GLI1 Transcription Factor Affects Tumor Aggressiveness in Patients With Papillary Thyroid Cancers

Lee, Jandee,Jeong, Seonhyang,Lee, Cho Rok,Ku, Cheol Ryong,Kang, Sang-Wook,Jeong, Jong Ju,Nam, Kee-Hyun,Shin, Dong Yeob,Chung, Woong Youn,Lee, Eun Jig,Jo, Young Suk Williams & Wilkins Co 2015 Medicine Vol.94 No.25

<▼1><P>Supplemental Digital Content is available in the text</P></▼1><▼2><P><B>Abstract</B></P><P>A significant proportion of patients with papillary thyroid cancer (PTC) present with extrathyroidal extension (ETE) and lymph node metastasis (LNM). However, the molecular mechanism of tumor invasiveness in PTC remains to be elucidated.</P><P>The aim of this study is to understand the role of Hedgehog (Hh) signaling in tumor aggressiveness in patients with PTC.</P><P>Subjects were patients who underwent thyroidectomy from 2012 to 2013 in a single institution. Frozen or paraffin-embedded tumor tissues with contralateral-matched normal thyroid tissues were collected. Hh signaling activity was analyzed by quantitative RT-PCR (qRT-PCR) and immunohistochemical (IHC) staining. Datasets from Gene Expression Omnibus (GEO) (National Center for Biotechnology Information) were subjected to Gene Set Enrichment Analysis (GSEA). BRAFT1799A and telomerase reverse transcriptase promoter mutation C228T were analyzed by direct sequencing.</P><P>Among 137 patients with PTC, glioma-associated oncogene homolog 1 (<I>GLI1</I>) group III (patients in whom the ratio of <I>GLI1</I> messenger ribonucleic acid (mRNA) level in tumor tissue to <I>GLI1</I> mRNA level in matched normal tissue was in the upper third of the subject population) had elevated risk for ETE (odds ratio [OR] 4.381, 95% confidence interval [CI] 1.414–13.569, <I>P</I> = 0.01) and LNM (OR 5.627, 95% CI 1.674–18.913, <I>P</I> = 0.005). Glioma-associated oncogene homolog 2 (<I>GLI2</I>) group III also had elevated risk for ETE (OR 4.152, 95% CI 1.292–13.342, <I>P</I> = 0.017) and LNM (OR 3.924, 95% CI 1.097–14.042, <I>P</I> = 0.036). GSEA suggested that higher <I>GLI1</I> expression is associated with expression of the <I>KEGG</I> gene set related to axon guidance (<I>P</I> = 0.031, false discovery rate < 0.05), as verified by qRT-PCR and IHC staining in our subjects.</P><P><I>GLI1</I> and <I>GLI2</I> expressions were clearly related to aggressive clinicopathological features and aberrant activation of GLI1 involved in the axon guidance pathway. These results may contribute to development of new prognostic markers, as well as novel therapeutic targets.</P></▼2>

Optimal Cut-Off Values of Lymph Node Ratio Predicting Recurrence in Papillary Thyroid Cancer

Lee, Seul Gi,Ho, Joon,Choi, Jung Bum,Kim, Tae Hyung,Kim, Min Jhi,Ban, Eun Jeong,Lee, Cho Rok,Kang, Sang-Wook,Jeong, Jong Ju,Nam, Kee-Hyun,Jung, Sang Geun,Jo, Young Suk,Lee, Jandee,Chung, Woong Youn Wolters Kluwer Health 2016 Medicine Vol.95 No.5

