Plasma EBV DNA Load Research Articles

The plasma EBV DNA load with IL-6 and VEGF levels as predictive and prognostic biomarker in nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NPC) is often diagnosed at a very advanced stage due to its location and non-specific initial symptoms. Moreover, no clinically useful serological marker has been established so far for early detection of NPC. In this study, we have investigated the clinical significance of plasma Epstein–Barr virus DNA load along with interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF) levels to evaluate if these three all together can be useful as a strong serological marker for early detection and prediction of treatment response in patients with NPC. Plasma EBV DNA load, IL-6 level, VEGF expressions were measured in 24 patients with NPC at presentation and various time points during and after treatment. There was a positive correlation between high plasma EBV DNA load with higher IL-6 and VEGF expression, which was closely associated with therapeutic response as well. Persistent or recurrent plasma EBV load with higher IL-6 and VEGF levels can potentially predict disease progression and may be useful to select patients for additional therapy and longer follow-up.

Clinical utility of plasma Epstein-Barr virus DNA monitoring in pediatric Epstein‐Barr virus-associated hemophagocytic lymphohistiocytosis: a Chinese retrospective observational study

BackgroundEpstein-Barr virus DNA (EBV-DNA) is closely related to the pathogenesis and prognosis of EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH). The quantitative measurement of blood EBV-DNA is widely used in EBV-HLH, but there remains a lack of evidence to guide clinicians.MethodsA retrospective analysis was conducted on clinical manifestations, laboratory tests, 310 blood EBV-DNA loads, and prognosis of 51 pediatric patients diagnosed with EBV-HLH. Receiver operating characteristic (ROC) curves were utilized to determine the optimal cutoff values of EBV-DNA for predicting mortality and evaluating the active status of EBV-HLH.ResultsEBV-positive- lymphoma-HLH had higher initial plasma EBV-DNA load(1.10 × 106copies/ml) compared to the EBV-HLH group (1.98 × 104 copies/ml) (P = 0.006), and experienced recurrently elevated plasma EBV-DNA levels during treatment. The optimal cut-off value of initial plasma EBV-DNA load in predicting mortality was 2.68 × 105 copies/ml, with a sensitivity of 88.57% and a specificity of 56.25%. For determining the active status of HLH, the optimal cutoff value of PBMC EBV-DNA load during treatment was 2.95 × 105 copies/ml, with a sensitivity of 69.14% and a specificity of 64.71%. The cut-off value of plasma EBV-DNA for determining active status was 1.32 × 103 copies/ml, with a sensitivity of 84.34%, and a specificity of 87.67%. Patients with higher PBMC and plasma EBV-DNA at initial and those with repeated elevated plasma EBV-DNA during treatment had worse prognoses (P < 0.05).ConclusionDynamic monitoring of EBV-DNA is a valuable tool for assessing disease status and predicting the prognosis of EBV-HLH, with plasma EBV-DNA being more effective than PBMC EBV-DNA. Patients with high levels of PBMC and plasma EBV-DNA at initial and those with repeated elevated plasma EBV-DNA during treatment had worse prognoses.

Role of plasma EBV-DNA load and EBER status on newly diagnosed peripheral T-cell lymphoma.

To explore the prognostic and therapeutic role of Epstein-Barr Virus (EBV) on peripheral T-cell lymphoma (PTCL). Totally 262 newly diagnosed PTCL patients who were hospitalized from January 2014 to December 2022 were retrospectively enrolled. Molecular analysis included 31 eligible patients. EBV-encoded RNA (EBER) presence in tumor tissue and EBV DNA levels in patients at baseline (DNA1) and after 4 cycles of chemotherapy (DNA4) were assessed. Our findings revealed that the EBER-positive cohort exhibited significant differences compared to counterparts in overall survival (OS, P = 0.047) and progression-free survival (PFS, P = 0.009). Both DNA1 and DNA4 were significantly associated with inferior OS. Multivariate analysis demonstrated that DNA4 independently affected PTCL prognosis for OS (hazard ratio = 5.1617; 95% confidence interval 1.1017-24.1831; P = 0.037). Treatment with the cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) plus azacytidine regimen showed a better OS compared to CHOP or CHOP plus etoposide for patients with partially positive EBER and EBER positive statuses (P = 0.192), although the improvement was not statistically significant. This study delineated the genetic paradigm of PTCL, comparing genetic differences by EBV status and found that EBER partially positive plus positive patients were more likely to have DNMT3A (P = 0.002), RHOAG17V (P = 0.023), and TET2 mutations (P = 0.032). EBER, DNA1, and DNA4 emerged as sensitive markers for prognosis. CHOP plus azacytidine might present a preferable option for PTCL patients with DNA methylation due to EBV infection.

