bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 174: Functional characterization of ZNF750 in esophageal squamous cell carcinoma
Authors: Choi Sheyne SA , Yu Valen Zhuoyou , Ko Josephine Mun-Yee , Dai Wei , Lung Maria L. 
Published: August 2019

Abstract
Background: Esophageal squamous cell carcinoma (ESCC) is a lethal and aggressive malignancy that is a threat worldwide. It is of significant prevalence in regions of Asia including Iran, Korea, Japan and especially China. Epidemiological studies of ESCC reveal genetic variations as a risk-factor to ESCC incidence. Genomic profiling of ESCC primary tumors have already identified candidate genes involved in different cellular processes such as cell cycle and apoptosis. Data from our whole exome sequencing study of primary tumors and lymph node metastases also reveal mutations involved in these pathways; however, we also uncovered an interesting candidate involved in differentiation namely, ZNF750. ZNF750 has been reported to play important roles in other squamous cell carcinomas, and as a candidate tumor suppressor gene in ESCC.
Methods and Results: We exogenously upregulated ZNF750 expression by lentiviral system and used this model to investigate ZNF750 function in ESCC. By MTT 2D cell proliferation assay, we showed that in vitro cell proliferation in the ZNF750 group was hindered compared to controls. In addition, we set up 2D and 3D colony formation assays and found that the ZNF750 groups produced fewer colonies in 2D culture plates and produced significantly smaller colonies, when embedded in Matrigel. All of these findings are consistent with previous studies. We also studied the potential functional role of ZNF750 in 3D invasion/migration. Transwell permeable inserts were used to support a thick Matrigel culture with a nutrient gradient. Cells were embedded in Matrigel and moved along the gradient within the gel. After prolonged incubation (&gt;2 weeks), location and size of the colonies were examined by confocal microscopy scanning. Cells with upregulated ZNF750 were found to exhibit greater mobility compared to the control cells. We also subjected ZNF750 and control groups to cisplatin to test for chemoresistance in ESCC. We found that ZNF750 expression increases chemoresistance, as opposed to the control cells.
Conclusion: From these results, ZNF750 expression inhibits cell proliferation, while enhancing cell mobility and drug resistance, all of which are characteristics of cells undergoing epithelial-mesenchymal transition (EMT). We, therefore, hypothesize that ZNF750 functions in ESSC beyond that of a tumor suppressor, but as a putative EMT regulator.
Acknowledgements: Research Grants Council Collaborative Research Fund grant number 106150246 and Asian Cancer Research Fund to MLL.
Citation Format: Sheyne SA Choi, Valen Zhuoyou Yu, Josephine Mun-Yee Ko, Wei Dai, Maria L. Lung. Functional characterization of ZNF750 in esophageal squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 174.
Abstract 175: Methyltransferase protein A regulates colorectal cancer cell proliferation and metastasis through methylation of protein B
Authors: Kang Songhwa , Kwon Sang-Mo 
Published: August 2019

Abstract
Post-translational modifications (PTMs) contribute to abnormal cell proliferation, adhesion, and morphology in cancer progression. Methylation is one such PTM that affects many of the protein properties and has a substantial role in tumorigenesis. Here, we identified that the methyltransferase protein A was upregulated in human colorectal cancer (CRC) and was associated with CRC proliferation, migration, and invasion in vitro. Our study showed that Protein B was a novel target of methyltransferase Protein A and that methylated Protein B was upregulated in human CRC. High expression of Protein A and B resulted in a poor prognosis of CRC; moreover, CRC proliferation, migration, and invasion were inhibited when protein A and B were unable to bind through specific residue, as predicted by in silico protein-protein docking analysis. Furthermore, our study showed that methylation of Protein B by methyltransferase protein A plays an important role in CRC proliferation and invasion via enhancement of HIF-1α stabilization. In conclusion, our data suggested that methylated Protein B may be a potentially valuable clinical prognostic marker of CRC and that interference with the binding of Protein B to methyltransferase Protein A may be a new anticancer target in CRC.
Citation Format: Songhwa Kang, Sang-Mo Kwon. Methyltransferase protein A regulates colorectal cancer cell proliferation and metastasis through methylation of protein B [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 175.
Abstract 1751: The role of SIRT2 in BRCA1 regulation
Authors: Minten Elizabeth , Zhang Hui , Li Chunyang , Head PamelaSara , Yu David S. 
Published: August 2019

