125 results on '"Tian Li Wang"'
Search Results
2. A novel human endometrial epithelial cell line for modeling gynecological diseases and for drug screening
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Zheng Cheng Yu, Jin Gyoung Jung, Ie Ming Shih, Geoff Shimberg, Yeh Wang, Vamsi Parimi, Ryoichi Asaka, Yun Chen, Wei-Hung Jung, Stephanie Gaillard, Wenjing Shen, Alicja Tomaszewski, Tian Li Wang, and Youngran Park
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ARID1A ,Tumor suppressor gene ,Cell ,Drug Evaluation, Preclinical ,Biology ,Article ,Chromatin remodeling ,Cell Line ,Pathology and Forensic Medicine ,Endometrium ,Mice ,Carcinoma ,medicine ,Animals ,Humans ,Molecular Biology ,Epithelial Cells ,Cell Biology ,medicine.disease ,medicine.anatomical_structure ,Receptors, Estrogen ,PARP inhibitor ,Clear cell carcinoma ,Cancer research ,Female ,DNA mismatch repair - Abstract
Endometrium-related malignancies including uterine endometrioid carcinoma, ovarian clear cell carcinoma and ovarian endometrioid carcinoma are major types of gynecologic cancer, claiming more than 13,000 women’s lives annually in the United States. In vitro cell models that recapitulate “normal” endometrial epithelial cells and their malignant counterparts are critically needed to facilitate the studies of pathogenesis in endometrium-related carcinomas. To achieve this objective, we have established a human endometrial epithelial cell line, hEM3, through immortalization and clonal selection from a primary human endometrium culture. hEM3 exhibits stable growth in vitro without senescence. hEM3 expresses protein markers characteristic of the endometrial epithelium, and they include PAX8, EpCAM, cytokeratin 7/8, and ER. hEM3 does not harbor pathogenic germline mutations in genes involving DNA mismatch repair (MMR) or homologous repair (HR) pathways. Despite its unlimited capacity of in vitro proliferation, hEM3 cells are not transformed, as they are not tumorigenic in immunocompromised mice. The cell line is amenable for gene editing, and we have established several gene-specific knockout clones targeting ARID1A, a tumor suppressor gene involved in the SWI/SNF chromatin remodeling. Drug screening demonstrates that both HDAC inhibitor and PARP inhibitor are effective in targeting cells with ARID1A deletion. Together, our data support the potential of hEM3 as a cell line model for studying the pathobiology of endometrium-related diseases and for developing effective precision therapies. This study describes the characterization of a human endometrial epithelial cell line, hEM3. hEM3 is not transformed, can grow organoids, and does not harbor pathogenic mutations in genes involving DNA mismatch repair (MMR). hEM3 with ARID1A knockout has been generated for drug screening in an ARID1A-deficient and MMR-proficient context. In conclusion, hEM3 can be a model for studying endometrial pathogenesis.
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- 2021
3. Spatial Transcriptomic Analysis of Ovarian Cancer Precursors Reveals Reactivation of IGFBP2 during Pathogenesis
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Yeh Wang, Peng Huang, Brant G. Wang, Tricia Murdock, Leslie Cope, Fang-Chi Hsu, Tian-Li Wang, and Ie-Ming Shih
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Ovarian Neoplasms ,Cancer Research ,Carcinoma ,Article ,Cystadenocarcinoma, Serous ,Oncology ,Tumor Microenvironment ,Humans ,Fallopian Tube Neoplasms ,Female ,Tumor Suppressor Protein p53 ,Transcriptome ,Carcinoma in Situ ,Fallopian Tubes - Abstract
Elucidating the earliest pathogenic steps in cancer development is fundamental to improving its early detection and prevention. Ovarian high-grade serous carcinoma (HGSC), a highly aggressive cancer, mostly originates from the fallopian tube epithelium through a precursor stage, serous tubal intraepithelial carcinoma (STIC). In this study, we performed spatial transcriptomic analysis to compare STICs, carcinoma, and their matched normal fallopian tube epithelium. Several differentially expressed genes in STICs and carcinomas were involved in cancer metabolism and detected in a larger independent transcriptomic dataset of ovarian HGSCs. Among these, insulin-like growth factor binding protein-2 (IGFBP2) was found to undergo DNA hypomethylation and to be increased at the protein level in STICs. Pyrosequencing revealed an association of IGFBP2 expression with the methylation state of its proximal enhancer, and 5-azacytidine treatment increased IGFBP2 expression. In postmenopausal fallopian tubes, where most STICs are detected, IGFBP2 immunoreactivity was detected in all 38 proliferatively active STICs but was undetectable in morphologically normal tubal epithelia, including those with TP53 mutations. In premenopausal fallopian tubes, IGFBP2 expression was limited to the secretory epithelium at the proliferative phase, and estradiol treatment increased IGFBP2 expression levels. IGFBP2 knockdown suppressed the growth of IGFBP2-expressing tubal epithelial cells via inactivation of the AKT pathway. Taken together, demethylation of the proximal enhancer of IGFBP2 drives tumor development by maintaining the increased IGFBP2 required for proliferation in an otherwise estrogen-deprived, proliferation-quiescent, and postmenopausal tubal microenvironment. Significance: Molecular studies of the earliest precursor lesions of ovarian cancer reveal a role of IGFBP2 in propelling tumor initiation, providing new insights into ovarian cancer development.
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- 2022
4. Current and Emerging Methods for Ovarian Cancer Screening and Diagnostics: A Comprehensive Review
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Juliane M. Liberto, Sheng-Yin Chen, Ie-Ming Shih, Tza-Huei Wang, Tian-Li Wang, and Thomas R. Pisanic
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Cancer Research ,Oncology - Abstract
With a 5-year survival rate of less than 50%, ovarian high-grade serous carcinoma (HGSC) is one of the most highly aggressive gynecological malignancies affecting women today. The high mortality rate of HGSC is largely attributable to delays in diagnosis, as most patients remain undiagnosed until the late stages of -disease. There are currently no recommended screening tests for ovarian cancer and there thus remains an urgent need for new diagnostic methods, particularly those that can detect the disease at early stages when clinical intervention remains effective. While diagnostics for ovarian cancer share many of the same technical hurdles as for other cancer types, the low prevalence of the disease in the general population, coupled with a notable lack of sensitive and specific biomarkers, have made the development of a clinically useful screening strategy particularly challenging. Here, we present a detailed review of the overall landscape of ovarian cancer diagnostics, with emphasis on emerging methods that employ novel protein, genetic, epigenetic and imaging-based biomarkers and/or advanced diagnostic technologies for the noninvasive detection of HGSC, particularly in women at high risk due to germline mutations such as BRCA1/2. Lastly, we discuss the translational potential of these approaches for achieving a clinically implementable solution for screening and diagnostics of early-stage ovarian cancer as a means of ultimately improving patient outcomes in both the general and high-risk populations.
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- 2022
5. Abstract 2231: Epigenomic profiling uncovers distinct enrichment patterns of PBX1 super-enhancer among lung adenocarcinoma
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Douglas M. Mansell, Maya Fridrich, Feng Jiang, Benjamin Chen, Ayushi Patel, Tian Li-Wang, Charles A. Powell, and Hideo Watanabe
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Cancer Research ,Oncology - Abstract
Small cell lung cancers and squamous cell cancers can be epigenetically subtyped (Kong R et al 2022, Sato T et al, 2019) with focus on super-enhancer (SE) signals within or adjacent to transcription factors (TF). We reasoned that specific epigenomic subsets in lung adenocarcinomas (LUAD) operate with similar cellular programming showing TF dependency for lineage commitment and survival. We hypothesize that these dependencies on lineage programming are associated with a specific epigenomic landscape. Therefore, we investigated enrichment of SEs across the spectrum of LUADs to identify potential regulators for lineage commitment and survival. We obtained data for genome-wide H3K27ac enrichment by chromatin immunoprecipitation followed by sequencing (ChIP-seq) on a total of 53 LUAD cell lines. We performed unsupervised hierarchical clustering of signals on SEs at gene loci annotated with transcriptional regulation identified in two or more cell lines. To Uncover 3D contact information, we employed HiChIP, using antibody against SMC1, a cohesin complex, on NCI-H460. Three LUAD cell lines with distinct enhancer profiles were transfected with five enhancer segments of the SE at the PBX1 gene cloned into a Firefly luciferase plasmid. Lastly, four LUAD cell lines were treated with a PBX1 inhibitor, T417 for 72 hours in varying concentrations ranging from 20uM to 10nM and evaluated for sensitivity to the compound. Clustering analysis identified LUADs with differential enhancer enrichment for TFs. Among those, we focused on the PBX1 gene locus in LUADs harboring SEs proximal to its promoter. SMC1 HiChIP analysis on NCI-H460 suggest all enhancer segments are within a local topologically-associated domain with the PBX1 promoter. Luciferase reporter assay demonstrate activity of enhancer segments within the PBX1 SE, unique to each cell line (NCI-H460, PC-9 and NCI-H1437), with enhancers e4 and e5 rendered the highest level of activity while e2 being the most active in PC-9. The effects of these enhancer segments are congruent to H3K27ac enrichment patterns. Treatment with T417 showed reduction in cell proliferation at an IC50 of 2.5 uM and 0.97 uM for NCI-H460 and PC-9, respectively, whereas T417 did not affect cell viability in NCI-H1792 and NCI-H125. Data indicate a distinct subset of LUADs, enriched for the enhancer segments within the SE at the PBX1 gene locus, at which the HiChIP data displayed the enhancer segments lie within the same topologically-associated domain. The enhancer activity of enhancer segments correlated with H3K27ac enrichment pattern for each cell line, suggesting enrichment correlates with impact. Data with PBX1 T417 exposure suggests dependency of PBX1 in maintaining lineage state and survival for unique subsets of LUADs. Further investigation is needed to determine if T417 dependent inhibition is uniform to all PBX1 active LUADs and if other LUAD subsets are dependent on other TFs. Citation Format: Douglas M. Mansell, Maya Fridrich, Feng Jiang, Benjamin Chen, Ayushi Patel, Tian Li-Wang, Charles A. Powell, Hideo Watanabe. Epigenomic profiling uncovers distinct enrichment patterns of PBX1 super-enhancer among lung adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2231.
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- 2023
6. Methylomic Landscapes of Ovarian Cancer Precursor Lesions
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Yeh Wang, Thomas R. Pisanic, Ie Ming Shih, Tza-Huei Wang, Tian Li Wang, Hanru Sun, Michael Considine, and Lihong Li
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Adult ,0301 basic medicine ,Cancer Research ,Biology ,03 medical and health sciences ,0302 clinical medicine ,Ovarian carcinoma ,Biomarkers, Tumor ,medicine ,Fallopian Tube Neoplasms ,Humans ,Aged ,Retrospective Studies ,Ovarian Neoplasms ,Serous Tubal Intraepithelial Carcinoma ,Methylation ,DNA Methylation ,Middle Aged ,Prognosis ,medicine.disease ,Cystadenocarcinoma, Serous ,Gene Expression Regulation, Neoplastic ,Serous fluid ,030104 developmental biology ,Differentially methylated regions ,medicine.anatomical_structure ,Oncology ,Case-Control Studies ,030220 oncology & carcinogenesis ,DNA methylation ,Cancer research ,Female ,Ovarian cancer ,Precancerous Conditions ,Follow-Up Studies ,Fallopian tube - Abstract
Purpose: The current paradigm in the development of high-grade serous ovarian carcinoma (HGSC) proposes that the majority of HGSCs arise from precursor serous tubal intraepithelial carcinoma (STIC) lesions of the fallopian tube. Here we survey genome-wide methylation in HGSC precursor lesions to identify genomic regions that exhibit high-specificity differential hypermethylation for potential use as biomarkers for detecting STIC and HGSC at stages when curative intervention likely remains feasible. Experimental Design: We first identified quality control criteria for performing reliable methylomic analysis of DNA-limited tubal precursor lesions with the Illumina Infinium MethylationEPIC array. We then used this platform to compare genome-wide methylation among 12 STICs with paired adjacent-normal epithelia, one p53 signature lesion and two samples of concurrent HGSC. The resulting methylomic data were analyzed by unsupervised hierarchical clustering and multidimensional analysis. Regions of high-confidence STIC-specific differential hypermethylation were identified using selective bioinformatic criteria and compared with published MethylationEPIC data from 23 HGSC tumors and 11 healthy fallopian tube mucosae. Results: Unsupervised analysis showed that STICs largely clustered with HGSCs, but were clearly distinct from adjacent-normal fallopian tube epithelia. Forty-two genomic regions exhibited high-confidence STIC-specific differential hypermethylation, of which 17 (40.5%) directly overlapped with HGSC-specific differentially methylated regions. Methylation at these shared loci was able to completely distinguish STIC and HGSC samples from normal and adjacent-normal specimens. Conclusions: Our results suggest that most STICs are epigenetically similar to HGSCs and share regions of differential hypermethylation that warrant further evaluation for potential use as biomarkers for early detection of ovarian HGSC. See related commentary by Ishak and De Carvalho, p. 6083
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- 2020
7. Genome‐wide mutation analysis in precancerous lesions of endometrial carcinoma
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Ting Tai Yen, Guangwen Yuan, Qianqian Song, Anna Beavis, Pinli Yue, Lihong Li, Yan Song, Amanda N. Fader, Shiho Asaka, Ie Ming Shih, Tian Li Wang, and Yuchen Jiao
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Adult ,0301 basic medicine ,DNA Copy Number Variations ,DNA Mutational Analysis ,Gene Dosage ,medicine.disease_cause ,Atypical hyperplasia ,Pathology and Forensic Medicine ,03 medical and health sciences ,0302 clinical medicine ,Exome Sequencing ,Biomarkers, Tumor ,medicine ,Carcinoma ,Humans ,PTEN ,Genetic Predisposition to Disease ,Aged ,Endometrial intraepithelial neoplasia ,Mutation ,biology ,Cancer ,Middle Aged ,medicine.disease ,Endometrial Neoplasms ,Cell Transformation, Neoplastic ,Phenotype ,030104 developmental biology ,Tumor progression ,Beijing ,030220 oncology & carcinogenesis ,Baltimore ,Disease Progression ,biology.protein ,Cancer research ,Female ,Microsatellite Instability ,DNA mismatch repair ,Carcinoma, Endometrioid ,Precancerous Conditions ,Genome-Wide Association Study - Abstract
Clinicopathological evidence supports endometrial atypical hyperplasia (AH) or endometrial intraepithelial neoplasia as the precursor of uterine endometrioid carcinoma (EC), the most common gynecologic malignancy. However, the pathogenic progression from AH to EC remains unclear. Here, we employed whole-exome sequencing to identify somatic mutations and copy number changes in micro-dissected lesions from 30 pairs of newly diagnosed AH and EC. We found that all but one pair of AHs shared the same DNA mismatch repair status as their corresponding ECs. The percentage of common mutations between AH lesions and corresponding ECs varied significantly, ranging from 0.1% to 82%. Microsatellite stable AHs had fewer cancer driver mutations than ECs (5 versus 7, p = 0.017), but among microsatellite unstable AHs and ECs there was no difference in mutational numbers (36 versus 38, p = 0.65). As compared to AH specimens, 19 (79%) of 24 microsatellite stable EC tumors gained new cancer driver mutations, most of which involved PTEN, ARID1A, PIK3CA, CTNNB1, or CHD4. Our results suggest that some AH lesions are the immediate precursor of ECs, and progression depends on acquisition of additional cancer driver mutations. However, a complex clonal relationship between AH and EC can also be appreciated, as in some cases both lesions diverge very early or arise independently, thus co-developing with distinct genetic trajectories. Our genome-wide profile of mutations in AH and EC shines new light on the molecular landscape of tumor progression. © 2020 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
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- 2020
8. Inhibition of the MYC-Regulated Glutaminase Metabolic Axis Is an Effective Synthetic Lethal Approach for Treating Chemoresistant Ovarian Cancers
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Alicja Tomaszewski, Angelo M. DeMarzo, Tian Li Wang, Ting Tai Yen, Ryoichi Asaka, Jiaxin Hong, Shiho Asaka, Stephanie Gaillard, Yohan Suryo Rahmanto, Ie Ming Shih, Xi Chen, Fang-Chi Hsu, Yao An Shen, Ben Davidson, Jin Gyoung Jung, Anne Le, Yu Wei Chen, Nabeel Attarwala, Cissy Zhang, and Chi Mu Chuang
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0301 basic medicine ,Cancer Research ,Cell Survival ,Glutamine ,Benzeneacetamides ,Mice, Nude ,Poly(ADP-ribose) Polymerase Inhibitors ,Article ,Piperazines ,Olaparib ,Proto-Oncogene Proteins c-myc ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Glutaminase ,Cell Line, Tumor ,Antineoplastic Combined Chemotherapy Protocols ,Thiadiazoles ,Animals ,Humans ,Ovarian Neoplasms ,Gene knockdown ,Glutaminolysis ,Oncogene ,Chemistry ,Glutathione ,Xenograft Model Antitumor Assays ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Oncology ,Drug Resistance, Neoplasm ,Cell culture ,030220 oncology & carcinogenesis ,PARP inhibitor ,Cancer cell ,Cancer research ,Phthalazines ,Female - Abstract
Amplification and overexpression of the MYC oncogene in tumor cells, including ovarian cancer cells, correlates with poor responses to chemotherapy. As MYC is not directly targetable, we have analyzed molecular pathways downstream of MYC to identify potential therapeutic targets. Here we report that ovarian cancer cells overexpressing glutaminase (GLS), a target of MYC and a key enzyme in glutaminolysis, are intrinsically resistant to platinum-based chemotherapy and are enriched with intracellular antioxidant glutathione. Deprivation of glutamine by glutamine-withdrawal, GLS knockdown, or exposure to the GLS inhibitor CB-839 resulted in robust induction of reactive oxygen species in high GLS-expressing but not in low GLS-expressing ovarian cancer cells. Treatment with CB-839 rendered GLShigh cells vulnerable to the poly(ADP-ribose) polymerase (PARP) inhibitor, olaparib, and prolonged survival in tumor-bearing mice. These findings suggest consideration of applying a combined therapy of GLS inhibitor and PARP inhibitor to treat chemoresistant ovarian cancers, especially those with high GLS expression. Significance: Targeting glutaminase disturbs redox homeostasis and nucleotide synthesis and causes replication stress in cancer cells, representing an exploitable vulnerability for the development of effective therapeutics.
