Distant Metastasis In Nasopharyngeal Carcinoma Research Articles

Arenobufagin induces cell apoptosis by modulating the cell cycle regulator claspin and the JNK pathway in nasopharyngeal carcinoma cells

ABSTRACT Background The high recurrence rate and incidence of distant metastasis of nasopharyngeal carcinoma (NPC) result in poor prognosis. It is necessary to identify natural compounds that can complement combination radiation therapy. Arenobufagin is commonly used for heart diseases and liver cancer, but its effectiveness in NPC is unclear. Study Design and Methods The effect of arenobufagin-induced apoptosis was measured by a cell viability assay, tumorigenic assay, fluorescence assay, and Western blot assay through NPC-039 and NPC-BM cell lines. The protease array, Western blot assay, and transient transfection were used to investigate the underlying mechanism of arenobufagin-induced apoptosis. An NPC xenograft model was established to explore the antitumor activity of arenobufagin in vivo. Results Our findings indicated that arenobufagin exerted cytotoxic effects on NPC cells, inhibiting proliferation through apoptosis activation. Downregulation of claspin was confirmed in arenobufagin-induced apoptosis. Combined treatment with arenobufagin and mitogen-activated protein kinase inhibitors demonstrated that arenobufagin induced NPC apoptosis through the c-Jun N-terminal kinases (JNK) pathway inhibition. Furthermore, arenobufagin suppressed NPC tumor proliferation in vivo. Conclusion Our results revealed the antitumor effect of arenobufagin in vitro and in vivo. Arenobufagin may have clinical utility in treating NPC due to its suppression of claspin and inhibition of the JNK pathway.

KLF5 regulates actin remodeling to enhance the metastasis of nasopharyngeal carcinoma.

Transcription factors (TFs) engage in various cellular essential processes including differentiation, growth and migration. However, the master TF involved in distant metastasis of nasopharyngeal carcinoma (NPC) remains largely unclear. Here we show that KLF5 regulates actin remodeling to enhance NPC metastasis. We analyzed the msVIPER algorithm-generated transcriptional regulatory networks and identified KLF5 as a master TF of metastatic NPC linked to poor clinical outcomes. KLF5 regulates actin remodeling and lamellipodia formation to promote the metastasis of NPC cells in vitro and in vivo. Mechanistically, KLF5 preferentially occupies distal enhancer regions of ACTN4 to activate its transcription, whereby decoding the informative DNA sequences. ACTN4, extensively localized within actin cytoskeleton, facilitates dense and branched actin networks and lamellipodia formation at the cell leading edge, empowering cells to migrate faster. Collectively, our findings reveal that KLF5 controls robust transcription program of ACTN4 to modulate actin remodeling and augment cell motility which enhances NPC metastasis, and provide new potential biomarkers and therapeutic interventions for NPC.

Double fatal consequences of distance metastasis in nasopharyngeal carcinoma after a completed chemoradiation in pregnancy: A case report.

Distant metastasis in nasopharyngeal carcinoma (NPC) patients is one of the reasons for the decreased life expectancy with the most common metastasis spreads are to the bone, liver, and lung. Hepatoma is the most frequent liver malignancy and is one of the highest causes of cancer death worldwide and this can be as a result of NPC metastasis. The aim of this case report was to present a patient with hepatoma in pregnancy as a result of NPC metastasis. A 34-year-old pregnant female at 24-25 weeks of gestation presented with a chief complaint of heartburn and unbearable pain radiating to the back. Previous medical history reported that the patient had a liver enlargement. The patient was G4P2A1 with a single living intrauterine fetus and active fetal movements. The patient has a history of NPC and received a completed chemoradiation one month prior to hospital admission. Physical examination showed bilateral rales and palpable diffuse multiple nodule masses in the upper right abdominal quadrant. Laboratory examination revealed anemia, thrombocytopenia, negative hepatitis B surface antigen (HBsAg), and elevated liver markers. Abdominal ultrasonography results showed multiple diffuse nodules in the liver. The patient was diagnosed with a metastatic hepatoma based on the clinical and imaging findings. During hospitalization, the patient repeatedly experienced pleural effusion with suspicion metastases. A few days later, the fetal movements stopped and the ultrasonography indicated negative fetal heart rate. After experiencing respiratory distress for hours, the patient expired the day after. This case highlights that due to the potential adverse effects of chemotherapy and radiotherapy, the initiation of these therapies should be carefully decided to avoid adverse effects to mother and fetus.

