Culturomics and 16 S rDNA sequencing were applied to identify lung tumor-resident microorganisms. In vitro characterization revealed potential functions of these cancer-associated microorganisms. Eighteen clinical lung cancer (LC) tissues samples were collected. All the samples were cultured by culturomics, and analyzed with 16 S rDNA sequencing. Four isolated isolates belonging to the genus Staphylococcus were detected to find out the possible functions in vitro. A549 cells were infected with supernatant of these bacteria, and cell index which reflected cell proliferation was determined by Real-Time Cell Analysis (RTCA). ELISA was employed to detect levels of proinflammatory cytokines (TNF-α, IL-1β, and IL-6) in THP-1 cells stimulated with bacterial culture supernatants. A total of 12 bacteria were cultured and identified, most of which belonged to the Staphylococcus genus. Bacillus was detected through both methods. In vitro experiment, the cultured strains promoted cellular proliferation, and enhanced the levels of proinflammatory cytokines (TNF-α, IL-1β, and IL-6) as well. Bacteria from LC tissues was isolated and identified. Lung tumor-resident microbiota was described. LC tumor-resident bacteria cause tumor development mediated by enhancing tumor cell proliferation and proinflammatory cytokines releasing by macrophages in the tumor microenvironment.

BACKGROUND:Immune checkpoint inhibitors (ICIs) are known for their durable efficacy and favorable tolerance profiles. However, elderly patients are often underrepresented in clinical trials. The manifestation of immune-related adverse events (irAEs) in elderly patients with different tumor types remains unclear. This study aimed to compare irAEs in elderly patients with gastrointestinal and lung cancers receiving ICIs and to provide real-world safety data. METHODS:This retrospective study included elderly patients (≥70 years) with gastrointestinal (GI) tumors or lung cancer who received ≥2 cycles of ICIs across multiple departments within a single academic medical center between January 2016 and February 2022. A 2:1 propensity score matching was used to balance baseline characteristics between the cohorts. The primary endpoint was the incidence of any-grade irAEs between GI tumors and lung cancer, whereas secondary analyses focused on organ-specific toxicities. Statistical comparisons were performed using the chi-squared test. RESULTS:The overall incidence of any-grade irAEs was higher in elderly patients with lung cancer than in those with GI tumors (61.0% vs . 47.9%; P = 0.013). After matching for baseline characteristics, the lung cancer group still showed a trend toward a higher incidence of irAEs (61.7% vs . 50.8%; P = 0.056). Notably, patients with GI tumors had a higher incidence of skin toxicity (28.7% vs . 15.1%; P = 0.002), whereas patients with lung cancer had a higher incidence of thyroid dysfunction (28.1% vs . 11.9%, P <0.001). CONCLUSIONS:Elderly patients with lung cancer were more likely to experience irAEs during immunotherapy than those with GI tumors. However, dermatological toxicities were more common in elderly patients with GI tumors, whereas thyroid dysfunction was more frequently observed in those with lung cancer.

e20502 Background: The HER2-targeted therapy trastuzumab deruxtecan (T-DXd) was approved for use in China in 2024, providing a novel option for patients with HER2-mutant non-small cell lung cancer (NSCLC). Despite its approval, data on treatment patterns and clinical outcomes in real-world settings remain scarce. This study aimed to analyze real-world treatment patterns and outcomes in HER2-mutant NSCLC, providing a foundation for optimizing HER2-targeted therapies in clinical practice. Methods: This retrospective analysis included clinical data from HER2-mutant NSCLC patients treated across four medical centers (Peking University Cancer Hospital & Institute, Peking University Cancer Hospital (Inner Mongolia Campus)/Affiliated Cancer Hospital of Inner Mongolia Medical University, The second affiliated hospital of Dalian medical university and The Affiliated Cancer Hospital of Zhengzhou University & Henan Cancer Hospital) between December 2015 and April 30, 2025. Statistical analyses were performed using SPSS software and included log-rank tests, Kaplan-Meier survival analyses, univariate and multivariate models, and Clopper-Pearson interval estimation. Results: 404 patients with HER2-altered NSCLC were included, of whom 49.75% (201/404) had HER2 exon 20 insertion mutations. Among patients harboring HER2 mutations (with or without amplification) receiving first-line therapy, trastuzumab deruxtecan (T-DXd) monotherapy achieved a median real-world progression-free survival (rwPFS) of 19.40 months (N = 11), which was significantly longer than chemotherapy (N = 34, 5.90 months), immunotherapy-based regimens (N = 106, 8.20 months), non-T-DXd HER2-targeted therapy (N = 25, 6.07 months), and anti-angiogenic based therapy (N = 44, 8.47 months) (P = 0.04). For second-line therapy, the median rwPFS of T-DXd monotherapy (N = 25, 10.13 months) was numerically longer than chemotherapy (N = 11, 3.90 months) and IO-based therapy (N = 28, 6.03 months) (P = 0.25). Conclusions: This is the largest real-world cohort study of HER2-mutant NSCLC to date. Real-world treatment strategies for HER2-mutant NSCLC patients are highly variable. T-DXd demonstrated the most favorable outcomes in both the first-line and second-line settings, showing superior effectiveness compared to other available treatments. These findings underscore the potential of T-DXd as a key therapeutic option for HER2-mutant NSCLC patients. Future studies are warranted to further validate these real-world outcomes and explore optimal therapeutic strategies.

