Tumor cell-derived exosomal miR-193b-3p promotes tumor-associated macrophage activation to facilitate nasopharyngeal cancer cell invasion and radioresistances.

Background: Communication between cancer cells and tumor-associated macrophages (TAMs) in the tumor microenvironment (TME) plays a crucial role in accelerating nasopharyngeal cancer (NPC) metastasis and radioresistance. However, the mechanisms through which NPC cells regulate the properties and activation of TAMs during NPC progression are not yet fully understood. Methods: A high-metastatic NPC subclone (HMC) and a low-metastatic NPC subclone (LMC) were screened from the CNE-2 cell line and exosomes were collected from HMCs and LMCs, respectively. The effects of HMC- and LMC-derived exosomes (HMC-Exos and LMC-Exos) on the regulation of TAM activation were evaluated by assessing the levels of inflammation-related or immunosuppression-related genes. The role of miRNA-193b-3p (miR-193b) in mediating communication between NPCs and TAMs was assessed using real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR), Western blot analysis, Transwell assays, and clonogenic survival assays. Results: HMCs and HMC-Exos exhibited a greater capacity to facilitate macrophage protumorigenic activation than LMCs and LMC-Exos. miR-193b levels derived from HMC-Exos were higher than those from LMC-Exos, and miR-193b levels were higher in metastatic NPC tissue-derived TAMs than in non-metastatic NPC tissue-derived TAMs. The upregulated miR-193b was packaged into exosomes and transferred to macrophages. Functionally, miR-193b up-regulation accelerated TAM activation by directly targeting mitogen-activated protein/ERK kinase kinase 3 (MEKK3). As a result, miR-193b-overexpressed macrophages facilitated NPC cell invasion and radioresistance. Conclusions: These data revealed a critical role for exosomal miR-193b in mediating intercellular communication between NPC cells and macrophages, providing a potential target for NPC treatment.

Prediction of Drug-Drug Interactions with Ensartinib as a Time-Dependent CYP3A Inhibitor Using Physiologically Based Pharmacokinetic Model.

Ensartinib (X-396) is a second-generation anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitor (TKI) indicated for the treatment of ALK-positive patients with locally advanced or metastatic non-small cell lung cancer. Although in vitro experiments and molecular docking suggested its potential as a cytochrome P450 inhibitor, no further investigation or clinical trials have been conducted to assess its drug-drug interaction (DDI) risk. In this study, we conducted a series of in vitro experiments to elucidate the inhibition mechanism of ensartinib. Furthermore, a physiologically-based pharmacokinetic (PBPK) model was developed based on in vitro, in silico, and in vivo parameters, verified using clinical data, and applied to predict the clinical DDI mediated by ensartinib. The in vitro incubation experiments suggested that ensartinib exhibited strong time-dependent inhibition. Simulation results from the PBPK model indicated a significant increase in the exposure of CYP3A substrates in the presence of ensartinib, with the maximal plasma concentration and area under the plasma concentration-time curve increasing up to 12-fold and 29-fold for sensitive substrates. Based on these findings, it is evident that co-administration of ensartinib and CYP3A substrates requires careful regulatory consideration. SIGNIFICANCE STATEMENT: Ensartinib was found to be a strong time-dependent inhibitor of CYP3A for the first time based on in vitro experiments, but there was no research conducted to estimate the risk of drug-drug interaction (DDI) of ensartinib in clinic. Therefore, the first ensartinib physiologically based pharmacokinetic model was developed and applied to predict various untested scenarios. The simulation result indicated that the exposure of CYP3A substrate increased significantly and urged the further clinical DDI study.

Prognostic value of circulating Epstein-Barr virus DNA level post-induction chemotherapy for patients with nasopharyngeal carcinoma: A recursive partitioning risk stratification analysis.

