70 results on '"Ylaya K"'
Search Results
2. Over-expression of BMI1 is associated with favorable prognosis in cervical cancer
- Author
-
Kim, B. W., Cho, H., Ylaya, K., Chung, J. -Y, Stephen Hewitt, and Kim, J. -H
3. Evidence of SARS-CoV-2-specific T-cell-mediated myocarditis in a MIS-A case
- Author
-
Vannella, KM, primary, Oguz, C, additional, Stein, SR, additional, Pittaluga, S, additional, Dikoglu, E, additional, Kanwal, A, additional, Ramelli, SC, additional, Briese, T, additional, Su, L, additional, Wu, X, additional, Ramos-Benitez, MJ, additional, Perez-Valencia, LJ, additional, Babyak, A, additional, Cha, NR, additional, Chung, JY, additional, Ylaya, K, additional, Madathil, RJ, additional, Saharia, KK, additional, Scalea, TM, additional, Tran, QK, additional, Herr, DL, additional, Kleiner, DE, additional, Hewitt, SM, additional, Notarangelo, LD, additional, Grazioli, A, additional, and Chertow, DS, additional
- Full Text
- View/download PDF
4. Identification of unique expression signatures and therapeutic targets in esophageal squamous cell carcinoma
- Author
-
Yan Wusheng, Shih Joanna H, Rodriguez-Canales Jaime, Tangrea Michael A, Ylaya Kris, Hipp Jason, Player Audrey, Hu Nan, Goldstein Alisa M, Taylor Philip R, Emmert-Buck Michael R, and Erickson Heidi S
- Subjects
Medicine ,Biology (General) ,QH301-705.5 ,Science (General) ,Q1-390 - Abstract
Abstract Background Esophageal squamous cell carcinoma (ESCC), the predominant histological subtype of esophageal cancer, is characterized by high mortality. Previous work identified important mRNA expression differences between normal and tumor cells; however, to date there are limited ex vivo studies examining expression changes occurring during normal esophageal squamous cell differentiation versus those associated with tumorigenesis. In this study, we used a unique tissue microdissection strategy and microarrays to measure gene expression profiles associated with cell differentiation versus tumorigenesis in twelve cases of patient-matched normal basal squamous epithelial cells (NB), normal differentiated squamous epithelium (ND), and squamous cell cancer. Class comparison and pathway analysis were used to compare NB versus tumor in a search for unique therapeutic targets. Results As a first step towards this goal, gene expression profiles and pathways were evaluated. Overall, ND expression patterns were markedly different from NB and tumor; whereas, tumor and NB were more closely related. Tumor showed a general decrease in differentially expressed genes relative to NB as opposed to ND that exhibited the opposite trend. FSH and IgG networks were most highly dysregulated in normal differentiation and tumorigenesis, respectively. DNA repair pathways were generally elevated in NB and tumor relative to ND indicating involvement in both normal and pathological growth. PDGF signaling pathway and 12 individual genes unique to the tumor/NB comparison were identified as therapeutic targets, and 10 associated ESCC gene-drug pairs were identified. We further examined the protein expression level and the distribution patterns of four genes: ODC1, POSTN, ASPA and IGF2BP3. Ultimately, three genes (ODC1, POSTN, ASPA) were verified to be dysregulated in the same pattern at both the mRNA and protein levels. Conclusions These data reveal insight into genes and molecular pathways mediating ESCC development and provide information potentially useful in designing novel therapeutic interventions for this tumor type.
- Published
- 2012
- Full Text
- View/download PDF
5. Membranous expression of Her3 is associated with a decreased survival in head and neck squamous cell carcinoma
- Author
-
Hong Seung-Mo, Moskaluk Christopher A, Ylaya Kris, Cho Hanbyoul, Chung Joon-Yong, Xie Ran, Takikita Mikiko, and Hewitt Stephen M
- Subjects
Medicine - Abstract
Abstract Background Head and neck squamous cell carcinoma (HNSCC) still remains a lethal malignancy benefiting from the identification of the new target for early detection and/or development of new therapeutic regimens based on a better understanding of the biological mechanism for treatment. The overexpression of Her2 and Her3 receptors have been identified in various solid tumors, but its prognostic relevance in HNSCC remains controversial. Methods Three hundred eighty-seven primary HNSCCs, 20 matching metasis and 17 recurrent HNSCCs were arrayed into tissue microarrays. The relationships between Her2 and Her3 protein expression and clinicopathological parameters/survival of HNSCC patients were analyzed with immunohistochemistry. Results Her3 is detected as either a cytoplasmic or a membranous dominant expression pattern whereas Her2 expression showed uniform membranous form. In primary tumor tissues, high membranous Her2 expression level was found in 104 (26.9%) cases while positive membranous and cytoplasmic Her3 expression was observed in 34 (8.8%) and 300 (77.5%) samples, respectively. Membranous Her2 expression was significantly associated with histological grade (P = 0.021), as grade 2 tumors showed the highest positive expression. Membranous Her3 over-expression was significantly prevalent in metastatic tissues compared to primary tumors (P = 0.003). Survival analysis indicates that membranous Her3 expression is significantly associated with worse overall survival (P = 0.027) and is an independent prognostic factor in multivariate analysis (hazard ratio, 1.51; 95% confidence interval, 1.01-2.23; P = 0.040). Conclusions These results suggest that membranous Her3 expression is strongly associated with poor prognosis of patients with HNSCC and is a potential candidate molecule for targeted therapy.
- Published
- 2011
- Full Text
- View/download PDF
6. Expression of POU2F3 Transcription Factor and POU2AF2, POU2F3 Coactivator, in Tuft Cell-like Carcinoma and Other Tumors.
- Author
-
Kaczorowski M, Ylaya K, Chłopek M, Lasota J, and Miettinen M
- Abstract
Epithelial chemosensory cells in hollow organs, also known as tuft cells, were implicated in tumorigenesis, including a tuft cell-like small cell lung carcinoma. Expression of the POU2F3 transcription factor is a marker of tuft cell lineage. However, tuft cell development, differentiation, and proliferation are controlled by the expression of the complex formed by POU2F3 and POU2AF2 or POU2AF3 transcriptional coactivators. A cohort of epithelial (n=6064) and mesenchymal/neuroectodermal (n=2730) tumors was screened for POU2F3 expression by immunohistochemistry. Variable immunoreactivity ranging from diffuse to scattered positive cells was found in ∼12.4% of epithelial and 4.6% of mesenchymal/neuroectodermal tumors. Cases with predominantly diffuse or patchy POU2F3 positivity representing various types of malignant tumors (n=43) were selected for further study, including POU2AF2 immunohistochemistry. Thirteen of 15 tumors with neuroendocrine differentiation originating from the lung, colon, head and neck, skin, and bladder revealed diffuse POU2F3 positivity. Most of those tumors (n=9) co-expressed POU2AF2, usually extensively. Seven squamous and basal cell carcinomas from the oral cavity, skin, lung, and thymus with diffuse POU2F3 immunostaining except one, lacked POU2AF2 expression. Other variably POU2F3-positive carcinomas (n=13) from the colon, pancreas, liver, kidney, testis, endometrium, ovary, and breast lacked POU2AF2 immunoreactivity. All POU2F3-positive mesenchymal and neuroectodermal tumors (n=8), including synovial sarcoma, solitary fibrous tumor, glioblastoma, Wilms tumor, and melanoma were POU2AF2-negative. POU2F3 expression is a highly sensitive but nonspecific indicator of tuft cell differentiation. Co-expression of POU2F3 and POU2AF2 appears to be a more specific marker, although it may not pinpoint tumors driven by the POU2F3-POU2AF3 complex., Competing Interests: Conflicts of Interest and Source of Funding: Supported as a part of NCI’s intramural research program. The authors have disclosed that they have no significant relationships with, or financial interest in, any commercial companies pertaining to this article., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
7. Immunohistochemical Evaluation of Schlafen 11 (SLFN11) Expression in Cancer in the Search of Biomarker-Informed Treatment Targets: A Study of 127 Entities Represented by 6658 Tumors.
- Author
-
Kaczorowski M, Ylaya K, Chłopek M, Taniyama D, Pommier Y, Lasota J, and Miettinen M
- Abstract
Schlafen 11 (SLFN11), a DNA/RNA helicase, acts as a regulator of cellular response to replicative stress and irreversibly triggers replication block and cell death. Several preclinical in vitro studies and clinical trials established that SLFN11 expression predicts outcomes in patients with advanced cancer treated with DNA-damaging chemotherapeutics and more recently with poly(ADP-ribose) polymerase inhibitors. SLFN11 expression status remains unknown in many cancer types, especially in mesenchymal tumors. This study evaluated a cohort of well characterized 3808 epithelial and 2850 mesenchymal and neuroectodermal tumors for SLFN11 expression using immunohistochemistry. Nuclear SLFN11 expression was rare in some of the most common carcinomas, for example, hepatocellular (1%), prostatic (2%), colorectal (5%), or breast (16%) cancers. In contrast, other epithelial tumors including mesotheliomas (92%), clear cell renal cell carcinomas (79%), small cell lung cancers (76%), squamous cell carcinomas of the tonsil (89%) and larynx (71%), or ovarian serous carcinomas (69%) were mostly SLFN11-positive. Compared with epithelial malignancies, SLFN11 expression was overall higher in neuroectodermal and mesenchymal tumors. Most positive entities included desmoplastic small round cell tumor (100%), Ewing sarcoma (92%), undifferentiated sarcoma (92%), solitary fibrous tumor (91%), dedifferentiated liposarcoma (89%), synovial sarcoma (86%), and malignant peripheral nerve sheath tumor (85%). Also, this study identifies tumors with potentially worse response to DNA-damaging drugs including antibody drug conjugates due to the absence of SLFN11 expression. Such entities may benefit from alternative treatments or strategies to overcome SLFN11 deficiency-related drug resistance. Our approach and results should serve as a foundation for future biomarker-associated clinical trials., Competing Interests: Conflicts of Interest and Source of Funding: The authors have disclosed that they have no significant relationships with, or financial interest in, any commercial companies pertaining to this article., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
8. Glypican-3 deficiency in liver cancer upregulates MAPK/ERK pathway but decreases cell proliferation.
- Author
-
Chung JY, Lee W, Lee OW, Ylaya K, Nambiar D, Sheehan-Klenk J, Fayn S, Hewitt SM, Choyke PL, and Escorcia FE
- Abstract
Glypican-3 (GPC3) is overexpressed in hepatocellular carcinomas and hepatoblastomas and represents an important therapeutic target but the biologic importance of GPC3 in liver cancer is unclear. To date, there are limited data characterizing the biological implications of GPC3 knockout (KO) in liver cancers that intrinsically express this target. Here, we report on the development and characterization of GPC3-KO liver cancer cell lines and compare to them to parental lines. GPC3-KO variants were established in HepG2 and Hep3B liver cancer cell lines using a lentivirus-mediated CRISPR/Cas9 system. We assessed the effects of GPC3 deficiency on oncogenic properties in vitro and in murine xenograft models. Downstream cellular signaling pathway changes induced by GPC3 deficiency were examined by RNAseq and western blot. To confirm the usefulness of the models for GPC3-targeted drug development, we evaluated the target engagement of a GPC3-selective antibody, GC33, conjugated to the positron-emitting zirconium-89 (
89 Zr) in subcutaneous murine xenografts of wild type (WT) and KO liver cancer cell lines. Deletion of GPC3 significantly reduced liver cancer cell proliferation, migration, and invasion compared to the parental cell lines. Additionally, the tumor growth of GPC3-KO liver cancer xenografts was significantly slower compared with control xenografts. RNA sequencing analysis also showed GPC3-KO resulted in a reduction in the expression of genes associated with cell cycle regulation, invasion, and migration. Specifically, we observed the downregulation of components in the AKT/NFκB/WNT signaling pathways and of molecules related to cell cycle regulation with GPC3-KO. In contrast, pMAPK/ERK1/2 was upregulated, suggesting an adaptive compensatory response. KO lines demonstrated increased sensitivity to ERK (GDC09994), while AKT (MK2206) inhibition was more effective in WT lines. Using antibody-based positron emission tomography (immunoPET) imaging, we confirmed that89 Zr-GC33 accumulated exclusively in GPC3-expression xenografts but not in GPC3-KO xenografts with high tumor uptake and tumor-to-liver signal ratio. We show that GPC3-KO liver cancer cell lines exhibit decreased tumorigenicity and altered signaling pathways, including upregulated pMAPK/ERK1/2, compared to parental lines. Furthermore, we successfully distinguished between GPC3+ and GPC3- tumors using the GPC3-targeted immunoPET imaging agent, demonstrating the potential utility of these cell lines in facilitating GPC3-selective drug development., Competing Interests: None., (AJCR Copyright © 2024.)- Published
- 2024
- Full Text
- View/download PDF
9. Spatial proximity of tumor-immune interactions predicts patient outcome in hepatocellular carcinoma.
- Author
-
Maestri E, Kedei N, Khatib S, Forgues M, Ylaya K, Hewitt SM, Wang L, Chaisaingmongkol J, Ruchirawat M, Ma L, and Wang XW
- Subjects
- Humans, Ecosystem, Prognosis, Gene Expression Profiling, Tumor Microenvironment, CD8-Positive T-Lymphocytes, Carcinoma, Hepatocellular pathology, Liver Neoplasms pathology
- Abstract
Background and Aims: The fitness and viability of a tumor ecosystem are influenced by the spatial organization of its cells. We aimed to study the structure, architecture, and cell-cell dynamics of the heterogeneous liver cancer tumor microenvironment using spatially resolved multiplexed imaging., Approach and Results: We performed co-detection by indexing multiplexed immunofluorescence imaging on 68 HCC biopsies from Thai patients [(Thailand Initiative in Genomics and Expression Research for Liver Cancer (TIGER-LC)] as a discovery cohort, and then validated the results in an additional 190 HCC biopsies from Chinese patients [Liver Cancer Institute (LCI)]. We segmented and annotated 117,270 and 465,632 cells from the TIGER-LC and LCI cohorts, respectively. We observed 4 patient groups of TIGER-LC (IC1, IC2, IC3, and IC4) with distinct tumor-immune cellular interaction patterns. In addition, patients from IC2 and IC4 had much better overall survival than those from IC1 and IC3. Noticeably, tumor and CD8 + T-cell interactions were strongly enriched in IC2, the group with the best patient outcomes. The close proximity between the tumor and CD8 + T cells was a strong predictor of patient outcome in both the TIGER-LC and the LCI cohorts. Bulk transcriptomic data from 51 of the 68 HCC cases were used to determine tumor-specific gene expression features of our classified subtypes. Moreover, we observed that the presence of immune spatial neighborhoods in HCC as a measure of overall immune infiltration is linked to better patient prognosis., Conclusions: Highly multiplexed imaging analysis of liver cancer reveals tumor-immune cellular heterogeneity within spatial contexts, such as tumor and CD8 + T-cell interactions, which may predict patient survival., (Copyright © 2023 American Association for the Study of Liver Diseases.)