Pattern of CD8+ tissue resident memory T cells between local recurrence and distant metastasis in nasopharyngeal carcinoma.

e15161 Background: CD8+ T lymphocytes are believed to be the major anti-tumor immune subsets. Consequently, most cancer immunotherapeutic approaches seek to amplify cytotoxic T lymphocytes specific to malignant cells. A recently identified subpopulation of memory CD8+ T cells, named tissue-resident memory T (Trm) cells, persists in non-lymphoid tissues, including solid tumors. This T-cell subset is considered an independent memory T-cell lineage with cytotoxic features. However, their function, spatial location and influence on disease-related outcomes (local recurrence [LR], distant metastasis [DM] in nasopharyngeal carcinoma (NPC), a disease entity characterized by substantial immune infiltrate, remain poorly understood. We aim to close this gap through investigation on the abundance and spatial localization of CD8+ Trm in relation to tumor site (primary or recurrence [LR and DM]). Methods: This retrospective cohort analysis based on 20 NPC patient samples: primary (initial diagnosis; [I; n = 19], recurrence ([LR; n = 14] and [DM; n = 10]). We accessed the spatial localization in tumor tissues within tissue-segmented classifications of either cancer islands (pan-cytokeratin–positive [CK+] areas) or stromal areas (CK– areas). Tumor infiltrating lymphocytes (TILs) among patients from I, LR and DM group was quantified through multiplexed 5-colour opal staining using biomarkers against CD8, CD103, IL17, CK plus DAPI (nuclear cell counterstain). Results: While total CD8+ TILs (total tumor) as well as their spatially localized cell densities (tumor islet [TI] vs. tumor stroma [TS]) did not show any significant changes among I, LR and DM patient groups, a higher proportion of CD8+ TILs in the TI co-express the CD103 molecule, an indication of CD8+ Trm (CD103+CD8+) infiltration (mean CD103+CD8+/CD8+ in TI: 32.1% in I group, 44.8% in LR group, 46.0% in DM group; mean CD103+CD8+/CD8+ in TS: 19.0% in I group, 26.2% in LR group 26.2% in DM group). Notably, we observed that such augmentation only reached statistically significant within TI from patient in LR group (p = 0.0451). Furthermore, we found that CD8+ Trm in the LR group expressed higher level of cytokine IL-17, which was ~2.1-fold (p = 0.0788) and 3.6-fold upregulated (p = 0.0285) as compared to I and DM, respectively. Conclusions: Cancer islet–localized CD8+ TILs are composed of CD103+ Trms, which make up nearly 45% the total CD8+ T cell population within LR tumors of NPC patients, highlighting spatial localization of CD103+CD8+ Trms as a key phenotype of CD8+ TIL subset. The CD8+ Trm expressed with increased IL17 suggests that enrichment of this T-cell subset might exerts cytotoxic functions. Our results demonstrate that CD8+ Trm had a greater likelihood as an indicator of LR risk for NPC patients and help identify patient cohorts likely to benefit from immunotherapy.

High proportion of circulating CD8 + CD28- senescent T cells is an independent predictor of distant metastasis in nasopharyngeal carcinoma after radiotherapy

BackgroundNasopharyngeal carcinoma (NPC) is a kind of epithelial carcinoma that is common in East and Southeast Asia. Distant metastasis after radiotherapy remains the main cause of treatment failure and preradiotherapy immune system function can influence prognosis. Our study aimed to identify immune-related prognostic factors for NPC after radiotherapy and establish a prognostic model to predict progression-free survival (PFS) and distant metastasis-free survival (DMFS).MethodsWe enrolled NPC patients and divided them into training and validation cohorts with follow-up. We collected clinical information and investigated immune cells, EBV DNA and cytokines in the peripheral blood of NPC patients before radiotherapy and EBV DNA after radiotherapy. Among these immune cells, we included CD8+CD28− T cells, which are a unique T-cell immunosenescent subset that increases in human peripheral blood with increasing age and declining immune function. Based on the detection results and clinical information, we utilized Cox regression and least absolute shrinkage and selection operator (LASSO) regression to screen the PFS and DMFS prognostic factors and build nomograms to predict the PFS and DMFS of NPC. We also verified the results in the validation set.ResultsThree factors associated with PFS were selected: proportion of CD8+CD28− T cells posttreatment EBV and N stage. Three factors associated with DMFS were screened: proportion of CD8+CD28− T cells, posttreatment EBV and N stage. CD8+CD28− T cells are correlated with systemic inflammation and posttreatment immunosuppression. The C-indexes were 0.735 and 0.745 in the training and validation cohorts for predicting PFS. For DMFS, the C-indexes were 0.793 and 0.774 in the training and validation cohorts.ConclusionsThe pretreatment proportion of CD8+CD28− T cells is a candidate prognostic biomarker for NPC after radiotherapy. The constructed nomogram models based on CD8+CD28− T cells have good predictive value.

