Development and clinical validation of a microfluidic-based platform for CTC enrichment and downstream molecular analysis.

Background: Although many CTC isolation and detection methods can provide information on cancer cell counts, downstream gene and protein analysis remain incomplete. Therefore, it is crucial to develop a technology that can provide comprehensive information on both the number and profile of CTC. Methods: In this study, we developed a novel microfluidics-based CTC separation and enrichment platform that provided detailed information about CTC. Results: This platform exhibits exceptional functionality, achieving high rates of CTC recovery (87.1%) and purification (∼4 log depletion of WBCs), as well as accurate detection (95.10%), providing intact and viable CTCs for downstream analysis. This platform enables successful separation and enrichment of CTCs from a 4 mL whole-blood sample within 15 minutes. Additionally, CTC subtypes, selected protein expression levels on the CTC surface, and target mutations in selected genes can be directly analyzed for clinical utility using immunofluorescence and real-time polymerase chain reaction, and the detected PD-L1 expression in CTCs is consistent with immunohistochemical assay results. Conclusion: The microfluidic-based CTC enrichment platform and downstream molecular analysis together provide a possible alternative to tissue biopsy for precision cancer management, especially for patients whose tissue biopsies are unavailable.

Development and validation of web-based dynamic nomograms predictive of disease-free and overall survival in patients who underwent pneumonectomy for primary lung cancer.

Background: The tumour-node-metastasis (TNM) staging system is insufficient to precisely distinguish the long-term survival of patients who underwent pneumonectomy for primary lung cancer. Therefore, this study sought to identify determinants of disease-free (DFS) and overall survival (OS) for incorporation into web-based dynamic nomograms. Methods: The clinicopathological variables, surgical methods and follow-up information of 1,261 consecutive patients who underwent pneumonectomy for primary lung cancer between January 2008 and December 2018 at Sun Yat-sen University Cancer Center were collected. Nomograms for predicting DFS and OS were built based on the significantly independent predictors identified in the training cohort (n = 1,009) and then were tested on the validation cohort (n = 252). The concordance index (C-index) and time-independent area under the receiver-operator characteristic curve (AUC) assessed the nomogram's discrimination accuracy. Decision curve analysis (DCA) was applied to evaluate the clinical utility. Results: During a median follow-up time of 40.5 months, disease recurrence and death were observed in 446 (35.4%) and 665 (52.7%) patients in the whole cohort, respectively. In the training cohort, a higher C-reactive protein to albumin ratio, intrapericardial pulmonary artery ligation, lymph node metastasis, and adjuvant therapy were significantly correlated with a higher risk for disease recurrence; similarly, the independent predictors for worse OS were intrapericardial pulmonary artery and vein ligation, higher T stage, lymph node metastasis, and no adjuvant therapy. In the validation cohort, the integrated DFS and OS nomograms showed well-fitted calibration curves and yielded good discrimination powers with C-index of 0.667 (95% confidence intervals CIs [0.610-0.724]) and 0.697 (95% CIs [0.649-0.745]), respectively. Moreover, the AUCs for 1-year, 3-year, and 5-year DFS were 0.655, 0.726, and 0.735, respectively, and those for 3-year, 5-year, and 10-year OS were 0.741, 0.765, and 0.709, respectively. DCA demonstrated that our nomograms could bring more net benefit than the TNM staging system. Conclusions: Although pneumonectomy for primary lung cancer has brought encouraging long-term outcomes, the constructed prediction models could assist in precisely identifying patients at high risk and developing personalized treatment strategies to further improve survival.

Anatomical variation analysis of left upper pulmonary blood vessels and bronchi based on three-dimensional reconstruction of chest CT.

