wang Zhaoling , Zhu Guoxiong , Wu Gaoyi , Guo Qingyuan , Liu Hongchen , Chen Lei
Published: September 2020

Osteoradionecrosis of the jaw (ORNJ) is an infrequent yet potentially devastating complication of head and neck radiation therapy. Low-intensity pulsed ultrasound (LIPUS) has been widely accepted as a promising method for the successful management of ORNJ, but the mechanism remains unclear. In this study, the effects of LIPUS on cytoskeletal reorganization, cell viability, and osteogenic differentiation capacity of rat mandible-derived bone marrow mesenchymal stem cells (M-BMMSCs) induced by radiation were determined by immunofluorescence staining, CCK-8 cell proliferation assay, quantification of alkaline phosphatase (ALP) activity, alizarin red staining, and real-time RT-PCR, respectively. Moreover, the involvement of the RhoA/ROCK signaling pathway underlying this process was investigated via western blot analysis. We found that radiation induced significant damage to the cytoskeleton, cell viability, and osteogenic differentiation capacity of M-BMMSCs and downregulated their expression of RhoA, ROCK, and vinculin while increasing FAK expression. LIPUS treatment effectively rescued the disordered cytoskeleton and redistributed vinculin. Furthermore, the cell viability and osteogenic differentiation capacity were also significantly recovered. More importantly, it could reverse the aberrant expression of the key molecules induced by radiation. Inhibition of RhoA/ROCK signaling remarkably aggravated the inhibitory effect of radiation and attenuated the therapeutic effect of LIPUS. In the light of these findings, the RhoA/ROCK signaling pathway might be a promising target for modifying the therapeutic effect of LIPUS on osteoradionecrosis.
Characterization of Novel Edible Films and Coatings for Food Preservation Based on Gum Cordia
Authors: Hashemi Gahruie Hadi , Safdarianghomsheh Reza , Zamanifar Parisa , Salehi Shahriyar , Niakousari Mehrdad , Hosseini Seyed Mohammad Hashem
Published: September 2020

As a pharmaceutical component, gum Cordia has been applied to improve crop resistance against many diseases. A large amount of gum appears around the fruit after soaking in an aqueous system. The mucilage possesses outstanding technofunctional properties as an emulsifier, thickening agent, and binding and stabilizing component in food and drug industries. The backbone of gum Cordia is composed of (1-2)-linked L-arabinofuranosyl and (1–6)-linked D-glucopyranosyl residues. This manuscript reviews the technofunctional properties and applications of gum Cordia in food systems. Particularly, our focus has been given to its application as a natural source for the formation of edible films and coatings for increasing the shelf life of food products and for the food preservation as a potential ingredient in formulation. The future research perspectives are also highlighted.
Adoption of Machine Learning in Intelligent Terrain Classification of Hyperspectral Remote Sensing Images
Authors: Li Yanyi , Wang Jian , Gao Tong , Sun Qiwen , Zhang Liguo , Tang Mingxiu
Published: September 2020

To overcome the difficulty of automating and intelligently classifying the ground features in remote-sensing hyperspectral images, machine learning methods are gradually introduced into the process of remote-sensing imaging. First, the PaviaU, Botswana, and Cuprite hyperspectral datasets are selected as research subjects in this study, and the objective is to process remote-sensing hyperspectral images via machine learning to realize the automatic and intelligent classification of features. Then, the basic principles of the support vector machine (SVM) and extreme learning machine (ELM) classification algorithms are introduced, and they are applied to the datasets. Next, by adjusting the parameter estimates using a restricted Boltzmann machine (RBM), a new terrain classification model of hyperspectral images that is based on a deep belief network (DBN) is constructed. Next, the SVM, ELM, and DBN classification algorithms for hyperspectral image terrain classification are analysed and compared in terms of accuracy and consistency. The results demonstrate that the average detection accuracies of ELM on the three datasets are 89.54%, 96.14%, and 96.28%, and the Kappa coefficient values are 0.832, 0.963, and 0.924; the average detection accuracies of SVM are 88.90%, 92.11%, and 91.68%, and the Kappa coefficient values are 0.768, 0.913, and 0.944; the average detection accuracies of the DBN classification model are 92.36%, 97.31%, and 98.84%, and the Kappa coefficient values are 0.883, 0.944, and 0.972. The results also demonstrate that the classification accuracy of the DBN algorithm exceeds those of the previous two methods because it fully utilizes the spatial and spectral information of hyperspectral remote-sensing images. In summary, the DBN algorithm that is proposed in this study has high application value in object classification for remote-sensing hyperspectral images.
Privacy-Enhancing Preferential LBS Query for Mobile Social Network Users
Authors: Siddula Madhuri , Li Yingshu , Cheng Xiuzhen , Tian Zhi , Cai Zhipeng
Published: September 2020

While social networking sites gain massive popularity for their friendship networks, user privacy issues arise due to the incorporation of location-based services (LBS) into the system. Preferential LBS takes a user’s social profile along with their location to generate personalized recommender systems. With the availability of the user’s profile and location history, we often reveal sensitive information to unwanted parties. Hence, providing location privacy to such preferential LBS requests has become crucial. However, the current technologies focus on anonymizing the location through granularity generalization. Such systems, although provides the required privacy, come at the cost of losing accurate recommendations. Hence, in this paper, we propose a novel location privacy-preserving mechanism that provides location privacy through k-anonymity and provides the most accurate results. Experimental results that focus on mobile users and context-aware LBS requests prove that the proposed method performs superior to the existing methods.
An Improved Broadcast Authentication Protocol for Wireless Sensor Networks Based on the Self-Reinitializable Hash Chains
Authors: Huang Haiping , Huang Qinglong , Xiao Fu , Wang Wenming , Li Qi , Dai Ting
Published: September 2020