- Published
- 2024
- Full Text
- View/download PDF
10. ATOH1, TFAP2B, and CEACAM6 as Immunohistochemical Markers to Distinguish Merkel Cell Carcinoma and Small Cell Lung Cancer.
- Author
-
Vilasi SM, Nguyen J, Wang CJ, Miao L, Daily K, Eid M, Song JS, Jiang H, Ylaya K, Busam KJ, Gaiser MR, Hewitt SM, and Brownell I
- Abstract
Merkel cell carcinoma (MCC) and small cell lung cancer (SCLC) can be histologically similar. Immunohistochemistry (IHC) for cytokeratin 20 (CK20) and thyroid transcription factor 1 (TTF-1) are commonly used to differentiate MCC from SCLC; however, these markers have limited sensitivity and specificity. To identify new diagnostic markers, we performed differential gene expression analysis on transcriptome data from MCC and SCLC tumors. Candidate markers included atonal BHLH transcription factor 1 (ATOH1) and transcription factor AP-2β (TFAP2B) for MCC, as well as carcinoembryonic antigen cell adhesion molecule 6 (CEACAM6) for SCLC. Immunostaining for CK20, TTF-1, and new candidate markers was performed on 43 MCC and 59 SCLC samples. All three MCC markers were sensitive and specific, with CK20 and ATOH1 staining 43/43 (100%) MCC and 0/59 (0%) SCLC cases and TFAP2B staining 40/43 (93%) MCC and 0/59 (0%) SCLC cases. TTF-1 stained 47/59 (80%) SCLC and 1/43 (2%) MCC cases. CEACAM6 stained 49/59 (83%) SCLC and 0/43 (0%) MCC cases. Combining CEACAM6 and TTF-1 increased SCLC detection sensitivity to 93% and specificity to 98%. These data suggest that ATOH1, TFAP2B, and CEACAM6 should be explored as markers to differentiate MCC and SCLC.
- Published
- 2024
- Full Text
- View/download PDF
11. Histopathology and SARS-CoV-2 Cellular Localization in Eye Tissues of COVID-19 Autopsies.
- Author
-
Sen HN, Vannella KM, Wang Y, Chung JY, Kodati S, Ramelli SC, Lee JW, Perez P, Stein SR, Grazioli A, Dickey JM, Ylaya K, Singh M, Yinda KC, Platt A, Ramos-Benitez MJ, Zerbe C, Munster VJ, de Wit E, Warner BM, Herr DL, Rabin J, Saharia KK, Kleiner DE, Hewitt SM, Chan CC, and Chertow DS
- Subjects
- Humans, SARS-CoV-2, Autopsy, RNA, Viral analysis, Inflammation, COVID-19
- Abstract
Ophthalmic manifestations and tissue tropism of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been reported in association with coronavirus disease 2019 (COVID-19), but the pathology and cellular localization of SARS-CoV-2 are not well characterized. The objective of this study was to evaluate macroscopic and microscopic changes and investigate cellular localization of SARS-CoV-2 across ocular tissues at autopsy. Ocular tissues were obtained from 25 patients with COVID-19 at autopsy. SARS-CoV-2 nucleocapsid gene RNA was previously quantified by droplet digital PCR from one eye. Herein, contralateral eyes from 21 patients were fixed in formalin and subject to histopathologic examination. Sections of the droplet digital PCR-positive eyes from four other patients were evaluated by in situ hybridization to determine the cellular localization of SARS-CoV-2 spike gene RNA. Histopathologic abnormalities, including cytoid bodies, vascular changes, and retinal edema, with minimal or no inflammation in ocular tissues were observed in all 21 cases evaluated. In situ hybridization localized SARS-CoV-2 RNA to neuronal cells of the retinal inner and outer layers, ganglion cells, corneal epithelia, scleral fibroblasts, and oligodendrocytes of the optic nerve. In conclusion, a range of common histopathologic alterations were identified within ocular tissue, and SARS-CoV-2 RNA was localized to multiple cell types. Further studies will be required to determine whether the alterations observed were caused by SARS-CoV-2 infection, the host immune response, and/or preexisting comorbidities., (Copyright © 2023 American Society for Investigative Pathology. All rights reserved.)
- Published
- 2023
- Full Text
- View/download PDF
12. Quality Assessment of Proteins and RNA Following Storage in Archival Formalin-Fixed Paraffin-Embedded Human Breast Cancer Tissue Microarray Sections.
- Author
-
Kim K, Ylaya K, Perry C, Lee MY, Kim JW, Chung JY, and Hewitt SM
- Subjects
- Humans, Female, Immunohistochemistry, Ki-67 Antigen genetics, Paraffin Embedding, Formaldehyde, Cadherins genetics, Breast Neoplasms genetics
- Abstract
Although the immunogenicity of formalin-fixed paraffin-embedded tissue sections can decrease during storage and transport, the exact mechanism of antigenic loss and how to prevent it are not clear. Herein, we investigated changes in the expression of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER-2), E-cadherin, and Ki-67 in human breast tissue microarray (TMA) tissue sections stored for up to 3 months in dry and wet conditions. The positive rates of ER and PR expression were minimally changed after 3 months of storage, but the Allred scores of ER and PR stored in humid conditions decreased remarkably in comparison to fresh-cut tissue. The HER-2 antigenicity and RNA integrity of breast TMA sections stored in dry conditions diminished gradually with storage time, whereas the immunoreactivity and RNA quality of HER-2 in humid conditions decreased sharply as storage length increased. The area and intensity of E-cadherin staining in tissue sections stored in dry conditions did not change significantly and were minimally changed after 3 months, respectively. In contrast, the area and intensity of E-cadherin staining in tissue sections stored in humid conditions decreased significantly as storage length increased. Finally, the Ki-67 labeling index of tissue sections stored for 3 months in dry (9% decrease) and wet (31.9% decrease) conditions was decreased in comparison to fresh sections. In conclusion, these results indicate that water is a crucial factor for protein and RNA degradation in stored tissue sections, and detailed guidelines are required in the clinic.
- Published
- 2023
- Full Text
- View/download PDF
13. Tumor biology and immune infiltration define primary liver cancer subsets linked to overall survival after immunotherapy.
- Author
-
Budhu A, Pehrsson EC, He A, Goyal L, Kelley RK, Dang H, Xie C, Monge C, Tandon M, Ma L, Revsine M, Kuhlman L, Zhang K, Baiev I, Lamm R, Patel K, Kleiner DE, Hewitt SM, Tran B, Shetty J, Wu X, Zhao Y, Shen TW, Choudhari S, Kriga Y, Ylaya K, Warner AC, Edmondson EF, Forgues M, Greten TF, and Wang XW
- Subjects
- Humans, Immune Checkpoint Inhibitors pharmacology, Immune Checkpoint Inhibitors therapeutic use, Retrospective Studies, Immunotherapy, Genomics, Carcinoma, Hepatocellular therapy, Liver Neoplasms therapy
- Abstract
Primary liver cancer is a rising cause of cancer deaths in the US. Although immunotherapy with immune checkpoint inhibitors induces a potent response in a subset of patients, response rates vary among individuals. Predicting which patients will respond to immune checkpoint inhibitors is of great interest in the field. In a retrospective arm of the National Cancer Institute Cancers of the Liver: Accelerating Research of Immunotherapy by a Transdisciplinary Network (NCI-CLARITY) study, we use archived formalin-fixed, paraffin-embedded samples to profile the transcriptome and genomic alterations among 86 hepatocellular carcinoma and cholangiocarcinoma patients prior to and following immune checkpoint inhibitor treatment. Using supervised and unsupervised approaches, we identify stable molecular subtypes linked to overall survival and distinguished by two axes of aggressive tumor biology and microenvironmental features. Moreover, molecular responses to immune checkpoint inhibitor treatment differ between subtypes. Thus, patients with heterogeneous liver cancer may be stratified by molecular status indicative of treatment response to immune checkpoint inhibitors., Competing Interests: Declaration of interests The authors declare no competing interests., (Published by Elsevier Inc.)
- Published
- 2023
- Full Text
- View/download PDF
14. The Application of Guanidinium to Improve Biomolecule Quality in Fixed, Paraffin-embedded Tissue.
- Author
-
Chung JY, Kim K, Ylaya K, Walker-Bawa KE, Perry C, Star RA, and Hewitt SM
- Subjects
- Fixatives, Guanidine, Paraffin Embedding, Paraffin, Proto-Oncogene Proteins c-akt, Formaldehyde, RNA analysis, Tissue Fixation, Proteomics, Nucleic Acids
- Abstract
Neutral buffered formalin (NBF) is the most common fixative in clinical applications. However, NBF damages proteins and nucleic acids, limiting the quality of proteomic and nucleic acid-based assays. Prior studies have demonstrated that BE70, a fixative of buffered 70% ethanol, has many benefits over NBF but the degradation of proteins and nucleic acids in archival paraffin blocks remain a challenge. Thus, we evaluated the addition of guanidinium salts to BE70 with the hypothesis that this may protect RNA and protein. Guanidinium salt supplemented BE70 (BE70G)-fixed tissue is comparable with that of BE70 via histology and immunohistochemistry. Western blot analysis also revealed that HSP70, AKT, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression signals in BE70G-fixed tissue were higher than those in BE70-fixed tissue. The quality of nucleic acids extracted from BE70G-fixed, paraffin-embedded tissue was also superior, and BE70G provides improved protein and RNA quality at shorter fixation times than its predecessors. The degradation of proteins, AKT and GAPDH, in archival tissue blocks is also decreased with the addition of guanidinium salt to BE70. In conclusion, BE70G fixative improves the quality of molecular analysis with more rapid fixation of tissue and enhanced long-term storage of paraffin blocks at room temperature for evaluation of protein epitopes.
- Published
- 2023
- Full Text
- View/download PDF
15. SARS-CoV-2 infection and persistence in the human body and brain at autopsy.
- Author
-
Stein SR, Ramelli SC, Grazioli A, Chung JY, Singh M, Yinda CK, Winkler CW, Sun J, Dickey JM, Ylaya K, Ko SH, Platt AP, Burbelo PD, Quezado M, Pittaluga S, Purcell M, Munster VJ, Belinky F, Ramos-Benitez MJ, Boritz EA, Lach IA, Herr DL, Rabin J, Saharia KK, Madathil RJ, Tabatabai A, Soherwardi S, McCurdy MT, Peterson KE, Cohen JI, de Wit E, Vannella KM, Hewitt SM, Kleiner DE, and Chertow DS
- Subjects
- Humans, RNA, Viral analysis, Virus Replication, Time Factors, Respiratory System pathology, Respiratory System virology, Autopsy, Brain virology, COVID-19 virology, SARS-CoV-2 genetics, SARS-CoV-2 isolation & purification, SARS-CoV-2 pathogenicity, SARS-CoV-2 physiology, Organ Specificity
- Abstract
Coronavirus disease 2019 (COVID-19) is known to cause multi-organ dysfunction
1-3 during acute infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), with some patients experiencing prolonged symptoms, termed post-acute sequelae of SARS-CoV-2 (refs.4,5 ). However, the burden of infection outside the respiratory tract and time to viral clearance are not well characterized, particularly in the brain3,6-14 . Here we carried out complete autopsies on 44 patients who died with COVID-19, with extensive sampling of the central nervous system in 11 of these patients, to map and quantify the distribution, replication and cell-type specificity of SARS-CoV-2 across the human body, including the brain, from acute infection to more than seven months following symptom onset. We show that SARS-CoV-2 is widely distributed, predominantly among patients who died with severe COVID-19, and that virus replication is present in multiple respiratory and non-respiratory tissues, including the brain, early in infection. Further, we detected persistent SARS-CoV-2 RNA in multiple anatomic sites, including throughout the brain, as late as 230 days following symptom onset in one case. Despite extensive distribution of SARS-CoV-2 RNA throughout the body, we observed little evidence of inflammation or direct viral cytopathology outside the respiratory tract. Our data indicate that in some patients SARS-CoV-2 can cause systemic infection and persist in the body for months., (© 2022. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)- Published
- 2022
- Full Text
- View/download PDF
16. Single-cell biology uncovers apoptotic cell death and its spatial organization as a potential modifier of tumor diversity in HCC.