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- 2020
9. Systems medicine dissection of chromosome 1q amplification reveals oncogenic regulatory circuits and informs targeted therapy in cancer
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Irene Roberts, Tian Li Wang, Pierangela Sabbattini, Luca Magnani, Xiaolin Xiao, Keren Keren, Kikkeri N. Naresh, Richard Szydlo, Anastasios Karadimitris, Valentina S. Caputo, Bien Bergonia, Kanagaraju Ponnusamy, Nikolaos Trasanidis, Ioannis Kostopoulos, Aristeidis Chaidos, Paudel Reema, Holger W. Auner, Yao-An Shen, and Alexia Katsarou
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business.industry ,medicine.medical_treatment ,Cancer ,Disease ,Gene signature ,medicine.disease ,Targeted therapy ,Systems medicine ,Cancer cell ,Cancer research ,medicine ,FOXM1 ,Epigenetics ,business - Abstract
Understanding the biological and clinical impact of copy number aberrations (CNA) in cancer remains an unmet challenge. Genetic amplification of chromosome 1q (chr1q-amp) is a major CNA conferring adverse prognosis in several cancers, including the blood cancer, multiple myeloma (MM). Although several chr1q genes portend high-risk MM disease, the underpinning molecular aetiology remains elusive. Here we integrate patient multi-omics datasets with genetic variables to identify 103 adverse prognosis genes in chr1q-amp MM. Amongst these, the transcription factor PBX1 is ectopically expressed by genetic amplification and epigenetic activation of its own preserved 3D regulatory domain. By binding to reprogrammed super-enhancers, PBX1 directly regulates critical oncogenic pathways, whilst in co-operation with FOXM1, activates a proliferative gene signature which predicts adverse prognosis across multiple cancers. Notably, pharmacological disruption of the PBX1-FOXM1 axis, including with a novel PBX1 inhibitor is selectively toxic against chr1q-amp cancer cells. Overall, our systems medicine approach successfully identifies CNA-driven oncogenic circuitries, links them to clinical phenotypes and proposes novel CNA-targeted therapy strategies in cancer.SignificanceWe provide a comprehensive systems medicine strategy to unveil oncogenic circuitries and inform novel precision therapy decisions against CNA in cancer. This first clinical multi-omic analysis of chr1q-amp in MM identifies a central PBX1-FOXM1 regulatory axis driving high-risk prognosis, as a novel therapeutic target against chr1q-amp in cancer.
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- 2021
10. Protein kinase RNA-activated controls mitotic progression and determines paclitaxel chemosensitivity through B-cell lymphoma 2 in ovarian cancer
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Kerry J. Rodabaugh, Ling Yin, Adam R. Karpf, Renya Zeng, Jixin Dong, Yuanhong Chen, Fang Yu, Yongji Zeng, Tian Li Wang, and Amarnath Natarajan
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Cancer Research ,Paclitaxel ,viruses ,Mice, Nude ,Mitosis ,Apoptosis ,Biology ,environment and public health ,chemistry.chemical_compound ,Mice ,eIF-2 Kinase ,Cell Movement ,Genetics ,medicine ,Biomarkers, Tumor ,Tumor Cells, Cultured ,Animals ,Humans ,Kinase activity ,Protein kinase A ,Molecular Biology ,Cell Proliferation ,Ovarian Neoplasms ,Cyclin-dependent kinase 1 ,Kinase ,virus diseases ,Cancer ,biochemical phenomena, metabolism, and nutrition ,medicine.disease ,Protein kinase R ,Antineoplastic Agents, Phytogenic ,Xenograft Model Antitumor Assays ,Gene Expression Regulation, Neoplastic ,enzymes and coenzymes (carbohydrates) ,chemistry ,Proto-Oncogene Proteins c-bcl-2 ,Cancer research ,Female ,Ovarian cancer - Abstract
Anti-tubulin agents, such as paclitaxel, have been used extensively for treatment of several types of cancer, including ovarian, lung, breast, and pancreatic cancers. Despite their wide use in cancer treatment, however, patient response is highly variable and drug resistance remains a major clinical issue. Protein kinase RNA-activated (PKR) plays a critical role in immune response to viral infection. We identified PKR as a phospho-protein in response to anti-tubulin agents and this phosphorylation occurs independent of its own kinase activity. PKR is phosphorylated by cyclin-dependent kinase 1 (CDK1) during anti-tubulin treatment and unperturbed mitosis and that PKR regulates mitotic progression in a phosphorylation-dependent manner. Furthermore, inactivation of PKR confers resistance to paclitaxel in ovarian and breast cancer cells in vitro and in vivo. PKR expression levels and activity are decreased in chemotherapeutic recurrent ovarian cancer patients. Mechanistically, our findings suggest that PKR controls paclitaxel chemosensitivity through repressing Bcl2 expression. Pharmacological inhibition of Bcl2 with FDA-approved agent venetoclax overcomes paclitaxel resistance in preclinical animal models of ovarian cancer. Our results suggest that PKR is a critical determinant of paclitaxel cytotoxicity and that PKR-Bcl2 axis as a potential therapeutic target for the treatment of recurrent drug-resistant ovarian tumors.
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- 2021
11. A Novel ZIP4-HDAC4-VEGFA Axis in High-Grade Serous Ovarian Cancer
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Tian Li Wang, Robert E. Emerson, Qipeng Fan, Lihong Li, and Yan Xu
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0301 basic medicine ,Cancer Research ,medicine.medical_treatment ,medicine.disease_cause ,Article ,cancer stem cell (CSC) ,03 medical and health sciences ,0302 clinical medicine ,Cancer stem cell ,medicine ,endothelial growth factor A (VEGFA) ,RC254-282 ,Cisplatin ,ZIP4 ,Gene knockdown ,Chemistry ,Growth factor ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,HDAC4 ,Vascular endothelial growth factor A ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,high-grade serous ovarian cancer (HGSOC) ,Cancer research ,histone deacetylase 4 (HDAC4) ,Histone deacetylase ,Carcinogenesis ,medicine.drug - Abstract
Simple Summary Despite tremendous research efforts, epithelial ovarian cancer (EOC) remains one of the most difficult cancers to detect early and treat successfully for >5-year survival. We have recently shown that ZIP4, a zinc transporter, is a novel cancer stem cell (CSC) marker and a therapeutic target for EOC. The current work focuses on developing new strategies to target ZIP4 and inhibit its CSC activities in EOC. We found that cells expressing high levels of ZIP4 were supersensitive to a group of inhibitors called HDACis. One of the major targets of these inhibitors is a protein called HDAC4. We revealed the new molecular bases for the ZIP4-HDAC4 axis and tested the efficacies of targeting this axis in the lab and in mouse models. Our study provides a new mechanistic-based targeting strategy for EOC. Abstract We have recently identified ZIP4 as a novel cancer stem cell (CSC) marker in high-grade serous ovarian cancer (HGSOC). While it converts drug-resistance to cisplatin (CDDP), we unexpectedly found that ZIP4 induced sensitization of HGSOC cells to histone deacetylase inhibitors (HDACis). Mechanistically, ZIP4 selectively upregulated HDAC IIa HDACs, with little or no effect on HDACs in other classes. HDAC4 knockdown (KD) and LMK-235 inhibited spheroid formation in vitro and tumorigenesis in vivo, with hypoxia inducible factor-1 alpha (HIF1α) and endothelial growth factor A (VEGFA) as functional downstream mediators of HDAC4. Moreover, we found that ZIP4, HDAC4, and HIF1α were involved in regulating secreted VEGFA in HGSOC cells. Furthermore, we tested our hypothesis that co-targeting CSC via the ZIP4-HDAC4 axis and non-CSC using CDDP is necessary and highly effective by comparing the effects of ZIP4-knockout/KD, HDAC4-KD, and HDACis, in the presence or absence of CDDP on tumorigenesis in mouse models. Our results showed that the co-targeting strategy was highly effective. Finally, data from human HGSOC tissues showed that ZIP4 and HDAC4 were upregulated in a subset of recurrent tumors, justifying the clinical relevance of the study. In summary, our study provides a new mechanistic-based targeting strategy for HGSOC.
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- 2021
12. Loss of ARID1A in Tumor Cells Renders Selective Vulnerability to Combined Ionizing Radiation and PARP Inhibitor Therapy
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Youngran Park, Zheng Cheng Yu, Ayse Ayhan, Tian Li Wang, Raghavendra A. Shamanna, Ie Ming Shih, Sonia Franco, Akila N. Viswanathan, Michael M. Seidman, Vilhelm A. Bohr, M. Herman Chui, Marina A. Bellani, Stephanie Gaillard, Yohan Suryo Rahmanto, and Anthony K.L. Leung
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0301 basic medicine ,Cancer Research ,DNA End-Joining Repair ,DNA Repair ,Cell Survival ,DNA repair ,DNA damage ,Poly ADP ribose polymerase ,Cell Cycle Proteins ,Mice, Transgenic ,Poly(ADP-ribose) Polymerase Inhibitors ,Models, Biological ,Radiation Tolerance ,Article ,Chromatin remodeling ,Olaparib ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cell Line, Tumor ,Animals ,Humans ,DNA Breaks, Double-Stranded ,Chromatin ,DNA-Binding Proteins ,Disease Models, Animal ,030104 developmental biology ,Oncology ,chemistry ,Drug Resistance, Neoplasm ,Gene Knockdown Techniques ,030220 oncology & carcinogenesis ,PARP inhibitor ,Cancer cell ,Cancer research ,DNA Damage ,Transcription Factors - Abstract
Purpose: Somatic inactivating mutations in ARID1A, a component of the SWI/SNF chromatin remodeling complex, are detected in various types of human malignancies. Loss of ARID1A compromises DNA damage repair. The induced DNA damage burden may increase reliance on PARP-dependent DNA repair of cancer cells to maintain genome integrity and render susceptibility to PARP inhibitor therapy. Experimental Design: Isogenic ARID1A−/− and wild-type cell lines were used for assessing DNA damage response, DNA compactness, and profiling global serine/threonine phosphoproteomic in vivo. A panel of inhibitors targeting DNA repair pathways was screened for a synergistic antitumor effect with irradiation in ARID1A−/− tumors. Results: ARID1A-deficient endometrial cells exhibit sustained levels in DNA damage response, a result further supported by in vivo phosphoproteomic analysis. Our results show that ARID1A is essential for establishing an open chromatin state upon DNA damage, a process required for recruitment of 53BP1 and RIF1, key mediators of non-homologous end-joining (NHEJ) machinery, to DNA lesions. The inability of ARID1A−/− cells to mount NHEJ repair results in a partial cytotoxic response to radiation. Small-molecule compound screens revealed that PARP inhibitors act synergistically with radiation to potentiate cytotoxicity in ARID1A−/− cells. Combination treatment with low-dose radiation and olaparib greatly improved antitumor efficacy, resulting in long-term remission in mice bearing ARID1A-deficient tumors. Conclusions: ARID1A-deficient cells acquire high sensitivity to PARP inhibition after exposure to exogenously induced DNA breaks such as ionizing radiation. Our findings suggest a novel biologically informed strategy for treating ARID1A-deficient malignancies.
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- 2019
13. T cell-inflamed phenotype and increased Foxp3 expression in infiltrating T-cells of mismatch-repair deficient endometrial cancers
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Ie Ming Shih, Stephanie Gaillard, Tian Li Wang, Ting Tai Yen, and Shiho Asaka
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Adult ,0301 basic medicine ,Pathology ,medicine.medical_specialty ,medicine.medical_treatment ,T cell ,Cell ,chemical and pharmacologic phenomena ,Article ,Pathology and Forensic Medicine ,03 medical and health sciences ,Lymphocytes, Tumor-Infiltrating ,0302 clinical medicine ,Immune system ,Germline mutation ,Neoplastic Syndromes, Hereditary ,T-Lymphocyte Subsets ,Humans ,Medicine ,Aged ,Aged, 80 and over ,Inflammation ,Brain Neoplasms ,business.industry ,Endometrial cancer ,FOXP3 ,Forkhead Transcription Factors ,Immunotherapy ,Middle Aged ,medicine.disease ,Endometrial Neoplasms ,Phenotype ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Cancer research ,Female ,DNA mismatch repair ,Colorectal Neoplasms ,business - Abstract
Mismatch repair-deficient endometrial cancers have a high somatic mutation burden, suggesting that patients with these tumors may benefit from immunotherapy. Elucidating the immune suppressive mechanisms of mismatch repair-deficient endometrial cancers is fundamental to developing future immune-based interventions. This study aimed to determine the immune cell populations associated with mismatch repair-deficient endometrial cancers, especially focusing on targetable regulatory pathways of the immune response. A total of 76 endometrial cancer hysterectomy specimens were evaluated for tumor-infiltrating immune cells by immunohistochemistry. Immune specific markers were used to evaluate each specimen for the number of CD8 + cytotoxic T lymphocytes, forkhead-box P3 (FoxP3) + regulatory T cells, CD68 + tumor-associated macrophages, as well as programmed death-1 (PD-1) + immune cells, and the percentage of programmed death ligand-1 (PD-L1) + immune cells. Mismatch repair-deficient tumors exhibited a significantly higher number of CD8 + cytotoxic T lymphocytes (p = 0.0006), FoxP3 + regulatory T cells (p = 0.0003), PD-1 + immune cells (p = 0.0069), and a higher percentage of PD-L1 + immune cells (p = 0.0007) occupying the tumor compared to mismatch repair-proficient endometrial cancers. There was no significant difference in CD68 + tumor-associated macrophages infiltration between the two groups. Endometrial cancers with tumor PD-L1 expression also showed significantly increased infiltration of CD8 + cytotoxic T lymphocytes (p = 0.0002), FoxP3 + regulatory T cells (p = 0.0003), PD-1 + immune cells (p < 0.0001), and PD-L1 + immune cells (p < 0.0001). Endometrial cancers showing mismatch repair-deficiency and PD-L1 expression in tumor cells exhibit a prominent T cell-inflamed phenotype. More importantly, the increased number of FoxP3 + regulatory T cells in mismatch repair-deficient endometrial cancers suggests that combination therapy by targeting both regulatory T cells and immune checkpoints may be warranted to improve clinical efficacy.