BACKGROUND: To evaluate the prognostic value of plasma Epstein-Barr virus (EBV) DNA level post-induction chemotherapy (IC) for patients with nasopharyngeal carcinoma (NPC). METHODS: A total of 893 newly diagnosed NPC patients treated with IC were retrospectively reviewed. The recursive partitioning analysis (RPA) was performed to construct a risk stratification model. The receiver operating characteristic (ROC) analysis was applied to determine the optimal cut-off value of post-IC EBV DNA. RESULTS: Post-IC EBV DNA levels and overall stage were independent predictors for distant metastasis-free survival (DMFS), overall survival (OS), and progression-free survival (PFS). The RPA model base on post-IC EBV DNA and overall stage categorized the patients into three distinct risk groups: RPA I (low-risk: stage II-III and post-IC EBV DNA < 200 copies/mL), RPA II (median-risk: stage II-III and post-IC EBV DNA ≥ 200 copies/mL, or stage IVA and post-IC EBV DNA < 200 copies/mL), and RPA III (high-risk: stage IVA and post-IC EBV DNA ≥ 200 copies/mL), with 3-year PFS of 91.1%, 82.6%, and 60.2%, respectively (p < 0.001). The DMFS and OS rates in different RPA groups were also distinct. The RPA model showed better risk discrimination than either the overall stage or post-RT EBV DNA alone. CONCLUSIONS: Plasma EBV DNA level post-IC was a robust prognostic biomarker for NPC. We developed an RPA model that provides improved risk discrimination over the 8th edition of the TNM staging system by integrating the post-IC EBV DNA level and the overall stage.

Applying multisequence MRI radiomics of the primary tumor and lymph node to predict HPV-related p16 status in patients with oropharyngeal squamous cell carcinoma.

Background: Human papillomavirus (HPV)-related oropharyngeal squamous cell carcinoma (OPSCC) is currently rising in incidence; however, a noninvasive approach to predicting HPV status that strongly correlates with p16 expression is lacking. This study aimed to develop a radiomics model based on multisequence magnetic resonance imaging (MRI) of primary tumor (PT) and lymph node (LN)-fused imaging features for the prediction of p16 status in OPSCC. Methods: In this retrospective study, 141 patients (comprising 116 patients in the training cohort and 25 patients in the testing cohort) with histopathologically confirmed (OPSCC) were enrolled consecutively from Fudan University Shanghai Cancer Center and Shanghai Ninth People's Hospital between January 2011 and December 2020. HPV status was determined by p16 immunohistochemistry analysis. A total of 2092 radiomics features were initially computed and extracted from the 3D-segmented PT and largest LN based on contrast-enhanced T1-weighted imaging (CE-T1WI) and T2-weighted imaging (T2WI). A support vector machine classifier was employed to build the machine learning-based classification models dependent on p16 status. The models were validated in the testing cohort. The area under the receiver operating characteristic curve (AUC) was computed to assess the performance of each model. This diagnostic study was not registered on the clinical trial platform. Results: In the testing cohort, fusion models yielded better performance (AUC) compared with models based on a sole PT/LN [CE-T1WI: 0.80 (95% CI: 0.55-0.94) vs. 0.71 (95% CI: 0.47-0.88)/0.73 (95% CI: 0.48-0.90); T2WI: 0.74 (95% CI: 0.51-0.95) vs. 0.64 (95% CI: 0.38-0.85)/0.71 (95% CI: 0.48-0.88)]. Models based on multisequence imaging outperformed single CE-T1WI/T2WI models [PT: 0.74 (95% CI: 0.46-0.91) vs. 0.71 (95% CI: 0.47-0.88)/0.64 (95% CI: 0.38-0.85); LN: 0.78 (95% CI: 0.55-0.75) vs. 0.73 (95% CI: 0.48-0.90)/0.71 (95% CI: 0.48-0.88)]. Finally, the PT-LN fusion model based on multisequencing yielded the best classification performance with the highest AUC value of 0.91 (95% CI: 0.72-0.98) for the prediction of p16 expression. The differences between the performance of the final model and the other 8 models were significant (all P values <0.05). Conclusions: The results demonstrated that (I) the PT-LN fusion radiomics models improved the classification performance of the sole use of PT or LN for the prediction of p16 status, (II) the radiomics models based on multisequences outperformed the single-sequence models in the prediction of p16 status, and (III) the PT-LN fusion model based on multisequence MRI radiomics features could serve as a noninvasive method for acquiring the molecular information of patients with OPSCC, potentially assisting oncologists with their clinical decision-making.

CD161 Characterizes an Inflamed Subset of Cytotoxic T Lymphocytes Associated with Prolonged Survival in Human Papillomavirus-Driven Oropharyngeal Cancer.

