GO-74 : MICA/B and ULBP1 NKG2D ligands are independent predictors of good prognosis in cervical cancer

( Han Byoul Cho ),( Joon Yong Chung ),( Till Braunschweig ),( Stephen M Hewitt ),( Jae Hoon Kim ) 대한산부인과학회 2012 대한산부인과학회 학술대회 Vol.99 No.-

NKG2D (natural killer group 2, member D) binds to a diverse array of cellular ligands of the MIC and ULBP/RAET family and is thought to participate in anticancer immune response. In this study, we investigated the clinical significance of NKG2D in the pathogenesis of cervical cancer. We assessed the expression of all MICA/B, ULBP1, ULBP2, ULBP3, RAET1E, and RAET1G proteins in archival tumor tissue specimens from 200 cervical cancers, 327 high-grade cervical intraepithelial neoplasias (CINs), 99 low-grade CINs, and 541 matched nonadjacent normal cervical epithelial tissues by immunohistochemical staining. MICA/B, ULBP1, and RAET1E expression were higher in cervical cancer than in low grade CIN (p<0.001, p=0.012, p=0.013, respectively) and normal cervix (all p<0.001). Among these markers, expression of ULBP1 was significantly different in FIGO stage (p=0.010) and tumor size (p=0.045). ULBP1 expression was correlated with MICA/B (p<0.001) and ULBP2 expression (p=0.002) in cervical cancer. MICA/B+ or ULBP1+ showed improved disease-free survival time (p=0.027 and p=0.009, respectively) than low expression group, whereas RAET1E+ or RAET1G+ showed shorter survival time (p=0.018 and p=0.029, respectively). In case of overall survival, ULBP1+ group showed significantly longer survival time (p=0.009). By using multivariate analysis, MICA/B+/ULBP1+ (HR=0.16, p=0.015), ULBP1+ (HR=0.31, p=0.024), tumor stage (HR=3.60, p=0.010), and lymph node metastasis (HR=2.71, p=0.032) were found to be independent predictors of prognosis for disease-free survival in cervical cancer. We demonstrate that MICA/B and ULBP1 expression is significantly increased in cervical cancer, which is consistent with immunoediting mechanism that select selects tumor cells that have lost or reduced expression of NKG2D ligands. Furthermore, the combination of MICA/B and ULBP1 was found to be the independent predictor of survival, suggesting clinical values in clinical assessment.

The expression of makorin ring finger protein 1 (MKRN1), pAKT, pmTOR, and PTEN in cervical neoplasias

( Han Byoul Cho ),( Joon Yong Chung ),( Mikiko Takikita ),( Stephen M Hewitt ),( Jae Hoon Kim ) 대한산부인과학회 2012 대한산부인과학회 학술대회 Vol.98 No.-

The protein kinase B (AKT) pathway plays a key role in the regulation of cell death and survival, and has been proposed as a central signaling event in carcinogenesis. We investigated the expression of MKRN1, which is a transcriptional co-regulator and an E3 ligase, and sought to define the role of AKT pathway in cervical neoplasias. One hundred eighty three cervical cancer, 415 cervical intraepithelial neoplasia, and 400 matched nonadjacent normal tissues were arrayed into tissue microarrays. MKRN1, phosphorylated AKT (pAKT), phosphorylated mammalian target of rapamycin (pmTOR), and total phosphatase and tensin homolog deleted on chromosome 10 (PTEN) protein expressions were assessed by immunohistochemistry (IHC) and the relationship between MKRN1 expression and clinicopathologic parameters including survival data was studied. MKRN1, pAKT, and pmTOR expressions were significantly increased in cervical cancer cases compared with normal epitheliums (p<0.001 for all markers), and this increased expression of MKRN1 was significantly associated with tumor stage (p=0.018) and tumor grade (p<0.001). Expression of MKRN1 was positively associated with pAKT expression (p=0.023), while negatively associated with PTEN expression (p=0.016) in cervical cancer specimens. In multivariate analysis, MKRN1+/pAKT+ (HR = 4.76 [1.66-13.58], p=0.004), pAKT+ (HR = 2.97 [1.02-8.06], p=0.045), and lymph node metastasis (HR = 3.94 [1.07-14.45], p=0.039) were independent prognostic factors for overall survival. This study provides evidence of an association between AKT pathway and cervical carcinogenesis and shows that MKRN1/pAKT expression predicts poor prognosis in cervical cancer. Our findings also suggest that future research assessing its clinical usefulness would be worthwhile.

Nanog signaling in cancer promotes stem-like phenotype and immune evasion.