Epithelial-mesenchymal transition classification of circulating tumor cells predicts clinical outcomes in progressive nasopharyngeal carcinoma.

BackgroundLiquid biopsy facilitates the enrichment and isolation of circulating tumor cells (CTCs) in various human cancers, including nasopharyngeal carcinoma (NPC). Characterizing CTCs allows observation of the evolutionary process of single tumor cells undergoing blood-borne dissemination, such as epithelial-mesenchymal transition. However, the prognostic value of phenotypic classification of CTCs in predicting the clinical outcomes of NPC remains poorly understood.Patients and methodsA total of 92 patients who met the inclusion criteria were enrolled in the present study. The CanPatrol™ CTC technology platform was employed to isolate CTCs, and an RNA in situ hybridization-based system was used for phenotypic classification. Kaplan–Meier survival curves were used for univariate survival analysis, and the log-rank test was performed for between-group comparisons of the survival curves.ResultsCTCs were detected in 88.0% (81/92) of the enrolled patients with NPC. The total CTC number did not vary between the T and N stages or between Epstein–Barr virus DNA-positive and -negative cases. The numbers of total CTCs and epithelial/mesenchymal (E/M) hybrid CTCs decreased significantly at 3 months post concurrent chemoradiotherapy (P=0.008 and P=0.023, respectively), whereas the numbers of epithelial or mesenchymal CTCs did not decrease. E/M hybrid-predominant cases had lower disease-free survival (P=0.043) and distant metastasis-free survival (P=0.046) rates than non-E/M hybrid-predominant cases.ConclusionCTC classification enables a better understanding of the cellular phenotypic alterations responsible for locoregional invasion and distant metastasis in NPC. E/M hybrid-predominant CTC distribution predicts unfavorable clinical outcomes in patients with progressive NPC.

Radiomics based on pretreatment MRI for predicting distant metastasis of nasopharyngeal carcinoma: A preliminary study

ObjectiveTo explore the feasibility of predicting distant metastasis (DM) of nasopharyngeal carcinoma (NPC) patients based on MRI radiomics model.MethodsA total of 146 patients with NPC pathologically confirmed, who did not exhibit DM before treatment, were retrospectively reviewed and followed up for at least one year to analyze the DM risk of the disease. The MRI images of these patients including T2WI and CE-T1WI sequences were extracted. The cases were randomly divided into training group (n=116) and validation group (n=30). The images were filtered before radiomics feature extraction. The least absolute shrinkage and selection operator (LASSO) regression was used to develop the dimension of texture parameters and the logistic regression was used to construct the prediction model. The ROC curve and calibration curve were used to evaluate the predictive performance of the model, and the area under curve (AUC), accuracy, sensitivity, and specificity were calculated.Results72 patients had DM and 74 patients had no DM. The AUC, accuracy, sensitivity and specificity of the model were 0. 80 (95% CI: 0.72~0. 88), 75.0%, 76.8%, 73.3%. and0.70 (95% CI: 0.51~0.90), 66.7%, 72.7%, 63.2% in training group and validation group, respectively.ConclusionThe radiomics model based on logistic regression algorithm has application potential for evaluating the DM risk of patients with NPC.

Utilization of the lymph node-to-primary tumor ratio of PET standardized uptake value and circulating Epstein–Barr virus DNA to predict distant metastasis in nasopharyngeal carcinoma

Background and purposeTo determine the clinical impact of integrating Epstein–Barr virus (EBV) DNA and lymph node-to-primary tumor ratio (NTR) of positron emission tomography (PET) standardized uptake value (SUV) in predicting distant metastasis, such as distant metastasis-free survival (DMFS), in patients with nasopharyngeal carcinoma (NPC). Materials and methodsWe retrospectively reviewed patients diagnosed with non-disseminated NPC between 2010 and 2017. The optimal cut-off values of EBV DNA and SUV NTR were determined using receiver operating characteristic analysis. The prognostic values of SUV NTR and EBV DNA on DMFS and overall survival were evaluated using the Kaplan–Meier method. Univariate and multivariable analyses were performed using the Wald Chi-squared test and Cox proportional hazards regression, respectively. ResultsA total of 488 patients were included in the analysis. The median follow-up period was 61.6 months. The optimal cut-off values of EBV DNA and SUV NTR were 3377.5 copies per mL and 0.64, respectively. The five-year DMFS for patients with high vs low EBV DNA and SUV NTR levels were 64.9% vs 86.6% (p < 0.001) and 78.7% vs 87.4% (p = 0.021), respectively. In subgroup analysis, the high-risk group with high levels of pretreatment EBV DNA and SUV NTR had worse DMFS in either American Joint Committee on Cancer (AJCC) stage I–III or IVA–B (p = 0.001 and <0.001, respectively). Univariate and multivariable analyses showed the statistical significance of EBV DNA, SUV NTR, and their composite in DMFS (p < 0.001 for EBV DNA; p = 0.022 for SUV NTR; p < 0.001 for their composite). ConclusionThis study showed that EBV DNA and SUV NTR have independent and additive values as prognosticators for distant metastasis in patients with NPC, suggesting that these two individual factors, except the AJCC staging system, should be included in future studies.