- Author
-
Khatib SA, Ma L, Dang H, Forgues M, Chung JY, Ylaya K, Hewitt SM, Chaisaingmongkol J, Rucchirawat M, and Wang XW
- Subjects
- Apoptosis, Caspase 3 metabolism, Cell Line, Tumor, Humans, Carcinoma, Hepatocellular genetics, Carcinoma, Hepatocellular pathology, Liver Neoplasms genetics, Liver Neoplasms pathology
- Abstract
Background and Aims: HCC is a highly aggressive and heterogeneous cancer type with limited treatment options. Identifying drivers of tumor heterogeneity may lead to better therapeutic options and favorable patient outcomes. We investigated whether apoptotic cell death and its spatial architecture is linked to tumor molecular heterogeneity using single-cell in situ hybridization analysis., Approach and Results: We analyzed 254 tumor samples from two HCC cohorts using tissue microarrays. We developed a mathematical model to quantify cellular diversity among HCC samples using two tumor markers, cyclin-dependent kinase inhibitor 3 and protein regulator of cytokinesis 1 as surrogates for heterogeneity and caspase 3 (CASP3) as an apoptotic cell death marker. We further explored the impact of potential dying-cell hubs on tumor cell diversity and patient outcome by density contour mapping and spatial proximity analysis. We also developed a selectively controlled in vitro model of cell death using CRISPR/CRISPR-associated 9 to determine therapy response and growth under hypoxic conditions. We found that increasing levels of CASP3
+ tumor cells are associated with higher tumor diversity. Interestingly, we discovered regions of densely populated CASP3+ , which we refer to as CASP3+ cell islands, in which the nearby cellular heterogeneity was found to be the greatest compared to cells farther away from these islands and that this phenomenon was associated with survival. Additionally, cell culture experiments revealed that higher levels of cell death, accompanied by increased CASP3 expression, led to greater therapy resistance and growth under hypoxia., Conclusions: These results are consistent with the hypothesis that increased apoptotic cell death may lead to greater tumor heterogeneity and thus worse patient outcomes., (© 2022 American Association for the Study of Liver Diseases. This article has been contributed to by US Government employees and their work is in the public domain in the USA.)- Published
- 2022
- Full Text
- View/download PDF
17. Evidence of SARS-CoV-2-Specific T-Cell-Mediated Myocarditis in a MIS-A Case.
- Author
-
Vannella KM, Oguz C, Stein SR, Pittaluga S, Dikoglu E, Kanwal A, Ramelli SC, Briese T, Su L, Wu X, Ramos-Benitez MJ, Perez-Valencia LJ, Babyak A, Cha NR, Chung JY, Ylaya K, Madathil RJ, Saharia KK, Scalea TM, Tran QK, Herr DL, Kleiner DE, Hewitt SM, Notarangelo LD, Grazioli A, and Chertow DS
- Subjects
- Adult, COVID-19 immunology, COVID-19 pathology, Humans, Male, Myocarditis immunology, RNA, Viral analysis, SARS-CoV-2, Systemic Inflammatory Response Syndrome immunology, COVID-19 complications, Myocarditis pathology, Myocarditis virology, Systemic Inflammatory Response Syndrome pathology, T-Lymphocytes immunology
- Abstract
A 26-year-old otherwise healthy man died of fulminant myocarditis. Nasopharyngeal specimens collected premortem tested negative for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Histopathological evaluation of the heart showed myocardial necrosis surrounded by cytotoxic T-cells and tissue-repair macrophages. Myocardial T-cell receptor (TCR) sequencing revealed hyper-dominant clones with highly similar sequences to TCRs that are specific for SARS-CoV-2 epitopes. SARS-CoV-2 RNA was detected in the gut, supporting a diagnosis of multisystem inflammatory syndrome in adults (MIS-A). Molecular targets of MIS-associated inflammation are not known. Our data indicate that SARS-CoV-2 antigens selected high-frequency T-cell clones that mediated fatal myocarditis., Competing Interests: Authors CO, LS and XW were employed by company Leidos Biomedical Research, Inc. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Vannella, Oguz, Stein, Pittaluga, Dikoglu, Kanwal, Ramelli, Briese, Su, Wu, Ramos-Benitez, Perez-Valencia, Babyak, Cha, Chung, Ylaya, Madathil, Saharia, Scalea, Tran, Herr, Kleiner, Hewitt, Notarangelo, Grazioli and Chertow.)
- Published
- 2021
- Full Text
- View/download PDF
18. Notch signaling and efficacy of PD-1/PD-L1 blockade in relapsed small cell lung cancer.
- Author
-
Roper N, Velez MJ, Chiappori A, Kim YS, Wei JS, Sindiri S, Takahashi N, Mulford D, Kumar S, Ylaya K, Trindade C, Manukyan I, Brown AL, Trepel JB, Lee JM, Hewitt S, Khan J, and Thomas A
- Subjects
- Antineoplastic Agents, Immunological therapeutic use, Biomarkers, Tumor genetics, Biomarkers, Tumor metabolism, Cell Line, Tumor, Humans, Lung Neoplasms genetics, Lung Neoplasms pathology, Neoplasm Recurrence, Local, Receptor, Notch1 metabolism, Small Cell Lung Carcinoma genetics, Small Cell Lung Carcinoma pathology, Treatment Outcome, Exome Sequencing methods, Immune Checkpoint Inhibitors therapeutic use, Lung Neoplasms drug therapy, Receptor, Notch1 genetics, Signal Transduction genetics, Small Cell Lung Carcinoma drug therapy
- Abstract
Immune checkpoint blockade (ICB) benefits only a small subset of patients with small cell lung cancer (SCLC), yet the mechanisms driving benefit are poorly understood. To identify predictors of clinical benefit to ICB, we performed immunogenomic profiling of tumor samples from patients with relapsed SCLC. Tumors of patients who derive clinical benefit from ICB exhibit cytotoxic T-cell infiltration, high expression of antigen processing and presentation machinery (APM) genes, and low neuroendocrine (NE) differentiation. However, elevated Notch signaling, which positively correlates with low NE differentiation, most significantly predicts clinical benefit to ICB. Activation of Notch signaling in a NE human SCLC cell line induces a low NE phenotype, marked by increased expression of APM genes, demonstrating a mechanistic link between Notch activation, low NE differentiation and increased intrinsic tumor immunity. Our findings suggest Notch signaling as a determinant of response to ICB in SCLC.
- Published
- 2021
- Full Text
- View/download PDF
19. IGF-1 Receptor Signaling Regulates Type II Pneumocyte Senescence and Resulting Macrophage Polarization in Lung Fibrosis.
- Author
-
Chung EJ, Kwon S, Reedy JL, White AO, Song JS, Hwang I, Chung JY, Ylaya K, Hewitt SM, and Citrin DE
- Subjects
- Alveolar Epithelial Cells radiation effects, Animals, Carcinoma, Non-Small-Cell Lung pathology, Carcinoma, Non-Small-Cell Lung therapy, Cellular Senescence radiation effects, Chemoradiotherapy, Gene Deletion, Humans, Hydroxyproline analysis, Lung metabolism, Lung radiation effects, Lung Neoplasms pathology, Lung Neoplasms therapy, Macrophage Activation, Macrophages, Alveolar radiation effects, Mice, Mice, Inbred C57BL, Pulmonary Fibrosis pathology, Radiation Injuries, Experimental metabolism, Radiation Injuries, Experimental physiopathology, Radiation Injuries, Experimental prevention & control, Receptor, IGF Type 1 deficiency, Receptor, IGF Type 1 genetics, Alveolar Epithelial Cells physiology, Cell Polarity, Cellular Senescence physiology, Macrophages, Alveolar physiology, Pulmonary Fibrosis etiology, Receptor, IGF Type 1 metabolism
- Abstract
Purpose: Type II pneumocyte (alveolar epithelial cells type II [AECII]) senescence has been implicated in the progression of lung fibrosis. The capacity of senescent cells to modulate pulmonary macrophages to drive fibrosis is unexplored. Insulin-like growth factor-1 receptor (IGF-1R) signaling has been implicated as a regulator of senescence and aging., Methods and Materials: Mice with an AECII-specific deletion of IGF-1R received thoracic irradiation (n ≥ 5 per condition), and the effect of IGF-1R deficiency on radiation-induced AECII senescence and macrophage polarization to an alternatively activated phenotype (M2) was investigated. IGF-1R signaling, macrophage polarization, and senescence were evaluated in surgically resected human lung (n = 63)., Results: IGF-1R deficient mice demonstrated reduced AECII senescence (senescent AECII/field; intact: 7.25% ± 3.5% [mean ± SD], deficient: 2.75% ± 2.8%, P = .0001), reduced accumulation of M2 macrophages (intact: 24.7 ± 2.2 cells/field, deficient: 15.5 ± 1.2 cells/field, P = .0086), and fibrosis (hydroxyproline content; intact: 71.9 ± 21.7 μg/lung, deficient: 31.7 ± 7.9, P = .0485) after irradiation. Senescent AECII enhanced M2 polarization in a paracrine fashion (relative Arg1 mRNA, 0 Gy: 1.0 ± 0.4, 17.5 Gy: 7.34 ± 0.5, P < .0001). Evaluation of surgical samples from patients treated with chemoradiation demonstrated increased expression of IGF-1 (unirradiated: 10.2% ± 4.9% area, irradiated: 15.1% ± 11.5%, P = .0377), p21 (unirradiated: 0.013 ± 0.02 histoscore, irradiated: 0.084 ± 0.09 histoscore, P = .0002), IL-13 (unirradiated: 13.7% ± 2.8% area, irradiated: 21.7% ± 3.8%, P < .0001), and M2 macrophages in fibrotic regions relative to nonfibrotic regions (unirradiated: 11.4 ± 12.2 CD163 + cells/core, irradiated: 43.1 ± 40.9 cells/core, P = .0011), consistent with findings from animal models of lung fibrosis., Conclusions: This study demonstrates that senescent AECII are necessary for the progression of pulmonary fibrosis and serve as a targetable, chronic stimuli for macrophage activation in fibrotic lung., (Published by Elsevier Inc.)
- Published
- 2021
- Full Text
- View/download PDF
20. Prognostic implication of SOX2 expression in small intestinal adenocarcinoma.
- Author
-
Kim JW, Chung JY, Ylaya K, Park Y, Jun SY, Hong SM, and Hewitt SM
- Subjects
- Adenocarcinoma diagnosis, Adenocarcinoma pathology, Adult, Aged, Aged, 80 and over, Biomarkers, Tumor analysis, Colorectal Neoplasms pathology, Duodenal Neoplasms diagnosis, Duodenal Neoplasms pathology, Female, Humans, Male, Middle Aged, Neoplastic Stem Cells metabolism, Neoplastic Stem Cells pathology, Prognosis, Adenocarcinoma metabolism, Colorectal Neoplasms metabolism, Duodenal Neoplasms metabolism, SOXB1 Transcription Factors metabolism
- Abstract
The presence of KRAS mutation enhances the stem cell features of colorectal carcinoma cells containing mutant adenomatous polyposis coli (APC). However, their potential role in small intestinal adenocarcinoma remains elusive. Here, we aimed to investigate the clinical significance of cancer stem cell markers expression in the context of small intestinal adenocarcinoma with the KRAS genotype. SOX2, NANOG, and OCT4 expression were assessed by immunohistochemistry and digital image analysis, and their potential association with KRAS was further examined in 185 Korean patients with small intestinal adenocarcinomas, which were collected from 22 institutions in South Korea. Positive expression of SOX2, NANOG, and OCT4 was detected in 65 (35.1%), 94 (50.8%), and 82 (44.3%) of patients, respectively. Patients with high SOX2 (SOX2+) expression displayed worse overall survival compared to those with low SOX2 (SOX2-) expression (P < 0.001). Patients with SOX2+/mutant KRAS (KRAS
MT ) (11.1 months) had significantly shorter overall survival than those with SOX2-/KRASWT (53.6 months) (P < 0.001). In multivariate analysis, SOX2+, distal location, high pT and pN categories, microsatellite stable, and absence of predisposing diseases were independent prognostic factors for worse overall survival. These results suggest that SOX2 expression has the potential to predict clinical outcomes in patients with small intestinal adenocarcinomas.- Published
- 2021
- Full Text
- View/download PDF
21. Clinical Significance of Tumor Infiltrating Lymphocytes in Association with Hormone Receptor Expression Patterns in Epithelial Ovarian Cancer.
- Author
-
Han GH, Hwang I, Cho H, Ylaya K, Choi JA, Kwon H, Chung JY, Hewitt SM, and Kim JH
- Subjects
- B7-H1 Antigen genetics, Carcinoma, Ovarian Epithelial metabolism, Carcinoma, Ovarian Epithelial pathology, Computational Biology methods, Databases, Genetic, Disease Susceptibility, Female, Gene Expression, Humans, Immunohistochemistry, Lymphocytes, Tumor-Infiltrating pathology, Ovarian Neoplasms metabolism, Ovarian Neoplasms pathology, Programmed Cell Death 1 Receptor genetics, Receptors, Steroid metabolism, Tumor Microenvironment genetics, Tumor Microenvironment immunology, Biomarkers, Tumor, Carcinoma, Ovarian Epithelial etiology, Lymphocytes, Tumor-Infiltrating immunology, Lymphocytes, Tumor-Infiltrating metabolism, Ovarian Neoplasms etiology, Receptors, Steroid genetics
- Abstract
Hormone receptor expression patterns often correlate with infiltration of specific lymphocytes in tumors. Specifically, the presence of specific tumor-infiltrating lymphocytes (TILs) with particular hormone receptor expression is reportedly associated with breast cancer, however, this has not been revealed in epithelial ovarian cancer (EOC). Therefore, we investigated the association between hormone receptor expression and TILs in EOC. Here we found that ERα, AR, and GR expression increased in EOC, while PR was significantly reduced and ERβ expression showed a reduced trend compared to normal epithelium. Cluster analysis indicated poor disease-free survival (DFS) in AR+/GR+/PR+ subgroup (triple dominant group); while the Cox proportional-hazards model highlighted the triple dominant group as an independent prognostic factor for DFS. In addition, significant upregulation of FoxP3+ TILs, PD-1, and PD-L1 was observed in the triple dominant group compared to other groups. NanoString analyses further suggested that tumor necrosis factor (TNF) and/or NF-κB signaling pathways were activated with significant upregulation of RELA , MAP3K5 , TNFAIP3 , BCL2L1 , RIPK1 , TRAF2 , PARP1 , and AKT1 in the triple dominant EOC group. The triple dominant subgroup correlates with poor prognosis in EOC. Moreover, the TNF and/or NF-κB signaling pathways may be responsible for hormone-mediated inhibition of the immune microenvironment.