Human papillomavirus (HPV)-driven oropharyngeal carcinoma (OPSCC) is distinct from tobacco- or alcohol-associated OPSCC and has a unique immune landscape. Studies have supported the heterogeneity of T cells, accompanied by a broad repertoire of T-cell responses, within tumors driven by HPV infection. However, the phenotype and function of these HPV-related T cells remain unclear. Using a combination of single-cell RNA sequencing, flow cytometry, pharmacologic inhibition, and immunofluorescence staining, we explored the prognostic implication of HPV-related T cells and further validated our findings in two independent cohorts. Cytotoxic T lymphocytes (CTL) within OPSCC displayed a spectrum of transcriptional signatures. Among which, we identified CD161 receptor, encoded by KLRB1, as a potential marker to distinguish the CTL subsets in HPV-positive OPSCC with a divergent evolutionary trajectory. In-depth analysis revealed that CD161+ CTLs exhibited a more robust immune response over the CD161- counterparts and a T cell-inflamed phenotype that could be further reinvigorated by immune-checkpoint blockade. Despite the high expression of exhaustion markers, reinforcement of CD161+ CTL reactivity was expected to boost immune responses, considering their functional reversibility. We further confirmed that the high level of intratumoral CD161+ CTLs associated with a favorable treatment response and prolonged overall survival. Therefore, our research not only provides an insight into the immune landscape of HPV-driven OPSCC but also sheds light on a special subset of CTLs with prognostic and therapeutic significance.

Response to induction chemotherapy predicts survival outcomes in oropharyngeal cancer.

BACKGROUND: The role of induction chemotherapy (IC) in oropharyngeal squamous cell carcinoma (OPSCC) remains controversial. Its interpretation can be confounded by heterogeneity in chemosensitivity and human papillomavirus (HPV) status. This study aimed to investigate the prognostic impact of IC response in HPV-positive and -negative OPSCC. METHODS: Patients with OPSCC who underwent IC and concurrent chemoradiotherapy (CCRT) were retrospectively analyzed. Radiologic response to IC by ≥50% was defined as IC-sensitive (IC-s), while lesser response was deemed as IC-resistant (IC-r). Progression-free survival (PFS) and overall survival (OS) were compared between subgroups. RESULTS: A total of 51 HPV-positive and 57 HPV-negative patients were included. IC-s patients accounted for 55.6%, 62.7%, and 49.1% in the entire cohort, HPV-positive, and HPV-negative subgroup, respectively. Compared with IC-r subgroup, IC-s was associated with better clinical outcomes either in the entire cohort (3y-PFS 91.7%vs.43.7%, P < 0.001; 3y-OS 98.3% vs. 67.4%, P = 0.002), the HPV-positive subgroup (3-year PFS 94.7% vs. 47.9%, P < 0.001; 3-year OS 100% vs. 73.5%, P = 0.055) or the HPV-negative subgroup (3-year PFS 88.2% vs. 40.9%, P = 0.001; 3-year OS 96.4% vs. 63.1%, P = 0.026). Multivariate analysis demonstrated that response to IC represents an independent prognosticator for 3-year PFS (HR, 0.088; 95% CI, 0.027-0.289; P < 0.001) and 3-year OS (HR, 0.100; 95% CI, 0.021-0.477; P = 0.004). CONCLUSIONS: Response to IC exerts a critical predictive effect on prognosis of both HPV-positive and -negative OPSCC. Personalized treatment strategy based on IC response is worthy of further exploration in the future.

A nomogram involving immune-inflammation index for predicting distant metastasis-free survival of major salivary gland carcinoma following postoperative radiotherapy.

BACKGROUND: Postoperative radiotherapy (PORT) is beneficial in the improvement of local-regional control and overall survival (OS) for major salivary gland carcinomas (SGCs), and distant metastasis remained the main failure pattern. This study was designed to develop a nomogram model involving immune-inflammation index to predict distant metastasis-free survival (DMFS) of major SGCs. PATIENTS AND METHODS: A total of 418 patients with major SGCs following PORT were randomly divided into a training (n = 334) and validation set (n = 84). The pre-radiotherapy neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR) were calculated and transformed as continuous variables for every patient. Associations between DMFS and variables were performed by univariate and multivariable analysis using Log-rank and Cox regression methods. A nomogram was constructed based on the prognostic factors identified by the Cox hazards model. The decision curve analysis (DCA) was conducted with the training and validation set. RESULTS: The estimated 3-, 5-, and 10-year DMFS were 79.4%, 71.8%, and 59.1%, respectively. The multivariate analysis revealed that age (p = 0.033), advanced T stage (p = 0.003), positive N stage (p < 0.001), high-risk pathology (p = 0.011), and high PLR (p = 0.001) were significantly associated with worse DMFS. The nomogram showed good calibration and discrimination in the training (AUC = 80.9) and validation set (AUC = 87.9). Furthermore, the DCA demonstrated favorable applicability, and a significant difference (p < 0.001) was observed for the DMFS between the subgroups based on the nomogram points. CONCLUSION: The nomogram incorporating clinicopathological features and PLR presented accurate individual prediction for DMFS of the patients with major SGCs following PORT. Further external validation of the model is warranted for clinical utility.