The Correlations among Circulating Tumor Cells, Epstein-Barr Virus Status, and Epidemiology in Patients with Nasopharyngeal Carcinoma.

Studies have shown that circulating tumor cells (CTCs) can be detected in nasopharyngeal carcinoma (NPC). However, the relationship between CTCs and tumor stage is still controversial. This study aims to investigate the correlations among CTCs, Epstein-Barr virus (EBV) status, clinicopathologic features, and epidemiological risk factors in patients with NPC. Three hundred and thirty primary NPC patients with complete clinical data and epidemiology information were collected. Analysis of CTCs was performed using the CTCBIOPSY system. The plasma EBV DNA load was detected by quantitative real-time PCR. Detection of VCA-IgA and EA-IgA antibodies titers was conducted by immunoenzymatic assay. EBNA1-IgA and Zta-IgA were measured using an enzyme-linked immunosorbent assay. The presence of CTCs was associated with high EBV DNA load (p < 0.05). The positive rate of CTCs was correlated with T and M classifications of NPC (T: 13.2% vs. 22.9%; M: 17.9% vs. 34.8%, p < 0.05). Compared with never and former smokers, current smokers exhibited a higher positive rate of EBNA1-IgA (83.3% and 81.0% vs. 92.5%, p < 0.05); the patients with pack-years of smoking ≥ 15 displayed a significantly higher positive rate of EBNA1-IgA than those with pack-years of smoking < 15 (98.0% and 92.5% vs. 81.0%, p < 0.05). CTCs positivity was closely associated with tumor burden and distant metastasis of NPC. Smoking status and smoking cumulative dose of NPC patients might be correlated with EBV activation.

Platelet-derived extracellular vesicles inhibit ferroptosis and promote distant metastasis of nasopharyngeal carcinoma by upregulating ITGB3.

Nasopharyngeal carcinoma (NPC) is a malignancy with high metastatic and invasive nature. Distant metastasis contributes substantially to treatment failure and mortality in NPC. Platelets are versatile blood cells and the number of platelets is positively associated with the distant metastasis of tumor cells. However, the role and underlying mechanism of platelets responsible for the metastasis of NPC cells remain unclear. Here we found that the distant metastasis of NPC patients was positively correlated with the expression levels of integrin β3 (ITGB3) in platelet-derived extracellular vesicles (EVs) from NPC patients (P-EVs). We further revealed that EVs transfer occurred from platelets to NPC cells, mediating cell-cell communication and inducing the metastasis of NPC cells by upregulating ITGB3 expression. Mechanistically, P-EVs-upregulated ITGB3 increased SLC7A11 expression by enhancing protein stability and activating the MAPK/ERK/ATF4/Nrf2 axis, which suppressed ferroptosis, thereby facilitating the metastasis of NPC cells. NPC xenografts in mouse models further confirmed that P-EVs inhibited the ferroptosis of circulating NPC cells and promoted the distant metastasis of NPC cells. Thus, these findings elucidate a novel role of platelet-derived EVs in NPC metastasis, which not only improves our understanding of platelet-mediated tumor distant metastasis, but also has important implications in diagnosis and treatment of NPC.

A comprehensive model based on temporal dynamics of peripheral T cell repertoire for predicting post-treatment distant metastasis of nasopharyngeal carcinoma.