- Published
- 2021
- Full Text
- View/download PDF
22. Intratumoral γδ T-Cell Infiltrates, Chemokine (C-C Motif) Ligand 4/Chemokine (C-C Motif) Ligand 5 Protein Expression and Survival in Patients With Hepatocellular Carcinoma.
- Author
-
Zhao N, Dang H, Ma L, Martin SP, Forgues M, Ylaya K, Hewitt SM, and Wang XW
- Subjects
- Animals, Carcinoma, Hepatocellular mortality, Carcinoma, Hepatocellular pathology, Female, Humans, Liver metabolism, Liver pathology, Liver Neoplasms mortality, Liver Neoplasms pathology, Male, Mice, Mice, Inbred C57BL, Middle Aged, Proportional Hazards Models, Survival Analysis, Tumor Microenvironment, Carcinoma, Hepatocellular metabolism, Chemokine CCL4 metabolism, Chemokine CCL5 metabolism, Intraepithelial Lymphocytes pathology, Liver Neoplasms metabolism, Lymphocytes, Tumor-Infiltrating pathology
- Abstract
Background and Aims: Hepatocellular carcinoma (HCC) is an aggressive malignancy which is often associated with a complex tumor microenvironment attributable to etiology-induced cellular inflammation. γδ T cells are known to detect and react to chronic inflammation, which is linked to cancer development, progression, and metastasis. Our recent genomic study revealed an increased infiltration of several immune cell types, including γδ T cells, in tumor microenvironments of a Thai HCC subtype associated with a good prognosis., Approach and Results: Here, we quantified the amount of γδ T cells using a γδ T-cell-specific gene signature in 247 Chinese HCC patients. We also validated the γδ T-cell signature in American HCC patients. Additionally, such an association was only found in tumor transcriptomic data, but not in adjacent nontumor transcriptomic data, suggesting a selective enrichment of γδ T cells in the tumor microenvironment. Moreover, the γδ T-cell signature was positively correlated with the expression of natural killer cell receptor genes, such as NKG2D and cytolytic T-cell genes granzymes and perforin, suggesting a stronger T-cell-mediated cytotoxic activity. Furthermore, we found that the γδ T-cell-specific gene expression is positively correlated with the expression of chemokine (C-C motif) ligand 4 (CCL4)/chemokine (C-C motif) ligand 5 (CCL5) and C-C chemokine receptor type 1 (CCR1)/C-C chemokine receptor type 5 (CCR5), the receptors for γδ T cells. We validated these results using immunohistochemical analysis of formalin-fixed, paraffin-embedded tumor biopsies from 182 HCC patients. Moreover, we found evidence of CCL4/CCL5-mediated recruitment of γδ T cells both in vitro and in a murine orthotopic Hepa1-6 HCC model., Conclusions: We propose that CCL4/CCL5 may interact with their receptor, CCR1/CCR5, which may facilitate the recruitment of γδ T cells from peripheral blood or peritumor regions to the tumor regions. Consequently, an increasing infiltration of γδ T cells in tumors may enhance antitumor immunity and improve patients' prognosis., (© 2020 by the American Association for the Study of Liver Diseases.)
- Published
- 2021
- Full Text
- View/download PDF
23. Tumor-associated macrophage, angiogenesis and lymphangiogenesis markers predict prognosis of non-small cell lung cancer patients.
- Author
-
Hwang I, Kim JW, Ylaya K, Chung EJ, Kitano H, Perry C, Hanaoka J, Fukuoka J, Chung JY, and Hewitt SM
- Subjects
- Humans, Lymphangiogenesis, Prognosis, Tumor Microenvironment, Tumor-Associated Macrophages, Carcinoma, Non-Small-Cell Lung, Lung Neoplasms
- Abstract
Background: The tumor microenvironment (TME) is a critical player in tumor progression, metastasis and therapy outcomes. Tumor-associated macrophages (TAMs) are a well-recognized core element of the TME and generally characterized as M2-like macrophages. TAMs are believed to contribute to tumor progression, but the mechanism behind this remains unclear. We aimed to investigate the clinical, angiogenic, and lymphangiogenic significance of TAMs in non-small cell lung cancer (NSCLC)., Methods: Utilizing combined immunohistochemistry and digital image analysis, we assessed CD68, CD163, VEGF-A, and VEGF-C expression in 349 patients with NSCLC. Subsequently, the potential association between M2 TAMs and angiogenic VEGF-A and/or lymphangiogenic VEGF-C was evaluated for its prognostic value. Furthermore, the effects of M2 TAMs on angiogenesis and lymphangiogenesis were explored via an in vitro co-culture system., Results: CD68 and CD163 expression were found to directly correlate with VEGF-A and/or VEGF-C expression (all p < 0.001). Furthermore, elevated M2 ratio (CD163+/CD68+) was significantly associated with poor overall survival (p = 0.023). Dual expression of M2 ratio
high and VEGF-Chigh (M2 ratiohigh VEGF-Chigh ) was correlated with worse overall survival (p = 0.033). Multivariate analysis revealed that M2 ratiohigh [HR (95% CI) = 1.53 (1.01-2.33), p = 0.046] and combined M2 ratiohigh VEGF-Chigh expression [HR (95% CI) = 2.01 (1.28-3.16), p = 0.003] were independent predictors of poor overall survival. Notably, we confirmed that M2 macrophages significantly enhanced the protein and mRNA expression of both VEGF-A and VEGF-C, while M1 macrophages induced only mRNA expression of VEGF-A in A549 cells., Conclusions: This study suggests that TAMs are significantly associated with angiogenesis and lymphangiogenesis, contributing to the progression of NSCLC. Furthermore, elevated M2 ratio, similar to combined high M2 ratio and high VEGF-C expression, is a strong indicator of poor prognosis in patients with NSCLC, providing insight for future TAM-based immunotherapy strategies.- Published
- 2020
- Full Text
- View/download PDF
24. 4-HNE Immunohistochemistry and Image Analysis for Detection of Lipid Peroxidation in Human Liver Samples Using Vitamin E Treatment in NAFLD as a Proof of Concept.
- Author
-
Podszun MC, Chung JY, Ylaya K, Kleiner DE, Hewitt SM, and Rotman Y
- Subjects
- Antioxidants administration & dosage, Cells, Cultured, Humans, Immunohistochemistry, Non-alcoholic Fatty Liver Disease blood, Vitamin E administration & dosage, Aldehydes analysis, Antioxidants pharmacology, Lipid Peroxidation drug effects, Liver drug effects, Liver metabolism, Non-alcoholic Fatty Liver Disease drug therapy, Optical Imaging, Vitamin E pharmacology
- Abstract
Lipid peroxidation is a common feature of liver diseases, especially non-alcoholic fatty liver disease (NAFLD). There are limited validated tools to study intra-hepatic lipid peroxidation, especially for small specimen. We developed a semi-quantitative, fully automated immunohistochemistry assay for the detection of 4-hydroxynoneal (4-HNE) protein adducts, a marker of lipid peroxidation, for adaptation to clinical diagnostics and research. We used Hep G2 cells treated with 4-HNE to validate specificity, sensitivity, and dynamic range of the antibody. Staining and semi-quantitative automated readout were confirmed in human needle-biopsy liver samples from subjects with NAFLD and normal liver histology. The ability to detect changes in lipid peroxidation was tested in paired liver biopsies from NAFLD subjects, obtained before and after 4 weeks of treatment with the antioxidant vitamin E (ClinicalTrials.gov NCT01792115, n =21). The cellular calibrator was linear and NAFLD patients had significantly higher levels of 4-HNE adducts compared to controls ( p =0.02). Vitamin E treatment significantly decreased 4-HNE ( p =0.0002). Our findings demonstrate that 4-HNE quantification by immunohistochemistry and automated image analysis is feasible and able to detect changes in hepatic lipid peroxidation in clinical trials. This method can be applied to archival and fresh samples and should be considered for use in assessing NAFLD histology.
- Published
- 2020
- Full Text
- View/download PDF
25. Loss of HES-1 Expression Predicts a Poor Prognosis for Small Intestinal Adenocarcinoma Patients.
- Author
-
Kim JW, Jun SY, Ylaya K, Chang HK, Oh YH, Hong SM, Chung JY, and Hewitt SM
- Abstract
Objective: Hairy and enhancer of split-1 (HES-1), which is a downstream target of the Notch signaling pathway, has been linked to KRAS mutations. HES-1 has been proposed as harboring oncogenic activity in colorectal cancer but has not been investigated in adenocarcinoma of the small intestine, where the drivers of oncogenesis are not as well-understood. Materials and Methods: To investigate the clinicopathologic and prognostic implications of HES-1, HES-1 immunohistochemical expression was analyzed in digital images along with clinicopathological variables, including survival and KRAS genotype, in 185 small intestinal adenocarcinomas. Results: The loss of HES-1 expression (HES-1
Loss ) was observed in 38.4% (71/185) of the patients, and was associated with higher pT category ( P = 0.018), pancreatic invasion ( P = 0.005), high grade ( P = 0.043), and non-tubular histology ( P = 0.004). Specifically, in tumors with mutant KRAS ( KRASMT ), HES-1Loss was related to proximal location ( P = 0.024), high T and N categories ( P = 0.005 and 0.047, respectively), and pancreatic invasion ( P = 0.004). Patients with HES-1Loss showed worse overall survival compared to those with intact HES-1 (HES-1Intact ) ( P = 0.013). Patients with HES-1Loss / KRASMT (median, 17.3 months) had significantly worse outcomes than those with HES-1Intact / KRASWT (39.9 months), HES-1Intact / KRASMT (47.6 month), and HES-1Loss / KRASWT (36.2 months; P = 0.010). By multivariate analysis, HES-1Loss (hazard ratio = 1.55, 95% confidence interval (CI), 1.07-2.26; P = 0.022) remained an independent prognostic factor. Conclusion: HES-1expression can be used as a potential prognostic marker and may aid in the management of patients with small intestinal adenocarcinomas., (Copyright © 2020 Kim, Jun, Ylaya, Chang, Oh, Hong, Chung and Hewitt.)- Published
- 2020
- Full Text
- View/download PDF
26. Deubiquitylation and stabilization of Notch1 intracellular domain by ubiquitin-specific protease 8 enhance tumorigenesis in breast cancer.
- Author
-
Shin S, Kim K, Kim HR, Ylaya K, Do SI, Hewitt SM, Park HS, Roe JS, Chung JY, and Song J
- Subjects
- Adult, Aged, Aged, 80 and over, Carcinogenesis, Cell Line, Tumor, Cell Proliferation, Down-Regulation, Female, Gene Deletion, Humans, Middle Aged, Protein Binding, Protein Domains, Protein Stability, Signal Transduction, Tumor Stem Cell Assay, Up-Regulation, Wound Healing, Breast Neoplasms metabolism, Breast Neoplasms pathology, Endopeptidases metabolism, Endosomal Sorting Complexes Required for Transport metabolism, Receptor, Notch1 chemistry, Receptor, Notch1 metabolism, Ubiquitin Thiolesterase metabolism, Ubiquitination
- Abstract
Notch, an essential factor in tissue development and homoeostasis, has been reported to play an oncogenic function in a variety of cancers. Here, we report ubiquitin-specific protease 8 (USP8) as a novel deubiquitylase of Notch1 intracellular domain (NICD). USP8 specifically stabilizes and deubiquitylates NICD through a direct interaction. The inhibition of USP8 downregulated the Notch signalling pathway via NICD destabilization, resulting in the retardation of cellular growth, wound closure, and colony forming ability of breast cancer cell lines. These phenomena were restored by the reconstitution of NICD or USP8, supporting the direct interaction between these two proteins. The expression levels of NICD and USP8 proteins were positively correlated in patients with advanced breast cancer. Taken together, our results suggest that USP8 functions as a positive regulator of Notch signalling, offering a therapeutic target for breast cancer.
- Published
- 2020
- Full Text
- View/download PDF
27. Melanoblast transcriptome analysis reveals pathways promoting melanoma metastasis.
- Author
-
Marie KL, Sassano A, Yang HH, Michalowski AM, Michael HT, Guo T, Tsai YC, Weissman AM, Lee MP, Jenkins LM, Zaidi MR, Pérez-Guijarro E, Day CP, Ylaya K, Hewitt SM, Patel NL, Arnheiter H, Davis S, Meltzer PS, Merlino G, and Mishra PJ
- Subjects
- Animals, Cell Line, Tumor, Endoplasmic Reticulum, Female, Gene Expression Regulation, Neoplastic, Gene Knockdown Techniques, Humans, Kangai-1 Protein genetics, Kangai-1 Protein metabolism, Lung pathology, Melanocytes metabolism, Melanoma pathology, Mice, Mice, Inbred C57BL, Neoplasm Metastasis genetics, Neoplasms, Second Primary pathology, Phenotype, Receptors, Peptide genetics, Receptors, Peptide metabolism, Skin Neoplasms pathology, Ubiquitin-Protein Ligases metabolism, Melanoma, Cutaneous Malignant, Gene Expression Profiling, Melanoma genetics, Melanoma metabolism, Skin Neoplasms genetics, Skin Neoplasms metabolism, Transcriptome
- Abstract
Cutaneous malignant melanoma is an aggressive cancer of melanocytes with a strong propensity to metastasize. We posit that melanoma cells acquire metastatic capability by adopting an embryonic-like phenotype, and that a lineage approach would uncover metastatic melanoma biology. Using a genetically engineered mouse model to generate a rich melanoblast transcriptome dataset, we identify melanoblast-specific genes whose expression contribute to metastatic competence and derive a 43-gene signature that predicts patient survival. We identify a melanoblast gene, KDELR3, whose loss impairs experimental metastasis. In contrast, KDELR1 deficiency enhances metastasis, providing the first example of different disease etiologies within the KDELR-family of retrograde transporters. We show that KDELR3 regulates the metastasis suppressor, KAI1, and report an interaction with the E3 ubiquitin-protein ligase gp78, a regulator of KAI1 degradation. Our work demonstrates that the melanoblast transcriptome can be mined to uncover targetable pathways for melanoma therapy.