BACKGROUND:SMARCA4-deficient thoracic tumors (SDTT) represent a newly defined and highly aggressive subtype of lung cancer for which no standard therapy has been established. Although immune checkpoint inhibitors (ICIs) have demonstrated potential clinical benefit, responses in SDTT are limited and heterogeneous, and the underlying immunologic mechanisms remain poorly understood. This study aimed to characterize the clinicopathologic features, survival outcomes, tumor immune microenvironment (TIME), and treatment responses of SDTT, as well as to explore potential therapeutic strategies. METHODS:In this retrospective, two-center study, 121 patients with SDTT and a comparative cohort of 132 patients with non-SDTT were analyzed. Clinicopathologic variables, treatment patterns, and survival outcomes were compared between groups. Multiplex immunohistochemistry (mIHC) was used to evaluate the immune contexture of SDTT. The efficacy of ICIs and anti-angiogenic therapy, administered as monotherapy or in combination, was also assessed. RESULTS:SDTT patients had significantly inferior survival compared with non-SDTT. Although ICIs conferred clinical benefit in SDTT patients, this benefit was attenuated compared with non-SDTT (median progression-free survival [PFS] 6.0 vs. 13.5 months; p = 0.002; median overall survival [OS], 21.1 months vs. not reached; p = 0.030). mIHC analysis showed a TIME characterized by stromal exclusion of CD8 + T cells and enrichment of TIM3 + exhausted T-cell subsets. A high density of CD8 + TIM3 + T cells was associated with a trend toward worse overall survival in the SDTT cohort. In exploratory analyses, combination therapy with ICIs and antiangiogenic agents was associated with longer progression-free survival than ICI monotherapy, along with a trend toward improved overall survival. CONCLUSION:Although ICIs provide clinical benefit in SDTT patients, their efficacy is attenuated compared with non-SDTT. Spatial mIHC analysis showed a CD8 + T cell-excluded, exhaustion-dominant immunosuppressive microenvironment. Exploratory findings suggest that combining antiangiogenic therapy with ICIs is associated with improved PFS; however, prospective validation is warranted.

Background:Micropapillary (MP) pattern has been identified as a negative prognostic factor in patients with lung adenocarcinoma, but it has not been recognized as a high-risk factor for patients with stage IB lung adenocarcinoma treated with adjuvant chemotherapy. This multicenter cohort study aimed to evaluate the prognostic value of histological subtypes for stage I lung adenocarcinoma and to determine the relative survival benefit of adjuvant chemotherapy for subgroups based on MP pattern. Methods:This retrospective study included 412 patients with stage I lung adenocarcinoma [according the eighth edition of the tumor-node-metastasis (TNM) classification] with MP pattern who underwent complete surgical resection between January 2010 and December 2019. Patients were classified into 3 groups based on the proportion of MP component (10% and 50% as the threshold): MP component >50% (n=8), 10-50% (n=273) and <10% (n=131). Results:Among the 412 patients, the median age was 63 years, and 73.4% (113/154) patients with MP component ≥10% and 63.8% (51/80) of those with MP component <10% had epidermal growth factor receptor (EGFR) mutations. Patients with MP component >50% had a shorter overall survival (OS) compared with those with MP components of 10-50% [10-50% vs. >50%: hazard ratio (HR) =0.293, 95% confidence interval (CI): 0.083-1.027; P=0.052] or <10% (<10% vs. >50%: HR =0.214, 95% CI: 0.056-0.816; P=0.02). Notably, in the univariate analysis, the factors associated with a worse recurrence-free survival (RFS) were spread-through-air-space (STAS) status (HR =2.131, 95% CI: 1.104-4.112; P=0.02), male sex (HR =1.693, 95% CI: 1.048-2.735; P=0.03), smoking history (HR =1.817, 95% CI: 1.126-2.931; P=0.01), and tumor size >2 cm (HR =1.832, 95% CI: 1.138-2.949; P=0.01). Conclusions:MP component and risk factors might be considered critical features for patients with stage I lung adenocarcinomas and may inform the selection of patients who may benefit from adjuvant chemotherapy although no randomized evidence is available.