Abstract
The most deleterious type of DNA damage a cell can sustain is the DNA double-stranded break (DSB). One of the major pathways responsible for the repair of DNA DSB breaks is the homologous recombination (HR) pathway, an error-free mechanism of repair that occurs primarily in the S and G2 phases of the cell cycle when a sister chromatid is available to be used as a repair template. Impairment of HR can result in genomic instability, which can increase the risk of tumorigenesis. However, many of the mechanisms by which defects in HR lead to an increased risk for developing cancer are unknown. Mutations or misregulation of the Breast Cancer 1 (BRCA1) protein, a major player in HR and an established tumor suppressor, have been linked to an increased lifetime risk for the development of cancer in both men and women, including colon, breast, ovarian, and pancreatic cancer. Yet, how BRCA1 is regulated in HR is not well understood and thus highlights a major a gap in our understanding of how deficiencies in HR contribute to the development of cancer. Our lab has discovered that the histone deacetylase and putative human tumor suppressor, SIRT2, has a crucial role in the repair of DNA DSBs. We have found that SIRT2 interacts with and deacetylates BRCA1, both in vitro and in cells, and that depletion of SIRT2 decreases BRCA1 protein levels, thus impairing HR. Our results show SIRT2 is a novel regulator of BRCA1 and is critical for the efficient repair of DNA DSBs through HR. These findings will provide invaluable insight into how to exploit the interplay between SIRT2 and BRCA1 as a novel therapeutic approach for the prevention and treatment of cancer.
Citation Format: Elizabeth Minten, Hui Zhang, Chunyang Li, PamelaSara Head, David S. Yu. The role of SIRT2 in BRCA1 regulation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1751.
Abstract 1758: Splicing component ISY1 interacts with APE1 and regulates base excision repair
Authors: Jaiswal Aruna , Williamson Elizabeth , Patel Bhavita , Srinivasan Gayathri , Kong Kimi , Lomelino Carrie , Narayan Satya , Hromas Robert 
Published: August 2019

Abstract
Cellular genome is continuously challenged by endogenous and exogenous DNA damaging agents. In the genome, base modifications are the most common form of DNA damage. Base excision DNA repair (BER) is the most important pathway for the removal of oxidized or alkylated DNA. If these base modifications continue to persist, the replisome may stall at DNA lesions, causing the replication fork to collapse, leading to genetic instability. While the main components of the BER pathway are well defined, its regulatory mechanism remains poorly understood. We report here that apurinic/apyrimidinic endonuclease 1 (APE1), a multifunctional protein interacts with the pre-mRNA splicing factor ISY1. However, this interaction is independent of DNA or RNA based upon our pull-down experimental results. ISY1 expression is induced by oxidative or DNA alkylation damage, which would then provide an immediate up-regulation of APE1 activity in vivo and enhanced BER of oxidized bases. This was supported from our results of in vitro reconstituted experiment suggesting that ISY1 can stimulate 5’-3’ endonuclease 1 activity of APE1 in both the short- and long-patch BER pathways and increase its binding to AP site DNA. The interaction between ISY1 and APE1 also establishes a connection between DNA damage repair and pre-mRNA splicing. Based upon these results, we propose that ISY1 is an important regulator of BER and enhances the ability of APE1 to efficiently recognize abasic sites in DNA. Role of ISY1 becomes important in aging tissues where decreased APE1 activity results in the accumulation of oxidative genomic damages. Therefore, ISY1 may work as an anti-aging factor. Induction of ISY1 in malignancies could constitute a mechanism to avoid apoptotic death after treatment with alkylating agents.
Citation Format: Aruna Jaiswal, Elizabeth Williamson, Bhavita Patel, Gayathri Srinivasan, Kimi Kong, Carrie Lomelino, Satya Narayan, Robert Hromas. Splicing component ISY1 interacts with APE1 and regulates base excision repair [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1758.
Abstract 176: Tetraspanins in oral squamous cell carcinoma
Authors: Reddi Kiran K. , Anderson Kenneth M. , Garcia-Godoy Franklin , Zhang Yanhui H. 
Published: August 2019