Broadcast authentication is a fundamental security primitive in wireless sensor networks (WSNs), which is a critical sensing component of IoT. Although symmetric-key-based μTESLA protocol has been proposed, some concerns about the difficulty of predicting the network lifecycle in advance and the security problems caused by an overlong long hash chain still remain. This paper presents a scalable broadcast authentication scheme named DH-μTESLA, which is an extension and improvement of μTESLA and Multilevel μTESLA, to achieve several vital properties, such as infinite lifecycle of hash chains, security authentication, scalability, and strong tolerance of message loss. The proposal consists of the t,n-threshold-based self-reinitializable hash chain scheme (SRHC-TD) and the d-left-counting-Bloom-filter-based authentication scheme (AdlCBF). In comparison to other broadcast authentication protocols, our proposal achieves more security properties such as fresh node’s participation and DoS resistance. Furthermore, the reinitializable hash chain constructed in SRHC-TD is proved to be secure and has less computation and communication overhead compared with typical solutions, and efficient storage is realized based on AdlCBF, which can also defend against DoS attacks.
Crosscultural Validation of the Community Integration Questionnaire–Revised in an Italian Population
Authors: Ioncoli Melissa , Berardi Anna , Tofani Marco , Panuccio Francescaroberta , Servadio Annamaria , Valente Donatella , Galeoto Giovanni
Published: September 2020

Objective. The aims of this study are the translation, cultural adaptation, and validation of the Community Integration Questionnaire–Revised (CIQ-R) in Italian in a group of individuals with no clinical evidence of disability. Methods. The test’s internal consistency and validity were assessed by following international guidelines. The test’s internal consistency was examined using Cronbach’s alpha (α) coefficient. Pearson’s correlation coefficient was calculated to assess the test’s concurrent validity compared with the Short Form-12 (SF-12) health survey. Results. The CIQ-R was administrated to 400 people with no clinical evidence of disease, impairment, or disability, aged between 18 and 64. Cronbach’s α reported a value of 0.82 in the home integration subscale. The test also showed a good test-retest reliability, with an Intraclass Correlation Coefficient of 0.78, and a significant correlation between the total score of the CIQ-R and the Physical Component Summary (PCS) of the SF-12 (r=0.118), between the “social integration” subscale’s score and PCS12 (r=0.121) and between the “Electronic Social Networking integration” subscale’s score and PCS12 (r=0.184), with p<0.05. Conclusion. This is the first study to report the results of the translation and validation of the CIQ-R in Italian. The CIQ-R is an important tool for Italian professionals and can be useful in both clinical practice and research for measuring the level of community integration among the healthy population.
Quality Visibility Improvement with Effort Alignment and Cost-Sharing Policies in a Food Supply Chain
Authors: Zhang Wenbo , Su Qin
Published: September 2020

Improving quality visibility along a food supply chain has been considered as a critical driver of quality risk mitigation, safety and security assurance, and performance sustainability. This paper explores the coordination mechanisms in a food supply chain, where the demand and costs are sensitive to the supply chain quality visibility that depends on an upstream supplier and a downstream retailer jointly, and the effort to improve quality visibility is increasingly expensive. After comparing the centralized and decentralized supply chain models to discover an opportunity for Pareto improvement, it is proved that a pure revenue-sharing contract fails to coordinate the supply chain, while the price discount contract with effort alignment policy or effort cost-sharing policy works. The two coordinating contracts’ boundary conditions of excluding deviated actions are presented. It is shown that the contract with effort alignment policy is cheaper but more rigid, whereas the cost-sharing one allows us to arbitrarily allocate the supply chain’s profits despite more information being collected. The models are applied to a fresh chicken supply chain in order to verify their effectiveness and robustness in reality. The impacts of several specific parameters on supply chain decisions and performances are analyzed, and the results reveal some meaningful managerial implications regarding supply chain quality visibility.
Predicting Politician’s Supporters’ Network on Twitter Using Social Network Analysis and Semantic Analysis
Authors: Khan Asif , Zhang Huaping , Shang Jianyun , Boudjellal Nada , Ahmad Arshad , Ali Asmat , Dai Lin
Published: September 2020

Politics is one of the hottest and most commonly mentioned and viewed topics on social media networks nowadays. Microblogging platforms like Twitter and Weibo are widely used by many politicians who have a huge number of followers and supporters on those platforms. It is essential to study the supporters’ network of political leaders because it can help in decision making when predicting their political futures. This study focuses on the supporters’ network of three famous political leaders of Pakistan, namely, Imran Khan (IK), Maryam Nawaz Sharif (MNS), and Bilawal Bhutto Zardari (BBZ). This is done using social network analysis and semantic analysis. The proposed method (1) detects and removes fake supporter(s), (2) mines communities in the politicians’ social network(s), (3) investigates the supporters’ reply network for conversations between supporters about each leader, and, finally, (4) analyses the retweet network for information diffusion of each political leader. Furthermore, sentiment analysis of the supporters of politicians is done using machine learning techniques, which ultimately predicted and revealed the strongest supporter network(s) among the three political leaders. Analysis of this data reveals that as of October 2017 (1) IK was the most renowned of the three politicians and had the strongest supporter’s community while using Twitter in a very controlled manner, (2) BBZ had the weakest supporters’ network on Twitter, and (3) the supporters of the political leaders in Pakistan are flexible on Twitter, communicating with each other, and that any group of supporters has a low level of isolation.
Methyl Esterification Combined with Gas Chromatography-Mass Spectrometry (GC-MS) for Determining the Contents of Lubricant to Evaluate the Compatibility of Chlorinated Butyl Rubber Stoppers with Liposome Injections
Authors: Gao Xun , Chen Kexin , Chi Miaomiao , Qin Kunming
Published: September 2020