Postoperative Chemoradiotherapy versus Radiotherapy Alone in Major Salivary Gland Cancers: A Stratified Study Based on the External Validation of the Distant Metastasis Risk Score Model.

BACKGROUND: The role of additional chemoradiotherapy (CRT) for distant metastasis (DM) on the resected malignancy of the major salivary gland (SGM) remained unknown. We conducted this study to externally validate a recently reported DM risk score model and compare the survival outcome between adjuvant CRT and RT alone. MATERIALS: We retrospectively reviewed the patients with SGM following postoperative radiotherapy (PORT). The cumulative incidence of DM was assessed using a competing risk method. Multivariate analysis was performed with Cox proportional-hazards regression to identify significant predictors for DM. Patients were classified as high- and low-risk subgroups with the cutoff value of the DM risk score model. The inverse probability of treatment weighting (IPTW) was conducted to minimize the bias of the groups. RESULTS: A total of 586 eligible patients were analyzed and 67 cases underwent adjuvant CRT. The 5-year incidence of DM was 19.5% (95% CI 16.0-23.0%). The model reasonably discriminated the DM risk between the high- and low-risk subgroup in our cohort, and the c-index was 0.75. No survival benefit was observed for the CRT group compared with RT alone in the entire cohort after IPTW (p = 0.095). After subgroup analysis, increased mortality was identified with the administration of CRT in the low-risk subset (p = 0.002) while no significant difference in OS was illustrated in the high-risk subgroup (p = 0.98). CONCLUSIONS: This external validation provides further exploration of the DM risk score model in major SGM. Our results demonstrated no support for the utility of additional chemotherapy to PORT in the major SGM, especially in the low-risk subgroup of patients with DM.

Failure pattern and suggestions for target volume delineation of carcinoma showing thymus-like differentiation treated with intensity-modulated radiotherapy.

BACKGROUND: To review our long-term clinical experience, analyze the failure patterns, and give suggestions for target volume delineation of carcinoma showing thymus-like differentiation (CASTLE) treated with intensity-modulated radiotherapy (IMRT). METHODS: From April 2008 to May 2019, 30 patients with CASTLE treated by postoperative or radical IMRT in our center were retrospectively reviewed. A total dose of 56-60 Gy in 28-30 fractions was prescribed to patients without residual disease and 66 Gy in 33 fractions for patients with residual or unresectable disease. Survival rates were calculated using the Kaplan-Meier method. Treatment-related toxicities were graded by National Cancer Institute Common Toxicity Criteria (NCI-CTC) version 4.0. RESULTS: Among the 30 patients, 12 (40%) received partial resection or biopsy. Lateral lymph node metastasis was observed in 7 (23.3%) patients. During follow-up, regional lymph node recurrence occurred in 2 patients and distant metastasis in 5 patients. With a median follow-up time of 63.5 months, the 5-year local recurrence-free survival (LRFS), regional recurrence-free survival (RRFS), distant metastasis-free survival (DMFS), overall survival (OS) and progression-free survival (PFS) rates were 100, 88.9, 78.9, 93.1 and 78.9%, respectively. For patients with no lateral neck node metastasis, prophylactic radiotherapy for lateral neck nodal regions failed to improve RRFS (p = 0.381) and OS (p = 0.153). CONCLUSION: Distant metastasis was the major failure pattern for CASTLE after surgery and IMRT. For patients with no lateral neck node metastasis, the omission of irradiation for lateral neck nodal regions seems to be safe and feasible.

Elevated preradiotherapy serum lactate dehydrogenase predicts distant metastasis for lymphoepithelial carcinoma of major salivary gland following postoperative radiotherapy.