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- 2019
14. Development of Small Molecule Inhibitors Targeting PBX1 Transcription Signaling as a Novel Cancer Therapeutic Strategy
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Yohan Suryo Rahmanto, Ie Ming Shih, Fang-Chi Hsu, Tian Li Wang, Jin Jung, Stephanie Gaillard, Geoffrey D. Shimberg, Chi-Mu Chuang, Jiaxin Hong, Yao An Shen, and Jürgen Bosch
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Multidisciplinary ,Science ,fungi ,Cell ,Cancer ,Biology ,medicine.disease ,medicine.disease_cause ,Small molecule ,Article ,Carboplatin ,Chemistry ,chemistry.chemical_compound ,medicine.anatomical_structure ,chemistry ,Transcription (biology) ,hemic and lymphatic diseases ,Chromosomal region ,Cancer cell ,medicine ,Cancer research ,Carcinogenesis ,Transcription factor - Abstract
Summary PBX1 is a transcription factor involved in diverse cellular functions including organ development, stem cell renewal, and tumorigenesis. PBX1 is localized at chr1q23.3, a frequently amplified chromosomal region, and it is overexpressed in many human malignancies. Cancer cells with elevated PBX1 signaling are particularly vulnerable to PBX1 withdrawal. We designed a series of small molecule compounds capable of docking to the interface between PBX1 and its cognate DNA target sequence. Among them, T417 is found to be a lead compound. In cell-based assays, T417 significantly suppressed self-renewal and proliferation of cancer cells expressing high levels of PBX1. T417 also re-sensitized platinum-resistant ovarian tumors to carboplatin. T417 did not affect healthy tissues likely due to their lower PBX1 expression levels. Therefore, targeting PBX-DNA interface can be a promising strategy for treating human tumors reliant on PBX1 for survival., Graphical abstract, Highlights • Developing small molecular compounds to interfere with PBX1 protein and DNA interaction • Lead compound, T417, is potent in affecting PBX1 transcription • T417 displays low in vivo toxicity and satisfactory in vivo anti-tumor potency, Chemistry; Cancer
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- 2021
15. Correction: BRAFV600E-mutated ovarian serous borderline tumors are at relatively low risk for progression to serous carcinoma
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Ie Ming Shih, Tian Li Wang, Kirsten Frederiksen, Susanne K. Kjaer, Charlotte Gerd Hannibal, M. Herman Chui, and Russell Vang
- Subjects
Serous fluid ,Oncology ,Serous carcinoma ,business.industry ,medicine ,Cancer research ,Correction ,medicine.disease ,business - Published
- 2021
16. The Origin of Ovarian Cancer Species and Precancerous Landscape
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Yeh Wang, Tian Li Wang, and Ie Ming Shih
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0301 basic medicine ,Serous carcinoma ,Carcinogenesis ,Review ,Carcinoma, Ovarian Epithelial ,Pathology and Forensic Medicine ,03 medical and health sciences ,0302 clinical medicine ,Genetic predisposition ,medicine ,Carcinoma ,Animals ,Humans ,Cystadenocarcinoma ,Fallopian Tubes ,business.industry ,Serous Tubal Intraepithelial Carcinoma ,medicine.disease ,female genital diseases and pregnancy complications ,Cystadenocarcinoma, Serous ,030104 developmental biology ,medicine.anatomical_structure ,Tumor progression ,030220 oncology & carcinogenesis ,Cancer research ,Disease Progression ,Female ,business ,Ovarian cancer ,Precancerous Conditions ,Carcinoma in Situ ,Fallopian tube - Abstract
Unlike other human cancers, in which all primary tumors arise de novo, ovarian epithelial cancers are primarily imported from either endometrial or fallopian tube epithelium. The prevailing paradigm in the genesis of high-grade serous carcinoma (HGSC), the most common ovarian cancer, posits to its development in fallopian tubes through stepwise tumor progression. Recent progress has been made not only in gathering terabytes of omics data but also in detailing the histologic-molecular correlations required for looking into, and making sense of, the tissue origin of HGSC. This emerging paradigm is changing many facets of ovarian cancer research and routine gynecology practice. The precancerous landscape in fallopian tubes contains multiple concurrent precursor lesions, including serous tubal intraepithelial carcinoma (STIC), with genetic heterogeneity providing a platform for HGSC evolution. Mathematical models imply that a prolonged time (decades) elapses from the development of a TP53 mutation, the earliest known molecular alteration, to an STIC, followed by a shorter span (6 years) for progression to an HGSC. Genetic predisposition accelerates the trajectory. This timeline may allow for the early diagnosis of HGSC and STIC, followed by intent-to-cure surgery. This review discusses the recent advances in this tubal paradigm and its biological and clinical implications, alongside the promise and challenge of studying STIC and other precancerous lesions of HGSC.
- Published
- 2020
17. Inhibition of ovarian tumor cell invasiveness by targeting SYK in the tyrosine kinase signaling pathway
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Denis Wirtz, Tian Li Wang, Jude M. Phillip, Stuart S. Martin, Chuan Hsiang Huang, Ie Ming Shih, Yu Yu, Michele Vitolo, Pei Hsun Wu, Stephanie Gaillard, Meng-Horng Lee, and Yohan Suryo Rahmanto
- Subjects
0301 basic medicine ,Cancer Research ,Syk ,macromolecular substances ,environment and public health ,Article ,Basement Membrane ,spleen tyrosine kinase ,03 medical and health sciences ,Ovarian tumor ,Cell Movement ,Cell Line, Tumor ,Genetics ,medicine ,Humans ,Syk Kinase ,Neoplasm Invasiveness ,Phosphorylation ,Molecular Biology ,F-actin dynamics ,biology ,Kinase ,phosphoproteomics ,hemic and immune systems ,cell invasion ,medicine.disease ,3. Good health ,enzymes and coenzymes (carbohydrates) ,ovarian cancer ,030104 developmental biology ,Tumor progression ,biology.protein ,Cancer research ,Signal transduction ,Ovarian cancer ,Tyrosine kinase ,Cortactin ,Signal Transduction - Abstract
Ovarian cancer cell motility and invasiveness are prerequisites for dissemination, and largely account for cancer mortality. We have identified an actionable kinase, spleen tyrosine kinase (SYK), which is keenly associated with tumor progression in ovarian cancer. Here, we report that active recombinant SYK directly phosphorylates cortactin and cofilin, which are critically involved in assembly and dynamics of actin filament through phosphorylation signaling. Enhancing SYK activity by inducing expression of a constitutively active SYK mutant, SYK130E, increased growth factor-stimulated migration and invasion of ovarian cancer cells, which was abrogated by cortactin knockdown. Similarly, SYK inhibitors significantly decreased invasion of ovarian cancer cells through basement membrane matrix in a real-time transwell assays and in a 3-D tumor spheroid model. SYK inactivation by gene knockout or by small molecule inhibition reduced actin polymerization. Collectively, the results reported here identify a new mechanism by which SYK signaling regulates ovarian cancer cell motility and invasiveness, and pinpoint a target-based strategy to prevent or suppress the advancement of ovarian malignancies.
- Published
- 2018
18. Tumor-associated macrophages and the tumor immune microenvironment of primary and recurrent epithelial ovarian cancer
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Ie Ming Shih, Leisha A. Emens, Amanda N. Fader, Alan K. Meeker, Tian Li Wang, Elizabeth D. Thompson, Ashley Cimino-Mathews, and Laureen S. Ojalvo
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Adult ,0301 basic medicine ,Programmed Cell Death 1 Receptor ,Disease ,Carcinoma, Ovarian Epithelial ,B7-H1 Antigen ,Pathology and Forensic Medicine ,03 medical and health sciences ,Lymphocytes, Tumor-Infiltrating ,0302 clinical medicine ,Immune system ,Tumor Microenvironment ,Humans ,Medicine ,Cytotoxic T cell ,Aged ,Ovarian Neoplasms ,Tumor microenvironment ,business.industry ,CD68 ,Macrophages ,FOXP3 ,Middle Aged ,Prognosis ,medicine.disease ,Progression-Free Survival ,Survival Rate ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer research ,Female ,Neoplasm Recurrence, Local ,business ,Ovarian cancer ,CD8 - Abstract
Tumor-infiltrating lymphocytes (TILs) are associated with better prognosis in newly diagnosed epithelial ovarian cancer (EOC), but clinical trials of immunotherapies in patients with heavily treated disease reveal limited activity. Understanding the tumor microenvironment (TME) of primary and recurrent EOC should guide future trials. Here, we evaluated the TME of paired primary and recurrent tumors (n = 17), and non-paired primary (n = 20) and recurrent (n = 15) tumors, for CD8+ T cells, FOXP3+ regulatory T cells (Tregs), CD68+ tumor-associated macrophages (TAMs), programmed cell death protein 1 (PD-1) and programmed cell death ligand 1 (PD-L1). CD8+ T cells were similar in primary and recurrent tumors, but Tregs were higher in recurrent tumors (P = .0210). Higher TAM density (≥5%) associated with higher Tregs (P = .001) and CD8+ T cells (P < .001) in recurrent tumors, but only with higher Tregs in primary tumors (P = .02). TAM-dense recurrent tumors expressed PD-L1 on tumor and immune cells, whereas TAM-dense primary tumors expressed PD-L1 predominantly on immune cells. In survival analyses, higher Tregs in primary tumors correlated with decreased time to first recurrence (17.0 versus 28.5 months, P = .022). Conversely, higher Tregs in recurrent tumors correlated with longer overall survival (OS) from recurrence (median not met versus 20.0 months, P = .022). TAM density did not affect patient survival. However, patients with increased TAMs at recurrence (n = 5) had longer OS from recurrence compared to patients without increased TAMs (n = 12) (56.0 versus 20.0 months); with the small sample size, this did not reach statistical significance (P = .074). Further characterization of the evolution of the TME is warranted.
- Published
- 2018
19. Repurposing Pan-HDAC Inhibitors for ARID1A-Mutated Ovarian Cancer
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Jose R. Conejo-Garcia, Timothy Nacarelli, Lin Zhang, Nail Fatkhutdinov, David W. Speicher, Takeshi Fukumoto, Shuai Wu, Tian Li Wang, Stephanie Jean, Andrew V. Kossenkov, Benjamin G. Bitler, Sergey Karakashev, Pyoung Hwa Park, Ie Ming Shih, and Rugang Zhang
- Subjects
0301 basic medicine ,ARID1A ,Mice, Transgenic ,macromolecular substances ,Biology ,medicine.disease_cause ,Article ,General Biochemistry, Genetics and Molecular Biology ,Chromatin remodeling ,Mice ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Humans ,lcsh:QH301-705.5 ,Ovarian Neoplasms ,Mutation ,EZH2 ,Drug Repositioning ,Nuclear Proteins ,Cancer ,medicine.disease ,Xenograft Model Antitumor Assays ,SWI/SNF ,3. Good health ,DNA-Binding Proteins ,Histone Deacetylase Inhibitors ,Mice, Inbred C57BL ,030104 developmental biology ,lcsh:Biology (General) ,Apoptosis ,030220 oncology & carcinogenesis ,Cancer research ,Female ,Ovarian cancer ,Transcription Factors - Abstract
SUMMARY ARID1A , a subunit of the SWI/SNF complex, is among the most frequently mutated genes across cancer types. ARID1A is mutated in more than 50% of ovarian clear cell carcinomas (OCCCs), diseases that have no effective therapy. Here, we show that ARID1A mutation confers sensitivity to pan-HDAC inhibitors such as SAHA in ovarian cancers. This correlated with enhanced growth suppression induced by the inhibition of HDAC2 activity in ARID1A-mutated cells. HDAC2 interacts with EZH2 in an ARID1A status-dependent manner. HDAC2 functions as a co-repressor of EZH2 to suppress the expression of EZH2/ARID1A target tumor suppressor genes such as PIK3IP1 to inhibit proliferation and promote apoptosis. SAHA reduced the growth and ascites of the ARID1A-inactivated OCCCs in both orthotopic and genetic mouse models. This correlated with a significant improvement of survival of mice bearing ARID1A-mutated OCCCs. These findings provided preclinical rationales for repurposing FDA-approved pan-HDAC inhibitors for treating ARID1A-mutated cancers., In Brief Fukumoto et al. show that ARID1A mutation confers sensitivity to pan-HDAC inhibitors such as SAHA in ovarian cancers. This correlated with enhanced growth suppression induced by the inhibition of HDAC2 activity in ARID1A-mutated cells. These findings provided preclinical rationales for repurposing FDA-approved pan-HDAC inhibitors for treating ARID1A-mutated cancers.
- Published
- 2018
20. High grade serous ovarian carcinomas originate in the fallopian tube
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Michaela Bowden, Jean-Christophe Tille, Michaël Noë, Lauren E. Schwartz, Rachel Karchin, Marian Novak, Carolyn Hruban, Tian Li Wang, Michelle S. Hirsch, Vilmos Adleff, Robert J. Kurman, Douglas I. Lin, Ayse Ayhan, Jillian Phallen, Ie Ming Shih, Robert B. Scharpf, Laura D. Wood, Noushin Niknafs, S. Intidhar Labidi-Galy, Dorothy Hallberg, Ronny Drapkin, Eniko Papp, Rohit Bhattacharya, Victor E. Velculescu, Siân Jones, and Cecile L. Maire
- Subjects
0301 basic medicine ,endocrine system ,animal structures ,DNA Copy Number Variations ,endocrine system diseases ,Science ,General Physics and Astronomy ,Laser Capture Microdissection ,ddc:616.07 ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,0302 clinical medicine ,Fallopian Tube Neoplasm ,Ovarian carcinoma ,PTEN ,Medicine ,Fallopian Tube Neoplasms ,Humans ,Cystadenocarcinoma ,lcsh:Science ,Alleles ,Fallopian Tubes ,ddc:616 ,Ovarian Neoplasms ,Multidisciplinary ,biology ,business.industry ,urogenital system ,Serous Tubal Intraepithelial Carcinoma ,General Chemistry ,medicine.disease ,Immunohistochemistry ,female genital diseases and pregnancy complications ,3. Good health ,Cystadenocarcinoma, Serous ,Serous fluid ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,Female ,lcsh:Q ,business ,Ovarian cancer ,Neoplasms, Cystic, Mucinous, and Serous ,Fallopian tube - Abstract
High-grade serous ovarian carcinoma (HGSOC) is the most frequent type of ovarian cancer and has a poor outcome. It has been proposed that fallopian tube cancers may be precursors of HGSOC but evolutionary evidence for this hypothesis has been limited. Here, we perform whole-exome sequence and copy number analyses of laser capture microdissected fallopian tube lesions (p53 signatures, serous tubal intraepithelial carcinomas (STICs), and fallopian tube carcinomas), ovarian cancers, and metastases from nine patients. The majority of tumor-specific alterations in ovarian cancers were present in STICs, including those affecting TP53, BRCA1, BRCA2 or PTEN. Evolutionary analyses reveal that p53 signatures and STICs are precursors of ovarian carcinoma and identify a window of 7 years between development of a STIC and initiation of ovarian carcinoma, with metastases following rapidly thereafter. Our results provide insights into the etiology of ovarian cancer and have implications for prevention, early detection and therapeutic intervention of this disease., It has previously been proposed that high-grade serous ovarian carcinoma (HGSOC) may originate from the fallopian tube. Here, the authors analyze genetic aberrances in fallopian tube lesions, ovarian cancers, and metastases from HGSOC patients and establish the evolutionary origins of HGSOC in the fallopian tube.