- Published
- 2020
- Full Text
- View/download PDF
28. Tumor Cell Biodiversity Drives Microenvironmental Reprogramming in Liver Cancer.
- Author
-
Ma L, Hernandez MO, Zhao Y, Mehta M, Tran B, Kelly M, Rae Z, Hernandez JM, Davis JL, Martin SP, Kleiner DE, Hewitt SM, Ylaya K, Wood BJ, Greten TF, and Wang XW
- Subjects
- Adult, Aged, Aged, 80 and over, Antineoplastic Agents, Immunological pharmacology, Antineoplastic Agents, Immunological therapeutic use, Bile Duct Neoplasms mortality, Bile Duct Neoplasms pathology, Bile Duct Neoplasms therapy, Bile Ducts, Intrahepatic pathology, Bile Ducts, Intrahepatic surgery, Biopsy, Carcinoma, Hepatocellular mortality, Carcinoma, Hepatocellular pathology, Carcinoma, Hepatocellular therapy, Cholangiocarcinoma mortality, Cholangiocarcinoma pathology, Cholangiocarcinoma therapy, DNA Copy Number Variations, Drug Resistance, Neoplasm genetics, Female, Gene Expression Regulation, Neoplastic, Genetic Variation, Hepatectomy, Humans, Liver pathology, Liver surgery, Liver Neoplasms mortality, Liver Neoplasms pathology, Liver Neoplasms therapy, Male, Middle Aged, Prognosis, Progression-Free Survival, RNA-Seq, Single-Cell Analysis, Tumor Microenvironment drug effects, Vascular Endothelial Growth Factor A genetics, Vascular Endothelial Growth Factor A metabolism, Bile Duct Neoplasms genetics, Carcinoma, Hepatocellular genetics, Cholangiocarcinoma genetics, Liver Neoplasms genetics, Tumor Microenvironment genetics
- Abstract
Cellular diversity in tumors is a key factor for therapeutic failures and lethal outcomes of solid malignancies. Here, we determined the single-cell transcriptomic landscape of liver cancer biospecimens from 19 patients. We found varying degrees of heterogeneity in malignant cells within and between tumors and diverse landscapes of tumor microenvironment (TME). Strikingly, tumors with higher transcriptomic diversity were associated with patient's worse overall survival. We found a link between hypoxia-dependent vascular endothelial growth factor expression in tumor diversity and TME polarization. Moreover, T cells from higher heterogeneous tumors showed lower cytolytic activities. Consistent results were found using bulk genomic and transcriptomic profiles of 765 liver tumors. Our results offer insight into the diverse ecosystem of liver cancer and its impact on patient prognosis., (Published by Elsevier Inc.)
- Published
- 2019
- Full Text
- View/download PDF
29. Immunophenotypic Characterization of Canine Splenic Follicular-Derived B-Cell Lymphoma.
- Author
-
Stein L, Bacmeister C, Ylaya K, Fetsch P, Wang Z, Hewitt SM, and Kiupel M
- Subjects
- Animals, Antigens, Differentiation, T-Lymphocyte immunology, Dog Diseases immunology, Dogs, Lymphoma, B-Cell immunology, Lymphoma, B-Cell pathology, Lymphoma, B-Cell, Marginal Zone immunology, Lymphoma, B-Cell, Marginal Zone pathology, Lymphoma, B-Cell, Marginal Zone veterinary, Lymphoma, Follicular immunology, Lymphoma, Follicular pathology, Lymphoma, Mantle-Cell immunology, Lymphoma, Mantle-Cell pathology, Lymphoma, Mantle-Cell veterinary, Retrospective Studies, Spleen immunology, Spleen pathology, Splenic Neoplasms immunology, Splenic Neoplasms pathology, Dog Diseases pathology, Immunophenotyping veterinary, Lymphoma, B-Cell veterinary, Lymphoma, Follicular veterinary, Splenic Neoplasms veterinary
- Abstract
Marginal zone lymphoma (MZL) and mantle cell lymphoma (MCL) belong to a subgroup of indolent B-cell lymphomas most commonly reported in the canine spleen. The goal of this study was to characterize the immunophenotype of splenic MZL and MCL in comparison to their human counterparts. Ten MCLs and 28 MZLs were selected based on morphology. A tissue microarray was generated, and expression of CD3, CD5, CD10, CD45, CD20, CD79a, Pax-5, Bcl-2, Bcl-6, cyclin D1, cyclin D3, MCL-1, MUM-1, and Sox-11 was evaluated. Neoplastic cells in all MCLs and MZLs were positive for CD5, CD20, CD45, CD79a, and BCL2 and negative for CD3, CD10, Bcl-6, cyclin D1, and cyclin D3. Positive labeling for Pax-5 was detected in 8 of 10 MCLs and 26 of 28 MZLs. Positive labeling for MUM-1 was detected in 3 of 10 MCLs, and 27 of 28 MZLs were positive for MUM-1. No MCLs but 8 of 24 MZLs were positive for MCL-1. Canine splenic MZL and MCL have a similar immunophenotype as their human counterparts. However, human splenic MCL overexpresses cyclin D1 due to a translocation. A similar genetic alteration has not been reported in dogs. In addition, in contrast to human MZL, canine splenic MZL generally expresses CD5. Following identification of B vs T cells with CD20 and CD3, a panel composed of BCL-2, Bcl-6, MUM-1, and MCL-1 combined with the histomorphological pattern can be used to accurately diagnose MZL and MCL in dogs. Expression of Bcl-2 and lack of MCL-1 expression in MCL may suggest a therapeutic benefit of BCL-2 inhibitors in canine MCL.
- Published
- 2019
- Full Text
- View/download PDF
30. Elevated expression of pancreatic adenocarcinoma upregulated factor (PAUF) is associated with poor prognosis and chemoresistance in epithelial ovarian cancer.
- Author
-
Choi CH, Kang TH, Song JS, Kim YS, Chung EJ, Ylaya K, Kim S, Koh SS, Chung JY, Kim JH, and Hewitt SM
- Subjects
- Adult, Aged, Cell Line, Tumor, Cell Proliferation, Epithelial Cells metabolism, Female, Gene Expression Regulation, Neoplastic genetics, Humans, Immunohistochemistry, Intercellular Signaling Peptides and Proteins, Lectins metabolism, Middle Aged, Ovarian Neoplasms genetics, Prognosis, Toll-Like Receptor 4 metabolism, Transcriptional Activation, Treatment Outcome, Carcinoma, Ovarian Epithelial genetics, Drug Resistance, Neoplasm genetics, Lectins genetics
- Abstract
Pancreatic adenocarcinoma upregulated factor (PAUF) is a ligand of toll-like receptors (TLRs) and has been reported to be involved in pancreatic tumor development. However, the significance of PAUF expression in epithelial ovarian cancer remains unclear. We aimed to investigate the possible clinical significance of PAUF in epithelial ovarian cancer. We examined the link between PAUF and TLR4 in ovarian cancer cell lines. Recombinant PAUF induced cell activation and proliferation in ovarian cancer cell lines, whereas PAUF knockdown inhibited these properties. Subsequently, we assessed PAUF and TLR4 expression by immunohistochemistry on tissue microarray of 408 ovarian samples ranging from normal to metastatic. PAUF expression positively correlated with TLR4 expression. Overexpression of PAUF was associated with high-grade tumor (p = 0.014) and chemoresistant tumor (p = 0.017). Similarly, high expression of TLR4 correlated with advanced tumor stage (p = 0.002) and chemoresistant tumor (p = 0.001). Multivariate analysis indicated that PAUF
high , TLR4high , and PAUFhigh /TLR4high expression are independent prognostic factor for progression-free survival, while TLR4high and PAUFhigh /TLR4high expression were independent prognostic factors for overall survival. Our results suggest that PAUF has a role in ovarian cancer progression and is a potential prognostic marker and novel chemotherapeutic target for ovarian cancer.- Published
- 2018
- Full Text
- View/download PDF
31. Aberrant tRNA processing causes an autoinflammatory syndrome responsive to TNF inhibitors.
- Author
-
Giannelou A, Wang H, Zhou Q, Park YH, Abu-Asab MS, Ylaya K, Stone DL, Sediva A, Sleiman R, Sramkova L, Bhatla D, Serti E, Tsai WL, Yang D, Bishop K, Carrington B, Pei W, Deuitch N, Brooks S, Edwan JH, Joshi S, Prader S, Kaiser D, Owen WC, Sonbul AA, Zhang Y, Niemela JE, Burgess SM, Boehm M, Rehermann B, Chae J, Quezado MM, Ombrello AK, Buckley RH, Grom AA, Remmers EF, Pachlopnik JM, Su HC, Gutierrez-Cruz G, Hewitt SM, Sood R, Risma K, Calvo KR, Rosenzweig SD, Gadina M, Hafner M, Sun HW, Kastner DL, and Aksentijevich I
- Subjects
- Adult, Anemia, Sideroblastic blood, Child, Child, Preschool, Cytokines blood, Cytokines genetics, Developmental Disabilities genetics, Female, Genetic Diseases, X-Linked blood, Humans, Immunophenotyping, Male, Pedigree, Phenotype, Tumor Necrosis Factor-alpha analysis, Exome Sequencing, Anemia, Sideroblastic genetics, Anti-Inflammatory Agents therapeutic use, Genetic Diseases, X-Linked genetics, Immunologic Deficiency Syndromes genetics, Mutation, Nucleotidyltransferases genetics, RNA, Transfer genetics, Tumor Necrosis Factor-alpha antagonists & inhibitors
- Abstract
Objectives: To characterise the clinical features, immune manifestations and molecular mechanisms in a recently described autoinflammatory disease caused by mutations in TRNT1 , a tRNA processing enzyme, and to explore the use of cytokine inhibitors in suppressing the inflammatory phenotype., Methods: We studied nine patients with biallelic mutations in TRNT1 and the syndrome of congenital sideroblastic anaemia with immunodeficiency, fevers and developmental delay (SIFD). Genetic studies included whole exome sequencing (WES) and candidate gene screening. Patients' primary cells were used for deep RNA and tRNA sequencing, cytokine profiling, immunophenotyping, immunoblotting and electron microscopy (EM)., Results: We identified eight mutations in these nine patients, three of which have not been previously associated with SIFD. Three patients died in early childhood. Inflammatory cytokines, mainly interleukin (IL)-6, interferon gamma (IFN-γ) and IFN-induced cytokines were elevated in the serum, whereas tumour necrosis factor (TNF) and IL-1β were present in tissue biopsies of patients with active inflammatory disease. Deep tRNA sequencing of patients' fibroblasts showed significant deficiency of mature cytosolic tRNAs. EM of bone marrow and skin biopsy samples revealed striking abnormalities across all cell types and a mix of necrotic and normal-appearing cells. By immunoprecipitation, we found evidence for dysregulation in protein clearance pathways. In 4/4 patients, treatment with a TNF inhibitor suppressed inflammation, reduced the need for blood transfusions and improved growth., Conclusions: Mutations of TRNT1 lead to a severe and often fatal syndrome, linking protein homeostasis and autoinflammation. Molecular diagnosis in early life will be crucial for initiating anti-TNF therapy, which might prevent some of the severe disease consequences., Competing Interests: Competing interests: None declared., (© Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.)
- Published
- 2018
- Full Text
- View/download PDF
32. Histomorphological and Molecular Assessments of the Fixation Times Comparing Formalin and Ethanol-Based Fixatives.
- Author
-
Chung JY, Song JS, Ylaya K, Sears JD, Choi L, Cho H, Rosenberg AZ, and Hewitt SM
- Subjects
- Animals, Buffers, DNA analysis, Female, Immunohistochemistry methods, Mice, Mice, Inbred BALB C, Paraffin Embedding methods, Proteins analysis, RNA analysis, Time Factors, Ethanol chemistry, Fixatives chemistry, Formaldehyde chemistry, Tissue Fixation methods
- Abstract
The lack of standardization of tissue handling and processing hinders the development and validation of new biomarkers in research and clinical settings. We compared the histomorphology and the quality and quantity of biomolecules in paraffin-embedded mouse tissues, followed by fixation with neutral buffered formalin (NBF), 70% ethanol, and buffered ethanol (BE70) fixative. The quality of the histomorphology and immunohistochemistry in BE70 was relatively time-independent, whereas those in NBF rapidly decreased after 1 week of fixation. Protein recovered from tissue fixed in 70% ethanol and BE70 was compatible with Western blot and protein array using AKT and GAPDH antibodies, regardless of the fixation time. In addition, the quality and quantity of RNA extracted from tissue in ethanol-based fixative showed minimal changes from 4 hr to 6 months, whereas NBF had a dramatic detrimental change in RNA quality after 1 week of fixation. Furthermore, ethanol-based fixative offers a superior DNA template for PCR amplification-based molecular assays than NBF. In conclusion, coagulative, ethanol-based fixatives show a broader time spectrum than the aldehyde crosslinking fixative NBF in their histomorphological features and the quantity and quality of the biomolecules from paraffin-embedded tissue, and they may facilitate the use of fixative-fixed paraffin-embedded tissues in research and clinical laboratories, avoiding overfixation.