BACKGROUND: To evaluate the predicting factors associated with distant metastasis (DM) for lymphoepithelial carcinoma of salivary gland (LECSG) following postoperative radiotherapy (PORT). METHODS: We retrospectively collected 160 eligible patients from two cancer institutions. The DM rate was evaluated using competing risk method. RESULTS: The median follow-up time was 65.6 months. Elevated preradiotherapy serum LDH (ratio >0.5) (p = 0.006) and N classification (N2-3) (p = 0.001) were independently associated with DM for the LECSG. After the risk stratification, the high-risk subgroup was defined as the patients presented higher risk score (score >0), whereas 5-year cumulative incidence of DM in the high- and low-risk group was 30.9% and 6.0%, respectively (p < 0.001). Moreover, a significantly worse overall survival (OS) was observed in the high-risk patients compared with the low-risk subgroup (5-year OS: 83.9% vs. 97.8%, p = 0.006). CONCLUSION: Elevated preradiotherapy serum LDH could serve as a predictive factor for DM in the LECSG following PORT.

Dissecting the heterogeneity of the microenvironment in primary and recurrent nasopharyngeal carcinomas using single-cell RNA sequencing.

Nasopharyngeal carcinoma (NPC) has a 10-15% recurrence rate, while no long term or durable treatment options are currently available. Single-cell profiling in recurrent NPC (rNPC) may aid in designing effective anticancer therapies, including immunotherapies. For the first time, we profiled the transcriptomes of ∼60,000 cells from four primary NPC and two rNPC cases to provide deeper insights into the dynamic changes in rNPC within radiation fields. Heterogeneity of both immune cells (T, natural killer, B, and myeloid cells) and tumor cells was characterized. Recurrent samples showed increased infiltration of regulatory T cells in a highly immunosuppressive state and CD8+ T cells in a highly cytotoxic and dysfunctional state. Enrichment of M2-polarized macrophages and LAMP3+ dendritic cells conferred enhanced immune suppression to rNPC. Furthermore, malignant cells showed enhanced immune-related features, such as antigen presentation. Elevated regulatory T cell levels were associated with a worse prognosis, with certain receptor-ligand communication pairs identified in rNPC. Even with relatively limited samples, our study provides important clues to complement the exploitation of rNPC immune environment and will help advance targeted immunotherapy of rNPC.

Survival impact of increasing time to IMRT initiation following induction chemotherapy in nasopharyngeal carcinoma: A propensity score-matched analysis.

PURPOSE: To explore the prognostic impact of waiting time for radiotherapy (RT) after induction chemotherapy (IC) for nasopharyngeal carcinoma (NPC). METHODS AND MATERIALS: A total of 648 NPC patients receiving IC between 2009 and 2011 were included. Propensity score matching (PSM) was performed to balance the variables. Survival outcomes were compared in subgroups based on time to RT (TTR) after IC. RESULTS: The optimal cutoff point for TTR was 28 days. A total of 330 patients were selected by 1:2 PSM. Stratified and dichotomized TTRs were both strongly correlated with prognosis. Patients with TTR > 28 days had significantly worse 5-year LRFS, DMFS, DFS and OS than those with TTR ≤ 28 days (P < 0.05). In multivariate analysis, TTR > 28 days was an independent predictor of worse LRFS [HR=2.08; 95% CI, 1.18-3.66; P = 0.011), DMFS (HR=1.65; 95% CI, 1.04-2.62; P = 0.033), DFS (HR=1.86; 95% CI, 1.35-2.62; P < 0.001) and OS (HR=1.90; 95% CI, 1.26-2.85; P < 0.001). High-risk patients with T4 or N2-3 disease were highly susceptible to RT delay with impaired DFS and OS. In high-risk patients with TTR > 28 days, concurrent chemotherapy yielded better DMFS (70.9% vs. 52.0%, P  =  0.041), DFS (52.5% vs. 34.3%, P = 0.039) and OS (70.3% vs. 53.2%, P = 0.048). CONCLUSIONS: Prolonged waiting is detrimental to survival in NPC, and it is strongly recommended to start RT within 28 days after IC. T4/N2-3 NPC has a higher risk of treatment failure with delayed RT. With potential protection against RT delay, concurrent chemotherapy should be performed in high-risk patients as salvage therapy.

The impact of body composition parameters on severe toxicities in patients with locoregionally advanced nasopharyngeal carcinoma undergoing neoadjuvant chemotherapy.