- Published
- 2017
21. Mutation of NRAS is a rare genetic event in ovarian low-grade serous carcinoma
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Felix Zeppernick, Ie Ming Shih, Yohan Suryo Rahmanto, Susanne K. Kjaer, Charlotte Gerd Hannibal, Deyin Xing, Russell Vang, and Tian Li Wang
- Subjects
Adult ,Proto-Oncogene Proteins B-raf ,0301 basic medicine ,Neuroblastoma RAS viral oncogene homolog ,endocrine system diseases ,Serous carcinoma ,Denmark ,DNA Mutational Analysis ,medicine.disease_cause ,Article ,GTP Phosphohydrolases ,Pathology and Forensic Medicine ,Young Adult ,03 medical and health sciences ,Exon ,0302 clinical medicine ,Mutation Rate ,Biomarkers, Tumor ,medicine ,Humans ,Genetic Predisposition to Disease ,Neoplasm Invasiveness ,neoplasms ,Aged ,Cell Proliferation ,Neoplasm Staging ,Aged, 80 and over ,Ovarian Neoplasms ,Mutation ,Oncogene ,business.industry ,Carcinoma ,Membrane Proteins ,Middle Aged ,medicine.disease ,digestive system diseases ,Serous fluid ,Phenotype ,030104 developmental biology ,030220 oncology & carcinogenesis ,Baltimore ,Cancer research ,Female ,KRAS ,Neoplasm Grading ,Neoplasms, Cystic, Mucinous, and Serous ,Ovarian cancer ,business - Abstract
Activating mutations involving the members of the RAS signaling pathway, including KRAS, NRAS, and BRAF, have been reported in ovarian low-grade serous carcinoma and its precursor lesion, serous borderline tumor (SBT). Whether additional genetic alterations in the RAS oncogene family accumulate during the progression of SBT to invasive low-grade serous carcinoma (LGSC) remains largely unknown. Although mutations of KRAS and BRAF occur at a very early stage of progression, even preceding the development of SBT, additional driving events, such as NRAS mutations, have been postulated to facilitate progression. In this study, we analyzed NRAS exon 3 mutational status in 98 cases that were diagnosed with SBT/atypical proliferative serous tumor, noninvasive LGSC, or invasive LGSC. Of the latter, NRAS Q61R (CAA to CGA) mutations were detected in only 2 of 56 (3.6%) cases. The same mutation was not detected in any of the SBTs (atypical proliferative serous tumors) or noninvasive LGSCs. Mutational analysis for hotspots in KRAS and BRAF demonstrated a wild-type pattern of KRAS and BRAF in one of the NRAS-mutated cases. Interestingly, another LGSC case with NRAS mutation harbored a concurrent BRAF V600L mutation. These findings indicate that, although recurrent NRAS mutations are present, their low prevalence indicates that NRAS plays a limited role in the development of LGSC. Further studies to identify other oncogenic events that drive LGSC progression are warranted.
- Published
- 2017
22. The novel ZIP4 regulation and its role in ovarian cancer
- Author
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Qipeng Fan, Qingchun Cai, Pengfei Li, Emily Gerry, Tian Li Wang, Jing Wang, Ie Ming Shih, Kenneth P. Nephew, Wenyan Wang, and Yan Xu
- Subjects
0301 basic medicine ,Context (language use) ,medicine.disease_cause ,03 medical and health sciences ,0302 clinical medicine ,Cancer stem cell ,RNA interference ,Medicine ,ZIP4 ,cancer stem cells (CSC) ,Gene knockdown ,business.industry ,Cancer ,medicine.disease ,3. Good health ,LPA ,030104 developmental biology ,ovarian cancer ,Oncology ,030220 oncology & carcinogenesis ,Immunology ,Cancer research ,Immunohistochemistry ,business ,Carcinogenesis ,Ovarian cancer ,Research Paper - Abstract
// Qipeng Fan 1 , Qingchun Cai 1 , Pengfei Li 1, 2 , Wenyan Wang 1, 3 , Jing Wang 1, 4 , Emily Gerry 5 , Tian-Li Wang 5 , Ie-Ming Shih 5 , Kenneth P. Nephew 6 and Yan Xu 1 1 Department of Obstetrics and Gynecology, Indiana University School of Medicine, Indianapolis, IN 46202, USA 2 Pharmaceutical Research Center, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, 100020, P.R. China 3 Department of Obstetrics and Gynecology, The Second Hospital of Anhui Medical University, Hefei City, 230601, P.R. China 4 MASDINO (Beijing) Medical Research Co. Ltd., Beijing, 100123, P.R. China 5 Department of Pathology, Johns Hopkins School of Medicine, Baltimore, MD 21231, USA 6 Medical Sciences, Indiana University School of Medicine, Jordan Hall 302, Bloomington, IN 47405, USA Correspondence to: Yan Xu, email: xu2@iupui.edu Keywords: ZIP4, LPA, ovarian cancer, cancer stem cells (CSC) Received: June 07, 2017 Accepted: August 26, 2017 Published: September 30, 2017 ABSTRACT Our RNAseq analyses revealed that ZIP4 is a top gene up-regulated in more aggressive ovarian cancer cells. ZIP4’s role in cancer stem cells has not been reported in any type of cancer. In addition, the role and regulation of ZIP4, a zinc transporter, have been studied in the context of extracellular zinc transporting. Factors other than zinc with ZIP4 regulatory effects are essentially unknown. ZIP4 expression and its regulation in epithelial ovarian cancer cells was assessed by immunoblotting, quantitative PCR, or immunohistochemistry staining in human ovarian tissues. Cancer stem cell-related activities were examined to evaluate the role of ZIP4 in human high-grade serous ovarian cancer cells in vitro and in vivo . RNAi and CRISPR techniques were used to knockdown or knockout ZIP4 and related genes. Ovarian cancer tissues overexpressed ZIP4 when compared with normal and benign tissues. ZIP4 knockout significantly reduced several cancer stem cell-related activities in EOC cells, including proliferation, anoikis-resistance, colony-formation, spheroid-formation, drug-resistance, and side-population in vitro . ZIP4-expressing side-population highly expressed known CSC markers ALDH1 and OCT4. ZIP4 knockout dramatically reduced tumorigenesis and ZIP4 overexpression increased tumorigenesis in vivo . In addition, the ZIP4-expressing side-population had the tumor initiating activity. Moreover, the oncolipid lysophosphatic acid effectively up-regulated ZIP4 expression via the nuclear receptor peroxisome proliferator-activated receptor gamma and lysophosphatic acid ’s promoting effects in cancer stem cell-related activities in HGSOC cells was at least partially mediated by ZIP4 in an extracellular zinc-independent manner. Our critical data imply that ZIP4 is a new and important cancer stem cell regulator in ovarian cancer. Our data also provide an innovative interpretation for the apparent disconnection between low levels of zinc and up-regulation of ZIP4 in ovarian cancer tissues.
- Published
- 2017
23. Structure-function analyses of candidate small molecule RPN13 inhibitors with antitumor properties
- Author
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Richard B.S. Roden, Ruey Shyang Soong, Palmer Foran, Ravi K. Anchoori, Shiwen Peng, Samarjit Das, Aliyah Algethami, Marietta Tan, Chenguang Wang, Ssu Hsueh Tseng, Chien Fu Hung, Tian Li Wang, and Hong Liang
- Subjects
0301 basic medicine ,Cancer Treatment ,Apoptosis ,Biochemistry ,Plasma Cell Disorders ,Hematologic Cancers and Related Disorders ,Mice ,0302 clinical medicine ,Spectrum Analysis Techniques ,Ubiquitin ,Neoplasms ,Medicine and Health Sciences ,Receptor ,Multidisciplinary ,biology ,Molecular Structure ,Cell Death ,Chemistry ,Intracellular Signaling Peptides and Proteins ,Hematology ,Flow Cytometry ,Small molecule ,Ubiquitinated Proteins ,3. Good health ,Ovarian Cancer ,Enzymes ,Myelomas ,Oncology ,Cell Processes ,Spectrophotometry ,030220 oncology & carcinogenesis ,Medicine ,Female ,Cytophotometry ,Pharmacophore ,Multiple Myeloma ,Oxidoreductases ,Proteasome Inhibitors ,Luciferase ,Protein Binding ,Research Article ,Cell Binding ,Proteasome Endopeptidase Complex ,Cell Physiology ,Science ,Antineoplastic Agents ,Research and Analysis Methods ,Benzylidene Compounds ,03 medical and health sciences ,Inhibitory Concentration 50 ,Structure-Activity Relationship ,Cell Line, Tumor ,medicine ,Structure–activity relationship ,Animals ,Humans ,Myelomas and Lymphoproliferative Diseases ,Ubiquitination ,Cancer ,Biology and Life Sciences ,Proteins ,Protein Complexes ,Proteasomes ,Cancers and Neoplasms ,Cell Biology ,medicine.disease ,Xenograft Model Antitumor Assays ,030104 developmental biology ,Proteasome ,Cancer research ,biology.protein ,Enzymology ,Gynecological Tumors - Abstract
We sought to design ubiquitin-proteasome system inhibitors active against solid cancers by targeting ubiquitin receptor RPN13 within the proteasome's 19S regulatory particle. The prototypic bis-benzylidine piperidone-based inhibitor RA190 is a michael acceptor that adducts Cysteine 88 of RPN13. In probing the pharmacophore, we showed the benefit of the central nitrogen-bearing piperidone ring moiety compared to a cyclohexanone, the importance of the span of the aromatic wings from the central enone-piperidone ring, the contribution of both wings, and that substituents with stronger electron withdrawing groups were more cytotoxic. Potency was further enhanced by coupling of a second warhead to the central nitrogen-bearing piperidone as RA375 exhibited ten-fold greater activity against cancer lines than RA190, reflecting its nitro ring substituents and the addition of a chloroacetamide warhead. Treatment with RA375 caused a rapid and profound accumulation of high molecular weight polyubiquitinated proteins and reduced intracellular glutathione levels, which produce endoplasmic reticulum and oxidative stress, and trigger apoptosis. RA375 was highly active against cell lines of multiple myeloma and diverse solid cancers, and demonstrated a wide therapeutic window against normal cells. For cervical and head and neck cancer cell lines, those associated with human papillomavirus were significantly more sensitive to RA375. While ARID1A-deficiency also enhanced sensitivity 4-fold, RA375 was active against all ovarian cancer cell lines tested. RA375 inhibited proteasome function in muscle for >72h after single i.p. administration to mice, and treatment reduced tumor burden and extended survival in mice carrying an orthotopic human xenograft derived from a clear cell ovarian carcinoma.
- Published
- 2019
24. Inactivation of Arid1a in the endometrium is associated with endometrioid tumorigenesis through transcriptional reprogramming
- Author
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Meng-Horng Lee, Geoffrey D. Shimberg, Yu Yu, Xu Shi, Michele Vitolo, Vivek Singh, Yohan Suryo Rahmanto, Xi Chen, Ryoichi Asaka, Jianhua Xuan, Zheng Cheng Yu, Ronny Drapkin, Ie Ming Shih, Stuart S. Martin, Wenjing Shen, Tian Li Wang, Denis Wirtz, and Tsutomu Miyamoto
- Subjects
0301 basic medicine ,ARID1A ,Carcinogenesis ,General Physics and Astronomy ,Pathogenesis ,Endometrium ,medicine.disease_cause ,Transcriptome ,0302 clinical medicine ,lcsh:Science ,Cells, Cultured ,Cancer ,Mice, Knockout ,Mutation ,Mice, Inbred BALB C ,Multidisciplinary ,Cellular Reprogramming ,DNA-Binding Proteins ,medicine.anatomical_structure ,Cell Transformation, Neoplastic ,030220 oncology & carcinogenesis ,Female ,Reprogramming ,Carcinoma, Endometrioid ,Cell biology ,Mice, 129 Strain ,Science ,Mice, Transgenic ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Chromatin remodeling ,Article ,03 medical and health sciences ,medicine ,Genetics ,Animals ,Humans ,Endometrial cancer ,Gene Expression Profiling ,General Chemistry ,medicine.disease ,Endometrial Neoplasms ,030104 developmental biology ,Cancer research ,lcsh:Q ,Transcription Factors - Abstract
Somatic inactivating mutations of ARID1A, a SWI/SNF chromatin remodeling gene, are prevalent in human endometrium-related malignancies. To elucidate the mechanisms underlying how ARID1A deleterious mutation contributes to tumorigenesis, we establish genetically engineered murine models with Arid1a and/or Pten conditional deletion in the endometrium. Transcriptomic analyses on endometrial cancers and precursors derived from these mouse models show a close resemblance to human uterine endometrioid carcinomas. We identify transcriptional networks that are controlled by Arid1a and have an impact on endometrial tumor development. To verify findings from the murine models, we analyze ARID1AWT and ARID1AKO human endometrial epithelial cells. Using a system biology approach and functional studies, we demonstrate that ARID1A-deficiency lead to loss of TGF-β tumor suppressive function and that inactivation of ARID1A/TGF-β axis promotes migration and invasion of PTEN-deleted endometrial tumor cells. These findings provide molecular insights into how ARID1A inactivation accelerates endometrial tumor progression and dissemination, the major causes of cancer mortality., ARID1A, which is often mutated in human endometrial cancer, is a component of the SWI/SNF chromatin remodelling complex. Here, the authors show that Arid1a mutations in the mouse endometrium and primary human endometrial epithelial cells cause widespread reprogramming of gene transcription and result in a loss of response to TGFβ.
- Published
- 2019
25. Cytomorphologic and molecular analyses of fallopian tube fimbrial brushings for diagnosis of serous tubal intraepithelial carcinoma
- Author
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Ie Ming Shih, Christopher J. VandenBussche, M. Herman Chui, Russell Vang, and Tian Li Wang
- Subjects
Cancer Research ,Pathology ,medicine.medical_specialty ,Serous carcinoma ,medicine.medical_treatment ,DNA Mutational Analysis ,Papanicolaou stain ,030209 endocrinology & metabolism ,Sensitivity and Specificity ,Article ,03 medical and health sciences ,Salpingectomy ,0302 clinical medicine ,Medicine ,Fallopian Tube Neoplasms ,Humans ,Fallopian Tubes ,Ovarian Neoplasms ,business.industry ,Histology ,Serous Tubal Intraepithelial Carcinoma ,Epithelial Cells ,medicine.disease ,Immunohistochemistry ,female genital diseases and pregnancy complications ,Cystadenocarcinoma, Serous ,medicine.anatomical_structure ,Oncology ,030220 oncology & carcinogenesis ,Mutation ,Female ,Cell Surface Extensions ,Tumor Suppressor Protein p53 ,business ,Ovarian cancer ,Immunostaining ,Carcinoma in Situ ,Fallopian tube - Abstract
Background The paradigm shift localizing the origin of ovarian high-grade serous carcinoma (HGSC) to the fallopian tube underscores the rationale for meticulous microscopic examination of salpingectomy specimens. The precursor, termed "serous tubal intraepithelial carcinoma," is often a focal lesion, which poses difficulties for histologic diagnosis. Methods The authors describe a method to examine exfoliated epithelial cells from fallopian tube fimbria by gentle brushing, thereby enabling thorough sampling of the mucosal surface. Fimbrial brushings were collected from 20 fresh salpingectomy specimens from 15 patients, including 5 who had pathologically confirmed ovarian HGSC. Samples taken only from tubes that were grossly negative for tumor were processed for Papanicolaou staining, p53 immunocytochemistry, and tumor protein 53 (TP53) mutation analysis. Results Cells with malignant cytomorphologic features were identified only in tubal brushings from patients with ovarian HGSC. In all cases, atypical/malignant cells on cytology corresponded to lesions with similar morphology and immunostaining pattern in permanent sections, demonstrating the sensitivity of the technique while providing reassurance that specimen integrity was not disrupted by the procedure. Targeted next-generation sequencing confirmed the presence of TP53 mutations in fimbrial brushings from HGSC, but not in benign samples, and demonstrated concordance with the immunostaining pattern. Identical mutations were observed in matched lesions microdissected from formalin-fixed tissue sections. Conclusions The described technique enables cytologic evaluation of the fallopian tube fimbria for a diagnosis of serous tubal intraepithelial carcinoma, serving as a complement to histology while offering distinct advantages with respect to the procurement of cellular material for ancillary testing and research.