Comparative Analysis of Primary and Reactivated EB Virus Infection Associated Hemophagocytic Lymphohistiocytosis

To compare the clinical characteristics of children with hemophagocytic lymphocytosis (HLH) associated with primary Epstein-Barr virus (EBV) infection and EBV reactivation, and explore the effects of different EBV infection status on the clinical indexes and prognosis of HLH. The clinical data of 51 children with EBV associated HLH treated in Henan Children's Hospital from June 2016 to June 2021 were collected. According to the detection results of plasma EBV antibody spectrum, they were divided into EBV primary infection-associated HLH group (18 cases) and EBV reactivation-associated HLH group (33 cases). The clinical features, laboratory indexes and prognosis of the two groups were analyzed and compared. There were no significant differences in age, gender, hepatomegaly, splenomegaly, lymphadenopathy, neutrophil count in peripheral blood, hemoglobin content, platelet count, plasma EBV-DNA load, lactate dehydrogenase, alanine aminotransferase, aspartate aminotransferase, albumin, fibrinogen, triglyceride, ferritin, hemophagocytosis in bone marrow, NK cell activity and sCD25 between the two groups(P>0.05). The central nervous system involvement and CD4/CD8 in EBV reactivation-associated HLH group were significantly higher than those in primary infection-associated HLH group, but the total bilirubin was significantly lower than that in primary infection-associated HLH group (P<0.05). After treatment according to HLH-2004 protocol, the remission rate, 5-year OS rate and 5-year EFS rate of patients in EBV reactivation-associated HLH group were significantly lower than those in EBV primary infection-associated HLH group (P<0.05). EBV reactivation-associated HLH is more likely to cause central nervous system involvement and the prognosis is worser than EBV primary infection-associated HLH, which requires intensive treatment.

Synergistic Association of Hepatitis B Surface Antigen and Plasma Epstein-Barr Virus DNA Load on Distant Metastasis in Patients With Nasopharyngeal Carcinoma

Hepatitis B surface antigen (HBsAg) reportedly increases the risk of distant metastasis among patients with nasopharyngeal carcinoma (NPC). However, the associated potential interaction and changes in hazard ratios (HRs) between HBsAg and different plasma Epstein-Barr (EBV) DNA levels are unknown. Moreover, the potential HBsAg-positive-associated NPC metastatic mechanism remains unclear. To investigate the prognostic value and biological associations of HBsAg and plasma EBV DNA levels on distant metastasis in patients with NPC. Retrospective cohort study performed at Sun Yat-sen University Cancer Center between January 2010 and January 2013. A total of 792 patients with nonmetastatic NPC were enrolled. The median (range) follow-up time was 62.1 (1.4-83.4) months. Of these patients, 17.8% presented with HBsAg positivity. Cytological experiments were performed to evaluate the role of HBsAg in the invasion and migration of EBV-positive NPC cells. Data analysis was performed from July 2020 to April 2021. The primary end point was distant metastasis-free survival. Association rules were used to identify new rules related to distant metastasis. Interaction plots, univariate and multivariate Cox regression analyses, stratification analysis, and quantification using HRs were conducted. Additionally, cell migration and invasion assays, as well as Western blotting, were performed in the cytological validation. Among the 792 patients, 576 (72.7%) were male, with a median (IQR) age of 45 (38-53) years. The HBsAg-positive group exhibited a significant interaction and increased risk of distant metastasis when plasma EBV DNA cutoff levels were 1.5 × 1000 copies/mL or greater. The HR was 9.16 (95% CI, 2.46-34.14) when the plasma EBV DNA load reached 6 × 1000 copies/mL, which was higher than that in patients with stage IV disease (HR, 2.01; 95% CI, 1.13-3.56; P = .02). In cytological experiments, HBsAg promoted epithelial-mesenchymal transition by upregulating vimentin and fibronectin in EBV-positive NPC cells in vitro, thereby promoting invasion and migration of EBV-positive NPC cells. In this cohort study, the observed synergistic association between HBsAg and plasma EBV DNA load represented a novel potential mechanism underlying the increased risk of distant metastasis in patients with NPC. Hence, attention should be paid to patients with NPC with HBsAg positivity, especially when the plasma EBV DNA level is 6 × 1000 copies/mL or greater. Consideration of this synergistic association will contribute to more accurate individualized management.

Clinical outcomes of non-nasopharyngeal lymphoepithelial carcinoma treated with a combined modality approach: A single-institution study.