Noh, Kyung Hee,Kim, Bo Wook,Song, Kwon-Ho,Cho, Hanbyoul,Lee, Young-Ho,Kim, Jin Hee,Chung, Joon-Yong,Kim, Jae-Hoon,Hewitt, Stephen M,Seong, Seung-Yong,Mao, Chih-Ping,Wu, T-C,Kim, Tae Woo American Society for Clinical Investigation 2012 The Journal of clinical investigation Vol.122 No.11

<P>Adaptation of tumor cells to the host is a major cause of cancer progression, failure of therapy, and ultimately death. Immune selection drives this adaptation in human cancer by enriching tumor cells with a cancer stem cell-like (CSC-like) phenotype that makes them resistant to CTL-mediated apoptosis; however, the mechanisms that mediate CSC maintenance and proliferation are largely unknown. Here, we report that CTL-mediated immune selection drives the evolution of tumor cells toward a CSC-like phenotype and that the CSC-like phenotype arises through the Akt signaling pathway via transcriptional induction of Tcl1a by Nanog. Furthermore, we found that hyperactivation of the Nanog/Tcl1a/Akt signaling axis was conserved across multiple types of human cancer. Inhibition of Nanog in a murine model of colon cancer rendered tumor cells susceptible to immune-mediated clearance and led to successful, long-term control of the disease. Our findings establish a firm link among immune selection, disease progression, and the development of a stem-like tumor phenotype in human cancer and implicate the Nanog/Tcl1a/Akt pathway as a central molecular target in this process.</P>

Identification of candidate susceptible genes associated with ovarian clear cell carcinoma using cis-eQTL analysis

( Jihye Kim ),( Joon-yong Chung ),( Tae-joong Kim ),( Jeong-won Lee ),( Byoung-gie Kim ),( Duk-soo Bae ),( Chel Hun Choi ),( Stephen M. Hewitt ) 대한산부인과학회 2018 대한산부인과학회 학술대회 Vol.104 No.-

Objective: Ovarian clear cell adenocarcinoma (Ov-CCA)has a significant ethnic difference between Western populations and Japanese. Understanding of specific genetic alterations which associated with ethnic differences could lead to understanding of pathogenesis in Ov-CCA. The purpose of this study is to investigate the single nucleotide polymorphisms (SNPs) and the associated genes which correlated with the Japanese Ov-CCA using expression quantitative trait loci (eQTL) analysis with the publicly available datasets. Methods: The differences of genotype between three races (Western, Chinese, and Japanese) were compared using eQTLs analysis with the HapMap3 and the 1000 genome project datasets. 4 SNPs and 7 associated genes were identified. To evaluating the clinical significance of 7 identified genes in clear cell histology, the survival analysis according to the gene expression were perfomred with kidney renal clear cell carcinoma (KIRC) and high grade serous ovarian carcinoma (HGSOC) datasets in TCGA database. Results: Of the 935 identified cis-eQTLs, 4 SNPs (rs4873815, rs12976454, rs11136002, and rs13259097) had the different frequencies of alternative allele among Western, Chinese and Japanese with statistically significant (all p < 0.001) and these 4 SNPs were associated with 7 transcripts (APBA3, C8orf58, KIAA1967, NAPRT1, RHOBTB2, TNFRSF10B and ZNF707). To investigate the clinical significance, survival analysis was done using TCGA dataset and the expressions of the 3 genes (ZNF707, TNFRSF10B and RHOBTB2) were correlated with overall survival in KIRC (p < 0.001, p < 0.001 and p = 0.007) which were not shown in HGSOC. Conclusion: In this study, we identified that the expression of ZNF707, TNFRSF10B and RHOBTB2 were correlated with overall survival in clear cell histology although the KIRC dataset were used. eQTL analysis using the publicly available datasets may facilitate the discovery of novel genetic alteration in Ov-CCA and provide the basis for the understanding of Ov-CCA pathogenesis.

Oncogene-induced senescence mediated by c-Myc requires USP10 dependent deubiquitination and stabilization of p14ARF

Ko, Aram,Han, Su Yeon,Choi, Chel Hun,Cho, Hanbyoul,Lee, Min-Sik,Kim, Soo-Youl,Song, Joon Seon,Hong, Kyeong-Man,Lee, Han-Woong,Hewitt, Stephen M.,Chung, Joon-Yong,Song, Jaewhan Nature Publishing Group UK 2018 Cell death and differentiation Vol.25 No.6