- Published
- 2019
26. PVRIG and PVRL2 Are Induced in Cancer and Inhibit CD8(+) T-cell Function
- Author
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Abha Soni, Benjamin Murter, Ofer Levy, Kyle Hansen, Sarah Whelan, Ling Leung, Sudipto Ganguly, Spencer Liang, David Bernados, Mark A. White, Sandeep Kumar, Ilan Vaknin, Tian Li Wang, Ie Ming Shih, Janis M. Taube, Amanda Nickles Fader, Eran Ophir, Liat Dassa, Drew M. Pardoll, and Maya F. Kotturi
- Subjects
0301 basic medicine ,Cancer Research ,CD96 ,medicine.medical_treatment ,Immunology ,Nectins ,Programmed Cell Death 1 Receptor ,Receptors, Cell Surface ,CD8-Positive T-Lymphocytes ,Lymphocyte Activation ,Article ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Lymphocytes, Tumor-Infiltrating ,TIGIT ,Neoplasms ,Medicine ,Cytotoxic T cell ,Animals ,Humans ,Receptors, Immunologic ,business.industry ,Cancer ,Immunotherapy ,medicine.disease ,Blockade ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Cytokine ,030220 oncology & carcinogenesis ,Cancer research ,business ,CD8 ,Protein Binding ,Signal Transduction - Abstract
Although checkpoint inhibitors that block CTLA-4 and PD-1 have improved cancer immunotherapies, targeting additional checkpoint receptors may be required to broaden patient response to immunotherapy. PVRIG is a coinhibitory receptor of the DNAM/TIGIT/CD96 nectin family that binds to PVRL2. We report that antagonism of PVRIG and TIGIT, but not CD96, increased CD8+ T-cell cytokine production and cytotoxic activity. The inhibitory effect of PVRL2 was mediated by PVRIG and not TIGIT, demonstrating that the PVRIG–PVRL2 pathway is a nonredundant signaling node. A combination of PVRIG blockade with TIGIT or PD-1 blockade further increased T-cell activation. In human tumors, PVRIG expression on T cells was increased relative to normal tissue and trended with TIGIT and PD-1 expression. Tumor cells coexpressing PVR and PVRL2 were observed in multiple tumor types, with highest coexpression in endometrial cancers. Tumor cells expressing either PVR or PVRL2 were also present in numbers that varied with the cancer type, with ovarian cancers having the highest percentage of PVR−PVRL2+ tumor cells and colorectal cancers having the highest percentage of PVR+PVRL2− cells. To demonstrate a role of PVRIG and TIGIT on tumor-derived T cells, we examined the effect of PVRIG and TIGIT blockade on human tumor-infiltrating lymphocytes. For some donors, blockade of PVRIG increased T-cell function, an effect enhanced by combination with TIGIT or PD-1 blockade. In summary, we demonstrate that PVRIG and PVRL2 are expressed in human cancers and the PVRIG–PVRL2 and TIGIT–PVR pathways are nonredundant inhibitory signaling pathways. See related article on p. 244
- Published
- 2019
27. Spleen tyrosine kinase activity regulates epidermal growth factor receptor signaling pathway in ovarian cancer
- Author
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Jianhua Xuan, Ie Ming Shih, Ben Davidson, Yu Yu, Ayse Ayhan, Tian Li Wang, Xu Shi, Stephanie Gaillard, Yohan Suryo Rahmanto, Laura Ardighieri, and Yao An Shen
- Subjects
0301 basic medicine ,Research paper ,Receptor, ErbB-2 ,Syk ,Lapatinib ,General Biochemistry, Genetics and Molecular Biology ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Biomarkers, Tumor ,medicine ,Animals ,Humans ,Syk Kinase ,Epidermal growth factor receptor ,Phosphorylation ,Protein Kinase Inhibitors ,Ovarian Neoplasms ,biology ,Kinase ,business.industry ,Gene Expression Profiling ,General Medicine ,Prognosis ,medicine.disease ,Immunohistochemistry ,3. Good health ,ErbB Receptors ,Disease Models, Animal ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer research ,biology.protein ,Female ,Signal transduction ,Transcriptome ,Ovarian cancer ,business ,Signal Transduction ,medicine.drug - Abstract
Background Spleen tyrosine kinase (SYK) is frequently upregulated in recurrent ovarian carcinomas, for which effective therapy is urgently needed. SYK phosphorylates several substrates, but their translational implications remain unclear. Here, we show that SYK interacts with EGFR and ERBB2, and directly enhances their phosphorylation. Methods We used immunohistochemistry and immunoblotting to assess SYK and EGFR phosphorylation in ovarian serous carcinomas. Association with survival was determined by Kaplan-Meier analysis and the log-rank test. To study its role in EGFR signaling, SYK activity was modulated using a small molecule inhibitor, a syngeneic knockout, and an active kinase inducible system. We applied RNA-seq and phosphoproteomic mass spectrometry to investigate the SYK-regulated EGF-induced transcriptome and downstream substrates. Findings Induced expression of constitutively active SYK130E reduced cellular response to EGFR/ERBB2 inhibitor, lapatinib. Expression of EGFRWT, but not SYK non-phosphorylatable EGFR3F mutant, resulted in paclitaxel resistance, a phenotype characteristic to SYK active ovarian cancers. In tumor xenografts, SYK inhibitor reduces phosphorylation of EGFR substrates. Compared to SYKWT cells, SYKKO cells have an attenuated EGFR/ERBB2-transcriptional activity and responsiveness to EGF-induced transcription. In ovarian cancer tissues, pSYK (Y525/526) levels showed a positive correlation with pEGFR (Y1187). Intense immunoreactivity of pSYK (Y525/526) correlated with poor overall survival in ovarian cancer patients. Interpretation These findings indicate that SYK activity positively modulates the EGFR pathway, providing a biological foundation for co-targeting SYK and EGFR. Fund Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine, NIH/NCI, Ovarian Cancer Research Foundation Alliance, HERA Women's Cancer Foundation and Roseman Foundation. Funders had no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript and eventually in the decision to submit the manuscript.
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- 2019
28. Ovarian Cancer Chemoresistance Relies on the Stem Cell Reprogramming Factor PBX1
- Author
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Ben Davidson, Emily Gerry, Tian Li Wang, Jin-Gyoung Jung, Licia Selleri, Tae Hoen Kim, Ie Ming Shih, Amanda N. Fader, Ayse Ayhan, Joon Tae Park, and Karen Handschuh
- Subjects
STAT3 Transcription Factor ,0301 basic medicine ,Cancer Research ,Biology ,Article ,Transcriptome ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cell Line, Tumor ,Proto-Oncogene Proteins ,hemic and lymphatic diseases ,medicine ,Humans ,Gene silencing ,STAT3 ,Ovarian Neoplasms ,Pre-B-Cell Leukemia Transcription Factor 1 ,Janus Kinase 2 ,Cellular Reprogramming ,medicine.disease ,Carboplatin ,DNA-Binding Proteins ,030104 developmental biology ,Oncology ,chemistry ,Drug Resistance, Neoplasm ,030220 oncology & carcinogenesis ,Cancer research ,biology.protein ,Female ,Stem cell ,Ovarian cancer ,Reprogramming ,Chromatin immunoprecipitation - Abstract
The evolution of chemoresistance is a fundamental characteristic of cancer that ultimately hampers its clinical management. However, it may be possible to improve patient outcomes significantly by a better understanding of resistance mechanisms, which cancers rely upon during the evolution to an untreatable state. Here we report an essential role of the stem cell reprogramming factor, PBX1, in mediating chemoresistance in ovarian carcinomas. In the clinical setting, high levels of PBX1 expression correlated with shorter survival in post-chemotherapy ovarian cancer patients. In tumor cells with low endogenous levels of PBX1, its enforced expression promoted cancer stem cell-like phenotypes, including most notably an increase in resistance to platinum-based therapy used most commonly for treating this disease. Conversely, silencing PBX1 in platinum-resistant cells that overexpressed PBX1 sensitized them to platinum treatment and reduced their stem-like properties. An analysis of published genome-wide chromatin immunoprecipitation data indicated that PBX1 binds directly to promoters of genes involved in stem cell maintenance and the response to tissue injury. We confirmed direct regulation of one of these genes, STAT3, demonstrating that the PBX1 binding motif at its promoter acted to positively regulate STAT3 transcription. We further demonstrated that a STAT3/JAK2 inhibitor could potently sensitize platinum-resistant cells to carboplatin and suppress their growth in vivo. Our findings offer a mechanistic rationale to target the PBX1/STAT3 axis to antagonize a key mechanism of chemoresistance in ovarian cancers and possibly other human cancers. Cancer Res; 76(21); 6351–61. ©2016 AACR.
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- 2016
29. Laminin C1 expression by uterine carcinoma cells is associated with tumor progression
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Tsutomu Miyamoto, Tian Li Wang, Ie Ming Shih, Abdulrahman K. Sinno, Yihong Wang, Tanri Shiozawa, Ren-Chin Wu, Hiroyasu Kashima, and Amanda N. Fader
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Pathology ,medicine.medical_specialty ,Databases, Factual ,Endometrium ,Article ,Atypical hyperplasia ,Cell Movement ,Laminin ,Cell Line, Tumor ,medicine ,Carcinoma ,Humans ,Neoplasm Invasiveness ,RNA, Messenger ,biology ,business.industry ,Obstetrics and Gynecology ,medicine.disease ,Immunohistochemistry ,Endometrial Neoplasms ,Up-Regulation ,Endometrial hyperplasia ,Gene Expression Regulation, Neoplastic ,Serous fluid ,medicine.anatomical_structure ,Oncology ,Tumor progression ,Gene Knockdown Techniques ,Endometrial Hyperplasia ,Clear cell carcinoma ,Disease Progression ,Cancer research ,biology.protein ,Female ,Neoplasm Grading ,Neoplasms, Cystic, Mucinous, and Serous ,business ,Carcinoma, Endometrioid ,Adenocarcinoma, Clear Cell - Abstract
Objectives Molecular markers associated with tumor progression in uterine carcinoma are poorly defined. In this study, we determine whether upregulation of LAMC1, a gene encoding extracellular matrix protein, laminin γ1, is associated with various uterine carcinoma subtypes and stages of tumor progression. Methods An analysis of the immunostaining patterns of laminin γ1 in normal endometrium, atypical hyperplasia, and a total of 150 uterine carcinomas, including low-grade and high-grade endometrioid carcinomas, uterine serous and clear cell carcinoma, was performed. Clinicopathological correlation was performed to determine biological significance. The Cancer Genome Atlas (TCGA) data set was used to validate our results. Results As compared to normal proliferative and secretory endometrium, for which laminin γ1 immunoreactivity was almost undetectable, increasing laminin C1 staining intensity was observed in epithelial cells from atypical hyperplasia to low-grade endometrioid to high-grade endometrioid carcinoma, respectively. Laminin γ1 expression was significantly associated with FIGO stage, myometrial invasion, cervical/adnexal involvement, angiolymphatic invasion and lymph node metastasis. Similarly, analysis of the endometrial carcinoma data set from TCGA revealed that LAMC1 transcript levels were higher in high-grade than those in low-grade endometrioid carcinoma. Silencing LAMC1 expression by siRNAs in a high-grade endometrioid carcinoma cell line did not affect its proliferative activity but significantly suppressed cell motility and invasion in vitro . Conclusions These data suggest that laminin γ1 may contribute to the development and progression of uterine carcinoma, likely through enhancing tumor cell motility and invasion. Laminin γ1 warrants further investigation regarding its role as a biomarker and therapeutic target in uterine carcinoma.
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- 2015
30. Abstract PR11: Dual blockade of BRD4 and the ATR/WEE1 pathway exploits ARID1A loss in clear-cell ovarian cancers
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Sarah B. Gitto, Erin George, Ronny Drapkin, Tian Li Wang, Gordon B. Mills, Yasuto Kinose, Lauren E. Schwartz, Haineng Xu, Margaret Whicker, Dorothy Hallberg, Hyoung Kim, Victor E. Velculescu, Fiona Simpkins, and Sergey Medvedev
- Subjects
Cancer Research ,Oncology ,ARID1A ,DNA repair ,DNA damage ,Cancer cell ,Cancer research ,Cell cycle ,Biology ,Mitotic catastrophe ,Chromatin remodeling ,Chromatin - Abstract
Purpose: Clear-cell ovarian cancer (CCOC) is often resistant to standard chemotherapy and has limited treatment options in the advanced or recurrent setting. ARID1A is the most prevalent mutation, with approximately 50% of all CCOC harboring this mutation. ARID1A, a member of the SWI/SNF family, regulates transcription and has a major role in the repair of DNA lesions, directly facilitating DNA accessibility on the chromatin or indirectly by facilitating the functions of DNA repair proteins. We propose that CCOCs unique genomic alterations (e.g., ARID1A frameshift or nonsense mutations) will increase dependency on chromatin remodeling and DNA repair (e.g., ATR/CHK1/WEE1) pathways for survival. We hypothesize that combination of low-dose small-molecule inhibitors of the BET family (BRD4i) and the DNA damage repair pathway (ATR/CHK1/WEE1), will especially target ARID1A mutant cancer cells promoting mitotic catastrophe, apoptosis, and tumor regression, sparing normal cells. Method: We compared the sensitivity of BRD4i + ATRi/WEE1i combinations in ARID1A knockdown and knockout cells compared with isogenic parental lines. The effects on transcription, kinase signaling, cell cycle, apoptosis, and DNA damage were evaluated. RNAseq/ATACseq and Reverse Phase Protein Array (RPPA) were also performed. A CCOC preclinical drug development platform was established and BRD4i combinations were evaluated in ARID1A mutant and ARID1A wild-type (ARID1AMUT and ARID1AWT) PDX models. Results: Low-dose BRD4i in combination with DNA damage repair inhibitors, ATRi/WEE1i (BDR4i-ATRi or BRD4i-WEE1i), were synergistic in decreasing survival and colony formation with an increase in apoptosis in ARID1AMUT cells compared to ARID1AWT. BRD4i combinations caused a robust G1 arrest with a decrease pCDC6 levels along with an increase in DNA double-strand breaks in the ARID1AMUT or knockout cells. Combination BRD4i-ATRi showed significant tumor regression and increased overall survival compared to standard chemotherapy or monotherapy in several ARID1AMUT PDX models but not in an ARID1AWT PDX. Among BRD4i-DDRi combinations, BRD4i-ATRi is superior to BRD4i-WEE1i in terms of antitumor effect and drug tolerability. Conclusion: Our studies identify a novel drug combination targeting a genetic alteration (e.g., ARID1A) common in CCOC that is highly effective and tolerable. BRD4i in combination with ATRi or WEE1i was synergistic, decreasing survival, colony formation with an increase in DNA damage and apoptosis in ARID1AMUT models. Using our novel CCOC drug development PDX platform, we demonstrated that BRD4i-ATRi combination therapy is more effective than standard chemotherapy or monotherapy alone with acceptable toxicity in ARID1AMUTPDXs. This abstract is also being presented as Poster B24. Citation Format: Yasuto Kinose, Hyoung Kim, Haineng Xu, Sergey Medvedev, Erin George, Sarah Gitto, Margaret Whicker, Dorothy Hallberg, Lauren Schwartz, Gordon Mills, Victor E. Velculescu, Tian-Li Wang, Ronny Drapkin, Fiona Simpkins. Dual blockade of BRD4 and the ATR/WEE1 pathway exploits ARID1A loss in clear-cell ovarian cancers [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research; 2019 Sep 13-16, 2019; Atlanta, GA. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(13_Suppl):Abstract nr PR11.