Abstract
Objectives: To (1) investigate the expression levels of several tetraspanins in oral cancer cells and tissues; (2) characterize the subpopulation of oral cancer cells with elevated CD82; and (3) investigate roles of the tetraspanins in oral cancer migration, invasion, and metastasis.
Methods: The protein expression levels of tetraspanins CD82(KAI1), CD9, CD81(TAPA), and their associate protein EWI-2(IGSF8) were quantified in normal human gingival epithelial cells (SG) and oral cancer cells (CAL27 and SCC-25 cells) using western blotting and immunofluorescence staining and imaging. CAL27 CD82 knockout cells (CD82KO) were created by CRISPR/CAS-9, sorted by Fluorescent Activated Cell Sorting, and validated by western blotting. A wound healing assay was used to study the role of CD82 in oral cancer cell migration. CD82 perturbation of the CAL27 genome and proteome were evaluated using Microarray (Affymetrix) and TRAQ/TMT Based Differential Protein Expression Analysis (Thermo, mass spectrometer).
Results: So far, we have found (1) CD82 is highly expressed in CAL27 cancer cells, but its expression is low in SG and SCC-25 cells; (2) CD82 is highly expressed in human tongue cancer tissues; (3) CD9 is highly expressed in CAL27 cells; (4) The expression levels of CD81 and IGSF8 are comparable in the three cancer cell lines; (5) CAL27 cancer cells migrate faster than normal SG cells; (6) Knockout of CD82 significantly decreases the migration rate of CAL27 cells, suggesting that CD82 promotes migration of these cancer cells.
Conclusions: CD82 has varied expression in different oral cancer cell lines. Its expression is very high in CAL27 cancer cells. Knockout of CD82 in CAL27 cells significantly inhibits migration of these cells. Our study suggests that CD82 might be used as a biomarker for a subpopulation of oral cancer cells and that CD82 could be a target for therapeutic interventions.
Acknowledgements:
Supported in part by the 2017 UTHSC CORNET Award.
Supported in part by the UTHSC College of Dentistry Alumni Endowment Fund and the Tennessee Dental Association Foundation.
Citation Format: Kiran K. Reddi, Kenneth M. Anderson, Franklin Garcia-Godoy, Yanhui H. Zhang. Tetraspanins in oral squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 176.
Abstract 1765: In vivo modeling of RIT1 function in development and lung cancer
Authors: McSharry Maria 
Published: August 2019

Abstract
Ras-like in most tissues 1 (RIT1) encodes a ubiquitously expressed RAS-family small GTPase. Recently, somatic RIT1 mutations were identified in lung adenocarcinoma and myeloid malignancies, and germline RIT1 mutations were found in. RIT1 is also implicated hematopoietic malignancies and in Noonan syndrome, a “Rasopathy” affecting craniofacial development and associated with congenital heart disease. Of a variety of RIT1 mutations discovered, the most recurrent mutation observed was M90I. In this study, murine models were used to determine whether expression of RIT1M90I mutation is sufficient for tumor initiation or developmental phenotypes observed in Noonan patients.
Mice were genetically engineered at the Rosa26 locus to harbor RIT1M90I under the control of a lox-Cre system (LSL-RIT1M90I). The Cre recombinase protein was introduced via intratracheal delivery of adenovirus to the lungs. At one-year post treatment, 7 of the 10 LSL-RIT1M90I mice examined exhibited the expected genetic recombination. Tumors were observed in 3 of these mice, one of which was wild-type for p53. No visible tumors were seen in mice lacking the RIT1M90I gene or in cases where recombination was not observed, regardless of p53 status.
Mouse embryonic fibroblasts (MEFs) were infected by adenovirus to recapitulate the in vivo studies and evaluate transformation. In vitro experiments conducted in RIT1M90I MEFs demonstrated faithful recombination of the transgene upon treatment with Cre. Protein analysis of these same cells reflected higher levels of RIT1 protein in transgenic cells treated with Cre, while in all other conditions, RIT1 was undetectable. Preliminary data from an immortalization assay suggested enhanced proliferation of MEFs expressing the RIT1M90I gene compared to wild-type counterparts.
Cre recombinase was expressed body-wide in tamoxifen (4OHT) inducible mice (UBC-Cre-ERT2) LSL-RIT1M90I animals crossed to the inducible UBC-Cre-ERT2 system exhibited faithful recombination of the transgene upon treatment with 4OHT.
To determine the consequences of constitutive expression of RIT1M90I, RIT1M90I sires were crossed to dams harboring constitutively active Cre recombinase or to a constitutively active Cre SRY-box promoter (Sox2-Cre). Of the 100 pups observed, none contained both RIT1M90I and Cre recombinase (p &lt; 0.0001), indicating embryonic lethality induced by RIT1 activation.
These data demonstrate the powerful effect of RIT1 activation on murine embryonic development and will enable future efforts on the characterization of the mechanism of RIT1-induced Noonan Syndrome and lung cancer.
Citation Format: Maria McSharry. In vivo modeling of RIT1 function in development and lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1765.
Abstract 1768: Biochemical and structural analysis of the Neurofibromin (NF1) protein and a potential role for protein destabilization in Rasopathy diseases
Authors: Sherekar Mukul , Han Sae-Won , Messing Simon , Drew Matthew , Gillette William , Lorenzo Claire , McCormick Frank , Esposito Dominic 
Published: August 2019