Many nasopharyngeal carcinoma (NPC) patients develop distant metastases after treatment, leading to poor outcomes. To date, there are no peripheral biomarkers suitable for all NPC patients to predict distant metastasis. Hence, we purposed to develop a noninvasive comprehensive model for predicting post-treatment distant metastasis of all NPC. Since T-cell receptor β chain (TCRB) repertoire has achieved prognostic prediction in many cancers, the clinical characteristics and parameters of TCRB repertoire of 71 cases of peripheral blood samples (pairwise pre-treatment and post-treatment samples from 40 NPC patients who without (nM, n = 21) or with (M, n = 19) post-treatment distant metastasis) were collected. The least absolute shrinkage and selection operator algorithm was used to construct a distant metastasis prediction model. In terms of TCRB repertoire parameters, the diversity of TCRB repertoire was significantly decreased in M group after treatment but not in nM group. Ascending TCRB diversity and higher similarity between pre- and post-treatment samples showed better distant metastasis-free survival (DMFS). The similarity still had robust DMFS prediction in patients with reduced TCRB diversity. More importantly, the 5-factor comprehensive model consisting of basic clinical characteristics and TCRB repertoire indices showed a higher prognostic accuracy than any one individual factor in DMFS predicting. In conclusion, treatment had different effects on the composition of TCRB repertoire in patients without and with post-treatment distant metastasis. The dynamics of TCRB diversity, the similarity of TCRB repertoires, and combinations of these factors with basic clinical characteristics could serve as noninvasive DMFS predictors for all NPC patients.

A PECULIAR CASE OF ISOLATED METASTATIC INGUINAL LYMPHADENOPATHY IN A YOUNG LADY – A CASE REPORT

Lymphadenopathy refers to the swelling of lymph nodes which can be secondary to bacterial, viral or fungal infections, autoimmune disease and malignancy. Lymphadenopathy can be localized or diffuse. About 75% of most lymphadenopathies are localized, and about 50% of those occur in the head and neck regions . Inguinal lymphadenopathy (3) occurs at the groin region and most common causes include infections of leg or foot, STDs, non Hodgkin's lymphoma, tuberculosis and pelvic malignancies. One of the rare causes includes distant metastasis of nasopharyngeal carcinoma (NPC) which is present in our case. Nasopharyngeal cancer is an uncommon squamous cell carcinoma in the head and neck region, in most parts of the world. It has a high propensity for lymphatic spread and is known for regional metastases with occult primary at presentation . The incidence of distant (1) metastasis at presentation ranges from 4.4 to 6%. The most common sites of metastasis are bone (70%–80%) followed by liver (30%), lungs (18%) and distant lymph nodes (axillary, mediastinal, pelvic and inguinal, in that order) . About 98% of them are discovered within 3 (2) years of treatment. As it is a highly chemo and radio-sensitive tumor, radiotherapy with concurrent chemotherapy is the mainstay in the management of local and advanced diseases. Here we are presenting a peculiar case of previously treated NPC presenting as isolated left inguinal metastatic lymphadenopathy in a young lady

Prediction of local recurrence and distant metastasis using radiomics analysis of pretreatment nasopharyngeal [18F]FDG PET/CT images.

To develop a radiomics signature to predict locoregional recurrence (LR) and distant metastasis (DM), as extracted from pretreatment 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG) positron emission tomography/X-ray computed tomography (PET/CT) images in locally advanced nasopharyngeal carcinoma (NPC). Eighty-five patients with Stage III-IVB NPC underwent pretreatment [18F]FDG PET/CT scans and received radiotherapy or chemoradiotherapy. 53 of them achieved disease control, and 32 of them failed after treatment (15: LR, 17: DM). A total of 114 radiomic features were extracted from PET/CT images. For univariate analysis, Wilcoxon test and Chi-square test were used to compare median values of features between different treatment outcomes and predict the risk of treatment failure, respectively. For multivariate analysis, all features were grouped into clusters based on Pearson correlation using hierarchical clustering, and the representative feature of each cluster was chosen by the Relief algorithm. Then sequential floating forward selection (SFFS) coupled with a support vector machine (SVM) classifier were used to derive the optimized feature set in terms of the area under receiver operating characteristic (ROC) curve (AUC). The performance of the model was evaluated by leave-one-out-cross-validation, fivefold cross-validation, tenfold cross-validation. Twenty features had significant differences between disease control and treatment failure. NPC patients with values of Compactness1, Compactness2, Coarseness_NGTDM or SGE_GLGLM above the median as well as patients with values of Irregularity, RLN_GLRLM or GLV_GLSZM below the median, showed a significant (p < 0.05) higher risk of treatment failure (about 50% vs. 25%). The derived radiomics signature consisted of 5 features with the highest AUC value of 0.8290 (sensitivity: 0.8438, specificity: 0.7736) using leave-one-out-cross-validation. Locoregional recurrence (LR) and DM of locally advanced NPC can be predicted using radiomics analysis of pretreatment [18F]FDG PET/CT. The SFFS feature selection coupled with SVM classifier can derive the optimized feature set with correspondingly highest AUC value for pretreatment prediction of LR and/or DM of NPC.