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- 2020
31. Reply to Haffner et al.: DNA hypomethylation renders tumors more immunogenic
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Vipul Bhargava, Lauren Murphy, Ie Ming Shih, Karla R. Wiehagen, Meredith L. Stone, Reiner Strick, Huili Li, Michael J. Topper, Tian Li Wang, Chien Fu Hung, Stephen B. Baylin, Kurtis E. Bachman, Pamela L. Strissel, Katherine B. Chiappinelli, Cynthia A. Zahnow, Meghan Travers, Dimitrios Mathios, Glenn S. Cowley, and Michael Lim
- Subjects
0301 basic medicine ,Tumor microenvironment ,Multidisciplinary ,Chemistry ,medicine.drug_class ,Histone deacetylase inhibitor ,medicine.disease ,DNA methyltransferase ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,0302 clinical medicine ,030220 oncology & carcinogenesis ,DNA methylation ,Cancer cell ,medicine ,Cancer research ,Ovarian cancer ,Givinostat ,DNA hypomethylation - Abstract
In the letter by Haffner et al. (1), they report that seminoma cell-intrinsic DNA hypomethylation is associated with endogenous retroviral expression, an IFN response, and lymphocytic infiltration. Their data complement and support our recent therapeutic study in a mouse model of ovarian cancer (2) and support our observations that low doses of the DNA methyltransferase (DNMT) inhibitor azacytidine (AZA), in combination with the histone deacetylase inhibitor, givinostat, activate type 1 IFN signaling in ovarian (2) and lung (3) cancer cells to increase numbers and activation of immune cells in the tumor microenvironment and to increase sensitivity to the … [↵][1]5To whom correspondence may be addressed. Email: zahnoci{at}jhmi.edu or sbaylin{at}jhmi.edu. [1]: #xref-corresp-1-1
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- 2018
32. Methylomic analysis of ovarian cancers identifies tumor-specific alterations readily detectable in early precursor lesions
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Kentaro Nakayama, Shiou Fu Lin, Ting Tai Yen, Leslie Cope, Ryoichi Asaka, Thomas R. Pisanic, Ie Ming Shih, Tza-Huei Wang, Tian Li Wang, Pornpat Athamanolap, and Amanda Nickles Fader
- Subjects
0301 basic medicine ,Cancer Research ,Pathology ,medicine.medical_specialty ,Biopsy ,Article ,03 medical and health sciences ,0302 clinical medicine ,Ovarian carcinoma ,medicine ,Biomarkers, Tumor ,Humans ,Early Detection of Cancer ,Neoplasm Staging ,Ovarian Neoplasms ,medicine.diagnostic_test ,business.industry ,Gene Expression Profiling ,Case-control study ,Computational Biology ,Serous Tubal Intraepithelial Carcinoma ,Methylation ,DNA Methylation ,Immunohistochemistry ,Serous fluid ,030104 developmental biology ,medicine.anatomical_structure ,Oncology ,ROC Curve ,Genetic Loci ,030220 oncology & carcinogenesis ,Case-Control Studies ,DNA methylation ,CpG Islands ,Female ,Neoplasm Grading ,business ,Transcriptome ,Fallopian tube - Abstract
Purpose: High-grade serous ovarian carcinoma (HGSOC) typically remains undiagnosed until advanced stages when peritoneal dissemination has already occurred. Here, we sought to identify HGSOC-specific alterations in DNA methylation and assess their potential to provide sensitive and specific detection of HGSOC at its earliest stages. Experimental Design: MethylationEPIC genome-wide methylation analysis was performed on a discovery cohort comprising 23 HGSOC, 37 non-HGSOC malignant, and 36 histologically unremarkable gynecologic tissue samples. The resulting data were processed using selective bioinformatic criteria to identify regions of high-confidence HGSOC-specific differential methylation. Quantitative methylation-specific real-time PCR (qMSP) assays were then developed for 8 of the top-performing regions and analytically validated in a cohort of 90 tissue samples. Lastly, qMSP assays were used to assess and compare methylation in 30 laser-capture microdissected (LCM) fallopian tube epithelia samples obtained from cancer-free and serous tubal intraepithelial carcinoma (STIC) positive women. Results: Bioinformatic selection identified 91 regions of robust, HGSOC-specific hypermethylation, 23 of which exhibited an area under the receiver-operator curve (AUC) value ≥ 0.9 in the discovery cohort. Seven of 8 top-performing regions demonstrated AUC values between 0.838 and 0.968 when analytically validated by qMSP in a 90-patient cohort. A panel of the 3 top-performing genes (c17orf64, IRX2, and TUBB6) was able to perfectly discriminate HGSOC (AUC 1.0). Hypermethylation within these loci was found exclusively in LCM fallopian tube epithelia from women with STIC lesions, but not in cancer-free fallopian tubes. Conclusions: A panel of methylation biomarkers can be used to accurately identify HGSOC, even at precursor stages of the disease.
- Published
- 2018
33. Fallopian Tube Lesions in Women at High Risk for Ovarian Cancer: A Multicenter Study
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Betty J. May, Kala Visvanathan, Douglas A. Levine, Russell Vang, Robert J. Kurman, Vinita Parkash, Harvey A. Risch, Steven A. Narod, Tian Li Wang, Patricia Shaw, Robert A. Soslow, Alpana Kaushiva, Ie Ming Shih, and Asli Bahadirli-Talbott
- Subjects
0301 basic medicine ,Adult ,Cancer Research ,medicine.medical_specialty ,Salpingo-oophorectomy ,Article ,03 medical and health sciences ,0302 clinical medicine ,Ovarian carcinoma ,medicine ,Biomarkers, Tumor ,Prevalence ,Fallopian Tube Neoplasms ,Humans ,Prospective Studies ,Prospective cohort study ,Cystadenocarcinoma ,Medical History Taking ,Fallopian Tubes ,Retrospective Studies ,Gynecology ,BRCA2 Protein ,Ovarian Neoplasms ,business.industry ,BRCA1 Protein ,Ovary ,Age Factors ,Cancer ,Serous Tubal Intraepithelial Carcinoma ,Middle Aged ,medicine.disease ,Prognosis ,female genital diseases and pregnancy complications ,Cystadenocarcinoma, Serous ,Serous fluid ,030104 developmental biology ,medicine.anatomical_structure ,Oncology ,030220 oncology & carcinogenesis ,Disease Progression ,Female ,Ovarian cancer ,business ,Precancerous Conditions ,Fallopian tube - Abstract
The prognosis of women diagnosed with invasive high-grade serous ovarian carcinoma (HGSC) is poor. More information about serous tubal intraepithelial carcinoma (STIC) and serous tubal intraepithelial lesions (STIL), putative precursor lesions of HGSC, could inform prevention efforts. We conducted a multicenter study to identify risk/protective factors associated with STIC/STILs and characterize p53 signatures in the fallopian tube. The fallopian tubes and ovaries of 479 high-risk women ≥30 years of age who underwent bilateral risk-reducing salpingo-oophorectomy were reviewed for invasive cancer/STICs/STILs. Epidemiologic data was available for 400 of these women. In 105 women, extensive sampling of the tubes for STICs/STILs/p53 signatures were undertaken. Descriptive statistics were used to compare groups with and without lesions. The combined prevalence of unique tubal lesions [invasive serous cancer (n = 6) /STICs (n = 14)/STILs (n = 5)] was 6.3% and this was split equally among BRCA1 (3.0%) and BRCA2 mutation carriers (3.3%). A diagnosis of invasive cancer was associated with older age but no risk/protective factor was significantly associated with STICs/STILs. Extensive sampling identified double the number of STICs/STILs (11.9%), many p53 signatures (27.0%), and multiple lesions in 50% of the cases. Women with p53 signatures in the fimbria were older than women with signatures in the remaining tube (P = 0.03). STICs/STILs may not share the protective factors that are associated with HGSC. It is plausible that these factors are only associated with STICs that progress to HGSC. Having multiple lesions in the fimbria may be an important predictor of disease progression. Cancer Prev Res; 11(11); 697–706. ©2018 AACR.
- Published
- 2018
34. Notch in Ovarian Cancer
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Emily Gerry, Tian Li Wang, and Vivek Singh
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JAG1 ,endocrine system diseases ,Angiogenesis ,Serous carcinoma ,business.industry ,Notch signaling pathway ,medicine.disease ,Notch proteins ,Cancer stem cell ,Cancer research ,medicine ,HES1 ,business ,Ovarian cancer - Abstract
Ovarian cancers are malignancies for which improved therapeutic approaches are urgently needed. The development of chemoresistance in ovarian high-grade serous carcinoma is almost inevitable, and researchers are constantly seeking new pathways to target in order to improve the dismal survival rates of women diagnosed with this disease. The Notch pathway in ovarian cancer represents a promising subject for research into new ovarian cancer treatment modalities. Over the last 12 years, the major Notch proteins (Notch1 and NOTCH3), prominent Notch ligands (JAG1 and DLL4), and downstream proteins (Hes1 and DLGAP5) have begun to be studied in ovarian cancers. The roles of Notch in conferring chemoresistance and acting in angiogenesis have also been demonstrated. Additionally, GSI and DLL4 inhibitors as well as Notch antibodies continue to be explored in both clinical and nonclinical settings. It is clear that future studies are needed in order to translate the results from these preclinical studies into practice. Most importantly, it is crucial to demonstrate the safety and efficacy of Notch-based therapy in ovarian cancer patients. There is still much work to be done in examining the pathways and proteins with which Notch may be associated as well as in developing more specific and more effective means of inhibiting Notch pathway components.
- Published
- 2018
35. Epigenetic therapy activates type I interferon signaling in murine ovarian cancer to reduce immunosuppression and tumor burden
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Lauren Murphy, Meghan Travers, Stephen B. Baylin, Dimitrios Mathios, Glenn S. Cowley, Meredith L. Stone, Kurtis E. Bachman, Karla R. Wiehagen, Huili Li, Tian Li Wang, Michael Lim, Katherine B. Chiappinelli, Ie Ming Shih, Cynthia A. Zahnow, Vipul Bhargava, Chien Fu Hung, Pamela L. Strissel, Reiner Strick, and Michael J. Topper
- Subjects
0301 basic medicine ,Tumor microenvironment ,Multidisciplinary ,business.industry ,medicine.medical_treatment ,Immunosuppression ,medicine.disease ,Immune checkpoint ,03 medical and health sciences ,030104 developmental biology ,Immune system ,PNAS Plus ,Interferon ,Cancer research ,Medicine ,business ,Ovarian cancer ,CD8 ,Epigenetic therapy ,medicine.drug - Abstract
Ovarian cancer is the most lethal of all gynecological cancers, and there is an urgent unmet need to develop new therapies. Epithelial ovarian cancer (EOC) is characterized by an immune suppressive microenvironment, and response of ovarian cancers to immune therapies has thus far been disappointing. We now find, in a mouse model of EOC, that clinically relevant doses of DNA methyltransferase and histone deacetylase inhibitors (DNMTi and HDACi, respectively) reduce the immune suppressive microenvironment through type I IFN signaling and improve response to immune checkpoint therapy. These data indicate that the type I IFN response is required for effective in vivo antitumorigenic actions of the DNMTi 5-azacytidine (AZA). Through type I IFN signaling, AZA increases the numbers of CD45+ immune cells and the percentage of active CD8+ T and natural killer (NK) cells in the tumor microenvironment, while reducing tumor burden and extending survival. AZA also increases viral defense gene expression in both tumor and immune cells, and reduces the percentage of macrophages and myeloid-derived suppressor cells in the tumor microenvironment. The addition of an HDACi to AZA enhances the modulation of the immune microenvironment, specifically increasing T and NK cell activation and reducing macrophages over AZA treatment alone, while further increasing the survival of the mice. Finally, a triple combination of DNMTi/HDACi plus the immune checkpoint inhibitor α-PD-1 provides the best antitumor effect and longest overall survival, and may be an attractive candidate for future clinical trials in ovarian cancer.
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- 2017
36. Mevalonate Pathway Antagonist Suppresses Formation of Serous Tubal Intraepithelial Carcinoma and Ovarian Carcinoma in Mouse Models
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Jinghua Gu, Jianhua Xuan, Jin Gyoung Jung, Ie Ming Shih, Hiroyasu Kashima, Kala Visvanathan, Tian Li Wang, Ren-Chin Wu, Yusuke Kobayashi, Jen Chun Kuan, and Lori J. Sokoll
- Subjects
DNA Replication ,Cancer Research ,medicine.medical_specialty ,Statin ,Serous carcinoma ,medicine.drug_class ,Mevalonic Acid ,Antineoplastic Agents ,Biology ,Article ,Mice ,Ovarian tumor ,Cell Line, Tumor ,Ovarian carcinoma ,Internal medicine ,medicine ,Animals ,Humans ,Lovastatin ,Cell Proliferation ,Ovarian Neoplasms ,Cancer ,Serous Tubal Intraepithelial Carcinoma ,medicine.disease ,Disease Models, Animal ,Endocrinology ,Oncology ,Cancer research ,Female ,lipids (amino acids, peptides, and proteins) ,Tumor Suppressor Protein p53 ,Ovarian cancer ,Carcinoma in Situ ,Signal Transduction ,medicine.drug - Abstract
Purpose: Statins are among the most frequently prescribed drugs because of their efficacy and low toxicity in treating hypercholesterolemia. Recently, statins have been reported to inhibit the proliferative activity of cancer cells, especially those with TP53 mutations. Because TP53 mutations occur in almost all ovarian high-grade serous carcinoma (HGSC), we determined whether statins suppressed tumor growth in animal models of ovarian cancer. Experimental Design: Two ovarian cancer mouse models were used. The first one was a genetically engineered model, mogp-TAg, in which the promoter of oviduct glycoprotein-1 was used to drive the expression of SV40 T-antigen in gynecologic tissues. These mice spontaneously developed serous tubal intraepithelial carcinomas (STICs), which are known as ovarian cancer precursor lesions. The second model was a xenograft tumor model in which human ovarian cancer cells were inoculated into immunocompromised mice. Mice in both models were treated with lovastatin, and effects on tumor growth were monitored. The molecular mechanisms underlying the antitumor effects of lovastatin were also investigated. Results: Lovastatin significantly reduced the development of STICs in mogp-TAg mice and inhibited ovarian tumor growth in the mouse xenograft model. Knockdown of prenylation enzymes in the mevalonate pathway recapitulated the lovastatin-induced antiproliferative phenotype. Transcriptome analysis indicated that lovastatin affected the expression of genes associated with DNA replication, Rho/PLC signaling, glycolysis, and cholesterol biosynthesis pathways, suggesting that statins have pleiotropic effects on tumor cells. Conclusions: The above results suggest that repurposing statin drugs for ovarian cancer may provide a promising strategy to prevent and manage this devastating disease. Clin Cancer Res; 21(20); 4652–62. ©2015 AACR.
- Published
- 2015
37. Increased proliferation in atypical hyperplasia/endometrioid intraepithelial neoplasia of the endometrium with concurrent inactivation of ARID1A and PTEN tumour suppressors
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Tsui Lien Mao, Tian Li Wang, Felix Zeppernick, Yohan Suryo Rahmanto, Hiroshi Ogawa, Ayse Ayhan, Ie Ming Shih, and Ren-Chin Wu
- Subjects
PTEN ,Pathology ,medicine.medical_specialty ,tumour suppressor ,ARID1A ,proliferation ,Endometrium ,Atypical hyperplasia ,Pathology and Forensic Medicine ,Carcinoma ,medicine ,Gene silencing ,biology ,Original Articles ,endometrioid intraepithelial neoplasia ,medicine.disease ,atypical hyperplasia ,in vitro cell culture model ,medicine.anatomical_structure ,Cell culture ,immunohistochemistry ,biology.protein ,Cancer research ,Immunohistochemistry ,Original Article ,co‐silencing - Abstract
Uterine endometrioid carcinoma is the most common neoplastic disease in the female genital tract and develops from a common precursor lesion, atypical hyperplasia/endometrioid intraepithelial neoplasia (AH/EIN). Although the genomic landscape of endometrioid carcinoma has been recently revealed, the molecular alterations that contribute to tumour progression from AH/EIN to carcinoma remain to be elucidated. In this study, we used immunohistochemistry to determine if loss of expression of two of the most commonly mutated tumour suppressors in endometrioid carcinoma, PTEN and ARID1A, was associated with increased proliferation in AH/EIN. We found that 80 (70%) of 114 cases exhibited decreased or undetectable PTEN and 17 (15%) of 114 cases had focal loss of ARID1A staining. ARID1A loss was focal, while PTEN loss was diffuse, and all specimens with ARID1A loss had concurrent PTEN loss (p = 0.0003). Mapping the distribution of PTEN and ARID1A staining in the same specimens demonstrated that all AH/EIN areas with ARID1A loss were geographically nested within the areas of PTEN loss. A significant increase in the proliferative activity was observed in areas of AH/EIN with concurrent loss of PTEN and ARID1A compared to immediately adjacent AH/EIN areas showing only PTEN loss. In a cell culture system, co‐silencing of ARID1A and PTEN in human endometrial epithelial cells increased cellular proliferation to a greater degree than silencing either ARID1A or PTEN alone. These results suggest an essential gatekeeper role for ARID1A that prevents PTEN inactivation from promoting cellular proliferation in the transition of pre‐cancerous lesions to uterine endometrioid carcinoma.