Abstract
Neurofibromin is the 320 kilodalton protein product of the NF1 gene which is mutated in the Rasopathy disease Neurofibromatosis Type I. Defects in NF1 lead to abrogation of the GTPase activating activity of the protein, leading to aberrant signaling through the RAS-MAPK pathway. Very little is known about the function of the majority of the NF1 protein—to date only the GAP domain and a region containing a Sec-PH motif have detailed structural information. To better understand the role of this large protein, we have carried out a series of biochemical and biophysical studies which clearly show that full length NF1 protein exists primarily as a dimer. Data from in vivo experiments confirmed that this dimerization also occurs in cells, and presumably has biological implications. A model was generated from negative-stain EM data which suggests that NF1 monomers form both intermolecular contacts to form dimers and intramolecular contacts which stabilize the individual monomers. Deletion analysis permitted us to map these two interacting domains to different parts of the NF1 protein, and crosslinking studies coupled with mass spectrometry helped to identify regions of possible interaction. In addition, introduction of point mutations observed in patients with NF diseases caused a dramatic decrease in the stability of proteins both in vitro and in vivo, potentially arguing for a rationale for loss of NF1 activity by these single mutations
Citation Format: Mukul Sherekar, Sae-Won Han, Simon Messing, Matthew Drew, William Gillette, Claire Lorenzo, Frank McCormick, Dominic Esposito. Biochemical and structural analysis of the Neurofibromin (NF1) protein and a potential role for protein destabilization in Rasopathy diseases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1768.
Abstract 1780: iSCREAM - an unbiased pipeline to screen for activating kinase mutations
Authors: Chakroborty Deepankar , Kurppa Kari J. , Paatero Ilkka , Elo Laura L. , Elenius Klaus 
Published: August 2019

Abstract
Cancer tissues harbor thousands of mutations, and a given oncogene may be mutated at hundreds of sites. Most of these somatic mutations are expected to be inconsequential passenger mutations that reflect the general instability of the tumors. The discovery of most of the currently known driver mutations has been facilitated by their accumulation in mutation hot-spots within their respective genes. However, a vast majority of mutations in cancer tissues are rare and no information is currently available about their functional significance. Several lines of in vitro and in vivo clinical evidence also indicate that there is a significant number of, as yet unidentified, activating driver mutations that could serve as predictive markers in oncology.
An unbiased iSCREAM (in vitro screen for activating mutations) platform was developed to functionally characterize thousands of variants of a kinase oncogene in a single assay. The functional genetics screen was based on expressing random variants of a cDNA encoding a tyrosine kinase in a cell line in which the activating mutations provide growth-advantage. Targeted next-generation sequencing of the cDNA inserts was used to quantitatively analyze variants that provided growth-advantage.
iSCREAM is able to identify activating mutations from a library of thousands of random mutations of a tyrosine kinase. The pipeline can also be modified to identify mutations conferring resistance to a tyrosine kinase inhibitor.
Citation Format: Deepankar Chakroborty, Kari J. Kurppa, Ilkka Paatero, Laura L. Elo, Klaus Elenius. iSCREAM - an unbiased pipeline to screen for activating kinase mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1780.
Abstract 1782: ANKRD52 inhibited tumor metastasis through dephosphorylation of PAK1 in lung adenocarcinomas
Authors: Lee Ting-Fang , Liu Yin-Pu , Lin Yen-Fan , Hsu Chong-Fang , Wu Cheng-Wen 
Published: August 2019

