Lin Feng (Sun Yat-sen University Cancer Center, China)
–”Nasopharyngeal Carcinoma Susceptibility Gene TNFRSF19 Inhibits TGFβ Signaling and Promotes Tumorigenesis”
- TNFRSF19 susceptibility gene. Nat Genet 2010. Bei JX et al
- A orphan receptor , no ligand found yet
- by IHC, TFNRSF19 is highly expressed in NPC tissues, possibly an oncogene
- Survival TNF receptors noncanonical TNFR of TNFRSF19
- Identification of TNFRSF19 as
- Constitutive association of SF19 with TbR1
- TbR1/Smad2/3 is blocked by SF19
- TGF signal transduction without SF19 – late response
- SF19 is mainly involved in TGFb pathway
- Reverse correlation between SF10 and TGFb in NPC biopsy
- TGFb and EBV reactivation
- TGFb induces latent to lytic switch of EBV by activating BZLF1
- Smads-responsive element
- First hit (SF19, HLA, p16)
- Pre-cancer TGFb signaling decreases, Epithelial cell proliferation increases, permissive for EBV latent infection
- after EBV infection. NF-KB, PI3K-KAT signaling, cell survival, anti-apoptosis, genomic instability
- in Clinics. TGFBRi. Used in in idiopathic ___ fibrosis
Miao Xu (Sun Yet-sen University Cancer Center, China)
–”Genome Sequencing Analysis Identifies Epstein-Barr Virus Subtypes Associated with the Risk of Nasopharyngeal Carcinoma”
- EBV subtypes
- High risk? -cancers
- Low-risk? -not associated with cancers
- vaccine development for prevention
- Risk prediction for early detection
- sequenced 269 EBV isolates from…
- EBV whole-genome targeted combined phylogenetic analysis of 230 newly-sequenced and 97 published EBV isolates
- The association of functional variants with NPC risk (in one coding region)
- 1st SNP is wild spreaded in all forms of EBV-associated malignancies
- 2nd SNPs is private for NPC samples
- Function of the three coding variants
- High-risk variants suppress EBNV lytic replication, thereby promote viral latency
- The amount of EBV DNA in saliva from NPC-endemic population.
- EBV genome replication
- High risk SNP suppresses EBV lytic replication, compares to that of the normal control saliva from NPC-endemic populations
- EBV lytic cycle reactivation as a treatment strategy (2004 J Virol Feng and Kenney]
- Cytolytic virus activation therapy for EBV tumors (CCR Wilderman and Greijer, Germany group)
- A pan-HDAC inhibitor (SAHS) induces EBV lytic cycle and apoptosis in NPC cells (Hsu and Chiang at al In H cancer 2012)
- – Class I HDAC inhibitors (romidepsin) induce higher percentage of cells into lytic cycle but hater is still a proportion of refractory cells
- – High trough put screening of mall organic o pounds in inducing lytic cycle of EBV-associated epithelial malignancies (Choi and China’s AK eat al,m PlosOne 2015)
- Primary screening AGX-Bx1 + compound library —> C7 cod can induce lytic cycle
- C7 induces EBV lyti ccycle i various NPC and GC cell lines
- A (HONE1-EBV B C666-1 C (is a more real cell line, SNU-719?, AGS-BX1
- C7 shares a similar structure wiht Dp44mT which is known to chelate idol. (C7 can bind transitoina metal, Dp44mT can bind iron)
- C7-1, C-3 and C7-5 analogous reactivation EBV lytic cycle strongly
- C7, C7-1, C7-3, C7-5 bound strongly to iron, suggesting a role of iron chelation in EBV lytic reactivation
- clinical iron cheaters can also reactivate EBV lytic cycle
- intracelluar iron chelation by C7 and other cheaters for EBVlytic reactivation
- C7 (10 uM) induces the hypoxia signaling pathway that associates with Zta, Rta, expression
- C7 sensitized EBV-positive GC cells to gain I lover
- Conclusions