Nomogram for the prediction of primary distant metastasis of nasopharyngeal carcinoma to guide individualized application of FDG PET/CT.

This study aimed to establish an effective nomogram to predict primary distant metastasis (DM) in patients with nasopharyngeal carcinoma (NPC) to guide the application of PET/CT. In total, 3591 patients with pathologically confirmed NPC were consecutively enrolled. The nomogram was constructed based on 1922 patients treated between 2007 and 2014. Multivariate logistical regression was applied to identify the independent risk factors of DM. The predictive value of the nomogram was evaluated using the concordance index (C-index), calibration curve, probability density functions (PDFs), and clinical utility curve (CUC). The results were validated in 1669 patients enrolled from 2015 to 2016. Net reclassification improvement (NRI) was applied to compare performances of the nomogram with other clinical factors. The best cut-off value of the nomogram chosen for clinical application was analyzed. A total of 355 patients showed primary DM among 3591 patients, yielding an incidence rate of 9.9%. Sex, N stage, EBV DNA level, lactate dehydrogenase level, and hemoglobin level were independent predictive factors for primary DM. C-indices in the training and validation cohort were 0.796 (95% CI, 0.76-0.83) and 0.779 (95% CI, 0.74-0.81), respectively. The NRI indices demonstrated that this model had better predictive performance than plasma EBV DNA level and N stage. We advocate for a threshold probability of 3.5% for guiding the application of PET/CT depending on the clinical utility analyses. This nomogram is a useful tool to predict primary DM of NPC and guide the clinical application of PET/CT individually at the initial staging.

Extranodal Expansion Of The Lymph Nodes Is An Effective Predictor Of Distant Metastasis Of Nasopharyngeal Carcinoma After Radical Radiotherapy And Chemotherapy

Extranodal extension (ENE) of the lymph nodes is a criterion for advanced nodal staging of oropharyngeal and hypopharyngeal carcinoma. The purpose is to investigate whether ENE is a factor affecting distant metastasis of nasopharyngeal carcinoma after radical radiotherapy and chemotherapy. From January 2009 to December 2014, 318 patients of NPC after radical radiotherapy and chemotherapy were enrolled. All patients received radical intensity-modulated radiotherapy. Concurrent chemotherapy was cisplatin intravenous drip every 3 weeks for 4-6cycles.MRI was retrospectively reviewed in all patients and the ENE of the lymph nodes was evaluated in patients with metastatic lymph nodes. According to the tendency of disease metastasis, patients were divided into ascending and descending types. Ascending type (Type A,T3-4N0-1) was defined as advanced local disease and early-stage cervical lymph-node involvement; advanced lymph-node metastases but early -stage local disease was classified as descending type (Type D,T1-2N2-3). Kaplan-Meier methods and Cox proportional hazard models were used to evaluate the impact of different types on distant metastasis free survival. The 5-year distant metastasis-free survival (DMFS) rate of all patients was 85%. There were 143 patients with type A and 175 patients with type D. Compared with type A, type D had worse 5-year DMFS (95% vs 78%; P < 0.05). For type A, univariate analysis and multivariate analysis suggested that ENE was closely related to distant metastasis (P < 0.05). Compared with negative ENE, ENE positive had a higher probability of metastasis within 5 years (DMFS: 99% vs 66.7%; P < 0.05). The independent prognostic factors of type D distant metastasis are ENE and the number of positive lymph nodes. ENEpositive is more likely to have distant metastasis than ENEnegative (DMFS: 84% vs 62.7%). Compared with patients with more than 4 positive lymph nodes, patients with less than 4 positive lymph nodes have significantly better DMFS (83.7% vs 90.9%; P < 0.05 for all). Patients with type D was more likely to metastasize than type A, but no matter how it was typed, ENE was an independent factor affecting distant metastasis and could be considered as a new standard for NPC lymph node staging.

Radiomics Analysis and Correlation With Metabolic Parameters in Nasopharyngeal Carcinoma Based on PET/MR Imaging.