BACKGROUND: Neoadjuvant chemotherapy (NACT) treatment in locoregionally advanced nasopharyngeal carcinoma (LA-NPC) can lead to considerable toxicity. Loss of skeletal muscle mass showed relevance with increased chemotherapy-related toxicity and poor survival in various cancer types, but its significance in NPC remains unclear. This study aimed to investigate the relationship between body composition parameters and the incidence of NACT toxicity in LA-NPC patients. METHODS: Ninety-six LA-NPC patients were retrospectively enrolled. All patients had pre-treatment abdominal computed tomography (CT) images to exclude distant metastasis. Lean body mass (LBM, kg) was estimated based on cross-sectional muscle area at the third lumbar vertebra (L3) level on CT, and skeletal muscle index (SMI, cm2/m2) was calculated. Doses of chemotherapeutics were normalized as dose/LBM (mg/kg). Grade 3-4 toxicity was defined as severe. The associations between body composition parameters and severe toxicities were assessed using univariate and multivariate logistic regression analyses. Optimal cutoff points were obtained with a receiver operating characteristic (ROC) curve. RESULTS: Of the 96 patients, 81.2% received the docetaxel + cisplatin (TP) regimen, and the rest received the gemcitabine + cisplatin (GP) regimen. Males had more LBM and a higher SMI at baseline, and females received a markedly higher dose of docetaxel and gemcitabine per kg LBM (P<0.001). With a cutoff value of 52.7 cm2/m2, patients with higher SMI showed lower risk of severe toxicity. For TP regimen group, those presented with grade 3-4 neutropenia had a higher dose per kg LBM. Univariate and multivariate analyses showed that the LBM-adjusted dose was significantly associated with severe neutropenia in the TP regimen group (P<0.001). The LBM-normalized docetaxel cutoff value of 2.64 mg/kg was a prominent predictor of ≥ grade 3 neutropenia (P=0.003), but a higher dose of docetaxel per kg LBM did not provide a better objective response rate. CONCLUSIONS: LA-NPC patients with lower SMI and higher dose of docetaxel per kg LBM are more likely to suffer from severe treatment-related toxicity. Higher docetaxel dose per kg LBM is a prominent predictor for severe neutropenia, but not for NACT response. LBM showed good potential in toxicity risk prediction and dose determination.

Progression-Free Survival Prediction in Patients with Nasopharyngeal Carcinoma after Intensity-Modulated Radiotherapy: Machine Learning vs. Traditional Statistics.

BACKGROUND: The Cox proportional hazards (CPH) model is the most commonly used statistical method for nasopharyngeal carcinoma (NPC) prognostication. Recently, machine learning (ML) models are increasingly adopted for this purpose. However, only a few studies have compared the performances between CPH and ML models. This study aimed at comparing CPH with two state-of-the-art ML algorithms, namely, conditional survival forest (CSF) and DeepSurv for disease progression prediction in NPC. METHODS: From January 2010 to March 2013, 412 eligible NPC patients were reviewed. The entire dataset was split into training cohort and testing cohort in a ratio of 90%:10%. Ten features from patient-related, disease-related, and treatment-related data were used to train the models for progression-free survival (PFS) prediction. The model performance was compared using the concordance index (c-index), Brier score, and log-rank test based on the risk stratification results. RESULTS: DeepSurv (c-index = 0.68, Brier score = 0.13, log-rank test p = 0.02) achieved the best performance compared to CSF (c-index = 0.63, Brier score = 0.14, log-rank test p = 0.38) and CPH (c-index = 0.57, Brier score = 0.15, log-rank test p = 0.81). CONCLUSIONS: Both CSF and DeepSurv outperformed CPH in our relatively small dataset. ML-based survival prediction may guide physicians in choosing the most suitable treatment strategy for NPC patients.

Human papillomavirus (HPV) in Chinese oropharyngeal squamous cell carcinoma (OPSCC): A strong predilection for the tonsil.