- Demonstrated a novel mechanism of induction of lyti ccycle of EBV in EBV+ NOC and GC through intracellular iron chelation
- Combination of anti-EBV (gancyclovir) drug and iron-chelators (HIF1 signaling) could be useful in EBV-cytolytic therapy
- Kenney asked in huMice model hypoxia area is also associated with EBV lytic replication (and early lytic EBV replication is important for cytolysis, late genes are not that required for such therapy)
- JIm Mai (麥智明) PhD student of ___
- Vitamins D deficiency
- – common in the world
- – EBV et in the sunniest areas (eg Hong Kong)
- – Potential role of VitD in NOC prevention
- – Vit D (anti-inflammatory and immuno-modulators, EBV infection and viral
- Method (AoE NPC case-control study NPC incident cases: 815 and 1502 controls
- – measurement of 25-hydroxyvitamin D concentration (nmol/L). —> current status
- – Genetically lowered VitD
- – < 37.5 nmols/L => adjusted OR 1.94
- – Further stratification of “negative confounders)
- – Higher genetics predicted 25-by
- Lower circulating and genetically predicted 25-hydroxyvitamine D levels —> increase risks of NPC
- [Vit D and EBV]
- OR and 95% condolence interval s of IgA against EBV for circulating proved I tend 25
- Thus, the association of EBV and VitD deficiency is still unclear (Increase sample size, prospective cohort; and use EBV markers other than IgA antibody
Shibing Li (Sun Yat-sen University Cancer Center, China)
- ISNLx was identified by comparisons of
- NPEC1-EBV vs NPEC1
- NPEC2-EBV vs NPEC2
- ISLNx was highly expressed in NPC cell Lines and the plasma of patients (serves a good prognosis Parker of DFS, OS…
- ISLNx prompted NPC cell (CNE1 and CNE2) proliferation depending on its receptor GPCRx
- ISNLx promotes elevation of the gluciolytic pathway in ENC1 and CNE2
- Metabolomica analysis of CNE1, NE2-expressing ISLNx
- ISNLx promotes aerobic glycolysis
- Activation of STAT5 by ISNLx facilitates aerobic glycolysis
- Overexpression of ISNLx sensitized NOC cells to glycolysis
- Both ISNLx and GPCRx neutralized antibodies can be potential therapeutic strategy for NPC
- EBV encoded BZLF1 enhances ISNLx expression -> ISNLx served as novel biomarker for NOC diagnosis and prognosis -> ISNLx enhanced glycolysis contribute to tumor progression. -> ISNLx-GPCRx axis is the therapeutic target for NPC
“Epstein-Barr Virus-Encoded LMP1 Reprograms Glucose Metabolism to Drive Cell Motility Through mTORC2/PDHE1α-Mediated Histone Acetylation in Nasopharyngeal Cells”
- PhD student of Tsao SW
- Tsao et al J Pathol 2015 235:323-333 a review of LMP1 (involved in all hallmarks of cancer)
- Aerobic glycolysis provides many essential metabolites (e.g pyruvate for cell proliferation)
- EBV induction programs the metabolic profile in NPE cells
- NP460EBV/NP460 + NP550EBV/NP550. Fold cut = 3 fold
- KO LMP1 diminish glucose metabolism
- EBV-encoded LMP1 involves in the metabolic reprogramming
- ECAR/OCAR EBV-LMP1 enhances olic
- LMP1 expression positively correlated with GLUT1 expression
- LMP1 upregulates GLUT1 transcription through NFKB.
- JVI 2017 published (https://www.ncbi.nlm.nih.gov/pubmed/28053105)
- LMP1-enhanced glycolysis involves in cell motality
- EBV-LMP1 -> glycolysis (inhibited by 2DG)
- LMP1-induced mTROC2 activation mediates cell motility
- ShRictor
- PDHE1a is important for LMP1 induced cell migration and invasion (in NP69)
- PDHE1a ser293 is essential for its translocation into the nucleus
- NP69-PDHE1a-KD. -> Cell motility shut down (Via acetylation of snail expression)
- LMP1 expression associated conditional medium has growth advantage in vitro (in NP69).