The role of the lung microbiota in cancer remains unclear. Here, we reveal that Staphylococcus is selectively enriched in metastatic tumor lesions and is associated with tumor recurrence in lung cancer patients. Using patient-derived bacterial strains, we employ a combination of cell line, organoid, mouse allograft, and xenograft models to demonstrate that S. nepalensis and S. capitis promote the metastatic potential of lung cancer cells. Mechanistically, lactate secreted by S. nepalensis and S. capitis upregulates MCT1 expression in tumor cells, facilitating lactate uptake and activating pseudohypoxia signaling. These effects can be eliminated by knocking out the lactate-producing genes (D-lactate dehydrogenase [ddh]/L-lactate dehydrogenase [ldh]) in the bacterial strains. Furthermore, we show that inhibiting MCT1 attenuates Staphylococcus-induced tumor metastasis both in vitro and in vivo. Collectively, our results demonstrate that tumor-resident Staphylococcus species promote lung cancer metastasis by activating host pseudohypoxia signaling and further identify key regulators as potential targets for therapeutic development.

BACKGROUND:Due to the low incidence of BRAF mutations, limited data is available about their prevalence and clinical characteristics. Moreover, comparative real-world efficacy of dabrafenib combined with trametinib versus other treatment regimens, especially in Chinese patients, is also lacking. METHODS:Patients who had BRAF genetic testing from the Lung Cancer Big Data Precise Treatment Collaboration Group (LANDSCAPE) database were included as Cohort I. The LANDSCAPE database comprises next-generation sequencing (NGS) data of 175,336 patients with lung cancer, originating from 6 Chinese genetic testing institutions. Cohort II included patients with unresectable locally advanced or metastatic NSCLC with a primary BRAF mutation from 19 centres in China from December 2015 to September 2022. FINDINGS:In Cohort I, of patients with NSCLC, 6249 (3.56%, 95% CI: 3.48%-3.65%) were confirmed to harbour a BRAF mutation. BRAF V600E accounted for 24.6% (1539/6249) of all patients with BRAF-mutated NSCLC. In Cohort II, a total of 129 patients with locally advanced or metastatic BRAF-mutated NSCLC were included. Of 112 patients who received NGS testing, 80 (71.4%) patients had concomitant mutations. The median first-line real-world progression-free survival (rwPFS) of dabrafenib plus trametinib for patients with BRAF V600 mutations was 25.0 months (N = 37), which was numerically longer than first-line immunotherapy-based therapy (N = 12, 15.7 months), and chemotherapy (N = 17, 9.2 months). INTERPRETATION:This study indicates that dabrafenib plus trametinib could be considered as the optimal treatment option for Chinese patients with NSCLC harbouring BRAF V600 mutations. FUNDING:National Natural Science Foundation of China (82072583); Beijing Municipal Administration of Hospitals Incubating Program (PX2020044); Beijing Hospitals Authority Youth Programme (QML20231113); Science Foundation of Peking University Cancer Hospital (2022-17); Peking University Cancer Hospital Inner Mongolia Hospital Public Hospital Reform and High-Quality Development Demonstration Project (Gastrointestinal Cancer + Thoracic Cancer) Research Fund (2024YNYB006).