OBJECTIVE: Compared with Occident's data, the incidence of Human papillomavirus (HPV)-driven oropharyngeal squamous cell carcinomas (OPSCCs) had been reported as relatively low in Mainland China. The objective of this study was to report the integrated prevalence of HPV and Epstein-Barr virus (EBV), and further evaluate the different behaviors of HPV-positive and -negative OPSCCs in eastern China. METHODS: In a cohort of 170 nonmetastatic OPSCCs treated from January 2007 to July 2019, p16 protein expression, HPV genotypes, and Epstein-Barr virus-encoded RNA (EBER) were determined by immunohistochemistry (IHC) and in situ hybridization (ISH). The clinical and pathologic findings were further collected and analyzed to comprehensively reveal the behaviors of Chinese OPSCCs. RESULTS: Out of the 170 tumor tissues evaluated, 57.6% (98) samples had positive p16 expressions. A total of 65.1% (99/152) samples had positive HPV genotypes, besides HPV16 (92/152), HPV11, 18, 33, 53, and 58 were also detected. The positive rate of EBER was 7.2% (9/124), and the co-infection rate of EBV/HPV was 4.0%. Related to the unequal distributions of p16 expression, HPV-related tumors arisen from tonsillar and non-tonsillar accounted for 68.8% (75/109) and 37.7% (23/61) of their cases, respectively (P < .001). With a median follow-up time of 13.1 months, significant survival advantages of HPV-related OSPCC were observed; 1-year OS, PFS, RFS, and MFS were 83.2% vs 96.7% (P < .001), 71.6% vs 96.2% (P < .001), 77.7% vs 96.2% (P = .002), and 90.4% vs 100.0% (P = .024) in p16-negative and -positive cases, respectively. CONCLUSIONS: The relative percent of HPV-positive OPSCCs in this study is close to the positive rate in many Western countries and a strong predilection was discovered for the tonsillar. The EBV infection and co-infection of HPV/EBV were largely low. The prognosis of HPV-positive OSPCCs was more favorable than its negative counterpart.

Effect of COVID-19 Epidemic on Delay of Diagnosis and Treatment Path for Patients with Nasopharyngeal Carcinoma.

INTRODUCTION: 2019 novel coronavirus disease (COVID-19) outbreaks have been occurring in China and other countries in the world. To prevent further spread of the disease, restrictions of population flow from the government and measures to reduce virus transmission from hospitals may lead to the delay of diagnosis and treatment in patients with nasopharyngeal carcinoma (NPC). METHODS: All NPC patients with radiotherapy indications were included from 20 weekdays before (group A) and after (group B) January 31, 2020, when the institute began to take measures against COVID-19. The waiting intervals of each step and variation from the diagnosis and treatment path of NPC between two groups were compared. RESULTS: Significant differences were found between the group A and group B in the median waiting days for pathological biopsy (5 vs 15, P=0.012), radiotherapy immobilization and simulation (3.5 vs 16.5, P<0.001), validation of position and plan (20 vs 61, P<0.001) and initiation of radiotherapy (28 vs 36, P=0.005). During the waiting period of radiotherapy, 32.4% of the NPC patients received an additional one cycle of chemotherapy to the original treatment strategy. CONCLUSION: The prevalence of COVID-19 caused delay in the diagnosis and treatment of NPC patients to a certain extent. Additional chemotherapy could be considered to counteract the effect of treatment delay. More specific measures should be taken to balance the risk of delayed diagnosis and treatment of NPC and infection of COVID-19.

Long-term treatment results and prognostic factors of synchronous and metachronous squamous cell carcinoma of head and neck and esophagus.

BACKGROUND: The purpose was to investigate the prognosis of patients with synchronous and metachronous squamous cell carcinoma of head and neck (HNSCC) and esophagus (ESCC), and to evaluate the prognostic factors of these patients. METHODS: A retrospective review was performed on 70 patients with synchronous and metachronous HNSCC and ESCC treated in our institution from January 2005 to December 2016. Kaplan-Meier method and Cox proportional hazard model were used to evaluate overall survival (OS) and determine the prognostic factors associated with survival outcomes. RESULTS: The 1-, 2- and 3-year OS rates were 77.1%, 57.1% and 37.1% with the median survival time for 33.5 months. The univariate analysis results revealed that the patients with early-stage of ESCC, metachronous cancer, and receiving surgery for both cancer had better OS (P=0.003; P=0.035; P=0.002). The multivariate analysis showed that the clinical stage of ESCC and receiving surgery for both cancer or not were the independent prognostic factors for OS. CONCLUSIONS: The multidisciplinary treatment outcome is acceptable, especially for patients with early clinical stage ESCC and with chance to receiving surgery for both cancer.