This study presents a summary of the clinical characteristics of non-nasopharyngeal lymphoepithelial carcinoma (NNPLEC), effects of combined modality treatment and prognostic value of plasma Epstein-Barr virus (EBV) deoxyribonucleic acid (DNA) load, with the aim of providing a reference framework for optimizing treatment practices and outcomes. Patients with NNPLEC treated by our center between January 2000 and December 2020 were retrospectively reviewed. In total, 728 patients were included. The lung was identified as the most common primary tumor site (64.0%), followed by the salivary gland (19.2%). A total of 539 (74.0%) patients underwent surgery, 459 (63.0%) received chemotherapy, and 361 (49.6%) were subjected to radiotherapy. The median follow-up time was 45 months (range, 6-212 months) and 5-year overall survival (OS) was 79.1%. Increased plasma EBV-DNA load of >513.5 copies/mL was predictive of disease progression, with a specificity of 98.1% and a sensitivity of 98.9%. In multivariate Cox analysis, N stage, surgery, and radiotherapy were independent prognostic factors for both OS and PFS. Radiotherapy significantly improves OS in comparison with no radiotherapy group for salivary LEC, while surgery significantly improves OS for pulmonary LEC. Based on our analysis, surgery and radiotherapy are associated with better OS and PFS for NNPLEC. Radiotherapy could be recommended for salivary LEC, while surgery remains the primary treatment strategy for pulmonary LEC patients. An increased plasma EBV-DNA load of >513.5 copies/mL is strongly predictive of disease progression, supporting the importance of regular evaluation of plasma EBV-DNA as part of the diagnostic routine.

High plasma EBV-DNA load and positive EBER status associated with viral recurrence and persistent infection in early treatment of lymphoma.

The relationship between plasma EBV-DNA load (PEDL) and Epstein-Barr virus (EBV)-encoded small RNA (EBER) during the early treatment of lymphoma remains unclear. We explored discrepancies in PEDL and variables associated with EBER and evaluated the consistency between EBER and qualitative analysis of PEDL (qPEDL). Serial measurements of PEDL were performed to determine the dynamic changes of PEDL in early treatment of lymphoma. As a result, the median PEDL of non-Hodgkin's lymphoma NKT cell subtype (NHL-NKT) was higher than that of non-Hodgkin's lymphoma B cell subtype (NHL-B), the median PEDL of extranodal NK/T cell lymphoma (ENKTCL) was higher than that of diffuse large B cell lymphoma (DLBCL), and the median PEDL of EBER positive was higher than that of EBER negative. Age, Ki-67 ≧ 80%, Bcl-2 ≧ 80%, p53, and qPEDL were related to EBER. The PEDL could distinguish NHL-B, DLBCL, NHL-NKT, and ENKTCL from other lymphoma subtypes. EBER-positive patients spent more time with viral "turn negative (TN)" and "continuous positive (CP)" and less time with viral "continuous negative (CN)." The median PEDL of CP was higher than that of TN. In conclusion, although EBER affects the levels of PEDL in general, it has poor concordance with qPEDL. Our results show, for the first time, that high PEDL and positive EBER present a strong association with viral recurrence and persistent infection in the early treatment of lymphoma.

P1238: EFFICACY ANALYSIS OF PEGASPARGASE, GEMCITABINE, OXALIPLATIN, DEXAMETHASONE COMBINED WITH TISLELIZUMAB IN FIRST-LINE TREATMENT OF PLASMA EBV-DNA POSITIVE EXTRANODAL NK/T CELL LYMPHOMA