- Published
- 2018
- Full Text
- View/download PDF
33. Bcl-2-like Protein 11 (BIM) Expression Is Associated with Favorable Prognosis for Patients with Cervical Cancer.
- Author
-
Kim BW, Cho H, Ylaya K, Kitano H, Chung JY, Hewitt SM, and Kim JH
- Subjects
- Adult, Female, Humans, Immunohistochemistry, Kaplan-Meier Estimate, Multivariate Analysis, Prognosis, Survival Analysis, Bcl-2-Like Protein 11 metabolism, Uterine Cervical Neoplasms metabolism, Uterine Cervical Neoplasms pathology
- Abstract
Background/aim: Bcl-2-like protein 11 (BIM) is a pro-apoptotic member of the Bcl-2 protein family. BIM elicits cell death by binding to pro-survival Bcl-2 proteins. Even though the association of BIM expression with cell death has been investigated, its clinical survival significance in cervical cancer has not. In the current study, the prognostic significance of BIM in cervical cancer was investigated., Patients and Methods: The study included normal cervical tissues (n=254), cervical intraepithelial neoplasia (CIN) tissues (n=275), and invasive cervical cancer (n=164). In order to identify BIM expression, immunohistochemistry (IHC) was performed, and IHC scoring by quantitative digital image analysis was determined. Then, the association of BIM with prognostic factors was investigated., Results: BIM expression was higher in cervical cancer than normal cervical tissues (p<0.001). Well and moderate differentiation indicated higher BIM expression than did poor differentiation (p=0.001). Also, BIM expression was high in radiation-sensitive cervical cancer relative to radiation-resistant cancer (p=0.049). High BIM expression showed better 5-year disease-free survival (DFS) and overall survival (OS) rates (p=0.049 and π=0.030, respectively) than did low expression. In a multivariate analysis, BIM was shown to be an independent risk factor for DFS and OS in cervical cancer, with hazard ratios of 0.22 (p=0.006) and 0.46 (p=0.046), respectively., Conclusion: BIM is associated with favorable prognostic markers for prediction of DFS and OS in cervical cancer. High BIM expression is a potential prognostic marker as well as a chemotherapeutic target for cervical cancer., (Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)
- Published
- 2017
- Full Text
- View/download PDF
34. The anti-cancer effects of itraconazole in epithelial ovarian cancer.
- Author
-
Choi CH, Ryu JY, Cho YJ, Jeon HK, Choi JJ, Ylaya K, Lee YY, Kim TJ, Chung JY, Hewitt SM, Kim BG, Bae DS, and Lee JW
- Subjects
- Animals, Antineoplastic Agents pharmacology, Disease Models, Animal, Drug Synergism, Endothelial Cells physiology, Female, Heterografts, Itraconazole pharmacology, Mice, Neoplasm Transplantation, Paclitaxel pharmacology, Treatment Outcome, Antineoplastic Agents administration & dosage, Carcinoma, Ovarian Epithelial drug therapy, Cell Proliferation drug effects, Endothelial Cells drug effects, Itraconazole administration & dosage, Paclitaxel administration & dosage
- Abstract
We assessed the anti-proliferative activity of itraconazole using an EOC cell line (SKOV3ip1) and endothelial cell lines (HUVEC & SVEC4-10). We also examined angiogenesis (VEGFR2, p-ERK, p-PLCr1/2), hedgehog (Gli1, Ptch1, SMO), and mTOR (pS6K1) signaling pathways to determine the mechanism of action of itraconazole. Furthermore, we evaluated the synergistic effects of itraconazole and paclitaxel using orthotopic mouse models with established EOC cells (SKOV3ip1 or HeyA8) as well as patient-derived xenografts (PDXs). Itraconazole treatment inhibited proliferation of endothelial cells in a dose-dependent manner, but had no effect on EOC cells. The endothelial cell antiproliferative effect was associated with inhibition of hedgehog, and mTOR pathways and angiogenesis. In xenograft models of EOC using SKOV3ip1 or HeyA8, mice treated with the combination of itraconazole and paclitaxel had significantly decreased tumor weight than the control, paclitaxel-alone, or itraconazole-alone groups. Tissue derived from these tumors had significantly lower microvessel density than tissue from the other groups as well as hedgehog and mTOR pathway inhibition. We confirmed those effects in two EOC PDX models. These results suggest that itraconazole selectively inhibits endothelial cells rather than cancer cells by targeting multiple pathways including hedgehog, and mTOR pathways and angiogenesis.
- Published
- 2017
- Full Text
- View/download PDF
35. Common Molecular Subtypes Among Asian Hepatocellular Carcinoma and Cholangiocarcinoma.
- Author
-
Chaisaingmongkol J, Budhu A, Dang H, Rabibhadana S, Pupacdi B, Kwon SM, Forgues M, Pomyen Y, Bhudhisawasdi V, Lertprasertsuke N, Chotirosniramit A, Pairojkul C, Auewarakul CU, Sricharunrat T, Phornphutkul K, Sangrajrang S, Cam M, He P, Hewitt SM, Ylaya K, Wu X, Andersen JB, Thorgeirsson SS, Waterfall JJ, Zhu YJ, Walling J, Stevenson HS, Edelman D, Meltzer PS, Loffredo CA, Hama N, Shibata T, Wiltrout RH, Harris CC, Mahidol C, Ruchirawat M, and Wang XW
- Subjects
- Carcinoma, Hepatocellular diagnosis, Cholangiocarcinoma diagnosis, Cluster Analysis, Humans, Kaplan-Meier Estimate, Liver Neoplasms diagnosis, Prognosis, Transcriptome, Asian People genetics, Carcinoma, Hepatocellular genetics, Cholangiocarcinoma genetics, Liver Neoplasms genetics
- Abstract
Intrahepatic cholangiocarcinoma (ICC) and hepatocellular carcinoma (HCC) are clinically disparate primary liver cancers with etiological and biological heterogeneity. We identified common molecular subtypes linked to similar prognosis among 199 Thai ICC and HCC patients through systems integration of genomics, transcriptomics, and metabolomics. While ICC and HCC share recurrently mutated genes, including TP53, ARID1A, and ARID2, mitotic checkpoint anomalies distinguish the C1 subtype with key drivers PLK1 and ECT2, whereas the C2 subtype is linked to obesity, T cell infiltration, and bile acid metabolism. These molecular subtypes are found in 582 Asian, but less so in 265 Caucasian patients. Thus, Asian ICC and HCC, while clinically treated as separate entities, share common molecular subtypes with similar actionable drivers to improve precision therapy., (Published by Elsevier Inc.)
- Published
- 2017
- Full Text
- View/download PDF
36. A melanin-bleaching methodology for molecular and histopathological analysis of formalin-fixed paraffin-embedded tissue.
- Author
-
Chung JY, Choi J, Sears JD, Ylaya K, Perry C, Choi CH, Hong SM, Cho H, Brown KM, and Hewitt SM
- Subjects
- Formaldehyde, Humans, Paraffin Embedding, Histological Techniques, Hot Temperature, Hydrogen Peroxide, Melanins
- Abstract
Removal of excessive melanin from heavily pigmented formalin-fixed paraffin-embedded (FFPE) melanoma tissues is essential for histomorphological and molecular diagnostic assessments. Although there have been efforts to address this issue, current methodologies remain complex and time-consuming, and are not suitable for multiple molecular applications. Herein, we have developed a robust and rapid melanin-bleaching methodology for FFPE tissue specimens. Our approach is based on quick bleaching (15 min) at high temperature (80 °C) with 0.5% diluted hydrogen peroxide (H2O2) in Tris-HCl, PBS, or Tris/Tricine/SDS buffer. Immunostaining for Ki-67 and HMB45 was enhanced by bleaching with 0.5% H2O2 in Tris/Tricine/SDS and Tris-HCl, respectively. In addition to histopathological applications, our approach also facilitates recovery of protein and nucleic acid from archival melanin-rich FFPE tissue sections. Protein extracted from bleached FFPE tissues was compatible with western blotting using anti-human GAPDH and AKT antibodies. Our bleaching condition significantly improved RNA quality compared with unbleached tissues without compromising the yield. Notably, the RNA/DNA obtained from bleached tissues was suitable for end point PCR and real-time quantitative RT-PCR. In conclusion, this improved melanin-bleaching method enhances and simplifies immunostaining procedures, and facilitates the use of melanin-rich FFPE tissues for histomorphological and PCR amplification-based molecular assays.
- Published
- 2016
- Full Text
- View/download PDF
37. A Buffered Alcohol-Based Fixative for Histomorphologic and Molecular Applications.
- Author
-
Perry C, Chung JY, Ylaya K, Choi CH, Simpson A, Matsumoto KT, Smith WA, and Hewitt SM
- Subjects
- Animals, Buffers, DNA analysis, Histological Techniques, Kidney chemistry, Kidney cytology, Mice, Proteins analysis, RNA analysis, Tissue Fixation methods, Ethanol, Fixatives, Phosphates
- Abstract
Formalin-fixed paraffin-embedded (FFPE) tissue is the predominant preparation for diagnostic histopathological evaluation and increasingly the biospecimen on which molecular diagnostics are performed. However, formalin is carcinogenic and results in cross-linking of proteins and nicking and alterations of nucleic acids. Alternative fixatives, including 70% ethanol, improved biomolecular integrity; however, they have yet to replace neutral-buffered formalin (NBF). Herein, we describe the phosphate-buffered ethanol 70% (BE70) fixative. The histomorphology of BE70-fixed tissue is very similar to that of NBF; however, it is a non-cross-linking fixative and lacks the carcinogenic profile of formaldehyde-based fixatives. RNA isolated from tissue fixed in BE70 was of substantially higher quality and quantity than that was recovered from formalin-fixed tissue. Furthermore, the BE70 fixative showed excellent RNA and DNA integrity compared with that of NBF fixative based on real-time polymerase chain reaction analysis results. Immunohistochemical staining was similar for the antigen tested. In conclusion, BE70 is a non-cross-linking fixative that is superior to NBF and 70% ethanol with reference to biomolecule recovery and quality from paraffin-embedded tissue. Additional studies to compare the histomorphologic and immunohistochemical performance and utility in a clinical setting are required., (© 2016 The Histochemical Society.)
- Published
- 2016
- Full Text
- View/download PDF
38. Clinical significance of OCT4 and SOX2 protein expression in cervical cancer.
- Author
-
Kim BW, Cho H, Choi CH, Ylaya K, Chung JY, Kim JH, and Hewitt SM
- Subjects
- Adult, Aged, Aged, 80 and over, Biomarkers, Tumor analysis, Female, Humans, Immunohistochemistry, Middle Aged, Octamer Transcription Factor-3 analysis, Prognosis, SOXB1 Transcription Factors analysis, Uterine Cervical Neoplasms diagnosis, Uterine Cervical Neoplasms mortality, Young Adult, Uterine Cervical Dysplasia diagnosis, Uterine Cervical Dysplasia mortality, Octamer Transcription Factor-3 metabolism, SOXB1 Transcription Factors metabolism, Uterine Cervical Neoplasms metabolism, Uterine Cervical Dysplasia metabolism
- Abstract
Background: Cancer stem cell markers have become a major research focus because of their relationship with radiation or chemotherapy resistance in cancer therapy. Cancer stem cell markers including OCT4 and SOX2 have been found in various solid tumors. Here, we investigate the expression and clinical significance of OCT4 and SOX2 in cervical cancer., Methods: To define the clinical significance of OCT4 and SOX2 expression, we performed immunohistochemistry for OCT4 and SOX2 on 305 normal cervical epithelium samples, 289 cervical intraepithelial neoplasia samples, and 161 cervical cancer cases and compared the data with clinicopathologic factors, including survival rates of patients with cervical cancer., Results: OCT4 and SOX2 expression was higher in cervical cancer than normal cervix (both p < 0.001). OCT4 overexpression was associated with lymphovascular space invasion (p = 0.045), whereas loss of SOX2 expression was correlated with large tumor size (p = 0.015). Notably, OCT4 and SOX2 were significantly co-expressed in premalignant cervical lesions, but not in malignant cervical tumor. OCT4 overexpression showed worse 5-year disease-free and overall survival rates (p = 0.012 and p = 0.021, respectively) when compared to the low-expression group, while SOX2 expression showed favorable overall survival (p = 0.025). Cox regression analysis showed that OCT4 was an independent risk factor (hazard ratio = 11.23, 95 % CI, 1.31 - 95.6; p = 0.027) for overall survival while SOX2 overexpression showed low hazard ratio for death (hazard ratio = 0.220, 95 % CI, 0.06-0.72; p = 0.013)., Conclusions: These results suggest that OCT4 overexpression and loss of SOX2 expression are strongly associated with poor prognosis in patients with cervical cancer.
- Published
- 2015
- Full Text
- View/download PDF
39. Inhibition of Survivin with YM155 Induces Durable Tumor Response in Anaplastic Thyroid Cancer.
- Author
-
Mehta A, Zhang L, Boufraqech M, Liu-Chittenden Y, Zhang Y, Patel D, Davis S, Rosenberg A, Ylaya K, Aufforth R, Li Z, Shen M, and Kebebew E
- Subjects
- Adaptor Proteins, Signal Transducing chemistry, Animals, Cell Cycle, Cell Line, Tumor, Cell Proliferation, Cell Survival, Gene Expression Profiling, Gene Expression Regulation, Neoplastic, Genome-Wide Association Study, HeLa Cells, Humans, Immunohistochemistry, Inhibitory Concentration 50, Mice, Neoplasm Metastasis, RNA, Small Interfering metabolism, S Phase, Survivin, Treatment Outcome, Imidazoles chemistry, Inhibitor of Apoptosis Proteins antagonists & inhibitors, Naphthoquinones chemistry, Thyroid Carcinoma, Anaplastic drug therapy, Thyroid Carcinoma, Anaplastic immunology, Thyroid Neoplasms drug therapy, Thyroid Neoplasms immunology
- Abstract
Purpose: Anaplastic thyroid cancer (ATC) is a rare but lethal malignancy without any effective therapy. The aim of this study is to use a high-throughput drug library screening to identify a novel therapeutic agent that targets dysregulated genes/pathways in ATC., Experimentaldesign: We performed quantitative high-throughput screening (qHTS) in ATC cell lines using a compound library of 3,282 drugs. Dysregulated genes in ATC were analyzed using genome-wide expression analysis and immunohistochemistry in human ATC tissue samples and ATC cell lines. In vitro and in vivo studies were performed for determining drug activity, effectiveness of targeting, and the mechanism of action., Results: qHTS identified 100 active compounds in three ATC cell lines. One of the most active agents was the first-in-class survivin inhibitor YM155. Genome-wide expression analysis and immunohistochemistry showed overexpression of survivin in human ATC tissue samples, and survivin was highly expressed in all ATC cell lines tested. YM155 significantly inhibited ATC cellular proliferation. Mechanistically, YM155 inhibited survivin expression in ATC cells. Furthermore, YM155 treatment reduced claspin expression, which was associated with S-phase arrest in ATC cells. In vivo, YM155 significantly inhibited growth and metastases and prolonged survival., Conclusions: Our data show that YM155 is a promising anticancer agent for ATC and that its target, survivin, is overexpressed in ATC. Our findings support the use of YM155 in clinical trials as a therapeutic option in advanced and metastatic ATC., (©2015 American Association for Cancer Research.)