A simple, sensitive, and exact methyl esterification in combination with gas chromatography-mass spectrometry (GC-MS) method was developed to determine the contents of palmitic acid and stearic acid in the chlorinated butyl rubber stoppers and liposome injections in order to evaluate the compatibility of pharmaceutical packaging materials. In this experiment, palmitic acid and stearic acid were detected in the form of methyl hexadecanoate and methyl stearate in chlorinated butyl rubber stoppers and liposome injections. The results showed good linearities in the range of 0.50–10.00 µg·mL−1 for methyl hexadecanoate and 1.00–20.00 µg·mL−1 for methyl stearate, with the limits of detection (LOD) 11.94 ng·mL−1 and 11.90 ng·mL−1, respectively. The recoveries that ranged from 95.25% to 100.29% were satisfied, and the relative standard deviation (RSD) was no more than 7.16%. The developed method was successfully applied to evaluate the compatibility of chlorinated butyl rubber stoppers with liposome injections and the safety assessment.
IL2RA Promotes Aggressiveness and Stem Cell–Related Properties of Acute Myeloid Leukemia
Authors: Nguyen Chi Huu , Schlerka Angela , Grandits Alexander M. , Koller Elisabeth , van der Kouwe Emiel , Vassiliou George S. , Staber Philipp B. , Heller Gerwin , Wieser Rotraud
Published: September 2020

Overexpression of IL2RA, which encodes the alpha chain of the IL2 receptor, is associated with chemotherapy resistance and poor outcome in acute myeloid leukemia (AML). The clinical potential of anti-IL2RA therapy is, therefore, being explored in early-stage clinical trials. Notwithstanding, only very limited information regarding the biological function of IL2RA in AML is available. Using genetic manipulation of IL2RA expression as well as antibody-mediated inhibition of IL2RA in human cell lines, mouse models, and primary patient samples, we investigated the effects of IL2RA on AML cell proliferation and apoptosis, and on pertinent signaling pathways. The impact of IL2RA on the properties of leukemic stem cells (LSC) and on leukemogenesis were queried. IL2RA promoted proliferation and cell-cycle activity and inhibited apoptosis in human AML cell lines and primary cells. These phenotypes were accompanied by corresponding alterations in cell-cycle machinery and in pathways associated with cell survival and apoptosis. The biological roles of IL2RA were confirmed in two genetically distinct AML mouse models, revealing that IL2RA inhibits differentiation, promotes stem cell–related properties, and is required for leukemogenesis. IL2RA antibodies inhibited leukemic, but not normal, hematopoietic cells and synergized with other antileukemic agents in this regard. Collectively, these data show for the first time that IL2RA plays key biological roles in AML and underscore its value as a potential therapeutic target in this disease.

Significance:

This study identifies IL2RA as a potential therapeutic target in AML, where it is shown to regulate proliferation, differentiation, apoptosis, stem cell–related properties, and leukemogenesis.
Targeted Natural Killer Cell–Based Adoptive Immunotherapy for the Treatment of Patients with NSCLC after Radiochemotherapy: A Randomized Phase II Clinical Trial
Authors: Multhoff Gabriele , Seier Sophie , Stangl Stefan , Sievert Wolfgang , Shevtsov Maxim , Werner Caroline , Pockley A. Graham , Blankenstein Christiane , Hildebrandt Martin , Offner Robert , Ahrens Norbert , Kokowski Konrad , Hautmann Matthias , Rödel Claus , Fietkau Rainer , Lubgan Dorota , Huber Rudolf , Hautmann Hubert , Duell Thomas , Molls Michael , Specht Hanno , Haller Bernhard , Devecka Michal , Sauter Andreas , Combs Stephanie E.
Published: September 2020

Purpose:

Non–small cell lung cancer (NSCLC) is a fatal disease with poor prognosis. A membrane-bound form of Hsp70 (mHsp70) which is selectively expressed on high-risk tumors serves as a target for mHsp70-targeting natural killer (NK) cells. Patients with advanced mHsp70-positive NSCLC may therefore benefit from a therapeutic intervention involving mHsp70-targeting NK cells. The randomized phase II clinical trial (EudraCT2008-002130-30) explores tolerability and efficacy of ex vivo–activated NK cells in patients with NSCLC after radiochemotherapy (RCT).

Patients and Methods:

Patients with unresectable, mHsp70-positive NSCLC (stage IIIa/b) received 4 cycles of autologous NK cells activated ex vivo with TKD/IL2 [interventional arm (INT)] after RCT (60–70 Gy, platinum-based chemotherapy) or RCT alone [control arm (CTRL)]. The primary objective was progression-free survival (PFS), and secondary objectives were the assessment of quality of life (QoL, QLQ-LC13), toxicity, and immunobiological responses.