Background: Positive circulating EBV-DNA is associated with a very poor outcome in Natural killer/T cell lymphoma (NKTCL). Efforts to further improve treatment of plasma EBV positive NKTCL are urgently needed. Aims: To investigate the clinical efficacy, plasma EBV-DNA load and safety of P-GEMOXD combined with anti-programmed death 1 (anti-PD-1) antibody Tislelizumab in first-line treatment of plasma EBV-DNA positive NKTCL. Methods: In this prospective, single-arm, phase 2 trial, we recruited patients aged 18–70 years with newly diagnosed NKTCL who were EBV-DNA positive in plasma in Henan People’s Hospital. Patients received four courses of P-GEMOXD plus Tislelizumab treatment (pegaspargase 2500U/m2 [day3], gemcitabine 800mg/m2 [day1 and 8]; oxaliplatin100 mg/m2[day1], dexamethasone 20mg/d [day1-4]; Tislelizumab 200mg /d [day9] every 21days). Thirteen patients were enrolled from January 2018 to August 2021 December, the median follow-up was 26 (12-46) months. The short-term and long-term clinical efficacy, load of EBV-DNA and adverse reactions were observed. Results: Among the 13 patients, there were 9 males and 4 females, the median age was 54 years old (range 25-64). After 4 courses of treatment, 10 patients (76.9%) had complete response, 3 patients (23.1%) had partial response, and the objective response rate (ORR) was 100%. The 2-year progress-free survival (PFS) rate was 76.9%, and the 2-year overall survival (OS) rate was 85.7%. Among them, there were 6 patients with advanced stage (III and IV), 4 patients (66.7%) had complete response, 2 patients (33.3%) had partial response, 5 patients (83.3%) plasma EBV-DNA converted negative; the 2-year progression-free survival (PFS) rate was 66.7%, and the 2-year overall survival (OS) rate was 66.7%. Treatment-related AEs of any grade occurred in 84.6% (11/13) of all patients. The main adverse reactions were albumin reduction (8/13), fibrinogen reduction (9/13), autoimmune diseases (3 cases：1 case of hypopituitarism， 1 case of Hashimoto’s thyroiditis, and 1 case spondylitis). Summary/Conclusion: P-GEMOXD plus Tislelizumab can significantly improve the prognosis of patients with NKTCL, improve the rate of plasma EBV-DNA negative conversion, and the adverse reactions can be well tolerated.

Development of a Nomogram to Predict the Risk of Chronic Active Epstein-Barr Virus Infection Progressing to Hemophagocytic Lymphohistiocytosis.

BackgroundChronic active Epstein-Barr virus infection (CAEBV) disease is sometimes associated with an aggressive clinical course, such as hemophagocytic lymphohistiocytosis (HLH). To explore the risk factors and predict the risk of CAEBV infection progressing to HLH, a retrospective research study was conducted.MethodsWe retrospectively reviewed the medical records of 187 CAEBV-infected patients who were admitted to our center between January 2015 and December 2020. The patients were followed up until May 2021. The patients were divided into a progression-to-HLH group and a no-progression-to-HLH group. Demographic, clinical and laboratory data were collected for each patient.ResultsAmong the 121 CAEBV-infected patients who fulfilled the study's inclusion criteria, 48 (30.7%) patients did not progress to HLH, and 73 (60.3%) patients progressed to HLH. The median time from CAEBV infection to progression to HLH was 14 months, and the cumulative incidence rate of HLH increased as the duration of follow up increased (24.9, 47.3, 55.1, and 85.2% at 1, 3, 5, and 10 years, respectively). Multivariate analyses showed that the independent risk factors for CAEBV progression to HLH were plasma EBV-DNA load (OR = 3.239, 95% CI 1.219–8.603, P = 0.018), Platelet count (OR=0.991, 95%CI 0.985–0.998, P = 0.010), elevated alanine aminotransferase (OR=1.019, 95%CI 1.005–1.034, P = 0.009) and ≥2 of 3 lineages of cytopenia (OR=8.364, 95%CI 1.062–65.839, P = 0.044). The regression coefficients (β) from the multivariate logistic model were used to construct a model for estimating the risk of CAEBV infection progressing to HLH. The discriminatory ability of the model was good, and the area under the receiver operating characteristic (ROC) curve (AUC) was 0.925.Conclusionplasma EBV-DNA load, platelet count, elevated alanine aminotransferase and ≥ 2 of 3 lineages of cytopenia increase the risk of CAEBV infection progressing to HLH. A nomogram can be used to estimate the risk of CAEBV-infected patients progressing to HLH.

Risk factors and regularity of transaminase elevation associated with primary Epstein-Barr virus infection in children