<▼1><P>Supplemental Digital Content is available in the text</P></▼1><▼2><P><B>Abstract</B></P><P>Regional lymph node (LN) metastasis has a significant impact for prediction of recurrence in patients with papillary thyroid cancers (PTC); however, the prognostic value of the lymph node ratio (LNR), which is defined as the ratio of the number of metastatic LNs to the total number of investigated LNs, is controversial. In this study, we determined the optimal cut-off values of LNRs for the prediction of recurrence in PTC patients.</P><P>This large cohort study retrospectively evaluated 2294 patients who had undergone total thyroidectomy for PTC at a single institution from October 1985 to June 2009. The prediction probability of central LNR (cLNR, level VI) and total LNR (tLNR, levels II–VI) were estimated by binominal logistic regression analysis. Hazard ratios of the cut-off LNR values for cancer recurrence were calculated for relevant covariates using multivariate Cox regression analyses. Kaplan–Meier analyses were also utilized to assess the effects of estimated LNR cut-off values on recurrence-free survival (RFS).</P><P>Of the 2294 patients, 138 (6.0%) presented cancer recurrence during the follow-up period (median duration = 107.1 months). The prediction probability indicated that LNRs of 0.4 and 0.5 for central LN and total LN, respectively, are optimal cut-off values for precise prediction with minimization of outliers. Multivariate Cox regression analyses revealed that cLNR ≥0.4 was independently predictive of recurrence in patients with N0 and N1a PTCs (hazard ratio [HR]: 7.016, 95% confidence interval [CI]: 3.72–12.986, <I>P</I> < 0.001) and that tLNR ≥0.5 indicated a significantly increased risk of recurrence in patients with N1b PTCs (HR: 2.372, 95% CI: 1.458–3.860, <I>P</I> < 0.001). In addition, Kaplan–Meier analyses clearly demonstrated that these LNR cut-off values are precisely operational in RFS estimation.</P><P>The cut-off LNR values of 0.4 and 0.5 for cLNR and tLNR, respectively, were identified. Risk stratification combined with these LNR cut-off values may prove useful to determine treatment and follow-up strategies for PTC patients.</P></▼2>

Practical Performance of the 2015 American Thyroid Association Guidelines for Predicting Tumor Recurrence in Patients with Papillary Thyroid Cancer in South Korea

Lee, Seul Gi,Lee, Woo Kyung,Lee, Hye Sun,Moon, Jieun,Lee, Cho Rok,Kang, Sang Wook,Jeong, Jong Ju,Nam, Kee-Hyun,Chung, Woong Youn,Jo, Young Suk,Lee, Jandee Mary Ann Liebert 2017 Thyroid Vol.27 No.2

KSR1 is coordinately regulated with Notch signaling and oxidative phosphorylation in thyroid cancer

Lee, Jandee,Seol, Mi-Youn,Jeong, Seonhyang,Kwon, Hyeong Ju,Lee, Cho Rok,Ku, Cheol Ryong,Kang, Sang-Wook,Jeong, Jong Ju,Shin, Dong Yeob,Nam, Kee-Hyun,Lee, Eun Jig,Chung, Woong Youn,Jo, Young Suk Journal of Endocrinology (Ltd. by Guarantee) 2015 Journal of molecular endocrinology Vol.54 No.2

<P>Kinase suppressor of RAS1 (KSR1) is a scaffold protein implicated in RAS-mediated RAF activation. However, the molecular function of KSR in papillary thyroid cancer (PTC) is unknown. Thus, this study aimed to characterize the role of KSR1 in patients with PTC. qRT-PCR and immunohistochemistry (IHC) revealed inter-tumor heterogeneities in the expression of KSR1 in PTC tissues. Interestingly, BRAFV600E-positive PTC showed higher <I>KSR1</I> mRNA expression than BRAFV600E-negative PTC (<I>P</I><0.001). Gene Set Enrichment Analysis (GSEA) using public repositories showed that high KSR1 expression coordinately upregulated Notch signaling (nominal <I>P</I>=0.019, false discovery rate (FDR) <I>q</I>-value=0.165); this finding was supported by GeneNetwork analysis, indicating that <I>KSR1</I> expression is positively correlated with <I>NOTCH1</I> expression (<I>ρ</I>=0.677, <I>P</I>=6.15×10<SUP>−9</SUP>). siRNA against KSR1 (siKSR1) significantly decreased ERK phosphorylation induced by BRAFV600E, resulting in reduced expression of <I>NOTCH1</I> and <I>HES1</I>, targets of Notch signaling. GSEA revealed that high KSR1 expression was also associated with downregulation of genes related to oxidative phosphorylation (OxPhos). Consistent with this, electron microscopy showed that PTCs with high KSR1 expression exhibited structural defects of the mitochondrial cristae. Furthermore, siKSR1-transfected BCPAP and 8505C cells generated fewer colonies in colony-forming assays. In addition, GSEA showed that high expression of KSR2 and connector enhancer of KSR1 (CNKSR1) also coordinately upregulated Notch signaling (KSR2: nominal <I>P</I>=0.0097, FDR <I>q</I>-value=0.154 and CNKSR1: nominal <I>P</I><0.0001, FDR <I>q</I>-value=0.00554), and high CNKSR2 was associated with downregulation of the OxPhos gene set (nominal <I>P</I><0.0001, FDR <I>q</I>-value <0.0001). In conclusion, KSR1 is coordinately regulated with Notch signaling and OxPhos in PTC, because its scaffold function might be required to sustain the proliferative signaling and metabolic remodeling associated with this type of cancer.</P>