Results:

The NK-cell therapy after RCT was well tolerated, and no differences in QoL parameters between the two study arms were detected. Estimated 1-year probabilities for PFS were 67% [95% confidence interval (CI), 19%–90%] for the INT arm and 33% (95% CI, 5%–68%) for the CTRL arm (P = 0.36, 1-sided log-rank test). Clinical responses in the INT group were associated with an increase in the prevalence of activated NK cells in their peripheral blood.

Conclusions:

Ex vivo TKD/IL2-activated, autologous NK cells are well tolerated and deliver positive clinical responses in patients with advanced NSCLC after RCT.
CXCR3 and Cognate Ligands are Associated with Immune Cell Alteration and Aggressiveness of Pancreatic Ductal Adenocarcinoma
Authors: Cannon Andrew , Thompson Christopher M. , Maurer H. Carlo , Atri Pranita , Bhatia Rakesh , West Sean , Ghersi Dario , Olive Kenneth P. , Kumar Sushil , Batra Surinder K.
Published: September 2020

Purpose:

The cytokine milieu in pancreatic ductal adenocarcinoma (PDAC) promotes tumor progression and immune suppression, contributing to the dismal prognosis of patients with PDAC. The roles of many of these cytokines, however, have not been thoroughly investigated in PDAC.

Experimental Design:

PDAC microarray and The Cancer Genome Atlas datasets were analyzed to identify cytokines and cognate receptors overexpressed in PDAC and associated with survival. Pathway and CIBERSORT analyses were used to elucidate potential mechanisms of altered patient survival. Comparative analysis of cytokine expression in KPC (K-rasG12D; TP53R172H; Pdx-1cre) and KC (K-rasG12D; Pdx-1cre) PDAC models and multicolor immunofluorescence (IF) staining of human PDAC–resected samples were used to validate these findings.

Results:

CXCL9 and CXCL10 were among the most highly overexpressed cytokines by bioinformatics analyses, while their receptor, CXCR3, was significantly overexpressed by IHC analysis. Higher CXCR3 ligand expression was associated with shorter overall survival, while high CXCR3 expression was associated with better survival. The CXCR3 ligands, CXCL4, 9, and 10, were overexpressed in KPC compared with KC mice. Pathway analysis of CXCR3- and CXCR3 ligand–associated genes showed that CXCR3 is a marker of antitumor immunity, while its ligands may promote immunosuppression. CIBERSORT and IF studies of PDAC tissues demonstrated that high CXCR3 expression was associated with increased CD8+ T-cell and naïve B-cell signatures and loss of plasma cell signatures. CXCR3 ligand expression was associated with increased CD8+ T-cell signatures and loss of natural killer–cell signatures.

Conclusions:

CXCR3 ligands are overexpressed in PDAC and are associated with poor survival likely related to alterations in tumor immune infiltrate/activity.
Abstract IA28: Disparities in cardiovascular disease incidence in breast cancer survivors
Authors: Shi Zaixing , Greenlee Heather , Neugebauer Romain S. , Iribarren Carlos , Nguyen-huynh Mai N. , Rana Jamal S. , Hershman Dawn L. , Kushi Lawrence H. , Kwan Marilyn L.
Published: September 2020

Many breast cancer therapies have adverse effects on the cardiovascular health of patients. Breast cancer patients who are members of racial/ethnic minority and disadvantaged socioeconomic groups suffer disproportionately from cardiotoxicity during and after cancer treatment, leading to disparities in overall survival. These disparities may exist, in part, because of inequality in the management of pre-existing cardiovascular diseases and its risk factors such as hypertension and diabetes. Shared lifestyle risk factors for breast cancer and cardiovascular diseases, including obesity, poorer diet, and lack of physical activity may contribute to disparate outcomes in cardiotoxicity. In addition, patients of racial/ethnic minority and disadvantaged socioeconomic groups may have limited access to cardio-oncology care due to lack of insurance coverage and longer geographic distance from their health care provider. Patients living in disadvantaged neighborhoods may lack healthy food options and exercise facilities, thus are unable to follow a heathier lifestyle. We present evidence from prior studies on these potential mechanisms of disparities in cardiotoxicity and accompanying gaps in research. Finally, we introduce our ongoing Pathways Heart Study (R01 CA21 4057, mPI: Kwan, Greenlee), a large population-based study of nearly 15,000 breast cancer survivors and 75,000 noncancer controls who are members of Kaiser Permanente Northern California, which is an extension of the Pathways Study of 4,505 breast cancer survivors (U01 CA19 5565, mPI: Kushi, Ambrosone). Our study provides a unique opportunity to better understand the individual and health care system level contributors to risk of cardiotoxic outcomes.

Citation Format: Zaixing Shi, Heather Greenlee, Romain S. Neugebauer, Carlos Iribarren, Mai N. Nguyen-huynh, Jamal S. Rana, Dawn L. Hershman, Lawrence H. Kushi, Marilyn L. Kwan. Disparities in cardiovascular disease incidence in breast cancer survivors [abstract]. In: Proceedings of the Twelfth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2019 Sep 20-23; San Francisco, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl_2):Abstract nr IA28.
Abstract A08: Loss of noncanonical Hippo signaling in fusion-positive alveolar rhabdomyosarcoma increases invasiveness and a dedifferentiated phenotype associated with metastasis
Authors: Oristian Kristianne M. , Crose Lisa E.S. , Kuprasertkul Napasorn , Bentley Rex C. , Lin Yi-Tzu , Williams Nerissa , Kirsch David G. , Linardic Corinne M.
Published: September 2020