Objective: To explore the risk factors and regularity of pediatric primary Epstein-Barr virus (EBV) infection accompanied with elevated transaminase. Methods: Clinical data of 399 children diagnosed as primary EBV infection in the outpatient department, Children's Hospital of Fudan University from September 2016 to October 2017 were analyzed retrospectively. Logistic regression analysis was performed to determine the potential correlations between elevated alanine transaminase (ALT) or aspartate transaminase (AST) and age, gender, course of fever and plasma EBV-DNA load. The cumulative rates of elevated transaminase recovery to nomal at different times were caculated. Results: Among 399 children diagnosed with primary EBV infection, there were 219 males and 180 females. The age was (4.2±2.7) years. Among all cases, 51.9% (207/399) had elevated transaminase. In patients who had elevated ALT, 74.5% (149/200), 21.0% (42/200) and 4.5% (9/200) had mild (40-160 U/L), moderate (160-400 U/L) and severe (>400 U/L) elevation of ALT, respectively. In patients who had elevated AST, 83.8% (155/185), 11.9% (22/185) and 4.3% (8/185) had mild (40-160 U/L), moderate (160-400 U/L) and severe (>400 U/L) elevation of AST, respectively. Only age was correlated with the occurrence of elevated transaminase (OR=1.13, 1.10, both P<0.05). A total of 167 repeated tests were ordered in patients with elevated ALT and/or AST, including 113 cases with elevated ALT and 104 cases with elevated AST. The time of ALT and AST returned to normal were (24±13) days and (25±18) days respectively. The cumulative rates for ALT returned to normal within 1, 1-<4, 4-<8 weeks and more than 8 weeks were 2.7% (3/113), 54.0% (61/113), 79.6% (90/113) and 81.4% (92/113) respectively, and were 1.9% (2/104), 48.1% (50/104), 71.2% (74/104) and 74.0% (77/104) for AST. Conclusions: Age is a risk factor for transaminase elevation associated with primary EBV infection in children. The transaminases returned to normal within 3 weeks in half of the cases, and within 8 weeks in most cases.

A rare presentation recurrent nasopharyngeal carcinoma with axillary lymph node metastasis

Introduction: Nasopharyngeal carcinoma (NPC) has the highest rate of local and regional cervical lymph node re-currence amongst other head and neck epithelial malignant tumour. Distant recurrence is less common and usually occurs in bone, liver and lung. Recurrent NPC to axillary lymph node is rare.Case report: A 44-year-old male presented with two-month history of painless right axillary swelling. He was di-agnosed with nasopharyngeal carcinoma (NPC) stage IVA (T2N3M0) two years prior to presentation and had underwent neoadjuvant chemotherapy with 5-Fluorouracil and Cisplatin, and concurrent chemo-radiotherapy (CCRT) of total 70 Gy over 35 sessions with Cisplatin. He was on regular monthly surveillance reviews, with no signs of recurrence. Fine Needle Aspiration Cytology (FNAC) of the axillary swelling was reported as metastatic NPC and Positron Emission Tomography/ Com-puted tomography (PET-CT) scan showed foci of high FDG hypermetabolism at right axillary lymph node confirming the diagnosis of recurrent NPC.Conclusion: Recurrence usually occurs in the first two years after completion of treatment. Patients with an overall TNM stage IV or N3, high pre-treatment or persistently detectable post-treatment plasma EBV DNA (pEBV DNA) load are at greatest risk. Management of recurrent NPC depends on local, regional or distant recurrence. Both PET scan and pEBV DNA load can be used for relapse detection. The man-agement for each recurrent NPC case is unique and should be determined by a multidisciplinary team, local expertise and facilities. Knowledge of potential sites of recurrence is essential to both physicians and patients for early detection.

Prospective observation: Clinical utility of plasma Epstein-Barr virus DNA load in EBV-associated gastric carcinoma patients.

Epstein-Barr virus (EBV)-associated gastric carcinomas (EBVaGCs) may account for 8-9% of all gastric cancer (GC) patients. All previous reports on EBVaGC were retrospective. Prospective study is warranted to evaluate the exact role of EBV status in predicting the prognosis of GC. It is of special interest to figure out whether dynamic detection of plasma EBV-DNA load could be a feasible biomarker for the monitor of EBVaGC. From October 2014 to September 2017, we consecutively collected GC patients (n = 2,760) from Sun Yat-sen University Cancer Center for EBER examination. We detected EBV-DNA load in plasma and tissue samples of EBVaGC patients at baseline. Subsequently, plasma EBV-DNA load was dynamically monitored in EBVaGC patients. The overall prevalence of EBVaGC is 5.1% (140/2,760). The incidence rate of EBVaGC decreased with advanced AJCC 7th TNM stage (p < 0.001), with the corresponding percentages of 9.3, 9.9, 6.7 and 1.4% for Stage I, II, III and IV patients. EBVaGC patients were predominately young males with better histologic differentiation and earlier TNM stage than EBV-negative GC (EBVnGC) patients. EBVaGC patients were confirmed to had a favorable 3-year survival rate (EBVaGC vs. EBVnGC: 76.8% vs. 58.2%, p = 0.0001). Though only 52.1% (73/140) EBVaGC patients gained detectable EBV-DNA and 43.6% (61/140) reached a positive cutoff of 100 copies/ml, we found the plasma EBV-DNA load in EBVaGC decreased when patients got response, while it increased when disease progressed. Our results suggested that plasma EBV-DNA is a good marker in predicting recurrence and chemotherapy response for EBVaGC patients.