A Metabolic Phenotype Based on Mitochondrial Ribosomal Protein Expression as a Predictor of Lymph Node Metastasis in Papillary Thyroid Carcinoma

Lee, Jandee,Seol, Mi-Youn,Jeong, Seonhyang,Lee, Cho Rok,Ku, Cheol Ryong,Kang, Sang-Wook,Jeong, Jong Ju,Shin, Dong Yeob,Nam, Kee-Hyun,Lee, Eun Jig,Chung, Woong Youn,Jo, Young Suk Wolters Kluwer Health 2015 Medicine Vol.94 No.2

<▼1><P>Supplemental Digital Content is available in the text</P></▼1><▼2><P><B>Abstract</B></P><P>Metabolic reprogramming has been regarded as an essential component of malignant transformation. However, the clinical significance of metabolic heterogeneity remains poorly characterized.</P><P>The aim of this study was to characterize metabolic heterogeneity in thyroid cancers via the analysis of the expression of mitochondrial ribosomal proteins (MRPs) and genes involved in oxidative phosphorylation (OxPhos), and investigate potential prognostic correlations.</P><P>Gene set enrichment analysis (GSEA) verified by reverse transcription polymerase chain reaction and gene network analysis was performed using public repository data. Cross-sectional observational study was conducted to classify papillary thyroid cancer (PTC) by the expression of MRP L44 (MRPL44) messenger RNA (mRNA), and to investigate the clinicopathological features.</P><P>GSEA clearly showed that the expression of OxPhos and MRP gene sets was significantly lower in primary thyroid cancer than in matched normal thyroid tissue. However, 8 of 49 primary thyroid tumors (16.3%) in the public repository did not show a reduction in OxPhos mRNA expression. Remarkably, strong positive correlations between MRPL44 expression and those of OxPhos and MRPs such as reduced nicotinamide adenine dinucleotide dehydrogenase (ubiquinone) 1 α subcomplex, 5; succinate dehydrogenase complex, subunit D; cytochrome c, somatic; adenosine triphosphate synthase, H+ transporting, mitochondrial Fo complex, subunit C1 (subunit 9); and MRP S5 (MRPS5) (<I>P</I> < 0.0001) were clearly denoted, suggesting that MRPL44 is a representative marker of OxPhos and MRP expressions. In laboratory experiments, metabolic heterogeneity in oxygen consumption, extracellular acidification rates (ECARs), and amounts of OxPhos complexes were consistently observed in BCPAP, TPC1, HTH-7, and XTC.UC1 cell lines. In PTCs, metabolic phenotype according to OxPhos amount defined by expression of MRPL44 mRNA was significantly related to lymph node metastasis (LNM) (<I>P</I> < 0.001). Furthermore, multivariate analysis clearly indicated that expression of MRPL44 is associated with an increased risk of lateral neck LNM (odds ratio 9.267, 95% confidence interval 1.852–46.371, <I>P</I> = 0.007).</P><P>MRPL44 expression may be a representative marker of metabolic phenotype according to OxPhos amount and a useful predictor of LNM.</P></▼2>

Association Between Obesity and BRAFV600E Mutation Status in Patients with Papillary Thyroid Cancer

Lee, Jandee,Lee, Cho Rok,Ku, Cheol Ryong,Kang, Sang-Wook,Jeong, Jong Ju,Shin, Dong Yeob,Nam, Kee-Hyun,Jung, Sang Geun,Lee, Eun Jig,Chung, Woong Youn,Jo, Young Suk Springer - Society of Surgical Oncology 2015 Annals of Surgical Oncology Vol.22 No.suppl3