A hallmark of alveolar rhabdomyosarcoma (ARMS) is the presence of a chromosomal translocation encoding the PAX3-FOXO1 fusion oncogene (FP-ARMS). Patients presenting with FP-ARMS represent the subset of rhabdomyosarcoma patients with the worst prognosis, and when metastatic, survival for these children and adolescents is less than 10%. Nevertheless, standard of care has not changed in the clinic in decades due to a lack of novel effective therapeutics, and as a result, efforts are ongoing to develop better models of this disease in vivo and identify new and more predictive biomarkers. Recently, our group identified a role for Hippo/MST signaling in a genetically engineered mouse model (GEMM) of FP-ARMS that demonstrates Stk3F/F;Stk4F/F; Pax3PF/PF;Cdkn2aF/F;Myf6ICN/+ ARMS tumor-derived cells are less differentiated and more invasive in vitro as compared to Pax3PF/PF;Cdkn2aF/F; Myf6ICN/+ counterparts. Preliminary in vivo studies show that FP-ARMS GEMM tumor-derived cells form secondary metastases in both the lungs and livers of recipient animals, suggesting a new utility of the model for studies of metastatic potential. Studies of bone marrow metastasis, a lethal disease progression in PF-ARMS patients, are ongoing. Further, canonical signaling down the kinase cascade from MST1/2 to YAP1 is largely unchanged in this model, implicating instead noncanonical Hippo signaling and a potential role for MOB1, a Hippo pathway modulator implicated in mitotic exit and expressed highly in the bone marrow niche. Together, these data present an opportunity to interrogate the biology driving the predisposition of PF-RMS patients to develop metastases, and a foundation to identify novel preclinical targets.

Citation Format: Kristianne M. Oristian, Lisa E.S. Crose, Napasorn Kuprasertkul, Rex C. Bentley, Yi-Tzu Lin, Nerissa Williams, David G. Kirsch, Corinne M. Linardic. Loss of noncanonical Hippo signaling in fusion-positive alveolar rhabdomyosarcoma increases invasiveness and a dedifferentiated phenotype associated with metastasis [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr A08.
Abstract A16: The α-arrestin ARRDC3 functions as a metastasis suppressor by regulating GPCR activation of the Hippo pathway
Authors: Arakaki Aleena K.S. , Pan Wen-An , Wedegaertner Helen , Trejo JoAnn
Published: September 2020

G protein-coupled receptors (GPCRs) are key regulators of the Hippo pathway, a well-established mediator of tumorigenesis and cancer progression. The Hippo pathway is a promising therapeutic target, yet developing inhibitors for the core pathway components has proven difficult. As critical regulators of the Hippo pathway, GPCRs represent a potential upstream therapeutic target for cancer. However, despite the success and promise of GPCRs as therapeutic targets, there are currently no FDA-approved drugs targeting GPCRs for cancer. Protease-activated receptor-1 (PAR1) is a GPCR that has been shown to activate the Hippo pathway and also promote breast cancer progression. PAR1 is overexpressed in breast cancer patient tissue biopsies and in breast carcinoma cell lines and correlates with increased rates of metastasis and poor prognosis and increased invasion and metastatic potential, respectively. One mechanism that leads to PAR1 aberrant overexpression is defective lysosomal trafficking and degradation of the receptor, leading to persistent signaling. We recently showed that arrestin domain containing protein-3 (ARRDC3), an adaptor protein for E3 ubiquitin ligases, functions as a tumor suppressor by controlling proper PAR1 trafficking and degradation in highly aggressive, triple-negative breast carcinoma cells. The dysregulation of PAR1 trafficking due to loss of ARRDC3 expression leads to persistent signaling and promotes breast cancer invasion. However, the mechanism by which ARRDC3 specifically regulates PAR1-stimulated Hippo signaling to promote breast cancer metastasis remains unknown. Here, we show that ARRDC3 regulates GPCR activation of the Hippo pathway in basal-like breast cancer, where the transcriptional coactivators YAP and TAZ play differential roles in GPCR-mediated signaling and invasive capacity. ARRDC3 re-expression in invasive breast carcinoma cells attenuates PAR1-mediated activation of the Hippo pathway through regulation of TAZ but not YAP activation. Furthermore, siRNA knockdown of TAZ inhibits PAR1-stimulated Hippo signaling and invasion, whereas YAP siRNA knockdown has no effect. Our studies suggest a crucial role for ARRDC3 and TAZ in PAR1-Hippo pathway signaling in breast carcinoma invasion and metastasis. A better understanding of the mechanisms by which the Hippo pathway is regulated by GPCRs may lead to new potential therapeutic targets for the treatment or prevention of metastatic breast cancer.

Citation Format: Aleena K.S. Arakaki, Wen-An Pan, Helen Wedegaertner, JoAnn Trejo. The α-arrestin ARRDC3 functions as a metastasis suppressor by regulating GPCR activation of the Hippo pathway [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr A16.
Abstract A27: Increasing proximity triggers Mst2 autophosphorylation
Authors: Tran Thao , Kavran Jennifer M.
Published: September 2020