Objective: Accurate staging is of great importance in treatment selection for patients with nasopharyngeal carcinoma (NPC). The aims of this study were to construct radiomic models of NPC staging based on positron emission tomography (PET) and magnetic resonance (MR) images and to investigate the correlation between metabolic parameters and radiomic features.Methods: A total of 100 consecutive cases of NPC (70 in training and 30 in the testing cohort) with undifferentiated carcinoma confirmed pathologically were recruited. Metabolic parameters of the local lesions of NPC were measured. A total of 396 radiomic features based on PET and MRI images were calculated [including histogram, Haralick, shape factor, gray level co-occurrence matrix (GLCM), and run length matrix (RLM)] and selected [using maximum relevance and minimum redundancy (mRMR) and least shrinkage and selection operator (LASSO)], respectively. The logistic regression models were established according to these features. Finally, the relationship between the metabolic parameters and radiomic features was analyzed.Results: We selected the nine most relevant radiomic features (six from MR images and three from PET images) from local NPC lesions. In the PET model, the area under the receiver operating characteristic (ROC) curve (AUC), accuracy, sensitivity, and the specificity of the training group were 0.84, 0.75, 0.90, and 0.69, respectively. In the MR model, those metrics were 0.85, 0.83, 0.75, and 0.86, respectively. Pearson's correlation analysis showed that the metabolic parameters had different degrees of correlation with the selected radiomic features.Conclusion: The PET and MR radiomic models were helpful in the diagnosis of NPC staging. There were correlations between the metabolic parameters and radiomic features of primary NPC based on PET/MR. In the future, PET/MR-based radiomic models, with further improvement and validation, can be a more useful and economical tool for predicting local invasion and distant metastasis of NPC.

LASP2 functions as a potential prognostic factor and therapeutic target in nasopharyngeal carcinoma.

The purpose of this study was to investigate the potential effects of LIM and Src homology 3 (SH3) protein 2 (LASP2) on nasopharyngeal carcinoma (NPC) and the relevant mechanism. The expression of LASP2 in NPC patients and non-cancer patients in the control group was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The patients were divided into LASP2 high-expression group (n=30) and low-expression group (n=30), according to the median expression level of LASP2. Then, the expression of LASP2 was detected in the chosen cell lines by qRT-PCR. In qRT-PCR experiment, LASP2 was found up-expressed in NPC clinical samples and cell lines. Besides, LASP2 expression was associated with the clinical stage and distant metastasis of NPC. Next, the expression of LASP2 was downregulated by transfection of si-LASP using LipofectamineTM 3000 in 6-10B cells in vitro. The transfection effects of si-LASP2 were confirmed by qRT-PCR and Western-blot (WB) experiments. In supplementary experiments, decreased expression of LASP2 in cells could inhibit the cell biological functions, including invasion, migration, and epithelial-mesenchymal transition (EMT). This research discovers the promotion effect of LASP2 on NPC, suggesting that LASP2 could be used as a potential therapeutic target for NPC.

Prognostic Value of Nodal Matting on MRI in Nasopharyngeal Carcinoma Patients.

Nodal (N) stage is one of the most important predictors for distant metastasis in nasopharyngeal carcinoma (NPC) patients. It may ignore potentially useful nodal features, such as nodal matting (three or more lymph nodes abutting together with the absence of intervening fat planes). To explore the prognostic value of nodal matting in NPC patients and construct a nomogram with nodal matting for predicting distant metastasis-free survival (DMFS). Retrospective. In all, 792 NPC patients treated with intensity modulated radiation therapy from 2010 to 2013 were enrolled with 2:1 training (n = 527) and validation (n = 65) cohorts. T1 - and T2 -weighted imaging at 1.5 or 3.0T. Nodal matting and other nodal characteristics were assessed with MRI. MR images were evaluated separately by three radiologists. The association between nodal matting and DMFS was analyzed. Univariate and multivariate analyses were performed using the Cox proportional hazards regression model. Nomograms were constructed from a multivariate logistic regression model with and without nodal matting. The predictive accuracy and discriminative ability of the nomograms were determined by concordance index (C-index) and calibration curves. The results were validated using bootstrap resampling and validation cohort. The incidence of nodal matting was 24.6% (195/792) in all patients. In the training cohort, nodal matting was independently associated with DMFS (hazard ratio [HR] = 1.97 [1.05-3.69], P < 0.05). N1 patients with nodal matting had worse DMFS than N1 patients without (P < 0.05). However, no significant difference was observed when comparing N1 patients with nodal matting to N2 patients (P = 0.464). The C-index of the nomogram with nodal matting was higher than the nomogram without (0.717 vs. 0.699, P = 0.084). Nodal matting was an independent prognostic factor for DMFS in NPC patients. It may help to select patients at high risk of distant metastasis.