<P>Oncogene-induced senescence (OIS) is a critical tumor-suppressor mechanism, which prevents hyper-proliferation and transformation of cells. c-Myc promotes OIS through the transcriptional activation of p14ARF followed by p53 activation. Although the oncogene-mediated transcriptional regulation of p14ARF has been well addressed, the post-translational modification of p14ARF regulated by oncogenic stress has yet to be investigated. Here, we found that c-Myc increased p14ARF protein stability by inducing the transcription of ubiquitin-specific protease 10 (USP10). USP10, in turn, mediated the deubiquitination of p14ARF, preventing its proteasome-dependent degradation. USP10-null mouse embryonic fibroblasts and human primary cells depleted of USP10 bypassed c-Myc-induced senescence via the destabilization of p14ARF, and these cells displayed accelerated hyper-proliferation and transformation. Clinically the c-Myc-USP10-p14ARF axis was disrupted in non-small cell lung cancer patients, resulting in significantly worse overall survival. Our studies indicate that USP10 induced by c-Myc has a crucial role in OIS by maintaining the stability of key tumor suppressor p14ARF.</P>

Molecular Chaperone HSP90 Is Necessary to Prevent Cellular Senescence via Lysosomal Degradation of p14ARF

Han, Su Yeon,Ko, Aram,Kitano, Haruhisa,Choi, Chel Hun,Lee, Min-Sik,Seo, Jinho,Fukuoka, Junya,Kim, Soo-Youl,Hewitt, Stephen M.,Chung, Joon-Yong,Song, Jaewhan American Association for Cancer Research 2017 Cancer research Vol.77 No.2

<P>In elucidating how the tumor suppressor p14ARF is degraded, this study has implications for prognosis and treatment or p14ARF-expressing lung cancers.</P><P>The tumor suppressor function of p14ARF is regulated at a posttranslational level via mechanisms yet to be fully understood. Here, we report the identification of an unconventional p14ARF degradation pathway induced by the chaperone HSP90 in association with the E3 ubiquitin ligase C-terminus of HSP70-interacting protein (CHIP). The ternary complex of HSP90, CHIP, and p14ARF was required to induce the lysosomal degradation of p14ARF by an ubiquitination-independent but LAMP2A-dependent mechanism. Depletion of HSP90 or CHIP induced p14ARF-dependent senescence in human fibroblasts. Premature senescence observed in cells genetically deficient in CHIP was rescued in cells that were doubly deficient in CHIP and p14ARF. Notably, non–small cell lung cancer cells (NSCLC) positive for p14ARF were sensitive to treatment with the HSP90 inhibitor geldanamycin. Furthermore, overexpression of HSP90 and CHIP with a concomitant loss of p14ARF correlated with poor prognosis in patients with NSCLC. Our findings identify a relationship between p14ARF and its chaperones that suggest new therapeutic strategies in cancers that overexpress HSP90. <I>Cancer Res; 77(2); 343–54. ©2016 AACR</I>.</P>

Targeting Cyclin D-CDK4/6 Sensitizes Immune-Refractory Cancer by Blocking the SCP3–NANOG Axis

Oh, Se Jin,Cho, Hanbyoul,Kim, Suhyun,Noh, Kyung Hee,Song, Kwon-Ho,Lee, Hyo-Jung,Woo, Seon Rang,Kim, Suyeon,Choi, Chel Hun,Chung, Joon-Yong,Hewitt, Stephen M.,Kim, Jae-Hoon,Baek, Seungki,Lee, Kyung-Mi American Association for Cancer Research 2018 Cancer Research Vol.78 No.10

<P>These findings reveal cyclin D1-CDK4/6 inhibition as an effective strategy for controlling SCP3<SUP>high</SUP> immune-refractroy cancer.</P><P>Immunoediting caused by antitumor immunity drives tumor cells to acquire refractory phenotypes. We demonstrated previously that tumor antigen–specific T cells edit these cells such that they become resistant to CTL killing and enrich NANOG<SUP>high</SUP> cancer stem cell-like cells. In this study, we show that synaptonemal complex protein 3 (SCP3), a member of the Cor1 family, is overexpressed in immunoedited cells and upregulates NANOG by hyperactivating the cyclin D1–CDK4/6 axis. The SCP3–cyclin D1–CDK4/6 axis was preserved across various types of human cancer and correlated negatively with progression-free survival of cervical cancer patients. Targeting CDK4/6 with the inhibitor palbociclib reversed multiaggressive phenotypes of SCP3<SUP>high</SUP> immunoedited tumor cells and led to long-term control of the disease. Collectively, our findings establish a firm molecular link of multiaggressiveness among SCP3, NANOG, cyclin D1, and CDK4/6 and identify CDK4/6 inhibitors as actionable drugs for controlling SCP3<SUP>high</SUP> immune-refractory cancer.</P><P><B>Significance:</B> These findings reveal cyclin D1-CDK4/6 inhibition as an effective strategy for controlling SCP3<SUP>high</SUP> immune-refractroy cancer. <I>Cancer Res; 78(10); 2638–53. ©2018 AACR</I>.</P>

CDK7 is a reliable prognostic factor and novel therapeutic target in epithelial ovarian cancer