Role of pretreatment 18F-FDG PET/CT parameters in predicting outcome of non-endemic EBV DNA-related nasopharyngeal cancer (NPC) patients treated with IMRT and chemotherapy.

To evaluate the prognostic role of pretreatment 18F-FDG PET/CT metabolic parameters in non-endemic Epstein-Barr Virus (EBV DNA)-related nasopharyngeal cancer (NPC) patients treated with curative intensity-modulated radiation therapy (IMRT) with or without chemotherapy (CHT). We retrospectively reviewed clinical data of 160 consecutive non-metastatic NPC patients who received IMRT with or without CHT. Forty-nine out of 160 patients that underwent whole body 18F-FDG PET/CT at our institution for disease staging with a minimum follow-up to 12months were included in this study. We evaluated the relationship between maximum and mean standardized uptake values (SUVmax and SUVmean, respectively), metabolic tumor volume and total lesion glycolysis (TLG) of primary tumor and cervical lymph nodes with disease-free survival (DFS) and overall survival (OS). We also investigated the prognostic role of clinical variables such as age, disease stage, plasma EBV DNA load (copies/ml), gross tumor volume of primary tumor and lymph nodes. Median follow-up was 55months. Two- and 5-year OS were 95.8% and 90.5%, respectively, while DFS was 83.4% at both time points. SUVmax of primary tumor ≥ 18.8g/ml and primary tumor TLG ≥ 203.1g were significant prognostic factors of worse OS. Furthermore, stages IVB and EBV DNA load ≥ 3493 copies/ml were significantly associated with lower DFS. No correlation was found between PET parameters and plasma EBV DNA load. Even in a limited series, our data suggested that SUVmax, SUVmean and TLG of primary tumor could predict a poor outcome in NPC patients also in non-endemic area hypothesizing their use for refinement of prognostication.

Long-Term Prognostic Effects of Plasma EBV DNA Load after Induction Chemotherapy in Advanced Nasopharyngeal Carcinoma

Induction chemotherapy (IndCT) has become an option of initial treatment for advanced nasopharyngeal carcinoma (NPC) patients. We hypothesized that plasma EBV DNA (pEBV DNA) status afterIndCT could predict long-term treatment outcome. This retrospective study included 206 previously untreated, pathologically-proven NPC patients with stage II-IVB diseases and available pEBV DNA data after IndCT before radiotherapy (RT). All patients received a uniform protocol of ten weekly IndCT consisting of cisplatin 60 mg/m2 on day 1 and 5- fluorouracil 2500 mg/m2 + leucovorin 250 mg/m2 on day 8, repeated every 2 weeks for 5 cycles, followed by RT 70-74 Gy. After a median follow-up of 90 months, there were 71 recurrences and 85 deaths. Pretreatment pEBV DNA (median, 1550 copies/ml; interquantile range, 375-5688) was detectable in 95.1% (196/206) patients. Only 40.8% (84/206) patients had detectable pEBV DNA (> 0 copy/ml) after IndCT with very lower copy numbers (median, 18 copies/ml; interquantile range, 7-153, for these 84 patients). Patients with a higher N-classification (N3 vs. N0-2, P<0.0001) and overall stage (IV vs. II-III, P=0.0400) had a higher detectable pEBV DNA after IndCT. Forty-twor of 84 (50.0%) patients with detectable pEBV DNA (> 0 copy/ml) developed tumor relapse, whereas 23.8% (29/122) patients with undetectable pEBV DNA (= 0 copy/ml) had tumor relapse (P<0.0001). The patterns of failure revealed that patients with detectable post-IndCT pEBV DNA had more distant (39.3% vs. 17.2%, P=0.0004) and neck (7.1% vs. 1.6%, P=0.0650) failures but not local recurrence (11.9% vs. 9.8%, P=0.6366), compared with those of undetectable EBV DNA.The 5-year overall survival rates were 63.1% and 70.4% for patients with post-IndCT viral load > and = 0 copies/ml (P=0.0010). The corresponding figures of relapse-free survival rates were 54.4% and 78.4% (P<0.0001). Post-IndCT pEBV DNA status can predict long-term outcome for NPC patients. More intensive treatment strategy for these high-risk patients with persistently detectable pEBV DNA after IndCT should be studied further.

Impacts of plasma EBV DNA load during different time-points of induction chemotherapy plus radiotherapy for nasopharyngeal carcinoma.

6058Background: To investigate the prognostic impacts of plasma EBV DNA measured at different time-points in patients with advanced nasopharyngeal carcinoma who received induction chemotherapy (Ind...

Dynamic changes in plasma Epstein-Barr virus DNA load during treatment have prognostic value in nasopharyngeal carcinoma: a retrospective study.