Background:Early-stage lung adenocarcinoma has the potential to achieve long-term survival through radical surgery and appropriate adjuvant therapy. Despite rapid advancements in perioperative treatment strategies, postoperative adjuvant chemotherapy remains critically valuable. However, the optimal adjuvant chemotherapy regimen is yet to be established. Therefore, we initiated this study to evaluate the efficacy and safety of pemetrexed (PEM) plus platinum versus other platinum-based regimes as postoperative adjuvant chemotherapy in patients with stage II-IIIA lung adenocarcinoma after radical surgery to provide further clinical practice evidence. Methods:We performed a non-randomized, open-label, historical-control study. Seventy-one patients with pathologic stage II-IIIA lung adenocarcinoma who had undergone complete resection were prospectively enrolled, all of whom received pemetrexed combined with platinum adjuvant chemotherapy every 3 weeks for 4 cycles. In addition, we retrospectively collected the data from 209 patients recorded between January 2003 and December 2021 with pathologic stage II-IIIA lung adenocarcinoma who received radical surgery followed by adjuvant non-PEM (paclitaxel, gemcitabine and vinorelbine) plus platinum as historical control. The primary end point was disease-free survival (DFS). Univariable and multivariable Cox proportional hazards regression analyses were performed to identify independent prognostic factors for DFS. Propensity score matching (PSM) was applied to generate best-matched pairs for the two categories. All adverse events (AEs) were collected and compared. Results:In the prospective group, the median follow-up was 45.6 months (interquartile range, 40.3-55.7 months). Thirty-five of the 71 prospective patients developed disease progression, and 2-year DFS was 67.61%. The median DFS was not reached in the PEM group, compared with 18.9 months [95% confidence interval (CI): 15.6-22.0] in the non-PEM group. In the overall unmatched population, Kaplan-Meier analysis demonstrated a significant improvement in DFS with PEM (median DFS not reached vs. 18.9 months; P<0.001, hazard ratio =0.499, 95% CI: 0.345-0.72). In the PSM-matched cohort, the DFS benefit with PEM remained statistically significant (P=0.01). Besides, AEs of all grades were reported more frequently in the non-PEM plus platinum group than in PEM based group, such as neutropenia (76.1% vs. 25.4%), anemia (48.3% vs. 8.5%) and thrombocytopenia (23.4% vs. 5.6%). Conclusions:This study showed that PEM plus platinum outperforms non-PEM plus platinum as an adjuvant chemotherapy regimen for resected stage II-IIIA lung adenocarcinoma and with better tolerability.

Immune checkpoint inhibitors (ICIs) have changed the treatment landscape for patients with non-small cell lung cancer (NSCLC). In spite of durable responses in some patients, many patients develop early disease progression during the ICI treatment. Thus, early identification of patients with no durable benefit would facilitate the clinical decision for these patients. In this prospective, multicenter study, 101 non-EGFR/ALK patients who received ICI treatment were enrolled after screening 328 stage III-IV NSCLC patients. At the date of cutoff, 83 patients were eligible for ICI efficacy evaluation, with 56 patients having progress-free survival (PFS) over 6 months, which was defined as durable clinical benefit (DCB). A multimodal model was established by integrating normalized bTMB, early dynamic of ctDNA and the first RECIST response. This model could robustly predict DCB with area under the curve (AUC) of 0.878, sensitivity of 79.2% at 86.4% specificity (accuracy = 80.0%). This model was further validated in the independent cohort of the DIREct-On study with AUC of 0.887, sensitivity of 94.7% at 85.3% specificity (accuracy = 90.3%). Patients with higher predict scores had substantially longer PFS than those with lower scores (training cohort: median PFS 13.6 vs 4.2 months, P < 0.001, HR = 0.24; validation cohort: median PFS 11.0 vs 2.2 months, P < 0.001, HR = 0.17). Taken together, these results demonstrate that integrating early changes of ctDNA, normalized bTMB, and the first RECIST response can provide accurate, noninvasive, and early prediction of durable benefits for NSCLC patients treated with ICIs. Further prospective studies are warranted to validate these findings and guide clinical decision-making for optimal immunotherapy in NSCLC patients.