Abstract
PAK1 is an important oncogene involved in the progression of a variety of cancer types including lung cancer. Although PAK1 inhibition is shown to suppress tumor progression, specific PAK1 inhibitors are not available due to the complex structure and insufficient understanding of this protein. In an attempt to find TAZ (WW domain-containing transcription regulator 1) regulated tumor suppressor, a member of ankyrin repeat proteins, ankyrin repeat domain 52 (ANKRD52), is observed to suppress PAK1 phosphorylation. Ankyrin repeat proteins are known to function as a platform for protein-protein interactions, and ANKRD52 was shown to interact with protein phosphatase 6 (PP6). Immunohistochemistry staining in lung cancer tissue revealed prolonged disease-free survival in high ANKRD52 expressing cases. Using lung cancer cell lines, ANKRD52 knockdowns showed significantly increased cell mobility while forced expressed ANKRD52 decreased cell mobility. Using mass spectrometry analysis, p21-activated kinases (PAK1) was identified to interact with ANKRD52. Upregulated PAK1 phosphorylation was detected in the knockdowns of ANKRD52 or PP6. Our model demonstrated a novel anti-metastatic protein, ANKRD52, that regulates cell mobility through interactions with PP6 and dephosphorylation of PAK1.
Citation Format: Ting-Fang Lee, Yin-Pu Liu, Yen-Fan Lin, Chong-Fang Hsu, Cheng-Wen Wu. ANKRD52 inhibited tumor metastasis through dephosphorylation of PAK1 in lung adenocarcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1782.
Abstract 1783: A combination therapy of atypical protein kinase inhibitor and phosphatidylinositol-3- kinase inhibitor reduces multidrug resistance in renal cell carcinoma (RCC)
Authors: Khalid Khandker Mohammad 
Published: August 2019

Abstract
Renal Cell Carcinoma (RCC) is the most common type of kidney cancer (85%) of which 75% of the cases involve conventional clear cell RCC (ccRCC). Choice of treatments recommended for clear cell carcinoma and non-clear cell carcinoma are partial nephroctomy (Stage I), radical nephoroctomy (Stage II and III) and chemotherapy (stage IV). Phosphatidylinositol-3- kinase (PI3K) inhibitors are one of the most prospective candidate in the therapy of ccRCC as the PI3CA gene for p110α (a class of the catalytic sub unit of PI3K) is mutated in most of the cases. Hence, PI3K inhibitors like Alpelisib (BYL 719) can be a treatment. But current therapeutic prospects are limited to the fact that RCC is a chemotherapy-resistant cancer type that acts as a barrier for successful treatment with this inhibitor. In addition, of all the other aPKCs, PKC ί/λ and PKC ζ are reported to be frequently overexpressed in most cancer cells and to have different significant roles in tumor progression and carcinogenesis of different type of cancers. Caki-1 metastatic clear cells were treated with PKC ζ inhibitor [8-hydroxynaphthalene-1,3,6-trisulfonic acid] or ζ-stat, Alpelisib and combination of both ζ-stat and Alpelisib at different doses. The results of WST-1 shows that the combination of ζ-stat (7.5 µM) and Alpelisib (10.0 µM) has the most cytotoxicity compared to the single therapy of Alpelisib. Annexin-V/PI assay also suggested that the combination therapy caused greater percentage of apoptosis and cell death compared to the single therapy of Alpelisib. The efflux proteins, (p-glycoprotein (P-gp) and ATP-binding cassette sub-family G member 2 (ABCG2), are responsible for developing cell resistance in Caki-1 cells had reduced expression in the cells when treated with the combination treatment of Alpelisib and ζ-stat. Therefore, combination therapy of Alpelisib and an aPKC inhibitor for metastatic ccRCC can reduce expression of multi drug resistance (MDR) proteins/efflux transporter P-gp and ABCG2. Hence, this combination may be a possible treatment that might help eradicating MDR of ccRCC. Thus, a combination of Alpelisib and an aPKC inhibitor can be a two pronged approach reducing the cell proliferation and resistance.
Citation Format: Khandker Mohammad Khalid. A combination therapy of atypical protein kinase inhibitor and phosphatidylinositol-3- kinase inhibitor reduces multidrug resistance in renal cell carcinoma (RCC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1783.