BackgroundWith 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 methodsA 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.ResultsThe 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.ConclusionsThe 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.

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.

ObjectiveDue 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.MethodsPatients 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.ResultsA 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 &lt; 0.001), had smaller tumor size (P = 0.010), and less lymph nodes dissection (P &lt; 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.ConclusionsAlthough 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.

e20591 Background: Non–small cell lung cancer (NSCLC) is one of the main leading causes of cancer mortality in the world. NSCLC is incurable in most patients with potentially resectable locally advanced stage IIIB disease. Numerous studies suggest that advantageous chemoimmunotherapy is a promising treatment for resectable NSCLC. The determination of PD-L1, T-cell receptor (TCR) repertoire and tumor mutational burden (TMB) have been reported to predict the response to immunotherapy. However, the value of pathologic response determination in the context of neoadjuvant chemoimmunotherapy in resectable stage IIIB NSCLC is limited. Therefore, it is necessary to identify biomarkers for the prediction of the benefit from neoadjuvant chemoimmunotherapy in patients with potentially resectable stage IIIB NSCLC. Methods: A total of 15 patients with potentially resectable clinical stage IIIB NSCLC were enrolled into the study receiving neoadjuvant sintilimab (200mg IV Q3W) plus chemotherapy therapies for 4-6 weeks followed by surgery. All peripheral blood and tissue samples were collected and detected by using TCR repertoire sequencing before and after chemoimmunotherapy. The patients were divided into MPR group (n=5) and non-MPR group (n=10) according to MPR (<10% viable tumour at resection) as an index. Furthermore, TMB spectra of 1021-gene panels and PD-L1 immunohistochemical staining were performed in paraffin-embedded human NSCLC tissue before neoadjuvant chemoimmunotherapy. The Pearson correlation coefficient method was used to analyze correlation. The ROC analysis were performed using “pROC” R package. Results: In baseline tissue samples, the levels of TMB and NAD were higher in MPR compared with non-MPR groups (P=0.0328, P=0.015, respectively). The negative correlation was existed between pretreatment tissue TCR richness and TMB in non-MPR group (P=0.028, R=-0.66). In MPR group, TCR clones and evenness showed significant difference (P=0.00237, P=0.04658, respectively). In non-MPR group, the TCR repertoire of tissue in posttreatment showed significant difference compared with those of pretreatment, including clones (P=1.04x10-6), shannon’s diversity (P=0.00142), evenness (P=0.00433), richness (P=0.00591). The clinical data showed that MPR patients with DDR mutation had longer DFS after sintilimab plus chemotherapy therapies (18.4±5.41 months, X-squared=9.8, P=0.02034). DDR mutation, TMB and TCR richness revealed better sensitivity and specificity to classify patients achieving MPR after neoadjuvant chemoimmunotherapy (AUC=1.000, AUC=0.796, AUC=0.773, respectively). Conclusions: This study confirmed the superiority of neoadjuvant chemoimmunotherapy in patients with potentially resectable stage IIIB NSCLC in terms of MPR. TCR repertoire, TMB and DDR mutation were associated with pathologic response to neoadjuvant chemoimmunotherapy as biomarkers for effective anti-tumor immunity. Clinical trial information: ChiCTR2000040673 .

Circulating tumor cells (CTCs) are shed from primary or metastatic tumors into the peripheral blood circulation which carry a wealth of information for cancer diagnosis, treatment and prognosis. However, most of current CTCs isolation and detection methods provide only cancer cell counting information which is far from meeting clinical needs. In addition to the numbers of CTCs, the target proteins and gene mutations carried by CTCs can also be used for clinical diagnosis, disease monitoring and therapeutic selection. In this work, we develop a novel microfluidic-based CTCs separation and enrichment platform that enables the extraction of CTCs information, including cell number, epithelial-mesenchymal transition (EMT) subtypes, protein expression levels, and target gene mutations. The platform offers a high CTCs recovery rate (> 85%), high CTCs purification (∼104 enrichment) and intact viable CTCs for downstream analysis. This platform can successfully enrich tumor cells from a 4 mL blood sample within 15 minutes. CTCs were detected in clinical samples from cancer patients with a detection rate of 95.8%. Furthermore, the CTCs subtypes (epithelial, mesenchymal or mix type), the expression levels of selected proteins (PD-L1, HER2, VEGF), and the target mutations in selected genes (EGFR, KRAS, BRAF) could also be directly analyzed by immunofluorescence and digital PCR for clinical utility. PD-L1 expression detected in the CTCs was consistent with the immunohistochemical results. This microfluidic-based CTCs enrichment platform and downstream molecular analysis provide a possible alternative to tissue biopsy for precision cancer management, especially for patients whose tissue biopsies are unavailable.