Background: With its growing popularity and potential outcome, preoperative three-dimensional reconstruction of chest computed tomography (CT) has been widely used in video-assisted thoracic surgery (VATS) segmentectomy for treating non-small cell lung cancer (NSCLC). This study aimed to summarize the experience of anatomical variation analysis of left upper pulmonary blood vessels and bronchi based on the three-dimensional reconstruction of chest CT. Materials and methods: A total of 103 patients with early-stage NSCLC were chosen to undergo VATS segmentectomy based on preoperative three-dimensional reconstruction of chest CT in our institute from September 2019 to July 2022. Data such as clinical characteristics and variations in blood vessels and bronchi were reviewed in this study. Results: The branches of the left lingular pulmonary artery may mutate into the LS1 + 2 + 3. A1 + 2 has four subtypes. The distribution of variation is relatively balanced, and the most common variation is type I (35/103, 33.9%). Most lingular arteries originate from the oblique cleft side of the lingular bronchus (79/103,76.7%). Most V(1 + 2)c* are small developments (70/103, 68.0%). The venous return of the proper segment mainly depends on V(1 + 2)b + c. The variation in the left upper lobe bronchus is complex. The most common variant is the bifurcation type (type A to G, 92/103, 89.3%) and bifurcation type A (62/103, 60.2%). The posterior apical segment artery of the left upper lobe is not accompanied by its bronchus. Conclusions: The variation types of blood vessels and bronchus in the upper lobe of the left lung are complex. Preoperative CT-based three-dimensional reconstruction of pulmonary arteries, veins, and bronchi is of great significance. It can help understand the variations, accurately locate lesions before the surgery, and effectively plan surgeries.

Characterization of the Immune Cell Infiltration Landscape in Esophageal Squamous Cell Carcinoma.

Background: Immunotherapy has achieved remarkable efficacy in treating oesophageal squamous cell carcinoma (ESCC). However, this treatment has limited efficacy in some patients. An increasing number of evidence suggested that immune cells within the tumour microenvironment (TME) are strongly related to immunotherapy response and patient prognosis. Thus, the landscape of immune cell infiltration (ICI) in ESCC needs to be mapped. Methods: In the study, the ICI pattern in 206 cases of ESCC was characterised by two algorithms, namely, CIBERSORT and single-sample gene set enrichment analysis (ssGSEA). The ICI score of each specimen was calculated by principal component analysis (PCA) according to ICI signature genes A (ICISGA) and B (ICISGB). The prognostic difference was evaluated by using the Kaplan-Meier method. The related pathways of ICI score were investigated by applying gene set enrichment analysis (GSEA). The R packages of 'regplot', 'timeROC' and 'rms' were applied for the construction of nomogram model. Result: Three TME subtypes were identified with no prognostic implication. A total of 333 differentially expressed genes (DEGs) among immune subtypes were determined, among which ICISGA and ICISGB were identified. Finally, ICI scores were constructed, and the patients were grouped into high or low ICI score group. Compared with the low ICI score group, the high ICI score group had better prognosis. GSEA revealed that the high ICI score group referred to multiple signalling pathways, including B cell receptor, Fc gamma R-mediated phagocytosis, NOD-like receptor and TGF-ß signalling pathways. In addition, the nomogram model was constructed to evaluate 1-, 3- and 5-year probability of death in an ESCC patient. The ROC and calibration curves indicated that the model has a good discrimination ability. Conclusion: We depicted a comprehensive ICI landscape in ESCC. ICI score may be used as a predictor of survival rate, which may be helpful for guiding immunotherapy in the future.

Survival After Lobectomy vs. Sublobar Resection for Stage IA Large-Cell Neuroendocrine Carcinoma of the Lung: A Population-Based Study.

Objective: Due to the low incidence of pulmonary large cell neuroendocrine carcinoma (LCNEC), the survival analysis for comparing lobectomy and sublobar resection (SLR) for stage IA LCNEC remains scarce. Methods: Patients diagnosed with pathological stage IA LCNEC between 1998 and 2016 were extracted from the Surveillance, Epidemiology, and End Results (SEER) database. The oncological outcomes were cancer-specific survival (CSS) and overall survival (OS). Kaplan-Meier analysis and Cox multivariate analysis were used to identify the independent prognostic factors for OS and CSS. Furthermore, propensity score matching (PSM) was performed between SLR and lobectomy to adjust the confounding factors. Results: A total of 308 patients with stage IA LCNEC met the inclusion criteria: 229 patients (74.4%) received lobectomy and 79 patients (25.6%) received SLR. Patients who underwent SLR were older (P < 0.001), had smaller tumor size (P = 0.010), and less lymph nodes dissection (P < 0.001). The 5-year CSS and OS rates were 56.5 and 42.9% for SLR, and 67.8 and 55.7% for lobectomy, respectively (P = 0.037 and 0.019, respectively). However, multivariate analysis did not identify any differences between the SLR group and lobectomy group in CSS (P = 0.135) and OS (P = 0.285); and the PSM also supported these results. In addition, the age at diagnosis and laterality of tumor were identified as significant predictors for CSS and OS, whereas the number of lymph nodes dissection was a significant predictor for CSS. Conclusions: Although SLR is not inferior to lobectomy in terms of oncological outcomes for patients with stage IA LCNEC, more lymph nodes can be dissected or sampled during lobectomy. Lobectomy should still be considered as a standard procedure for patients with early-stage LCNEC who are able to withstand lobectomy.