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- 2015
38. Early and consistent overexpression of ADRM1 in ovarian high-grade serous carcinoma
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Deyin Xing, Ravi K. Anchoori, Jeffrey D. Seidman, Anna Yemelyanova, Rosie Jiang, Richard B.S. Roden, Jun Hamazaki, Tian Li Wang, and Shigeo Murata
- Subjects
0301 basic medicine ,medicine.medical_specialty ,endocrine system diseases ,ADRM1 ,Ovary ,Ovarian/fallopian tube cancer ,Biology ,lcsh:Gynecology and obstetrics ,Benzylidene Compounds ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,Internal medicine ,Ovarian carcinoma ,medicine ,Fallopian Tube Neoplasms ,Humans ,Proteasome inhibitor ,RNA, Messenger ,lcsh:RG1-991 ,RPN13 ,Aged ,Ovarian Neoplasms ,Membrane Glycoproteins ,Research ,Intracellular Signaling Peptides and Proteins ,Obstetrics and Gynecology ,Serous Tubal Intraepithelial Carcinoma ,Middle Aged ,medicine.disease ,female genital diseases and pregnancy complications ,Cystadenocarcinoma, Serous ,3. Good health ,030104 developmental biology ,medicine.anatomical_structure ,Endocrinology ,Oncology ,030220 oncology & carcinogenesis ,Fallopian tube cancer ,Cancer cell ,Cancer research ,Female ,Tumor Suppressor Protein p53 ,Ovarian cancer ,Proteasome Inhibitors ,STIC ,medicine.drug - Abstract
Background Ovarian carcinoma is highly dependent on the ubiquitin proteasome system (UPS), but its clinical response to treatment with the proteasome inhibitor bortezomib has been disappointing. This has driven exploration of alternate approaches to target the UPS in ovarian cancer. Recently, proteasome inhibitors targeting the 19S regulatory particle-associated RPN13 protein have been described, such as RA190. RPN13, which is encoded by ADRM1, facilitates the recognition by the proteasome of its polyubiquinated substrates. Inhibition of RPN13 produces a rapid, toxic accumulation of polyubiquitinated proteins in ovarian and other cancer cells, triggering apoptosis. Here, we sought to determine if RPN13 is available as a target in precursors of ovarian/fallopian tube cancer as well as all advanced cases, and the impact of increased ADRM1 gene copy number on sensitivity of ovarian cancer to RA190. Methods ADRM1 mRNA was quantified by RNAscope in situ hybridization and RPN13 protein detected by immunohistochemistry in high grade serous carcinoma (HGSC) of the ovary and serous tubal intraepithelial carcinoma (STIC). Amplification of ADRM1 and sensitivity to RA190 were determined in ovarian cancer cell lines. Results Here, we demonstrate that expression of ADRM1mRNA is significantly elevated in STIC and HGSC as compared to normal fallopian tube epithelium. ADRM1 mRNA and RPN13 were ubiquitously and robustly expressed in ovarian carcinoma tissue and cell lines. No correlation was found between ADRM1 amplification and sensitivity of ovarian cancer cell lines to RA190, but all were susceptible. Conclusions RPN13 can potentially be targeted by RA190 in both in situ and metastatic ovarian carcinoma. Ovarian cancer cell lines are sensitive to RA190 regardless of whether the ADRM1 gene is amplified. Electronic supplementary material The online version of this article (doi:10.1186/s13048-017-0347-y) contains supplementary material, which is available to authorized users.
- Published
- 2017
39. The emerging roles of ARID1A in tumor suppression
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Ie Ming Shih, Tian Li Wang, and Ren-Chin Wu
- Subjects
Cancer Research ,ARID1A ,Tumor suppressor gene ,Carcinogenesis ,Review ,Biology ,medicine.disease_cause ,Genomic Instability ,Chromatin remodeling ,Mutation Rate ,Neoplasms ,medicine ,Animals ,Humans ,Genes, Tumor Suppressor ,Transcription factor ,PI3K/AKT/mTOR pathway ,Pharmacology ,Nuclear Proteins ,SWI/SNF ,Chromatin ,DNA-Binding Proteins ,Oncology ,Mutation ,Cancer research ,Molecular Medicine ,Signal Transduction ,Transcription Factors - Abstract
ARID1A has emerged as a tumor suppressor gene, which is mutated in a broad spectrum of cancers, especially in those arising from ectopic or eutopic endometrium. As a subunit of SWI/SNF chromatin remodeler, ARID1A facilitates target-specific binding of SWI/SNF complexes to chromatin, thereby altering the accessibility of chromatin to a variety of nuclear factors. In human cancer, ARID1A possesses not only features of a gatekeeper, regulating cell cycle progression, but also features of a caretaker, preventing genomic instability. An increasing body of evidence suggests crosstalk between ARID1A and PI3K/Akt pathways, and between ARID1A and p53. In this review, we discuss the spectrum of ARID1A alterations in cancers, tumor suppression mechanisms of ARID1A, oncogenic pathways cooperating with ARID1A, and clinical implications of ARID1A mutation.
- Published
- 2014
40. Frequent somatic mutations of the telomerase reverse transcriptase promoter in ovarian clear cell carcinoma but not in other major types of gynaecological malignancy
- Author
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Ie Ming Shih, Ayse Ayhan, Blaise A. Clarke, Patricia Shaw, Kyu Rae Kim, Daichi Maeda, Michael Herman Chui, Ren-Chin Wu, Tian Li Wang, and Barry P. Rosen
- Subjects
Telomerase ,Somatic cell ,Biology ,medicine.disease_cause ,medicine.disease ,Pathology and Forensic Medicine ,Telomere ,Clear cell carcinoma ,Cancer research ,medicine ,Telomerase reverse transcriptase ,Carcinogenesis ,Ovarian cancer ,Clear cell - Abstract
Up-regulated expression of telomerase reverse transcriptase (TERT) and subsequent maintenance of telomere length are essential in tumour development. Recent studies have implicated somatic gain-of-function mutations at the TERT promoter as one of the mechanisms that promote transcriptional activation of TERT; however, it remains unclear whether this genetic abnormality is prevalent in gynaecological neoplasms. We performed mutational analysis in a total of 525 gynaecological cancers, and correlated TERT promoter mutations with clinicopathological features. With the exception of ovarian clear cell carcinomas, in which mutations were found in 37 (15.9%) of 233 cases, the majority of gynaecological malignancies were wild-type. TERT promoter mutation does not appear to be an early event during oncogenesis, as it was not detected in the contiguous endometriosis associated with ovarian clear cell carcinoma. Ovarian clear cell carcinoma cell lines with TERT promoter mutations exhibited higher TERT mRNA expression than those with wild-type sequences (p = 0.0238). TERT promoter mutation tended to be mutually exclusive with loss of ARID1A protein expression (p = 4.4 × 10(-9) ) and PIK3CA mutation (p = 0.0019) in ovarian clear cell carcinomas. No associations with disease-specific survival were observed for ovarian clear cell carcinoma. The above results, in conjunction with our previous report showing longer telomeres in ovarian clear cell carcinomas relative to other types of ovarian cancer, suggests that aberrations in telomere biology may play an important role in the pathogenesis of ovarian clear cell carcinoma.
- Published
- 2014
41. RNA-sequencing reveals immunotherapy targets in gynecological cancer
- Author
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Tian-Li Wang, Kunle Odunsi, Sean T. Glenn, Antonios Papanicolau-Sengos, Carl Morrison, Paul DePietro, Sarabjot Pabla, Mary Nesline, Felicia L. Lenzo, Ting-Tai Yen, Devin Dressman, and Jeffrey M. Conroy
- Subjects
Cancer Research ,endocrine system diseases ,business.industry ,medicine.medical_treatment ,RNA ,Microsatellite instability ,Immunotherapy ,medicine.disease ,Gynecological cancer ,female genital diseases and pregnancy complications ,Immune profiling ,03 medical and health sciences ,Serous fluid ,0302 clinical medicine ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,Medicine ,business ,030215 immunology - Abstract
8 Background: We performed microsatellite instability testing and expression immune profiling of endometrioid and serous carcinomas to identify immunotherapeutic targets and histologic attribute correlates. Methods: Microsatellite instability and RNA-seq of 395 immune-related genes were performed in 37 endometrioid and 53 serous carcinomas. Each gene was ranked against a reference population and subsequently compared to a separate clinical database reference of diverse neoplasms to perform Wilcoxon ranked sum test comparisons. Benjamin-Hochberg corrected p-values are reported. Results: VTCN1, IDO1, LAG3, and components of the mTOR/AKT1 pathway were overexpressed in both endometrioid and serous carcinomas compared to the clinical database population. CD276 and NT5E were uniquely overexpressed in the endometrioid set (Table). In the endometrioid set ADGRE5 was overexpressed in endometrioid primaries when compared to their metastatic counterparts. Serous metastases relatively overexpressed HLA-DPA1 compared to their primary counterparts. Ten of 26 endometrioid cancers with available microsatellite instability status were microsatellite unstable. All serous carcinomas were microsatellite stable. There was no correlation between microsatellite instability status and expression of any gene in either endometrioid or serous cancers. Conclusions: We identified high RNA-expression of VTCN1, IDO1, CD276, and LAG3 in endometrioid and serous carcinomas while NT5E (ADORA2A ligand) was found to be uniquely upregulated in endometrioid carcinomas. These markers may play a role in immune suppression in gynecological cancers and are potential immunotherapeutic targets. The upregulation of ADGRE5 in primary endometrioid cancers and upregulation of HLA-DPA1 in metastatic serous cancers suggests immunologic differences which may be relevant to the formation of metastatic disease.
- Published
- 2019
42. Loss of ARID1A Expression Correlates With Stages of Tumor Progression in Uterine Endometrioid Carcinoma
- Author
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Tsui Lien Mao, Kuan-Ting Kuo, Ie Ming Shih, Ayse Ayhan, Laura Ardighieri, Chen Hsuan Wu, and Tian Li Wang
- Subjects
Adult ,Pathology ,medicine.medical_specialty ,Time Factors ,endocrine system diseases ,ARID1A ,Biopsy ,Down-Regulation ,Biology ,Endometrium ,Article ,Atypical hyperplasia ,Pathology and Forensic Medicine ,Predictive Value of Tests ,Biomarkers, Tumor ,medicine ,Carcinoma ,Humans ,Uterine Neoplasm ,Aged ,Neoplasm Staging ,Aged, 80 and over ,Nuclear Proteins ,Middle Aged ,medicine.disease ,Immunohistochemistry ,female genital diseases and pregnancy complications ,DNA-Binding Proteins ,medicine.anatomical_structure ,Tumor progression ,Case-Control Studies ,Uterine Neoplasms ,Clear cell carcinoma ,Disease Progression ,Cancer research ,Female ,Surgery ,Neoplasm Grading ,Anatomy ,Carcinoma, Endometrioid ,Transcription Factors - Abstract
ARID1A is a recently identified tumor suppressor that functions in chromatin remodeling. Inactivating mutations of ARID1A and loss of its expression most frequently occur in ovarian clear cell carcinoma, ovarian endometrioid carcinoma, and uterine endometrioid carcinoma. In this study, we performed a detailed immunostaining analysis of ARID1A in 246 cases including benign endometrium and endometrioid carcinoma at different stages of progression. Special attention was paid to recording intratumoral heterogeneity of clonal loss of ARID1A immunoreactivity. All normal endometria (n= 51) and endometrial polyps (n= 14) retained ARID1A expression. Among complex atypical hyperplasia (n= 38), 16% exhibited clonal loss of ARID1A, but none showed complete loss. Among low-grade endometrioid carcinomas (n= 88), 25% exhibited complete loss and 24% exhibited clonal loss. In contrast, 44% of high-grade endometrioid carcinomas (n= 55) showed complete loss of ARID1A and 9% exhibited clonal loss. We found that 19 high-grade carcinomas also contained concurrent low-grade carcinomas. In the high-grade areas, 63% exhibited complete loss and 11% exhibited clonal loss, whereas in the low-grade areas, 37% exhibited complete loss and 42% clonal loss. In 5 of these 19 cases, progressive loss of ARID1A from retention or clonal loss to complete loss was observed between the low-grade and high-grade areas. Overall, the percentage of complete ARID1A loss increased from 0% in complex atypical hyperplasia, to 25% in low-grade endometrioid carcinoma, to 44% in high-grade endometrioid carcinoma. These findings suggest that loss of ARID1A expression, presumably due to mutation, plays an important role in tumor progression of uterine endometrioid carcinoma.
- Published
- 2013
43. Rsf-1, a chromatin remodelling protein, interacts with cyclin E1 and promotes tumour development
- Author
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Ming Tsung Lai, Jim Jinn-Chyuan Sheu, Bin Guan, Chun Hung Hua, Jung Hye Choi, Tian Li Wang, Ie Ming Shih, and Fuu Jen Tsai
- Subjects
Cyclin E1 ,Cyclin D1 ,Cyclin E ,biology ,Cyclin D ,Cyclin A ,Cyclin B ,biology.protein ,Cancer research ,Cyclin-dependent kinase complex ,Cyclin A2 ,Pathology and Forensic Medicine ,Cell biology - Abstract
Chromosome 11q13.5 containing RSF1 (HBXAP), a gene involved in chromatin remodeling, is amplified in several human cancers including ovarian carcinoma. Our previous studies demonstrated requirement of Rsf-1 for cell survival in cancer cells, which contributed to tumor progression; however, its role in tumorigenesis has not yet been elucidated. In this study, we co-immunoprecipitated proteins with Rsf-1 followed by nanoelectrospray mass spectrometry and identified cyclin E1, besides SNF2H, as one of the major Rsf-1 interacting proteins. Like RSF1, CCNE1 is frequently amplified in ovarian cancer, and both Rsf-1 and cyclin E1 were found co-upregulated in ovarian cancer tissues. Ectopic expression of Rsf-1 and cyclin E1 in non-tumorigenic TP53mut RK3E cells led to an increase in cellular proliferation and tumor formation by activating cyclin E1-associated kinase (CDK2). Tumorigenesis was not detected if either cyclin E1 or Rsf-1 was expressed, or they were expressed in a TP53wt background. Domain mapping showed that cyclin E1 interacted with the first 441 amino acids of Rsf-1. Ectopic expression of this truncated domain significantly suppressed G1/S-phase transition, cellular proliferation and tumor formation of RK3E-p53R175H/Rsf-1/cyclin E1 cells. The above findings suggest that Rsf-1 interacts and collaborates with cyclin E1 in neoplastic transformation and TP53 mutations are prerequisite for tumor-promoting functions of RSF/cyclin E1 complex.