- Published
- 2015
- Full Text
- View/download PDF
40. Modification of Occupational Exposures on Bladder Cancer Risk by Common Genetic Polymorphisms.
- Author
-
Figueroa JD, Koutros S, Colt JS, Kogevinas M, Garcia-Closas M, Real FX, Friesen MC, Baris D, Stewart P, Schwenn M, Johnson A, Karagas MR, Armenti KR, Moore LE, Schned A, Lenz P, Prokunina-Olsson L, Banday AR, Paquin A, Ylaya K, Chung JY, Hewitt SM, Nickerson ML, Tardón A, Serra C, Carrato A, García-Closas R, Lloreta J, Malats N, Fraumeni JF Jr, Chanock SJ, Chatterjee N, Rothman N, and Silverman DT
- Subjects
- Adult, Aged, Female, Gene Deletion, Genetic Predisposition to Disease, Germ-Line Mutation, Glucuronosyltransferase genetics, Glutathione Transferase genetics, Humans, Male, Metallurgy, Microtubule-Associated Proteins genetics, Middle Aged, Occupational Diseases genetics, Receptor, Fibroblast Growth Factor, Type 3 genetics, Risk Factors, Scotland epidemiology, Surveys and Questionnaires, Ubiquitin-Protein Ligases genetics, Urinary Bladder Neoplasms genetics, Gene-Environment Interaction, Occupational Diseases epidemiology, Occupational Diseases etiology, Occupational Exposure adverse effects, Polymorphism, Single Nucleotide, Urinary Bladder Neoplasms epidemiology, Urinary Bladder Neoplasms etiology
- Abstract
Few studies have demonstrated gene/environment interactions in cancer research. Using data on high-risk occupations for 2258 case patients and 2410 control patients from two bladder cancer studies, we observed that three of 16 known or candidate bladder cancer susceptibility variants displayed statistically significant and consistent evidence of additive interactions; specifically, the GSTM1 deletion polymorphism (P interaction ≤ .001), rs11892031 (UGT1A, P interaction = .01), and rs798766 (TMEM129-TACC3-FGFR3, P interaction = .03). There was limited evidence for multiplicative interactions. When we examined detailed data on a prevalent occupational exposure associated with increased bladder cancer risk, straight metalworking fluids, we also observed statistically significant additive interaction for rs798766 (TMEM129-TACC3-FGFR3, P interaction = .02), with the interaction more apparent in patients with tumors positive for FGFR3 expression. All statistical tests were two-sided. The interaction we observed for rs798766 (TMEM129-TACC3-FGFR3) with specific exposure to straight metalworking fluids illustrates the value of integrating germline genetic variation, environmental exposures, and tumor marker data to provide insight into the mechanisms of bladder carcinogenesis., (Published by Oxford University Press 2015. This work is written by (a) US Government employee(s) and is in the public domain in the US.)
- Published
- 2015
- Full Text
- View/download PDF
41. Expression of stanniocalcin 1 in thyroid side population cells and thyroid cancer cells.
- Author
-
Hayase S, Sasaki Y, Matsubara T, Seo D, Miyakoshi M, Murata T, Ozaki T, Kakudo K, Kumamoto K, Ylaya K, Cheng SY, Thorgeirsson SS, Hewitt SM, Ward JM, and Kimura S
- Subjects
- Adult, Aged, Animals, Cell Line, Tumor, Female, Gene Expression Regulation, Neoplastic, Gene Regulatory Networks, Glycoproteins genetics, Humans, Male, Mice, Inbred C57BL, Neoplastic Stem Cells pathology, RNA, Messenger metabolism, Side-Population Cells pathology, Thyroid Gland pathology, Thyroid Neoplasms genetics, Thyroid Neoplasms pathology, Up-Regulation, Glycoproteins metabolism, Neoplastic Stem Cells metabolism, Side-Population Cells metabolism, Thyroid Gland metabolism, Thyroid Neoplasms metabolism
- Abstract
Background: Mouse thyroid side population (SP) cells consist of a minor population of mouse thyroid cells that may have multipotent thyroid stem cell characteristics. However the nature of thyroid SP cells remains elusive, particularly in relation to thyroid cancer. Stanniocalcin (STC) 1 and 2 are secreted glycoproteins known to regulate serum calcium and phosphate homeostasis. In recent years, the relationship of STC1/2 expression to cancer has been described in various tissues., Method: Microarray analysis was carried out to determine genes up- and down-regulated in thyroid SP cells as compared with non-SP cells. Among genes up-regulated, stanniocalcin 1 (STC1) was chosen for study because of its expression in various thyroid cells by Western blotting and immunohistochemistry., Results: Gene expression analysis revealed that genes known to be highly expressed in cancer cells and/or involved in cancer invasion/metastasis were markedly up-regulated in SP cells from both intact as well as partial thyroidectomized thyroids. Among these genes, expression of STC1 was found in five human thyroid carcinoma-derived cell lines as revealed by analysis of mRNA and protein, and its expression was inversely correlated with the differentiation status of the cells. Immunohistochemical analysis demonstrated higher expression of STC1 in the thyroid tumor cell line and thyroid tumor tissues from humans and mice., Conclusion: These results suggest that SP cells contain a population of cells that express genes also highly expressed in cancer cells including Stc1, which warrants further study on the role of SP cells and/or STC1 expression in thyroid cancer.
- Published
- 2015
- Full Text
- View/download PDF
42. Cell-cycle protein expression in a population-based study of ovarian and endometrial cancers.
- Author
-
Felix AS, Sherman ME, Hewitt SM, Gunja MZ, Yang HP, Cora RL, Boudreau V, Ylaya K, Lissowska J, Brinton LA, and Wentzensen N
- Abstract
Aberrant expression of cyclin-dependent kinase (CDK) inhibitors is implicated in the carcinogenesis of many cancers, including ovarian and endometrial cancers. We examined associations between CDK inhibitor expression, cancer risk factors, tumor characteristics, and survival outcomes among ovarian and endometrial cancer patients enrolled in a population-based case-control study. Expression (negative vs. positive) of three CDK inhibitors (p16, p21, and p27) and ki67 was examined with immunohistochemical staining of tissue microarrays. Logistic regression was used to estimate adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for associations between biomarkers, risk factors, and tumor characteristics. Survival outcomes were only available for ovarian cancer patients and examined using Kaplan-Meier plots and Cox proportional hazards regression. Among ovarian cancer patients (n = 175), positive p21 expression was associated with endometrioid tumors (OR = 12.22, 95% CI = 1.45-102.78) and higher overall survival (log-rank p = 0.002). In Cox models adjusted for stage, grade, and histology, the association between p21 expression and overall survival was borderline significant (hazard ratio = 0.65, 95% CI = 0.42-1.05). Among endometrial cancer patients (n = 289), positive p21 expression was inversely associated with age (OR ≥ 65 years of age = 0.25, 95% CI = 0.07-0.84) and current smoking status (OR: 0.33, 95% CI 0.15, 0.72) compared to negative expression. Our study showed heterogeneity in expression of cell-cycle proteins associated with risk factors and tumor characteristics of gynecologic cancers. Future studies to assess these markers of etiological classification and behavior may be warranted.
- Published
- 2015
- Full Text
- View/download PDF
43. Prognostic Significance of AMP-Dependent Kinase Alpha Expression in Cervical Cancer.
- Author
-
Choi CH, Chung JY, Cho H, Kitano H, Chang E, Ylaya K, Chung EJ, Kim JH, and Hewitt SM
- Subjects
- Biomarkers, Tumor, Disease Progression, Disease-Free Survival, Female, Humans, Immunohistochemistry, Isoenzymes genetics, Isoenzymes metabolism, Phosphorylation, Prognosis, Uterine Cervical Neoplasms diagnosis, AMP-Activated Protein Kinases genetics, AMP-Activated Protein Kinases metabolism, Uterine Cervical Neoplasms enzymology, Uterine Cervical Neoplasms genetics
- Abstract
Objectives: Cervical cancer is one of the most common gynecological malignancies worldwide, and its association with the AMP-activated protein kinase (AMPK) is still unknown. We aimed to investigate the clinical correlation between AMPK expression and cervical cancer., Methods: The expression of AMPKα1, AMPKα2 and phosphorylated AMPKα (p-AMPKα) was determined immunohistochemically in 524 formalin-fixed, paraffin-embedded malignant and premalignant cervical tissues. Subsequently, associations with clinicopathological characteristics and patient survival were assessed., Results: AMPKα2 expression was observed in the cytoplasm and nucleus, while expression of AMPKα1 and p-AMPKα was mainly observed in the cytoplasm. p-AMPKα expression increased during the normal-to-tumor transition of cervical carcinoma (p < 0.001), but, once cancer developed, the expression of AMPKα2 and p-AMPKα decreased in large-sized tumors when compared to smaller tumors (36 vs. 68%, p = 0.004 and 39 vs. 64%, p = 0.029, respectively). Notably, AMPKα2 expression was significantly associated with better disease-free survival (HR 0.29, 95% CI 0.10-0.86, p = 0.026)., Conclusion: The AMPKα2 isoform showed potential as a favorable prognostic marker in cervical cancer. Therefore, additional studies are necessary to further clarify the complex contribution of AMPK isoforms and of phosphorylation status to cervical cancer progression and prognosis., (© 2015 S. Karger AG, Basel.)
- Published
- 2015
- Full Text
- View/download PDF
44. The 19q12 bladder cancer GWAS signal: association with cyclin E function and aggressive disease.
- Author
-
Fu YP, Kohaar I, Moore LE, Lenz P, Figueroa JD, Tang W, Porter-Gill P, Chatterjee N, Scott-Johnson A, Garcia-Closas M, Muchmore B, Baris D, Paquin A, Ylaya K, Schwenn M, Apolo AB, Karagas MR, Tarway M, Johnson A, Mumy A, Schned A, Guedez L, Jones MA, Kida M, Hosain GM, Malats N, Kogevinas M, Tardon A, Serra C, Carrato A, Garcia-Closas R, Lloreta J, Wu X, Purdue M, Andriole GL Jr, Grubb RL 3rd, Black A, Landi MT, Caporaso NE, Vineis P, Siddiq A, Bueno-de-Mesquita HB, Trichopoulos D, Ljungberg B, Severi G, Weiderpass E, Krogh V, Dorronsoro M, Travis RC, Tjønneland A, Brennan P, Chang-Claude J, Riboli E, Prescott J, Chen C, De Vivo I, Govannucci E, Hunter D, Kraft P, Lindstrom S, Gapstur SM, Jacobs EJ, Diver WR, Albanes D, Weinstein SJ, Virtamo J, Kooperberg C, Hohensee C, Rodabough RJ, Cortessis VK, Conti DV, Gago-Dominguez M, Stern MC, Pike MC, Van Den Berg D, Yuan JM, Haiman CA, Cussenot O, Cancel-Tassin G, Roupret M, Comperat E, Porru S, Carta A, Pavanello S, Arici C, Mastrangelo G, Grossman HB, Wang Z, Deng X, Chung CC, Hutchinson A, Burdette L, Wheeler W, Fraumeni J Jr, Chanock SJ, Hewitt SM, Silverman DT, Rothman N, and Prokunina-Olsson L
- Subjects
- Case-Control Studies, Cyclin E metabolism, Gene Expression, Gene Frequency, Genome-Wide Association Study, Haplotypes, HeLa Cells, Humans, Oncogene Proteins metabolism, Polymorphism, Single Nucleotide, Urinary Bladder metabolism, Urinary Bladder pathology, Urinary Bladder Neoplasms pathology, Chromosomes, Human, Pair 19 genetics, Cyclin E genetics, Oncogene Proteins genetics, Urinary Bladder Neoplasms genetics
- Abstract
A genome-wide association study (GWAS) of bladder cancer identified a genetic marker rs8102137 within the 19q12 region as a novel susceptibility variant. This marker is located upstream of the CCNE1 gene, which encodes cyclin E, a cell-cycle protein. We performed genetic fine-mapping analysis of the CCNE1 region using data from two bladder cancer GWAS (5,942 cases and 10,857 controls). We found that the original GWAS marker rs8102137 represents a group of 47 linked SNPs (with r(2) ≥ 0.7) associated with increased bladder cancer risk. From this group, we selected a functional promoter variant rs7257330, which showed strong allele-specific binding of nuclear proteins in several cell lines. In both GWASs, rs7257330 was associated only with aggressive bladder cancer, with a combined per-allele OR = 1.18 [95% confidence interval (CI), 1.09-1.27, P = 4.67 × 10(-5)] versus OR = 1.01 (95% CI, 0.93-1.10, P = 0.79) for nonaggressive disease, with P = 0.0015 for case-only analysis. Cyclin E protein expression analyzed in 265 bladder tumors was increased in aggressive tumors (P = 0.013) and, independently, with each rs7257330-A risk allele (P(trend) = 0.024). Overexpression of recombinant cyclin E in cell lines caused significant acceleration of cell cycle. In conclusion, we defined the 19q12 signal as the first GWAS signal specific for aggressive bladder cancer. Molecular mechanisms of this genetic association may be related to cyclin E overexpression and alteration of cell cycle in carriers of CCNE1 risk variants. In combination with established bladder cancer risk factors and other somatic and germline genetic markers, the CCNE1 variants could be useful for inclusion into bladder cancer risk prediction models., (©2014 American Association for Cancer Research.)