AbstractBackgroundUsage of immune checkpoint inhibitors (ICIs) has prolonged the overall survival (OS) of patients with extensive‐stage small‐cell lung cancer (ES‐SCLC). In clinical trials, males accounted for a large proportion, leading to the uncertainty of its efficacy in female patients. We therefore conducted this study to explore the efficacy and safety of using ICIs in female patients with ES‐SCLC.MethodsWe retrospectively enrolled female SCLC patients and subdivided them into two groups. Group A (n = 40) was defined as ES‐SCLC patients who received first‐line standard chemotherapy with or without ICIs. Group B (n = 47) included relapsed SCLC patients who were administered with second‐line therapies. Kaplan–Meier methodology was used to calculate survival analysis. Chi‐squared tests were used to analyze the incidence of adverse events (AEs).ResultsMedian progression‐free survival (PFS) and median OS favored the ICI‐contained cohorts (Group A PFS: 8.3 vs. 6.1 months; OS: not reached vs. 11.3 months; Group B PFS: 15.1 vs. 3.3 months; OS: 35.3 vs. 8.3 months), especially in those patients who received second‐line immunotherapies. Patients who received immunotherapy had a slightly higher incidence rate of grade ≥3 AEs (Group A: 71.4% vs. 46.2%; Group B: 44.5% vs. 13.2%). Those who developed grade ≥3 AEs in first‐line ICIs cohort had a more favorable survival (PFS: 8.3 vs. 3.2 months; OS: not reached vs. 5.1 months).ConclusionsOur study suggested that female ES‐SCLC patients treated with immunotherapy tended to achieve a relatively longer survival. The incidence of AEs (grade ≥3) was higher in women patients receiving ICIs, which requires monitoring more closely.

Accurate prediction of the efficacy of immunotherapy for cancer patients through the characterization of both genetic and phenotypic heterogeneity in individual patient cells holds great promise in informing targeted treatments, and ultimately in improving care pathways and clinical outcomes. Here, we describe the nanoplatform for interrogating living cell host-gene and (micro-)environment (NICHE) relationships, that integrates micro- and nanofluidics to enable highly efficient capture of circulating tumor cells (CTCs) from blood samples. The platform uses a unique nanopore-enhanced electrodelivery system that efficiently and rapidly integrates stable multichannel fluorescence probes into living CTCs for in situ quantification of target gene expression, while on-chip coculturing of CTCs with immune cells allows for the real-time correlative quantification of their phenotypic heterogeneities in response to immune checkpoint inhibitors (ICI). The NICHE microfluidic device provides a unique ability to perform both gene expression and phenotypic analysis on the same single cells in situ, allowing us to generate a predictive index for screening patients who could benefit from ICI. This index, which simultaneously integrates the heterogeneity of single cellular responses for both gene expression and phenotype, was validated by clinically tracing 80 non-small cell lung cancer patients, demonstrating significantly higher AUC (area under the curve) (0.906) than current clinical reference for immunotherapy prediction.

Background/aim: Elevated baseline body mass index (BMI) is independently correlated with the efficacy and prognosis of anti-tumour immunotherapy. But the effects of BMI change in patients undergoing PD-1/PD-L1 monoclonal antibody therapy have not been well studied. Method A retrospective analysis of patients who were consecutively receiving anti-PD-1/PD-L1 inhibitor treatment diagnosed with advanced NSCLC was conducted to investigate the effects of baseline and maximum variation in BMI within the first 12 weeks on objective response rate (ORR), progression-free survival (PFS), and overall survival (OS). Baseline BMI and change in BMI were both analysed as continuous and categorical variables. Results For baseline BMI, only the overweight group showed an improvement in OS ([HR] = 0.67, 95% CI:0.49–0.91, P = 0.011). BMI variation analysis showed significant differences among stable, increase, and decrease groups (P = 0.0033), and the increase group showed a significantly improved OS (HR = 0.38, 95%CI: 0.20–0.74, P = 0.004). Each 1% increase in BMI was associated with a 9% increase in survival time (HR = 0.91, 95%CI:0.87–0.96, P < 0.001). Also, patients in the increased group showed a significantly higher ORR (OR = 5.42, 95%CI:2.02–14.54, P < 0.001). Subsequent analysis revealed that the increase group showed a significant benefit in PFS (HR = 0.57, 95%CI:0.35–0.92, P = 0.022); and each percentage point increase in BMI was associated with a 9% improvement (HR = 0.93, 95%CI:0.89–0.96, P < 0.001). Conclusion Weight gain during treatment should be considered a potentially more potent predictive factor in immunotherapy compared to baseline body mass index (BMI).