Circulating plasma Epstein–Barr virus DNA (EBV DNA) is related to tumor recurrence and metastasis and has potential as a dynamic, sensitive, and specific marker in nasopharyngeal carcinoma (NPC). We investigated the clinical significance of assessing plasma EBV DNA load at various time points during treatment. Patients with NPC (n = 949) for whom plasma EBV DNA load was measured by real‐time quantitative polymerase chain reaction (RT‐qPCR) before treatment (pre‐EBV) and at midtreatment (mid‐EBV), end of treatment (end‐EBV), and 3 months after completing treatment (3 m‐EBV) were retrospectively assessed. Receiver operating characteristic (ROC) curve analysis was used to identify the optimal EBV DNA cutoff point for each time point. Overall survival (OS), distant metastasis‐free survival (DMFS), and progression‐free survival (PFS) were compared using Kaplan–Meier estimates. High pre‐EBV, high mid‐EBV, high end‐EBV, and high 3 m‐EBV were all associated with significantly poorer OS, DMFS, and PFS in the entire cohort. Detectable end‐EBV and 3 m‐EBV was associated with significantly poorer OS, DMFS, and PFS. Among patients with detectable end‐EBV, adjuvant therapy significantly improved OS (HR 2.419; 95% CI 1.297–4.51, P = 0.03) and DMFS (HR 2.45; 95% CI 1.243–4.828, P = 0.04), but not PFS (P = 0.17). EBV DNA represents a dynamic biomarker for monitoring treatment and predicting survival in NPC. Assessing plasma EBV DNA before, during, and after chemoradiotherapy could be clinically valuable and enable selection of patients most likely to benefit from additional therapy and improve assessment of treatment response and disease surveillance. Further multicenter prospective investigations are warranted.

Abstract 2262: Correlation between number and sites of metastases and differential pretreated EBV DNA load profiles and their effects on outcomes in patients with metastatic nasopharyngeal carcinoma

Abstract Differential overall survival of different organ site metastases in nasopharyngeal carcinoma (NPC) had been reported but the underlying mechanism was unclear. We try to find out the possible mechanism from plasma and tumor tissue markers. Total 178 patients of metastatic NPC were enrolled. Their pre-treatment plasma EBV DNA concentrations and cytokines and tissue macrophage, proliferation and apoptosis markers were determined. The overall patient response rate after treatment was 51.7% and median overall survival (OS) was 19 months. Single organ site metastasis had better outcome than multiple organs involved (median OS: 26 months vs. 16 months) with statistical significance. Among single organ site involved, pure lung metastasis had longer survival than bone or liver involved (median OS: 50 months vs. 21 months vs. 18 months) with P&amp;lt;0.001. Pretreatment plasma EBV DNA concentrations were lower in patients with lung metastasis than bone or liver metastasis in single organ involved group. Plasma IP-10 and MCP-1 expression level correlated with the differential single organ site metastasis OS and EBV DNA load. Liver metastatic tissue had higher macrophage infiltration density and higher proliferation index than lung metastatic group. In single organ site metastasis of NPC, lung involved patients had better outcome than bone or liver metastasis. Low pretreatment plasma EBV DNA load, cytokines expression such as IP-10 and MCP-1, tissue macrophage infiltration, and proliferation index may contribute these results. Citation Format: Cheng-Lung Hsu, Hung-Ming Wang, Tung-Liang Lin, Yung-Chia Kuo. Correlation between number and sites of metastases and differential pretreated EBV DNA load profiles and their effects on outcomes in patients with metastatic nasopharyngeal carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2262. doi:10.1158/1538-7445.AM2017-2262

Abstract 626: Application of patient-derived xenograft model in nasopharyngeal carcinoma research

Abstract Nasopharyngeal carcinoma (NPC) was an Epstein Barr virus (EBV)-related malignancy and tumor microenvironment had a pivotal role in tumor progression. Paucity of good NPC animal models hindered the research in this field. Recently, patient-derived xenograft (PDX) had been shown to be a good preclinical model for drug screening and cancer related research. We had developed two PDX mice lines from engrafting NPC metastatic tumors. Positive EBV-encoded small RNAs staining confirmed these tumors harboring EBV. Further gene expression profile analysis showed higher similarity of PDX to primary parent tumor than NPC cell line xenograft. In vivo drug screening in the PDX system demonstrated gemcitabine had the best antitumor effect among the tested drugs. In this PDX corresponding patient also showed excellent response to gemcitabine treatment. Combination of gemcitabine and valproic acid had synergistic antitumor effect. Further adding ganciclovir in this two combined regimen enhancing cytolytic viral activation had the best antitumor response among the tested regimens. This three combined regimen treated group had lower plasma EBV-DNA load and tumor viral concentration and less viable tumor cells than gemcitabine + valproic acid group. These promising results would open a new era for EBV-targeting therapy in NPC treatment. Citation Format: Cheng-Lung Hsu, Yung-Chia Kuo, Yenlin Huang, Yin-Cheng Huang, Kar-Wai Lui, Kai-Ping Chang, Tung-Liang Lin, Hsien-Chi Fan, An-Chi Lin, Chia-Hsun Hsieh, Li-Yu Lee, Hung-Ming Wang, Hsin-Pai Li, Angel Chao, Yu-Sun Chang. Application of patient-derived xenograft model in nasopharyngeal carcinoma research. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 626.