- Published
- 2014
- Full Text
- View/download PDF
45. HER-2 assessment in formalin-fixed paraffin-embedded breast cancer tissue by well-based reverse phase protein array.
- Author
-
Perry C, Conway CM, Ha JW, Braunschweig T, Morris J, Ylaya K, Cho H, Chung JY, and Hewitt SM
- Abstract
Background: The human epidermal growth factor receptor-2 (HER-2) expression level is a critical element for determining the prognosis and management of breast cancer. HER-2 targeted therapy in breast cancer depends on the reliable assessment of HER-2 expression status but current standard methods are lacking a rigorous quantitative assay. To address this challenge, we developed an assessment of HER-2 expression method by well-based reverse phase protein array (RPPA)., Results: Well-based RPPA is based on a robust protein isolation methodology paired with a novel electrochemiluminescence detection system. HER-2 value of well-based RPPA significantly correlated with dot blotting results (R(2) = 0.939). By well-based RPPA, we successfully detected HER-2 expression in 76 human breast formalin-fixed paraffin-embedded tissue samples. We observed 93.4% (71/76) concordance between well-based RPPA and current HER-2 immunohistochemical assessment guideline. When the cutoff level of HER-2 value was set to 0.689 (HER-2/GAPDH) on the basis of receiver-operating characteristic curve, the area under the curve was 0.975 (95% CI, 0.941-1.000). Sensitivity and specificity of well-based RPPA was 92.1% and 94.7%, respectively., Conclusions: HER-2 value by well-based RPPA was correlated with the current HER-2 status guideline, suggesting that this normalized HER-2 assessment may offer advantages over unnormalized current immunohistochemical assessment methods.
- Published
- 2014
- Full Text
- View/download PDF
46. microRNA alterations driving acute and late stages of radiation-induced fibrosis in a murine skin model.
- Author
-
Simone BA, Ly D, Savage JE, Hewitt SM, Dan TD, Ylaya K, Shankavaram U, Lim M, Jin L, Camphausen K, Mitchell JB, and Simone NL
- Subjects
- Animals, Fibrosis, Leg Length Inequality pathology, Mice, Mice, Inbred C3H, Muscle, Skeletal metabolism, Muscle, Skeletal pathology, Muscle, Skeletal radiation effects, Radiation Injuries, Experimental genetics, Radiation Injuries, Experimental pathology, Skin metabolism, Skin pathology, Subcutaneous Tissue metabolism, Subcutaneous Tissue pathology, Subcutaneous Tissue radiation effects, Leg Length Inequality etiology, MicroRNAs metabolism, Proto-Oncogene Proteins c-met metabolism, Radiation Injuries, Experimental metabolism, Skin radiation effects
- Abstract
Purpose: Although ionizing radiation is critical in treating cancer, radiation-induced fibrosis (RIF) can have a devastating impact on patients' quality of life. The molecular changes leading to radiation-induced fibrosis must be elucidated so that novel treatments can be designed., Methods and Materials: To determine whether microRNAs (miRs) could be responsible for RIF, the fibrotic process was induced in the right hind legs of 9-week old CH3 mice by a single-fraction dose of irradiation to 35 Gy, and the left leg served as an unirradiated control. Fibrosis was quantified by measurements of leg length compared with control leg length. By 120 days after irradiation, the irradiated legs were 20% (P=.013) shorter on average than were the control legs., Results: Tissue analysis was done on muscle, skin, and subcutaneous tissue from irradiated and control legs. Fibrosis was noted on both gross and histologic examination by use of a pentachrome stain. Microarrays were performed at various times after irradiation, including 7 days, 14 days, 50 days, 90 days, and 120 days after irradiation. miR-15a, miR-21, miR-30a, and miR-34a were the miRs with the most significant alteration by array with miR-34a, proving most significant on confirmation by reverse transcriptase polymerase chain reaction, c-Met, a known effector of fibrosis and downstream molecule of miR-34a, was evaluated by use of 2 cell lines: HCT116 and 1522. The cell lines were exposed to various stressors to induce miR changes, specifically ionizing radiation. Additionally, in vitro transfections with pre-miRs and anti-miRs confirmed the relationship of miR-34a and c-Met., Conclusions: Our data demonstrate an inverse relationship with miR-34a and c-Met; the upregulation of miR-34a in RIF causes inhibition of c-Met production. miRs may play a role in RIF; in particular, miR-34a should be investigated as a potential target to prevent or treat this devastating side effect of irradiation., (Published by Elsevier Inc.)
- Published
- 2014
- Full Text
- View/download PDF
47. AR-regulated TWEAK-FN14 pathway promotes prostate cancer bone metastasis.
- Author
-
Yin J, Liu YN, Tillman H, Barrett B, Hewitt S, Ylaya K, Fang L, Lake R, Corey E, Morrissey C, Vessella R, and Kelly K
- Subjects
- Animals, Bone Neoplasms genetics, Gene Silencing, Heterografts, Humans, Male, Mice, Mice, Nude, Neoplasm Metastasis, Prostatic Neoplasms, Castration-Resistant genetics, Receptors, Androgen genetics, Receptors, Tumor Necrosis Factor genetics, Signal Transduction, TWEAK Receptor, Tissue Array Analysis, Treatment Outcome, Bone Neoplasms metabolism, Bone Neoplasms secondary, Prostatic Neoplasms, Castration-Resistant metabolism, Prostatic Neoplasms, Castration-Resistant pathology, Receptors, Androgen metabolism, Receptors, Tumor Necrosis Factor metabolism
- Abstract
The recurrence of prostate cancer metastases to bone after androgen deprivation therapy is a major clinical challenge. We identified FN14 (TNFRSF12A), a TNF receptor family member, as a factor that promotes prostate cancer bone metastasis. In experimental models, depletion of FN14 inhibited bone metastasis, and FN14 could be functionally reconstituted with IKKβ-dependent, NFκB signaling activation. In human prostate cancer, upregulated FN14 expression was observed in more than half of metastatic samples. In addition, FN14 expression was correlated inversely with androgen receptor (AR) signaling output in clinical samples. Consistent with this, AR binding to the FN14 enhancer decreased expression. We show here that FN14 may be a survival factor in low AR output prostate cancer cells. Our results define one upstream mechanism, via FN14 signaling, through which the NFκB pathway contributes to prostate cancer metastasis and suggest FN14 as a candidate therapeutic and imaging target for castrate-resistant prostate cancers., (©2014 American Association for Cancer Research.)
- Published
- 2014
- Full Text
- View/download PDF
48. Apoptosis inhibitor-5 overexpression is associated with tumor progression and poor prognosis in patients with cervical cancer.
- Author
-
Cho H, Chung JY, Song KH, Noh KH, Kim BW, Chung EJ, Ylaya K, Kim JH, Kim TW, Hewitt SM, and Kim JH
- Subjects
- Cell Line, Tumor, Disease Progression, Female, Gene Expression Regulation, Neoplastic, HeLa Cells, Humans, Phosphorylation, Prognosis, Survival Analysis, Tissue Array Analysis, Uterine Cervical Neoplasms metabolism, Apoptosis Regulatory Proteins metabolism, Mitogen-Activated Protein Kinase 1 metabolism, Mitogen-Activated Protein Kinase 3 metabolism, Nuclear Proteins metabolism, Uterine Cervical Neoplasms pathology
- Abstract
Background: The apoptosis inhibitor-5 (API5), anti-apoptosis protein, is considered a key molecule in the tumor progression and malignant phenotype of tumor cells. Here, we investigated API5 expression in cervical cancer, its clinical significance, and its relationship with phosphorylated extracellular signal-regulated kinase 1 and 2 (pERK1/2) in development and progression of cervical cancer., Methods: API5 effects on cell growth were assessed in cervical cancer cell lines. API5 and pERK1/2 immunohistochemical staining were performed on a cervical cancer tissue microarray consisting of 173 primary cervical cancers, 306 cervical intraepithelial neoplasias (CINs), and 429 matched normal tissues., Results: API5 overexpression promoted cell proliferation and colony formation in CaSki cells, whereas API5 knockdown inhibited the both properties in HeLa cells. Immunohistochemical staining showed that API5 expression increased during the normal to tumor transition of cervical carcinoma (P < 0.001), and this increased expression was significantly associated with tumor stage (P = 0.004), tumor grade (P < 0.001), and chemo-radiation response (P = 0.004). API5 expression levels were positively associated with pERK1/2 in cervical cancer (P < 0.001) and high grade CIN (P = 0.031). In multivariate analysis, API5+ (P = 0.039) and combined API5+/pERK1/2+ (P = 0.032) were independent prognostic factors for overall survival., Conclusions: API5 expression is associated with pERK1/2 in a subset of cervical cancer patients and its expression predicts poor overall survival, supporting that API5 may be a promising novel target for therapeutic interventions.
- Published
- 2014
- Full Text
- View/download PDF
49. CLPTM1L promotes growth and enhances aneuploidy in pancreatic cancer cells.
- Author
-
Jia J, Bosley AD, Thompson A, Hoskins JW, Cheuk A, Collins I, Parikh H, Xiao Z, Ylaya K, Dzyadyk M, Cozen W, Hernandez BY, Lynch CF, Loncarek J, Altekruse SF, Zhang L, Westlake CJ, Factor VM, Thorgeirsson S, Bamlet WR, Hewitt SM, Petersen GM, Andresson T, and Amundadottir LT
- Subjects
- Aneuploidy, Animals, Carcinoma, Pancreatic Ductal genetics, Carcinoma, Pancreatic Ductal metabolism, Cell Growth Processes physiology, Cell Line, Tumor, Female, HEK293 Cells, Heterografts, Humans, Membrane Proteins genetics, Membrane Proteins metabolism, Mice, Mice, Nude, Myosin Type II metabolism, Neoplasm Proteins genetics, Neoplasm Proteins metabolism, Pancreatic Neoplasms genetics, Pancreatic Neoplasms metabolism, Subcellular Fractions metabolism, Carcinoma, Pancreatic Ductal pathology, Membrane Proteins biosynthesis, Neoplasm Proteins biosynthesis, Pancreatic Neoplasms pathology
- Abstract
Genome-wide association studies (GWAS) of 10 different cancers have identified pleiotropic cancer predisposition loci across a region of chromosome 5p15.33 that includes the TERT and CLPTM1L genes. Of these, susceptibility alleles for pancreatic cancer have mapped to the CLPTM1L gene, thus prompting an investigation of the function of CLPTM1L in the pancreas. Immunofluorescence analysis indicated that CLPTM1L localized to the endoplasmic reticulum where it is likely embedded in the membrane, in accord with multiple predicted transmembrane domains. Overexpression of CLPTM1L enhanced growth of pancreatic cancer cells in vitro (1.3-1.5-fold; PDAY7 < 0.003) and in vivo (3.46-fold; PDAY68 = 0.039), suggesting a role in tumor growth; this effect was abrogated by deletion of two hydrophilic domains. Affinity purification followed by mass spectrometry identified an interaction between CLPTM1L and non-muscle myosin II (NMM-II), a protein involved in maintaining cell shape, migration, and cytokinesis. The two proteins colocalized in the cytoplasm and, after treatment with a DNA-damaging agent, at the centrosomes. Overexpression of CLPTM1L and depletion of NMM-II induced aneuploidy, indicating that CLPTM1L may interfere with normal NMM-II function in regulating cytokinesis. Immunohistochemical analysis revealed enhanced staining of CLPTM1L in human pancreatic ductal adenocarcinoma (n = 378) as compared with normal pancreatic tissue samples (n = 17; P = 1.7 × 10(-4)). Our results suggest that CLPTM1L functions as a growth-promoting gene in the pancreas and that overexpression may lead to an abrogation of normal cytokinesis, indicating that it should be considered as a plausible candidate gene that could explain the effect of pancreatic cancer susceptibility alleles on chr5p15.33., (©2014 American Association for Cancer Research.)
- Published
- 2014
- Full Text
- View/download PDF
50. Profiling of phospho-AKT, phospho-mTOR, phospho-MAPK and EGFR in non-small cell lung cancer.
- Author
-
Kitano H, Chung JY, Ylaya K, Conway C, Takikita M, Fukuoka J, Doki Y, Hanaoka J, and Hewitt SM
- Subjects
- Aged, Biomarkers metabolism, Carcinoma, Non-Small-Cell Lung enzymology, Carcinoma, Non-Small-Cell Lung pathology, Cluster Analysis, Female, Humans, Lung Neoplasms enzymology, Lung Neoplasms pathology, Male, Middle Aged, Phosphorylation, Survival Analysis, Carcinoma, Non-Small-Cell Lung metabolism, ErbB Receptors metabolism, Lung Neoplasms metabolism, Mitogen-Activated Protein Kinases metabolism, Proto-Oncogene Proteins c-akt metabolism, TOR Serine-Threonine Kinases metabolism
- Abstract
Activation of numerous pathways has been documented in non-small cell lung cancer (NSCLC). Epidermal growth factor receptor (EGFR) has emerged as a common therapeutic target. The mitogen-activated protein kinase (MAPK) and AKT signaling pathways are downstream of EGFR and deregulated via genetic and epigenetic mechanisms in many human cancers. We evaluated selected markers in the EGFR pathway with reference to outcome. Tissues from 220 cases of NSCLC patients presented in a tissue microarray were assayed with immunohistochemistry for phosphorylated AKT, phosphorylated MAPK, phosphorylated mTOR, and EGFR and then quantified by automated image analysis. Individually, the biomarkers did not predict. Combined as ratios, p-mTOR/p-AKT, and p-MAPK/EGFR function as prognostic markers of survival (p=0.008 and p=0.029, respectively), however, no significance was found after adjustment (p=0.221, p=0.103). The sum of these ratios demonstrates a stronger correlation with survival (p<0.001) and remained statistically significant after adjustment (p=0.026). The algebraic combination of biomarkers offer the capacity to understand factors that predict outcome better than current approaches of evaluating biomarkers individually or in pairs. Our results show the sum of p-mTOR/p-AKT and p-MAPK/EGFR is a potential predictive marker of survival in NSCLC patients.
- Published
- 2014
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.