Purpose: This multicenter cohort study aimed to evaluate the prognostic effects of histological subtypes on stage I lung adenocarcinoma and to investigate whether adjuvant chemotherapy can contribute to survival benefits in different micropapillary (MP) pattern subgroups.Experimental Design: This retrospective study included 412 patients with stage I lung adenocarcinoma (the 8th TNM classification) with MP pattern who underwent complete surgical resection between January 2010 and December 2019. Patients were classified into four groups based on the proportion of MP components (10% as the threshold) and the presence or absence of solid components: Group 1 (n =108; MP < 10% & solid negative); Group 2 (n = 23; MP < 10% & solid positive); Group 3 (n = 211; MP ≥ 10% & solid negative) and Group 4 (n = 70; MP ≥ 10% & solid positive).Results: Among the 412 patients, the median age was 63 years, and 73.4% of patients (113/154) with MP ≥10% and 63.8% of patients (51/80) with MP <10% had epidermal growth factor receptor (EGFR) mutations. No significant differences in the recurrence- free survival (RFS) and overall survival (OS) were observed among the four groups. However, patients with MP >50% had shorter OS compared with those with MP components of 10%-50% (10%-50% vs >50%, HR=0.293, 95%CI: 0.083-1.027, P=0.055) or < 10% (< 10% vs >50%, HR=0.214, 95%CI: 0.056-0.816, P=0.024). Notably, univariate analysis found spread-through-air-space (STAS) (HR=2.131, 95%CI 1.104-4.112, P = 0.021), male (HR=1.693, 95%CI: 1.048-2.735, P=0.031), smoking history (HR=1.817, 95%CI: 1.126-2.931, P = 0.014) and tumor size > 2 cm (HR=1.832, 95%CI: 1.138-2.949, P=0.013) were associated with a worse RFS.Conclusions: MP component should be considered as an important factor for stage I lung adenocarcinomas and may contribute to the selection of potential candidates who may benefit from adjuvant chemotherapy.

目的 探讨白细胞介素10(IL-10)对皮肤角质形成细胞(HaCaT)增殖影响和对氯化钙(CaCl2)诱导的角质形成细胞分化标志物的表达影响及其可能的分子机制.方法 以不同浓度IL-10(0、3、10、30 ng/ml)处理HaCaT细胞不同时间(0、24、48、72 h),MTS分析细胞增殖,流式细胞仪检测细胞周期;IL-10(终浓度为 10 ng/ml)预处理HaCaT 1 h,加入或不加CaCl2(终浓度为 1.2 mmol/L)培养 24、48、72 h,Western blot检测 IL-10 对 HaCaT分化标志物表达影响;丝裂原蛋白激活激酶-胞外信号调节激酶(MAPKs-ERK1/2)特异性抑制剂PD98059 及磷脂酰肌醇激酶-丝氨酸/苏氨酸激酶(PI3K-AKT)特异性抑制剂 LY294002 预处理HaCaT细胞,分别提取细胞总 RNA 和蛋白,荧光定量 PCR(RT-qPCR)和Western blot检测 IL-10 对 HaCaT细胞分化标志物(Keratin1、Keratin5、Involucrin)表达影响.结果 MTS结果显示,在72h内,IL-10(30 ng/ml和较低浓度)对HaCaT细胞增殖无影响;流式细胞分析结果提示IL-10 不影响HaCaT细胞周期进程.Western blot分析显示,IL-10 上调HaCaT角质形成细胞角质细胞分化标志物Involucrin表达,而对Kera-tin1 及Keratin5 没有显著的影响.机制研究分析显示,IL-10能够活化 ERK1/2 和 AKT,增加其磷酸化水平;RT-qPCR和Western blot结果显示PD98059 及LY294002 部分阻断IL-10诱导的Involucrin的表达.结论 IL-10 在一定浓度范围内不影响 HaCaT 的增殖;IL-10 部分通过 MAPKs-ERK1/2 和PI3K-AKT 途径上调 HaCaT分化标志物Involucrin表达.