Whole Exome Sequencing Identifies a Novel Hedgehog-Interacting Protein G516R Mutation in Locally Advanced Papillary Thyroid Cancer

Lee, Woo Kyung,Lee, Seul Gi,Yim, Seung Hyuk,Kim, Daham,Kim, Hyunji,Jeong, Seonhyang,Jung, Sang Geun,Jo, Young Suk,Lee, Jandee MDPI 2018 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.19 No.10

<P>Locally advanced thyroid cancer exhibits aggressive clinical features requiring extensive neck dissection. Therefore, it is important to identify changes in the tumor biology before local progression. Here, whole exome sequencing (WES) using tissues from locally advanced papillary thyroid cancer (PTC) presented a large number of single nucleotide variants (SNVs) in the metastatic lymph node (MLN), but not in normal tissues and primary tumors. Among those MLN-specific SNVs, a novel HHIP G516R (G1546A) mutation was also observed. Interestingly, in-depth analysis for exome sequencing data from the primary tumor presented altered nucleotide ‘A’ at a very low frequency indicating intra-tumor heterogeneity between the primary tumor and MLN. Computational prediction models such as PROVEAN and Polyphen suggested that HHIP G516R might affect protein function and stability. In vitro, HHIP G516R increased cell proliferation and promoted cell migration in thyroid cancer cells. HHIP G516R, a missense mutation, could be a representative example for the intra-tumor heterogeneity of locally advanced thyroid cancer, which can be a potential future therapeutic target for this disease.</P>

10 Years Experience using Robot- Assisted Gasless Transaxillary Thyroidectomy for the Management of Graves’ Disease

Lee Cho Rok,Kwangsoon Kim,Jin Kyong Kim,Sang-Wook Kang,Jandee Lee,Jong Ju Jeong,Kee-Hyun Nam,Woong Youn Chung 대한외과학회 2018 대한외과학회 학술대회 초록집 Vol.2018 No.11

Distinct Features of Nonthyroidal Illness in Critically Ill Patients With Infectious Diseases

Lee, Woo Kyung,Hwang, Sena,Kim, Daham,Lee, Seul Gi,Jeong, Seonhyang,Seol, Mi-Youn,Kim, Hyunji,Ku, Cheol Ryong,Shin, Dong Yeop,Chung, Woong Youn,Lee, Eun Jig,Lee, Jandee,Jo, Young Suk Wolters Kluwer Health 2016 Medicine Vol.95 No.14

<▼1><P>Supplemental Digital Content is available in the text</P></▼1><▼2><P><B>Abstract</B></P><P>Nonthyroidal illness (NTI), often observed in critically ill patients, arises through diverse alterations in the hypothalamus-pituitary-thyroid (HPT) axis. However, the causal relationship between underlying disease and NTI diversity in critically ill patients is poorly understood.</P><P>The aim of this study was to examine NTI severity and adverse outcomes in critically ill patients with respect to their underlying disease(s).</P><P>The medical records of 616 patients admitted to the intensive care unit (ICU) between January 2009 and October 2014 were retrospectively reviewed. Patients with known diseases or taking medications that affect thyroid function were excluded. All-cause mortality (ACM) and length of stay (LOS) in the ICU were assessed as adverse outcomes.</P><P>The enrolled patients (n = 213) were divided into the following 4 groups according to the severity of NTI at the nadir of their thyroid function test (TFT): normal (n = 11, 5.2%), mild NTI (n = 113, 53.1%), moderate NTI (n = 78, 36.6%), and severe NTI (n = 11, 5.2%). There was no significant difference between the groups in terms of age and gender. NTI severity showed a significantly strong association with ACM (<I>P</I> < 0.0001) and a significant positive association with LOS in the ICU (<I>P</I> = 0.031). After adjusting for age, gender, and current medications affecting TFT, increasing NTI severity led to increased ACM (odds ratio = 3.101; 95% confidence interval = 1.711–5.618; <I>P</I> < 0.0001). Notably, the prevalence of moderate-to-severe NTI was markedly higher in patients with infectious disease than in those with noninfectious disease (<I>P</I> = 0.012). Consistent with this, serum C-reactive protein levels were higher in patients with moderate-to-severe NTI (<I>P</I> = 0.016).</P><P>NTI severity is associated with increased ACM, LOS, and underlying infectious disease. Future studies will focus on the biological and clinical implications of infectious disease on the HPT axis.</P></▼2>