Identifying the distant metastasis genes by next-generation sequencing panel in nasopharyngeal carcinoma.

e18547 Background: Even though local and regional controls have been substantially improved in nasopharyngeal carcinoma (NPC) in the contemporary era of intensity-modulated radiotherapy with extensive use of combined chemotherapy, the distant metastasis becomes the major cause of treatment failure and cancer-related death. To date, the genes contributed to metastasis of NPC is still unclear. The aim of this study was to identify the genes which lead to distant metastasis. Methods: A total of forty primary nonkeratinizing NPC patients were diagnosed at Shandong Cancer Hospital in this study. The formaldehyde-fixed paraffin embedded (FFPE) taken from primary sites or metastatic lymph nodes were performed next-generation sequencing (NGS) panel (Shanghai OrigiMed Co., Ltd.) to determine variated genes, such as single nucleotide variants (SNV), copy number variants (CNV) and rearrangement. These patients were followed up until Febr. 8, 2020. The genes related to distant metastasis were identified by logistic regression. Moreover, this study compared the frequency of mutated gene between our data and Catalog of Somatic Mutations in Cancer (COSMIC) database by the Chi-square test or Fisher’s exact test. Results: The study included 31 men and 9 women. The median age of the patients at diagnosis was 47 years (range 15–71 years). With the median follow-up of 10.6 months (range 16.8–72.3 months), 7 patients had distant metastasis and 1 undergone recurrent. Notably, EMSY and MCL1 variants were contributed to NPC distant metastasis (OR = 31, P = 0.049). The top eight SNV of genes in our study were CYLD, KMT2D, BAP1, EP300, TP53, ATM, NFKBIA and SPEN. When compared to COSMIC database, the mutant frequencies of CYLD, EP300 and BAP1 in our study were significantly higher than that of COSMIC database. However, the mutant frequencies of IDH2 and KMT2C were significantly lower than COSMIC database. Conclusions: This is the first study which suggests that EMSY and MCL1 variants were involved in the metastasis of NPC. The study identified 5 genes, which mutation frequency is significantly different from the COSMIC database. The study provided a molecular basis for a comprehensive understanding of, and exploring targeted therapies for nasopharyngeal carcinoma.

Abstract 3782: Tracking the evolution of nasopharyngeal carcinoma metastasis

Abstract Understanding the evolutionary history of distant metastasis in Nasopharyngeal carcinoma (NPC) is important for curing NPC and has not been systematically investigated to date. Here, we profiled whole exomes/genomes of 142 bulk samples, and transcriptomes of 60 bulk samples from 42 NPC patients with matched primary tumor, and regional lymph node/distant metastasis tumor tissues. Among them, 55755 single cells from 2 patients was also sequenced. In addition, we analyzed radiomic data from 600 NPC patients. We observed that NPC primary, regional lymph node metastasis and distant metastasis tumors had concordant mutation profiling and intratumor heterogeneity. Furthermore, we found the genetic events selected during metastasis were enriched in pathways related to metastasis, such angiogenesis, Notch and Wnt signaling. Some critical genetic events were identified, such as mutations in FAT1, RPLP1, TET2, NFKBIA, NOTCH1/3 and ARID1A, copy number variations in NOTCH2 and ARID1A. According to the mutation based phylogenetic tree and single cell RNA-Seq data, two distinct dissemination routes of distant metastases were observed, including lymphatic dissemination from regional lymph node metastases and hematogenous dissemination from primary tumors. We observed that the two different metastatic dissemination routes had different genetic mechanisms revealed by profound genomic and transcriptomic evidence. The hematogenous and lymphatic route had distinct mutation signatures, where hematogenous route was significantly enriched with higher DNA mismatch repair related mutation signature compared to lymphatic route. The bulk and single cell RNA-Seq data showed that the lymphatic route had higher activity of Epithelial-mesenchymal transition (EMT) signaling, while the hematogenous route was characterized by higher expression of INF-γ response genes and higher fraction of exhausted CD8+ T cells. Moreover, the two dissemination routes exhibited evidently different clinical outcomes. The radio image data showed that the hematogenous and lymphatic routes significantly differed in the size of primary lesion, and the range of involved lymph nodes. The radiomic data further indicated that the hematogenous and lymphatic routes could be clearly separated according to radiomic features. By applying the radiomic model to a larger cohort, we found hematogenous route had poorer survival outcome compared to lymphatic route. Finally, at cellular resolution, we observed the primary tumor cells could be separated into two distinct clones, including non-metastatic clone and metastatic-like clone. The metastatic-like clone was characterized by a significant reduction of a ribosomol protein, RPL22. We experimentally confirmed that the reduction of RPL22 could activate NFKB pathway, thereby promoting NPC cell metastasis. Citation Format: Ming Yuan Chen, Zhi-Xiang Zuo, Rui You, Xiao-Long Zhang, Mei Lin. Tracking the evolution of nasopharyngeal carcinoma metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3782.