Background Cancer immunotherapy has emerged as a promising approach for cancer treatment.1 However, the efficacy of current immunotherapies is limited by the immunosuppressive tumor microenvironment, novel strategies to further enhance anti-tumor activity of immunotherapy are still required. Recent studies reveal the profound impact of microbiomes on host immunity, influencing anti-tumor responses.2 Efforts to enhance immunotherapy involve microbial preparations with immunostimulatory properties, but these approaches remain in early stages.3 Methods In this pre-clinical research, we analyzed the metagenomic profiles of 24 non-small cell lung cancer (NSCLC) patients who had been treated with PD-1 inhibitors (11 responders and 13 non-responders) and identified an enrichment of the bacterium Deinococcus geothermalis (D. geothermalis) in responder's tumor tissues. To further investigate the effects of D. geothermalis on immunotherapy for NSCLC, we established patient-derived organoid (PDO) models from surgical samples of primary NSCLC tissues and assessed the activation of cytotoxic T cells in vitro. We evaluated the expression of TNF-α and IFN-γ using enzyme-linked immunosorbent assay (ELISA) and assessed cell viability in different treatment groups. Immunofluorescence staining was performed to detect the expression of Granzyme B (GZMB) and CD3, allowing us to evaluate the activation of cytotoxic T cells by calculating the ratio of GZMB-positive to CD3-positive cells. Results Our results showed that the combination therapy with D. geothermalis and PD-1 inhibitors significantly increased the expression of TNF-α (approximately 100-fold) and IFN-γ (50–60 fold) in the NSCLC PDO models and also led to 20–30% reduction in tumor cell viability compared to control group. Immunofluorescence staining revealed significant increase in the proportion of GZMB-positive cells in lung cancer PDOs treated with the combination therapy, indicating enhanced activation of cytotoxic T cells. These findings suggest that the combination of D. geothermalis and PD-1 inhibitors could increase the activation of cytotoxic T cells in the tumor microenvironment, leading to significant tumor growth inhibition. Conclusions Our study demonstrates the potential of combining D. geothermalis with immune checkpoint inhibitors as a novel strategy for cancer immunotherapy. This combination therapy has the potential to overcome the limitations of current immunotherapies and improve the clinical outcomes of cancer patients. Further studies are warranted to evaluate the safety and efficacy of this combination therapy in clinical trials. Acknowledgements This study was supported by the Science Foundation of Peking University Cancer Hospital (18–02), the General Program of National Natural Science Foundation of China (82073247), Beijing Municipal Administration of Hospitals Incubating Program (PX2019038), and Beijing Youth Program for Outstanding Talents (2018000021469G262). References Gandhi L, Rodríguez-Abreu D, Gadgeel S, et al. Pembrolizumab plus Chemotherapy in Metastatic Non-Small-Cell Lung Cancer. N Engl J Med. 2018;378(22):2078–2092. Matson V, Chervin CS, Gajewski TF. Cancer and the Microbiome-Influence of the Commensal Microbiota on Cancer, Immune Responses, and Immunotherapy. Gastroenterology. 2021;160(2):600–613. Sepich-Poore GD, Zitvogel L, Straussman R, Hasty J, Wargo JA, Knight R. The microbiome and human cancer. Science. 2021;371(6536):eabc4552. Ethics Approval The study was approved by Peking University Cancer Hospital and Institute Ethics Board, approval number 2018KT89. Consent Written informed consent was obtained from the patient for publication of this abstract and any accompanying images. A copy of the written consent is available for review by the Editor of this journal.

Objectives: Cardiac hypertrophy is the heart's compensatory response stimulated by various patho-physiological factors. However, prolonged cardiac hypertrophy poses a significant risk of progression to heart failure, lethal arrhythmias, and even sudden cardiac death. For this reason, it is crucial to effectively prevent the occurrence and development of cardiac hypertrophy. CMTM is a superfamily of human chemotaxis, which is involved in immune response and tumorigenesis. CMTM3 expressed widely in tissues, including the heart, but its cardiac function remains unclear. This research aims to explore the effect and mechanism of CMTM3 in the development of cardiac hypertrophy.Methods and results: We generated a Cmtm3 knockout mouse model (Cmtm3-/-) as the loss-of-function approach. CMTM3 deficiency induced cardiac hypertrophy and further exacerbated hypertrophy and cardiac dysfunction stimulated by Angiotensin II infusion. In Ang II-infusion stimulated hypertrophic hearts and phenylephrine-induced hypertrophic neonatal cardiomyocytes, CMTM3 expression signifi-cantly increased. However, adenovirus-mediated overexpression of CMTM3 inhibited the hypertrophy of rat neonatal cardiomyocytes induced by PE stimulation. In terms of mechanism, RNA-seq data revealed that Cmtm3 knockout-induced cardiac hypertrophy was related to MAPK/ERK activation. In vitro, CMTM3 overexpression significantly inhibited the increased phosphorylation of p38 and ERK induced by PE stimulation.Conclusions: CMTM3 deficiency induces cardiac hypertrophy and aggravates hypertrophy and impaired cardiac function stimulated by angiotensin II infusion. The expression of CMTM3 increases during cardiac hypertrophy, and the increased CMTM3 can inhibit further hypertrophy of cardiomyocytes by inhibiting MAPK signaling. Thus, CMTM3 plays a negative regulatory effect in the occurrence and development of cardiac hypertrophy.& COPY; 2023 Elsevier Inc. All rights reserved.