The Hippo pathway limits cell number and differentiation by blocking the transcriptional activity of YAP. The fidelity of signal transduction relies on the coordinated activity of a core kinase cassette that includes the kinases Mst1/2 and Lats1/2 and the accessory proteins Mob1 and hSalvador. Mst1/2 contains a kinase domain, linker region, and a C-terminal coiled-coil domain termed SARAH and is responsible for the phosphorylation of downstream components. Activation of Mst1/2 requires phosphorylation of the activation loop, a process believed to be a consequence of trans-autophosphorylation that is promoted by SARAH-domain mediated homodimerization. We aimed to understand what mechanistically triggers Mst1/2 autophosphorylation. Using purified proteins, we show that the kinase domain of Mst2 is sufficient for autophosphorylation but full-length Mst2 undergoes autophosphorylation faster than variants lacking the SARAH domain. This increased autophosphorylation could arise from either a specific contribution of a SARAH-domain mediated homodimer (allostery) or the increased proximity of kinase domains following homodimerization (effective local concentration). We investigated both possibilities using a series of biochemical assays and revealed that autophosphorylation of Mst2 variants lacking a SARAH domain could be stimulated by either chemically induced dimerization or increased localization following recruitment to the surface of a lipid-vesicle. Our results suggest that SARAH-domain mediated homodimerization is not the only event that promotes autophosphorylation of Mst1/2 and, instead, supports a more inclusive model that relies on colocalization. Any cellular event that increases the proximity of Mst1/2 could activate the kinase such as SARAH-domain mediated homodimerization, higher-order complex formation with hSalvador, or membrane recruitment.

Citation Format: Thao Tran, Jennifer M. Kavran. Increasing proximity triggers Mst2 autophosphorylation [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr A27.
Abstract A29: Genome-wide CRISPR/Cas9 screens for the identification of novel YAP1/TAZ modulators
Authors: Naujoks Jan , Potze Lisette , Kuehnlenz Julia , Kamburov Atanas , Nevedomskaya Ekaterina , Steffen Andreas , Luther Claudia , Anurin Anna , Buttgereit Anne , Prechtl Stefan , Bader Benjamin , Lesche Ralf , Staller Peter , Lange Martin , Nicke Barbara
Published: September 2020

Aberrant activation of the Hippo pathway effectors YAP1/TAZ promotes cell proliferation and tumorigenesis. To identify novel regulators of YAP1/TAZ in cancer, we established a FACS-based screening system monitoring YAP1/TAZ activity in MDA-MB-231 breast cancer cells. Using these cells, we performed pooled genome-wide CRISPR/Cas9 knockout and CRISPR activation/interference (a/i) screens. The list of hits included previously known YAP1/TAZ modulators such as LATS2, AJUBA, and TAZ itself, demonstrating the robustness of the screen. Moreover, we identified about 30 novel candidate genes with potential inhibitory activity on YAP1/TAZ and about 50 candidate genes that may play a role in YAP1/TAZ activation. These genes represent diverse cellular functions such as regulation of actin cytoskeleton, integrin signaling, and ER protein processing, among others. Modulation of endogenous YAP1/TAZ target genes was assessed by individual gene knockout using crRNAs. Functional characterization of the novel potential YAP1/TAZ modulators will aid to the further understanding of YAP1/TAZ biology in health and disease.

Citation Format: Jan Naujoks, Lisette Potze, Julia Kuehnlenz, Atanas Kamburov, Ekaterina Nevedomskaya, Andreas Steffen, Claudia Luther, Anna Anurin, Anne Buttgereit, Stefan Prechtl, Benjamin Bader, Ralf Lesche, Peter Staller, Martin Lange, Barbara Nicke. Genome-wide CRISPR/Cas9 screens for the identification of novel YAP1/TAZ modulators [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr A29.
Abstract A30: Super-enhancer-associated long noncoding RNA UCA1 interacts directly with AMOT to inhibit Hippo signaling pathway in epithelial ovarian cancer
Authors: Lin Xianzhi , Spindler Tassja J. , de Souza Fonseca Marcos A , Corona Rosario I. , Seo Ji-Heui , Dezem Felipe S , Li Lewyn , Lee Janet M. , Long Henry W. , Sellers Thomas A. , Karlan Beth Y. , Noushmehr Houtan , Freedman Matthew L. , Gayther Simon A. , Lawrenson Kate
Published: September 2020

Long noncoding RNAs (lncRNAs) have emerged as critical regulators of tumorigenesis and yet their mechanistic role remains challenging to characterize. Here, we present LncRNA Interpreter, a strategy integrating functional proteomics with interacting proteins to characterize lncRNAs. Its robustness is exemplified by lncRNA Urothelial Cancer Associated 1 (UCA1), a driver of ovarian cancer development. Reverse phase protein array (RPPA) analysis indicates that UCA1 inhibits Hippo signaling pathway and in vivo RNA antisense purification (iRAP) of UCA1 interacting proteins identified angiomotin (AMOT), a known YAP regulator, as direct binding partner. Loss-of-function experiments show that AMOT mediates Hippo signaling pathway inhibition by UCA1. UCA1 enhances the AMOT-YAP interaction to prevent YAP phosphorylation and facilitate its nuclear translocation. Together, our LncRNA Interpreter pipeline identified UCA1 as an lncRNA regulator of the Hippo signaling pathway and highlighted the UCA1-AMOT-YAP signaling axis in ovarian cancer development. LncRNA Interpreter can readily be applied to other lncRNAs implicated in complex diseases.