Radiation therapy-induced reactive oxygen species specifically eliminates CD19+IgA+ B cells in nasopharyngeal carcinoma.

PURPOSE: Nasopharyngeal carcinoma (NPC) is one of the most common head and neck cancers and is thought to be related to the mucosal immune system. Radiation therapy (RT) is the primary treatment for NPC due to the high radiosensitivity of cancer cells. However, little is known about the impact of RT on the mucosal immune system. PATIENTS AND METHODS: In this study, the expression of immune markers CD19, CD24, CD27, CD8, and IgA before and after RT, were analyzed using flow cytometry. Cytokines were assessed using the enzyme-linked immunosorbent assay. Reactive oxygen species (ROS) was assayed by flow cytometry and fluorescence staining using 2',7' -dichlorofluorescein diacetate. RESULTS: We found that primary NPC patients had a significant increase in CD19+CD138-IgA+ B cells, which was then decreased after RT. Interestingly, the changes in CD19+CD138-IgA+ B cell frequency was accompanied by corresponding frequency changes in cytotoxic T cells (CTL), which are powerful anti-tumor lymphocytes. Mechanistically, we found that ROS release during RT specifically eliminated CD19+CD138-IgA+ B cells. CONCLUSION: These findings suggest that RT may regulate the immune system and opens up new avenues for the utilization of immune-radiotherapy in NPC.

Serial Monitoring of Circulating Tumor DNA in Patients With Metastatic Colorectal Cancer to Predict the Therapeutic Response.

Early biomarkers of therapeutic responses can help optimize the treatment of metastatic colorectal cancers (mCRC). In this prospective exploratory study, we examined serial changes of plasma-circulating tumor DNA (ctDNA) in 41 mCRC patients receiving first-line chemotherapies and tested its association with treatment outcomes according to radiological assessments. Using next-generation sequencing technologies, we profiled somatic mutations in 50 cancer-related genes in ctDNA before each of the first four treatment cycles. We observed mutations in 95.7% of pre-treatment ctDNA samples. Using mutations of the maximal frequency in each pre-treatment plasma ctDNA sample as the candidate targets, we computed log2 fold changes of ctDNA levels between adjacent treatment cycles. We found that ctDNA reductions as early as prior to cycle 2 predicted responses after cycle 4. Log2 fold changes of ctDNA after cycle 1 (ctDNA log2 (C1/C0)) > -0.126 predicted progressive disease, with an accuracy of 94.6%. These patients also showed significantly worse progression-free survival than those with ctDNA log2 (C1/C0) ≤ -0.126 (median 2.0 vs. 9.0 months; P = 0.007). Together, the present exploratory study suggests that early changes in ctDNA levels detected via targeted sequencing are potential biomarkers of future treatment responses in mCRCs.

Dosimetry of the brain and hypothalamus predicting acute lymphopenia and the survival of glioma patients with postoperative radiotherapy.

BACKGROUND: The aim of this study was to investigate dosimetric factors for predicting acute lymphopenia and the survival of glioma patients with postoperative intensity-modulated radiotherapy (IMRT). METHODS: A total of 148 glioma patients were reviewed. Acute lymphopenia was defined as a peripheral lymphocyte count (PLC) lower than 1.0 × 109 /L during radiotherapy with a normal level at pretreatment. PLCs with the corresponding dates and dose volume histogram parameters were collected. Univariate and multivariate Cox regression analyses were constructed to assess the significance of risk factors associated with lymphopenia and overall survival (OS). RESULTS: Sixty-nine (46.6%) patients developed lymphopenia during radiotherapy. Multivariate analyses revealed that the risk increased with the maximal dose of the hypothalamus (HT Dmax) ≥56 Gy (58.9% vs 28.5%, P = 0.002), minimal dose of the whole brain (WB Dmin) ≥2 Gy (54.3% vs 33.9%, P = 0.006), or mean dose of the WB (WB Dmean) ≥34 Gy (56.0% vs 37.0%, P = 0.022). Patients with older age, high-grade glioma, development of lymphopenia, high HT Dmax, WB Dmin, and WB Dmean had significantly inferior OS in the multivariate analyses. CONCLUSIONS: HT Dmax, WB Dmin, and WB Dmean are promising indicators of lymphopenia and the survival of glioma patients undergoing postoperative IMRT. The necessity and feasibility of dosimetric constraints for HT and WB is warranted with further investigation.