- Published
- 2013
44. Abstract P4-08-07: Novel insight into the tumor 'flare' phenomenon and lapatinib resistance
- Author
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Shengli Zhao, Herbert Kim Lyerly, Tian Li Wang, Takuya Osada, Neil L. Spector, Leihua Liu, JA Piede, and Wenle Xia
- Subjects
Cancer Research ,Pathology ,medicine.medical_specialty ,Stromal cell ,business.industry ,Cancer ,Cell cycle ,medicine.disease ,Lapatinib ,Oncology ,Cell culture ,SKBR3 ,In vivo ,Cancer stem cell ,Cancer research ,medicine ,skin and connective tissue diseases ,business ,medicine.drug - Abstract
BACKGROUND: Resistance to lapatinib generally develops in approximately half of patients within one year of initiating treatment. When lapatinib is withdrawn, there is often a rapid progression of disease which is associated with high mortality rates. This “flare” phenomenon has also been observed upon discontinuation of other tyrosine kinase inhibitors (TKIs) in lung cancer, renal cell carcinoma, and gastrointestinal stromal tumors. METHODS: Using the HER2+ cell lines SKBR3 and BT474, we developed both in vitro and in vivo models demonstrating lapatinib resistance and the tumor “flare” phenomenon. Parental HER2+ cell lines were gradually exposed to increasing doses of lapatinib (100nM to 5uM) to develop a lapatinib-resistant form which was ultimately maintained in 1uM of lapatinib. Lapatinib-”released” cell lines were developed by removing lapatinib exposure from resistant cell lines and allowing the cells to grow for at least two weeks. Cell lines were grown in vitro using traditional monolayer cell culturing techniques and mammosphere technology. A comprehensive analysis of parental, lapatinib-resistant, and lapatinib-released cell lines was performed using microscopy, protein/phosphoprotein analysis, cell cycle, cancer stem cell markers by FACS, and invasion assays. Parental cells served as the control in all experiments. In vivo models were performed by injecting 10,000 cells of parental, resistant, and released cell lines in the mammary fat pads of SCID mice. Tumors were surgically resected after approximately sixty days and volume recorded. All experiments were repeated three times and calculated for statistical significance. RESULTS: Cell cycle analysis and proliferation assays demonstrate that lapatinib-resistant and released cell lines continue to proliferate despite the addition of 1uM lapatinib. Released cells also demonstrate less of a response to retreatment with lapatinib. Tumor volume of lapatinib-released cell lines were significantly larger than parental and resistant counterparts [parental: 39.9mm3, SD 9.48; resistant: 71.8mm3, SD 62.33; released: 943.4mm3, SD 100.1] and this difference was statistically significant (p < 0.01). After three series of passages, mammosphere forming efficiency (MFE) was higher among lapatinib-released cell lines [parental: 1.03%; resistant: 1.93% released: 7.23%] and was statistically significant (p CONCLUSIONS: We demonstrate the first in vitro and in vivo models of the tumor “flare” phenomenon using HER2+ breast cancer cell lines. This model demonstrates that lapatinib-resistant cells released from the exposure to lapatinib are propelled into a unique and more aggressive phenotype. This model has potential to elucidate new mechanisms of resistance to TKI therapy and provide novel preclinical data that may help in the understanding of disease progression. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-08-07.
- Published
- 2012
45. Functional Analysis of In-frame Indel ARID1A Mutations Reveals New Regulatory Mechanisms of Its Tumor Suppressor Functions
- Author
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Ie Ming Shih, Min Gao, Tian Li Wang, Bin Guan, and Chen Hsuan Wu
- Subjects
Regulation of gene expression ,Genetics ,0303 health sciences ,Cancer Research ,Nonsense mutation ,Biology ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,lcsh:RC254-282 ,03 medical and health sciences ,XPO1 ,0302 clinical medicine ,030220 oncology & carcinogenesis ,Nuclear protein ,Indel ,Nuclear export signal ,Transcription factor ,INDEL Mutation ,030304 developmental biology - Abstract
AT-rich interactive domain 1A (ARID1A) has emerged as a new tumor suppressor in which frequent somatic mutations have been identified in several types of human cancers. Although most ARID1A somatic mutations are frame-shift or nonsense mutations that contribute to mRNA decay and loss of protein expression, 5% of ARID1A mutations are in-frame insertions or deletions (indels) that involve only a small stretch of peptides. Naturally occurring in-frame indel mutations provide unique and useful models to explore the biology and regulatory role of ARID1A. In this study, we analyzed indel mutations identified in gynecological cancers to determine how these mutations affect the tumor suppressor function of ARID1A. Our results demonstrate that all in-frame mutants analyzed lost their ability to inhibit cellular proliferation or activate transcription of CDKN1A, which encodes p21, a downstream effector of ARID1A. We also showed that ARID1A is a nucleocytoplasmic protein whose stability depends on its subcellular localization. Nuclear ARID1A is less stable than cytoplasmic ARID1A because ARID1A is rapidly degraded by the ubiquitin-proteasome system in the nucleus. In-frame deletions affecting the consensus nuclear export signal reduce steady-state protein levels of ARID1A. This defect in nuclear exportation leads to nuclear retention and subsequent degradation. Our findings delineate a mechanism underlying the regulation of ARID1A subcellular distribution and protein stability and suggest that targeting the nuclear ubiquitin-proteasome system can increase the amount of the ARID1A protein in the nucleus and restore its tumor suppressor functions.
- Published
- 2012
46. NAC1 Is an Actin-Binding Protein That Is Essential for Effective Cytokinesis in Cancer Cells
- Author
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Tian Li Wang, Michelle M. Thiaville, Denis Wirtz, Ie Ming Shih, Natini Jinawath, Kai Lee Yap, Stephanie I. Fraley, Kentaro Nakayama, and Jian-Long Wang
- Subjects
Cancer Research ,Arp2/3 complex ,macromolecular substances ,Septin ,Article ,Profilins ,Neoplasms ,Humans ,Actin-binding protein ,Actin ,Cytokinesis ,biology ,Actin monomer binding ,Actins ,Neoplasm Proteins ,Cell biology ,Repressor Proteins ,HEK293 Cells ,Phenotype ,Oncology ,Profilin ,Cancer cell ,MCF-7 Cells ,biology.protein ,HeLa Cells - Abstract
NAC1 is a transcriptional corepressor protein that is essential to sustain cancer cell proliferation and migration. However, the underlying molecular mechanisms of NAC1 function in cancer cells remain unknown. In this study, we show that NAC1 functions as an actin monomer–binding protein. The conserved BTB protein interaction domain in NAC1 is the minimal region for actin binding. Disrupting NAC1 complex function by dominant-negative or siRNA strategies reduced cell retraction and abscission during late-stage cytokinesis, causing multinucleation in cancer cells. In Nac1-deficient murine fibroblasts, restoring NAC1 expression was sufficient to partially avert multinucleation. We found that siRNA-mediated silencing of the actin-binding protein profilin-1 in cancer cells caused a similar multinucleation phenotype and that NAC1 modulated the binding of actin to profillin-1. Taken together, our results indicate that the NAC1/actin/profilin-1 complex is crucial for cancer cell cytokinesis, with a variety of important biologic and clinical implications. Cancer Res; 72(16); 4085–96. ©2012 AACR.
- Published
- 2012
47. Endocervical-type Mucinous Borderline Tumors are Related to Endometrioid Tumors Based on Mutation and Loss of Expression of ARID1A
- Author
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Chen Hsuan Wu, Tsui Lien Mao, Ayse Ayhan, Russell Vang, Robert J. Kurman, Tian Li Wang, and Ie Ming Shih
- Subjects
Adult ,Pathology ,medicine.medical_specialty ,endocrine system diseases ,Endometriosis ,Polymerase Chain Reaction ,Article ,Pathology and Forensic Medicine ,Immunophenotyping ,Papillary Cystadenoma ,Carcinoma ,medicine ,Humans ,Genes, Tumor Suppressor ,Aged ,Retrospective Studies ,Aged, 80 and over ,Ovarian Neoplasms ,Endometrioid tumor ,business.industry ,Genetic Variation ,Nuclear Proteins ,Obstetrics and Gynecology ,DNA, Neoplasm ,Middle Aged ,medicine.disease ,Immunohistochemistry ,female genital diseases and pregnancy complications ,DNA-Binding Proteins ,Serous fluid ,Cancer research ,Female ,business ,Carcinoma, Endometrioid ,Clear cell ,Transcription Factors - Abstract
Nongastrointestinal-type mucinous borderline tumors have been described as displaying endocervical and serous differentiation and hence have been termed "endocervical-type" mucinous borderline tumors, "mixed-epithelial papillary cystadenoma of borderline malignancy of mullerian type," or "atypical proliferative seromucinous tumors." A striking feature of these tumors is their frequent association with endometriosis, which has been reported in a third to a half of cases. This is an unusual finding, as pure endocervical and serous tumors are not usually associated with endometriosis. ARID1A is a recently identified tumor suppressor, which frequently loses its expression and is mutated in endometrium-related carcinomas including ovarian clear cell, ovarian endometrioid, and uterine endometrioid carcinomas. Although ARID1A mutations and their expression have been studied in gynecologic cancer, the expression pattern of ARID1A has not been investigated in ovarian atypical proliferative (borderline) tumors. In this study, we analyzed ARID1A expression in serous, gastrointestinal-type and endocervical-type (seromucinous) mucinous, and endometrioid atypical proliferative (borderline) tumors using immunohistochemistry and performed mutational analysis in selected cases. We observed loss of ARID1A staining in 8 (33%) of 24 seromucinous tumors. In contrast, ARID1A staining was retained in all the other 32 tumors except in 1 endometrioid tumor (P
- Published
- 2012
48. Defining NOTCH3 Target Genes in Ovarian Cancer
- Author
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Jianhua Xuan, Tian Li Wang, Michelle M. Thiaville, Xu Chen, Ie Ming Shih, Alexander Stoeck, Li Chen, and Min Gao
- Subjects
Transcriptome ,Regulation of gene expression ,Cancer Research ,Oncology ,Cancer cell ,Notch signaling pathway ,Gene silencing ,Biology ,Cell cycle ,Gene ,Chromatin immunoprecipitation ,Molecular biology ,Cell biology - Abstract
NOTCH3 gene amplification plays an important role in the progression of many ovarian and breast cancers, but the targets of NOTCH3 signaling are unclear. Here, we report the use of an integrated systems biology approach to identify direct target genes for NOTCH3. Transcriptome analysis showed that suppression of NOTCH signaling in ovarian and breast cancer cells led to downregulation of genes in pathways involved in cell-cycle regulation and nucleotide metabolism. Chromatin immunoprecipitation (ChIP)-on-chip analysis defined promoter target sequences, including a new CSL binding motif (N1) in addition to the canonical CSL binding motif, that were occupied by the NOTCH3/CSL transcription complex. Integration of transcriptome and ChIP-on-chip data showed that the ChIP target genes overlapped significantly with the NOTCH-regulated transcriptome in ovarian cancer cells. From the set of genes identified, we showed that the mitotic apparatus organizing protein DLGAP5 (HURP/DLG7) was a critical target. Both the N1 motif and the canonical CSL binding motif were essential to activate DLGAP5 transcription. DLGAP5 silencing in cancer cells suppressed tumorigenicity and inhibited cellular proliferation by arresting the cell cycle at the G2–M phase. In contrast, enforced expression of DLGAP5 partially counteracted the growth inhibitory effects of a pharmacologic or RNA interference–mediated NOTCH inhibition in cancer cells. Our findings define direct target genes of NOTCH3 and highlight the role of DLGAP5 in mediating the function of NOTCH3. Cancer Res; 72(9); 2294–303. ©2012 AACR.
- Published
- 2012
49. Overexpression of a Chromatin Remodeling Factor, RSF-1/HBXAP, Correlates with Aggressive Oral Squamous Cell Carcinoma
- Author
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Fu-Min Fang, Tian Li Wang, I-Wen Chiu, Ie Ming Shih, Chien-Feng Li, Chih-Mei Chen, Jim Jinn-Chyuan Sheu, Hsuan-Ying Huang, Fuu Jen Tsai, and Ming-Tsong Lai
- Subjects
Adult ,Male ,Pathology ,medicine.medical_specialty ,Blotting, Western ,Chromatin Remodeling Factor ,Kaplan-Meier Estimate ,Biology ,Chromatin remodeling ,Pathology and Forensic Medicine ,Metastasis ,Biomarkers, Tumor ,Carcinoma ,medicine ,Humans ,RNA, Small Interfering ,Aged ,Proportional Hazards Models ,Aged, 80 and over ,Reverse Transcriptase Polymerase Chain Reaction ,Cell growth ,Nuclear Proteins ,Cancer ,Regular Article ,Middle Aged ,Cell cycle ,Chromatin Assembly and Disassembly ,Prognosis ,medicine.disease ,Immunohistochemistry ,Up-Regulation ,Chromatin ,stomatognathic diseases ,Tissue Array Analysis ,Carcinoma, Squamous Cell ,Trans-Activators ,Cancer research ,Female ,Mouth Neoplasms - Abstract
RSF-1, also known as hepatitis B X-antigen associated protein (HBXAP), is a subunit of an ISWI chromatin remodeling complex, remodeling and spacing factor (RSF). Recent studies have provided new evidence that chromatin remodeling participates in the pathogenesis of neoplastic diseases by altering cell cycle regulation and gene expression. In this report, we studied the biological roles of RSF-1 in oral squamous cell carcinoma (OSCC), a highly invasive neoplastic disease. Based on IHC and quantitative real-time PCR, we demonstrated that RSF-1 expression could be detected in the majority of OSCC cases, and the levels were significantly higher in OSCC cells than in their normal counterparts. Moreover, expression levels of RSF-1 significantly correlated with the presence of angiolymphatic invasion, abnormal mitoses, metastasis, tumor recurrence, and advanced stage disease at presentation. Univariate and multivariate analyses showed a significant association of RSF-1 overexpression and worse overall survival in OSCC patients. RSF-1 knockdown remarkably decreased cellular proliferation and induced apoptosis in OSCC cells with high RSF-1 expression levels, but not in those without. Taken together, our results suggest that RSF-1 up-regulation is associated with several clinicopathological features of disease aggressiveness in OSCC patients, and RSF-1 plays an important role in maintaining cellular growth and survival in OSCC.
- Published
- 2011
50. Somatic Mutations of PPP2R1A in Ovarian and Uterine Carcinomas
- Author
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Elisabetta Kuhn, Sian Jones, Ie Ming Shih, Tsui Lien Mao, Tian Li Wang, Robert J. Kurman, Kuan Tin Kuo, Pradeep K. Panuganti, and Victor E. Velculescu
- Subjects
Enzyme complex ,endocrine system diseases ,Somatic cell ,Short Communication ,DNA Mutational Analysis ,Biology ,medicine.disease_cause ,Pathology and Forensic Medicine ,Exon ,Carcinoma ,medicine ,Humans ,Protein Phosphatase 2 ,Codon ,Uterine Neoplasm ,Ovarian Neoplasms ,Mutation ,Exons ,medicine.disease ,Adenocarcinoma, Mucinous ,Immunohistochemistry ,female genital diseases and pregnancy complications ,Gene Expression Regulation, Neoplastic ,Serous fluid ,Uterine Neoplasms ,Cancer research ,Adenocarcinoma ,Female ,Adenocarcinoma, Clear Cell - Abstract
Exome sequencing of ovarian clear-cell carcinoma has identified somatic mutations in PPP2R1A, a subunit of protein phosphatase 2A. The present study was performed to determine the frequency of PPP2R1A mutations in exon 5, which harbors previously reported mutation hot spots, and adjacent exon 6, in 209 ovarian and 56 uterine tumors of various histologic subtypes. PPP2R1A mutations were demonstrated in 10 of 110 type I ovarian tumors (9.1%) including low-grade serous, low-grade endometrioid, clear-cell, and mucinous carcinomas. In contrast, none of 71 type II ovarian (high-grade serous) carcinomas exhibited PPP2R1A mutations. Moreover, PPP2R1A mutations were observed in 2 of 30 type I uterine (endometrioid) carcinomas (6.7%) and 5 of 26 type II uterine (serous) carcinomas (19.2%). Of the 18 mutations, 13 affected the R182 or 183, and there were 5 novel mutations including 3 involving S256, 1 involving W257, and 1 involving P179. All mutations were located in the α-helix repeats near the interface between the A subunit and the regulatory B subunit of the enzyme complex. These data provide new evidence that PPP2R1A somatic mutations occur in certain types of uterine and ovarian neoplastic lesions, especially uterine serous carcinomas, and suggest that mutation of PPP2R1A may participate in the pathogenesis of ovarian type I and uterine type II carcinomas.
- Published
- 2011
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