Citation Format: Xianzhi Lin, Tassja J. Spindler, Marcos A de Souza Fonseca, Rosario I. Corona, Ji-Heui Seo, Felipe S Dezem, Lewyn Li, Janet M. Lee, Henry W. Long, Thomas A. Sellers, Beth Y. Karlan, Houtan Noushmehr, Matthew L. Freedman, Simon A. Gayther, Kate Lawrenson. Super-enhancer-associated long noncoding RNA UCA1 interacts directly with AMOT to inhibit Hippo signaling pathway in epithelial ovarian cancer [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr A30.
Abstract A44: PR55α regulatory subunit of PP2A inhibits the MOB1/LATS cascade and activates YAP in pancreatic cancer cells
Authors: Hein Ashley L. , Brandquist Nichole D. , Ouellette Caroline Y. , Seshacharyulu Parthasarathy , Enke Charles A. , Ouellette Michel M. , Batra Surinder K. , Yan Ying
Published: September 2020

PP2A holoenzyme complexes constitute the majority of Ser/Thr phosphatase activities in human cells. Each PP2A consists of a catalytic subunit (C), a scaffold subunit (A), and a regulatory subunit (B). While the A and C subunits are highly conserved, a large number of B subunits share no homology, which determines PP2A substrate specificity and/or cellular localization. Although different PP2A complexes play distinct roles in cell signaling networks, PP2A has been defined only as a putative tumor suppressor, mostly based on loss-of-function studies using inhibitors (pharmacologic or biologic) for the highly conserved A or C subunit of PP2A. Many studies of specific pathways indicate that PP2A also possesses tumor-promoting function. We have reported an essential role of PR55α, a PP2A regulatory subunit, in supporting oncogenic phenotypes and in vivo tumorigenicity and metastasis of pancreatic cancer cells. In this study, we investigated a novel mechanism by which PR55α activates the YAP oncoprotein that plays an essential role in oncogenesis and metastasis of solid tumors including pancreatic cancer, resulting in its stabilization and nuclear retention. This mechanism involves PR55α’s inhibition of the MOB1-mediated LATS1/2 autoactivation, which otherwise would suppress YAP activation, and its direct activation of YAP itself. However, this mechanism does not involve the inhibition of upstream MST1/2 kinases and their subsequent T1079/T1041-phosphorylation of the LATS1/2 kinases. Furthermore, PR55α has the predominant effect on YAP activation, and the effect cannot be substituted by the depletion of either LATS1/2 or MOB1. Accordingly, PR55α is essential for YAP-promoted gene transcription, as well as for the anchorage-independent growth of cancer cells. In summary, current studies demonstrate a novel pancreatic cancer promoting mechanism through the PR55α-regulated PP2A.

Citation Format: Ashley L. Hein, Nichole D. Brandquist, Caroline Y. Ouellette, Parthasarathy Seshacharyulu, Charles A. Enke, Michel M. Ouellette, Surinder K. Batra, Ying Yan. PR55α regulatory subunit of PP2A inhibits the MOB1/LATS cascade and activates YAP in pancreatic cancer cells [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr A44.
Abstract B01: Understanding the LATS1 pro-apoptotic signaling network in melanoma
Authors: Garcia-Gutierrez Lucia , McKenna Stephanie , Badrock Andrew , Hurlstone Adam , Matallanas David
Published: September 2020

The tumor suppressor LATS1 is a core kinase of the MST2 pathway that regulates many cellular processes such as proliferation, cell growth, differentiation, and apoptosis. The MST2 pathway has been reported to interact with the MAPK pathway at the level of MST2 through RAF1, blocking MTS2 signaling. Our group has reported that, upon a proapoptotic signal, the tumor suppressor RASSF1A disrupts RAF1-MST2 complexes, leading to activation of MST2 and further LATS1-mediated phosphorylation of YAP, which triggers YAP translocation to the nucleus together with p73, leading to apoptosis. Apart from this mechanism, our group has found evidence showing that LATS1 could act as a signaling hub integrating proliferative and apoptotic signals and thus deciding cell fate. Several lines of evidence have shown that LATS1 crosstalks with several proteins of the apoptotic pathways including Mdm2, Omi, and p53. To get a better knowledge of the signaling network mediating LATS1 proapoptotic signaling, we have performed a proteomics screening. In this screening we have identified a cluster of proteins that are part of the intrinsic apoptotic pathway. Here we present validation experiments of these novel interactions and characterize the molecular mechanisms mediated by LATS1 proapoptotic network. We focus in melanoma cells, a cancer type where loss of function of the RASSF1A/MST2/LATS1 seems to be common. Thus, the main driving mutation of melanoma mutant BRAF has been shown to inhibit MST2; loss of expression of RASSF1A by promotor methylation is rather frequent and our analysis show that LAST1 is also downregulated. Our results indicate that the silencing of LATS1 crosstalk with the intrinsic pathway results in the evasion of apoptotic signals in melanocytes and can lead to cell transformation.

Citation Format: Lucia Garcia-Gutierrez, Stephanie McKenna, Andrew Badrock, Adam Hurlstone, David Matallanas. Understanding the LATS1 pro-apoptotic signaling network in melanoma [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr B01.
Abstract B03: Differential YAP expression in glioma cells induces cell competition and promotes tumorigenesis
Authors: Liu Zhijun , Yee Patricia P. , Wei Yiju , Liu Zhenqiu , Kawasawa Yuka Imamura , Li Wei
Published: September 2020

Intratumor heterogeneity associates with cancer progression and may account for a substantial portion of therapeutic resistance. Although extensive studies have focused on the origin of the heterogeneity, biologic interactions between heterogeneous malignant cells within a tumor are largely unexplored. Glioblastoma (GBM) is the most aggressive primary brain tumor. Here, we found that the expression of Yes-associated protein (YAP) is intratumorally heterogeneous in GBM. In a xenograft mouse model, differential YAP expression in glioma cells promotes tumorigenesis and leads to clonal dominance by cells expressing more YAP. Such clonal dominance also occurs in vitro when cells reach confluence in the two-dimensional culture condition or grow into tumor spheroids. During this process, growth of the dominant cell population is enhanced. In tumor spheroid, such enhanced growth is accompanied by increased apoptosis in cells expressing less YAP. The cellular interaction during clonal dominance appears to be reminiscent of cell competition. RNA-seq analysis suggested that the interaction induces expression of tumorigenic genes, which may contribute to the enhanced tumor growth. These results suggested that tumorigenesis benefits from competitive interactions between heterogeneous tumor cells.