Integrative analysis and experiments to explore angiogenesis regulators correlated with poor prognosis, immune infiltration and cancer progression in lung adenocarcinoma.

Angiogenesis is the process of capillary sprouting from pre-existing vessels and it plays a critical role in the carcinogenic process of lung adenocarcinoma (LUAD). However, the association of angiogenesis regulators with the prognosis and progression of LUAD needs to be further elucidated. In this study, we adopted differential expression analysis, Cox proportional hazards (PH) regression analysis and experimental validation to identify angiogenesis regulators correlated with a poor prognosis, immune infiltration and cancer progression in LUAD. These results showed that the diagnostic and prognostic models based on COL5A2 and EPHB2 served as independent biomarkers with superior predictive ability. The patients in the high-risk group exhibited a worse prognosis in the TCGA cohort (P < 0.001, HR = 1.72, 95% CI 1.28-2.30), GSE310210 cohort (P = 0.005, HR = 2.87, 95% CI 1.46-5.61), and GSE31019 cohort (P = 0.01, HR = 2.14, 95% CI 1.19-3.86) than patients in the low-risk group. The high prognostic risk patients had a higher TMB (P < 0.001); higher fractions of M0 macrophages, neutrophils, NK cells resting, and T cells CD4 memory activated (P < 0.05); and higher expression of immune checkpoints PD-1, PDL-1, PDL-2, and B7H3 (P < 0.001). Patients in the high-risk group were more sensitive to chemotherapeutic drugs and molecular targeted drugs such as cisplatin, doxorubicin, gefitinib, and bosutinib (P < 0.0001). In addition, inhibition of COL5A2 and EPHB2 effectively suppressed the proliferation and migration of LUAD cells. The current study identified angiogenesis regulators as potential biomarkers and therapeutic targets for LUAD and may help to further optimize cancer therapy.

Mediastinoscopy-assisted transhiatal esophagectomy versus thoraco-laparoscopic esophagectomy for esophageal cancer: a single-center initial experience.

BACKGROUND: We aimed to compare mediastinoscopy-assisted transhiatal esophagectomy (MATHE) with thoraco-laparoscopic esophagectomy (TLE) for patients with esophageal cancer in terms of the clinical effectiveness and perioperative complications. METHODS: In total, 98 patients who underwent esophagectomy consecutively for esophageal squamous cell carcinoma in our center from Jan. 2018 to Dec. 2019 were included in this study. Thirty patients underwent mediastinoscopy-assisted and laparoscopic transhiatal esophagectomy with cervical anastomosis (the MATHE group). The other sixty-eight patients received TLE (the TLE group). Each patient's general conditions and perioperative complications were recorded. RESULTS: Patients in the MATHE group were observed to have a higher incidence of postoperative hoarseness than those in the TLE group. There were no significant differences between the MATHE group and the TLE group in regards to the operation time, intraoperative blood loss, number of lymph nodes dissected or postoperative hospital stay. Similarly, no statistically significant differences were observed in the incidence of anastomotic fistula, respiratory complications, or chylothorax or in the conversion rate or in-hospital mortality rate between the two groups. CONCLUSIONS: The short-term efficacy in the MATHE group was similar to that in the TLE group, although patients in the MATHE group may have had a higher incidence of postoperative hoarseness. Therefore, MATHE may be a feasible and safe surgical procedure for appropriate patients with esophageal cancer.