Aberrant Expression of COT Is Related to Recurrence of Papillary Thyroid Cancer

Lee, Jandee,Jeong, Seonhyang,Park, Jae Hyun,Lee, Cho Rok,Ku, Cheol Ryong,Kang, Sang-Wook,Jeong, Jong Ju,Nam, Kee-Hyun,Shin, Dong Yeob,Lee, Eun Jig,Chung, Woong Youn,Jo, Young Suk Williams & Wilkins Co 2015 Medicine Vol.94 No.6

<▼1><P>Supplemental Digital Content is available in the text</P></▼1><▼2><P><B>Abstract</B></P><P>Aberrant expression of Cancer Osaka Thyroid Oncogene mitogen-activated protein kinase kinase kinase 8 (COT) (MAP3K8) is a driver of resistance to B-RAF inhibition. However, the de novo expression and clinical implications of COT in papillary thyroid cancer (PTC) have not been investigated.</P><P>The aim of this study is to investigate the expression of A-, B-, C-RAF, and COT in PTC (n = 167) and analyze the clinical implications of aberrant expression of these genes.</P><P>Quantitative polymerase chain reaction (qPCR) and immunohistochemical staining (IHC) were performed on primary thyroid cancers. Expression of COT was compared with clinicopathological characteristics including recurrence-free survival. Datasets from public repository (NCBI) were subjected to Gene Set Enrichment Analysis (GSEA).</P><P>qPCR data showed that the relative mRNA expression of <I>A-</I>, <I>B-</I>, <I>C-RAF</I> and <I>COT</I> of PTC were higher than normal tissues (all <I>P</I> < 0.01). In addition, the expression of COT mRNA in PTC showed positive correlation with A- (<I>r</I> = 0.4083, <I>P</I> < 0.001), B- (<I>r</I> = 0.2773, <I>P</I> = 0.0003), and C-RAF (<I>r</I> = 0.5954, <I>P</I> < 0.001). The mRNA expressions of A-, B,- and C-RAF were also correlated with each other (all <I>P</I> < 0.001). In IHC, the staining intensities of B-RAF and COT were higher in PTC than in normal tissue (<I>P</I> < 0.001). Interestingly, moderate-to-strong staining intensities of B-RAF and COT were more frequent in B-RAF<SUP>V600E</SUP>-positive PTC (<I>P</I> < 0.001, <I>P</I> = 0.013, respectively). In addition, aberrant expression of COT was related to old age at initial diagnosis (<I>P</I> = 0.045) and higher recurrence rate (<I>P</I> = 0.025). In multivariate analysis, tumor recurrence was persistently associated with moderate-to-strong staining of COT after adjusting for age, sex, extrathyroidal extension, multifocality, T-stage, N-stage, TNM stage, and B-RAF<SUP>V600E</SUP> mutation (odds ratio, 4.662; 95% confidence interval 1.066 − 21.609; <I>P</I> = 0.045). Moreover, moderate-to-strong COT expression in PTC was associated with shorter recurrence-free survival (mean follow-up duration, 14.2 ± 4.1 years; <I>P</I> = 0.0403). GSEA indicated that gene sets related to B-RAF-RAS (<I>P</I> < 0.0001, false discovery rate [FDR] <I>q</I>-value = 0.000) and thyroid differentiation (<I>P</I> = 0.048, FDR <I>q</I>-value = 0.05) scores were enriched in lower COT expression group and gene sets such as T-cell receptor signaling pathway and Toll-like receptor signaling pathway are coordinately upregulated in higher COT expression group (both, <I>P</I> < 0.0001, FDR <I>q</I>-value = 0.000).</P><P>Aberrant expression of A-, B-, and C-RAF, and COT is frequent in PTC; increased expression of COT is correlated with recurrence of PTC.</P></▼2>