Kim, Jihye,Cho, Young-Jae,Ryu, Ji-Yoon,Hwang, Ilseon,Han, Hee Dong,Ahn, Hyung Jun,Kim, Woo Young,Cho, Hanbyoul,Chung, Joon-Yong,Hewitt, Stephen M.,Kim, Jae-Hoon,Kim, Byoung-Gie,Bae, Duk-Soo,Choi, Chel Elsevier 2020 Gynecologic oncology Vol.156 No.1

<P><B>Abstract</B></P> <P><B>Objective</B></P> <P>Cyclin-dependent kinase 7 (CDK7) engages tumor growth by acting as a direct link between the regulation of transcription and the cell cycle. Here, we investigated the clinical significance of CDK7 expression and its potential as a therapeutic target in epithelial ovarian cancer (EOC).</P> <P><B>Methods</B></P> <P>CDK7 expression was examined in 436 ovarian tissues including normal to metastatic ovarian tumors using immunohistochemistry, and its clinical implications were analyzed. Furthermore, we performed <I>in vitro</I> and <I>in vivo</I> experiments using CDK7 siRNA or a covalent CDK7 inhibitor (THZ1) to elucidate the effect of CDK7 inhibition on tumorigenesis in EOC cells.</P> <P><B>Results</B></P> <P>The patient incidence of high CDK7 expression (CDK7<SUP>High</SUP>) gradually increased from normal ovarian epithelium to EOC (<I>P</I> < 0.001). Moreover, CDK7<SUP>High</SUP> was associated with an advanced stage and high-grade histology (<I>P</I> = 0.035 and <I>P</I> = 0.011, respectively) in EOC patients and had an independent prognostic significance in EOC recurrence (<I>P</I> = 0.034). CDK7 inhibition with siRNA or THZ1 decreased cell proliferation and migration, and increased apoptosis in EOC cells, and this anti-cancer mechanism is caused by G0/G1 cell cycle arrest. In <I>in vivo</I> therapeutic experiments using cell-line xenograft and PDX models, CDK7 inhibition significantly decreased the tumor weight, which was mediated by cell proliferation and apoptosis.</P> <P><B>Conclusion</B></P> <P>Mechanistic interrogation of CDK7 revealed that it is significantly associated with an aggressive phenotype of EOC, and it has independent prognostic power for EOC recurrence. Furthermore, CDK7 may be a potential therapeutic target for patients with EOC, whether platinum sensitive or resistant.</P> <P><B>Highlights</B></P> <P> <UL> <LI> CDK7 inhibition has an anti-cancer effect on platinum-sensitive EOC. </LI> <LI> CDK7 inhibition could induce responsiveness to platinum chemotherapy in platinum-resistant EOC. </LI> <LI> CDK7 overexpression had independent negative prognostic value for disease recurrence of EOC. </LI> <LI> CDK7 might play a critical role in EOC tumorigenesis, and it also serves as a possible therapeutic target in EOC. </LI> </UL> </P>

HDAC1 Upregulation by NANOG Promotes Multidrug Resistance and a Stem-like Phenotype in Immune Edited Tumor Cells

Song, Kwon-Ho,Choi, Chel Hun,Lee, Hyo-Jung,Oh, Se Jin,Woo, Seon Rang,Hong, Soon-Oh,Noh, Kyung Hee,Cho, Hanbyoul,Chung, Eun Joo,Kim, Jae-Hoon,Chung, Joon-Yong,Hewitt, Stephen M.,Baek, Seungki,Lee, Kyun American Association for Cancer Research 2017 Cancer Research Vol.77 No.18

<P>These findings link a regulator of cancer stem-like cells to epigenetic silencing events that may improve immunotherapy outcomes.</P><P>Cancer immunoediting drives the adaptation of tumor cells to host immune surveillance. Immunoediting driven by antigen (Ag)-specific T cells enriches NANOG expression in tumor cells, resulting in a stem-like phenotype and immune resistance. Here, we identify HDAC1 as a key mediator of the NANOG-associated phenotype. NANOG upregulated HDAC1 through promoter occupancy, thereby decreasing histone H3 acetylation on K14 and K27. NANOG-dependent, HDAC1-driven epigenetic silencing of cell-cycle inhibitors CDKN2D and CDKN1B induced stem-like features. Silencing of TRIM17 and NOXA induced immune and drug resistance in tumor cells by increasing antiapoptotic MCL1. Importantly, HDAC inhibition synergized with Ag-specific adoptive T-cell therapy to control immune refractory cancers. Our results reveal that NANOG influences the epigenetic state of tumor cells via HDAC1, and they encourage a rational application of epigenetic modulators and immunotherapy in treatment of NANOG<SUP>+</SUP> refractory cancer types. <I>Cancer Res; 77(18); 5039–53. ©2017 AACR</I>.</P>