Citation Format: Zhijun Liu, Patricia P. Yee, Yiju Wei, Zhenqiu Liu, Yuka Imamura Kawasawa, Wei Li. Differential YAP expression in glioma cells induces cell competition and promotes tumorigenesis [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr B03.
Abstract B24: Identification of YAP modulators using genome-wide gain-of-function screening
Authors: Cramer Paul , von Eyss Björn
Published: September 2020

The Hippo pathway is a signaling pathway that is crucial to regulate tissue homeostasis and organ growth. The core of the Hippo pathway consists of the MST1/2 and LATS1/2 kinase cascade that regulates the activity of the transcriptional coactivators YAP/TAZ by phosphorylation. Unphosphorylated YAP/TAZ display nuclear localization and interact with TEAD proteins to regulate gene expression. A tight regulation is crucial, since deregulated YAP/TAZ can lead to several organismal defects including cancer development. To ensure robustness of the signaling pathway to perturbations, many Hippo components are organized in a redundant manner, e.g., redundancy of LATS kinases. This feature aggravates the identification of new Hippo signaling components using loss-of-function screens since a knockout of one factor could potentially be compensated for by its redundant counterpart. To discover novel potential regulators of YAP, we performed a genome-scale gain-of-function CRISPR/Cas9 screening in MCF10A cells using the synergistic activation mediator (SAM) system. SAM is an adaptation of CRISPR/Cas9 to overexpress endogenous genes by targeting the proximal promoter with sgRNAs rather than knocking out target genes. Using the nuclear localization of YAP as a measure of YAP activity, we could identify several genes as positive regulators of YAP. One of the best screening hits is ISOC1 (Isochorismatase domain-containing protein 1). We validated the screening by using single sgRNAs to induce the endogenous ISOC1 locus by SAM. The expression of YAP-target genes as ANKRD1, CTGF, and IGFBP3 was increased upon ISOC1 overexpression. Main goals in the future will be to understand the biochemical mechanism by which ISOC1 modulates YAP activity and the biologic role of ISOC1. Since ISOC1 is reported to localize to peroxisomes and is induced upon dietary restriction in mice, we hypothesize that ISOC1 acts as a central hub to connect peroxisomal metabolism and YAP/TAZ activity.

Citation Format: Paul Cramer, Björn von Eyss. Identification of YAP modulators using genome-wide gain-of-function screening [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr B24.
Abstract B26: A tumor specific molecular network promotes tumor growth by enforcing a JNK-YKI feed forward loop
Authors: Kango-Singh Madhuri
Published: September 2020

Cancer cells expand rapidly in response to altered cell-cell interactions among tumor and their surrounding cells; however, the molecular mechanisms by which alterations in these interactions promote aggressive metastatic tumor growth remain unclear. Earlier studies showed that increased oncogenic signaling and deregulation of inflammatory cytokines may be underlying tumor growth and progression. To study the nature of these interactions, we made mosaic clones expressing oncogenic forms of Yorkie (Yki) or Ras in polarity deficient cells (RasV12, scrib- or Yki3SA, scrib-) that are marked with GFP in Drosophila imaginal discs. This mosaic system recapitulates the clonal origins of human cancer and provides a distinct advantage in analyzing cell-cell (intratumoral and tumor-surrounding cell) interactions affecting tumor growth. Using these models of oncogenic cooperation, we show that tumor cells grow by activation of a previously unidentified network comprising Wingless (Wg), Dronc (Drosophila Caspase 9 ortholog), Jun N-terminal kinase (JNK), and Yki. Further, Toll-like receptor (TLR) component Cactus is highly expressed in the tumor cells. We show that Wg, Dronc, JNK, and Yki signaling are all required for the growth of tumors (RasV12, scrib-, and other tumor models), and downregulating Cactus affects both tumor progression and invasion. It is notable that Wg, Dronc, JNK, and Yki play a signaling and patterning role in normal development, and act as regulators of cell-cell interactions like cell competition and compensatory proliferation. JNK was previously shown to undergo a paradoxical signaling switch from proapoptosis to proproliferation that leads to activation of Yki. We report that Yki not only regulates the TLR component Cactus, but also positively regulates JNK in the tumor cells to form a JNK-Yki self-reinforcing positive feedback signal-amplification loop downstream of Wg and Dronc. We further demonstrate that the Wg-Dronc-JNK-Yki signaling network promotes tumor growth in other contexts of oncogenic cooperation. Overall, the identification of the Wg-Dronc-Yki-JNK molecular network and the organization of the signaling elements provide novel insights into how altered cell-cell interactions and signaling promote tumorigenesis. Our findings suggest that identification of molecular networks may provide significant insights about the key biologically meaningful changes in signaling pathways and paradoxical signals that promote tumorigenesis.

Citation Format: Madhuri Kango-Singh. A tumor specific molecular network promotes tumor growth by enforcing a JNK-YKI feed forward loop [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr B26.