Diagnosis and Treatment of Nasopharyngeal Carcinoma in Children and Adolescents - Recommendations of the GPOH-NPC Study Group.

Nasopharyngeal carcinoma (NPC) is a rare malignant tumor arising from epithelial cells of the nasopharynx. Its incidence is highest in Southeast Asia. Age distribution of NPC is bimodal, with one peak in young adolescents and another in patients 55-59 years of age. EBV appears to be the primary etiologic agent in the pathogenesis, environmental factors such as nitrosamines and genetic factors are contributory. NPC is most commonly diagnosed in locally advanced stages, with lymph node metastases occurring in up to 90% of patients. About 5-10% of patients present with distant metastases. Diagnosis of NPC is made histologically, supported by an abnormal anti-EBV-VCA IgA titer and elevated plasma EBV-DNA load. Superior results in children and adolescents with advanced locoregional NPC, with overall and event-free survival rates>90%, have been achieved by neoadjuvant chemotherapy with 5-fluoruracil and cisplatin, followed by synchronous radiochemotherapy and subsequent maintenance therapy with interferon-ß as demonstrated by the 2 prospective studies GPOH-NPC-91 and -2003. Response to therapy can be assessed by PET-imaging and in patients with complete remission after neoadjuvant chemotherapy, the radiation dose to the primary tumor can be safely reduced from 59.4 to 54.4 Gy. Since the majority of long term sequalae such as xerostomia, skin and tissue fibrosis are caused by high radiation dosages, radiotherapy modalities such as intensity-modulated radiotherapy should be used to efficiently spare non-tumorous tissue. For patients with metastatic disease and relapse, survival chances are low. New treatment strategies, such as the application of EBV-specific T-lymphocytes should be considered for these patients.

Cigarette smoking complements the prognostic value of baseline plasma Epstein-Barr virus deoxyribonucleic acid in patients with nasopharyngeal carcinoma undergoing intensity-modulated radiation therapy: a large-scale retrospective cohort study.

We evaluated the combined prognostic value of cigarette smoking and baseline plasma Epstein-Barr virus deoxyribonucleic acid (EBV DNA) in patients with nasopharyngeal carcinoma (NPC) treated with intensity-modulated radiation therapy (IMRT). Of consecutive patients, 1501 with complete data were eligible for retrospective analysis. Smoking index (SI; cigarette packs per day times smoking duration [years]), was used to evaluate the cumulative effect of smoking. Primary endpoint was overall survival (OS); progression-free survival (PFS), distant metastasisfree survival (DMFS) and locoregional relapse-free survival (LRFS) were secondary end-points. Both cigarette smoking and baseline plasma EBV DNA load were associated with poorer survival (P<0.001). Patients were divided into four groups: low EBV DNA and light smoker (LL), low EBV DNA and heavy smoker (LH), high EBV DNA and light smoker (HL), and high EBV DNA and heavy smoker (HH). The respective 5-year survival rates were: OS (93.1%, 87.2%, 82.9%, and 76.3%, P<0.001), PFS (87.0%, 84.0%, 73.9%, and 64.6%, P<0.001), DMFS (94.1%, 92.1%, 82.4%, and72.5%, P<0.001), and LRFS (92.8%, 92.4%, 88.7%, and 84.0%, P=0.012).OS and PFS were significantly different between the LH and HL groups and HL and HH groups, but not LL and LH groups (pairwise comparisons). The combined risk stratification remained an independent prognostic factor for all endpoints (all Ptrend<0.001; multivariate analysis). Both cigarette smoking and baseline plasma EBV DNA were independent prognostic factors for survival outcomes. Combined interpretation of EBV DNA with smoking led to the refinement of the risks stratification for patient subsets, especially with improved risk discrimination in patients with high baseline plasma EBV DNA.