- LMP1 induces IGF-1 secretion activate mTORC2 signaling
- NCI launches PDX programs
- New cell lines in Broad Institutes
- The Georgetown method
- Feeder cells + chemical inhibitor The American J Pathol 2012
- Nat Protocols 2017, Liu X et al
- Deprogrammed cells retain lineage differentiation
- Use of reprogrammed cells to identify therapy for (NEJM)
- Patient-derived models of acquired resistance can identify effective drugs combination
- CRCs (cell for reprogramming)
- NGLV: Reduce significantly cost, workload
- Unique for many rare diseases
- Authentication of nasopharyngeal carcinoma
Laboratory Investigation 2018 May
Establishment of a nasopharyngeal carcinoma cell line capable of undergoing lytic Epstein-Barr virus reactivation.
Yip YL1, Lin W1, Deng W2, Jia L1, Lo KW3, Busson P4, Vérillaud B4, Liu X5,6, Tsang CM1, Lung ML7, Tsao SW8.
Epstein-Barr virus (EBV) infects more than 90% of the adult human population. Undifferentiated nasopharyngeal carcinoma (NPC) is common in Southeast Asia, with a particularly high incidence among southern Chinese. The EBV genome can be detected in practically all cancer cells in undifferentiated NPC. The role of EBV in pathogenesis of undifferentiated NPC remains elusive. NPC cell lines are known to be difficult to establish in culture. The EBV+ve NPC cell lines, even if established in culture, rapidly lost their EBV episomes upon prolonged propagation. At present, the C666-1 NPC cell line, which is defective in lytic EBV reactivation, is the only EBV+ve NPC cell line available for NPC and EBV research. The need to establish new and representative NPC cell lines is eminent for NPC and EBV research. In this study, we report the use of the Rho-associated kinase inhibitor (Y-27632) has facilitated the establishment of a new EBV+ve NPC cell line from an earlier established NPC xenograft, C17. The C17 cell line was tumorigenic in immune-deficient mice (NOD/SCID). It retained the EBV episomes and could be induced to undergo productive lytic reactivation of EBV to generate infectious virus particles. The C17 cell line represents a new investigative tool for NPC and EBV studies. The ability of C17 to undergo lytic reactivation is unique and opens up the opportunity to examine regulation of latent and lytic infection of EBV and their contributions to NPC pathogenesis.
Yu F1, Lu Y2, Tao L2, Jiang YY3, Lin DC3, Wang L3, Petersson F4, Yoshiyama H5, Koeffler PH3, Goh BC3, Loh KS2,6.
1 Department of Otolaryngology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore. entyf@nus.edu.sg.
2 Department of Otolaryngology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
3 Cancer Science Institute of Singapore, National University of Singapore, Singapore, Singapore.
4 Department of Pathology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
5 Department of Microbiology, Shimane University Faculty of Medicine, 89-1 Enyacho, Izumo City, Shimane, 693-8501, Japan.
6 Head & Neck Tumor group, National Cancer Institute of Singapore, Head & Neck Surgery, NUHS, Singapore, Singapore.
Abstract
Nasopharyngeal carcinoma (NPC) is an invasive cancer with particularly high incidence in Southern China and Southeast Asia. The study of NPC is greatly hampered by the lack of reliable cell lines due to the loss of EBV genome and HeLa cell contamination. Conditional reprogramming (CR) cell culture technique has been reported for rapid and efficient establishment of patient-derived normal and tumor cell cultures. The purpose of this study was to assess this method to culture NPC patient-derived primary tumor cells. Using CR protocol, we demonstrated that epithelial cells could be efficiently cultured from normal (70%) and cancerous nasopharyngeal (46%) biopsies. However, by comparing with original tumors in terms of mutation and methylation profiles, epithelial cells derived from cancerous biopsy represented non-malignant cells. Further, they exhibited stem-like characteristics based on their cell surface proteins and could differentiate into pseudostratified epithelium in an air-liquid interface culture system. We conclude that CR method is a highly selective and useful method for growing non-malignant nasopharyngeal epithelial cells.
Dewi Kartikawati Paramita (Universitas Gadjah Mada, Indonesia)
“NPC Strip G: Rapid Test Strip Detecting IgG Against EBV Antigens in NPC”
Selected Posters 筆記結束。