肺癌是全球所有恶性肿瘤中死亡率最高的恶性肿瘤,发病率紧随乳腺癌之后居第2位.近年来,以针对程序性死亡受体1(PD-1)/程序性死亡配体1(PD-L1)以及细胞毒性T淋巴细胞相关抗原4(CTLA4)为代表的免疫检查点抑制剂(ICI)的研发为肺癌的治疗带来了巨大的转变,显著延长了肺癌患者的生存时间.然而,仅不足30%的晚期肺癌患者可以从ICI治疗中长期获益,目前除PD-L1外尚缺乏其他明确的生物标志物来有效预测ICI的疗效.近些年针对肺癌ICI治疗新兴生物标志物的研究主要集中在肿瘤突变负荷、基因突变、错配修复基因、外泌体、肿瘤免疫微环境、转录组特征以及微生物组等方面,文章对这些新兴生物标志物的研究进展进行综述.

BACKGROUND:Early diagnosis of lung adenocarcinoma (LUAD), one of the most common types of lung cancer, is very important to improve the prognosis of patients. The current methods can't meet the requirements of early diagnosis. There is a pressing need to identify novel diagnostic biomarkers. Secretory proteins are the richest source for biomarker research. This study aimed to identify candidate secretory protein biomarkers for early diagnosis of LUAD by integrated bioinformatics analysis and clinical validation.METHODS:Differentially expressed genes (DEGs) of GSE31210, gene expression data of early stage of LUAD, were analyzed by GEO2R. Upregulated DEGs predicted to encode secreted proteins were obtained by taking the intersection of the DEGs list with the list of genes encoding secreted proteins predicted by the majority decision-based method (MDSEC). The expressions of the identified secreted proteins in the lung tissues of early-stage LUAD patients were further compared with the healthy control group in mRNA and protein levels by using the UALCAN database (TCGA and CPTAC). The selected proteins expressed in plasma were further validated by using Luminex technology. The diagnostic value of the screened proteins was evaluated by receiver operating characteristic (ROC) analysis. Cell counting kit-8 assay was carried out to investigate the proliferative effects of these screened proteins.RESULTS:A total of 2183 DEGs, including 1240 downregulated genes and 943 upregulated genes, were identified in the GSE31210. Of the upregulated genes, 199 genes were predicted to encode secreted proteins. After analysis using the UALCAN database, 16 molecules were selected for further clinical validation. Plasma concentrations of three proteins, Midkine (MDK), WAP four-disulfide core domain 2 (WFDC2), and C-X-C motif chemokine ligand 14 (CXCL14), were significantly higher in LUAD patients than in healthy donors. The area under the curve values was 0.944, 0.881, and 0.809 for MDK, WFDC2, and CXCL14, 0.962 when combined them. Overexpression of the three proteins enhanced the proliferation activity of A549 cells.CONCLUSIONS:MDK, WFDC2, and CXCL14 were identified as candidate diagnostic biomarkers for early-stage LUAD and might also play vital roles in tumorigenesis.

The relationship between lung microbiota and cancer has been explored based on DNA sequencing; however, culture-dependent approaches are indispensable for further studies on the lung microbiota. In this study, we applied a comprehensive approach combining culturomics and 16S rRNA gene amplicon sequencing to detect members of the microbiotas in saliva and BALF samples from patients with unilateral lobar masses.

BACKGROUND:Progressive cardiac fibrosis leads to ventricular wall stiffness, cardiac dysfunction, and eventually heart failure, but the underlying mechanism remains unexplored. PDCD5 (programmed cell death 5) ubiquitously expresses in tissues, including the heart; however, the role of PDCD5 in cardiac fibrosis is largely unknown. Therefore, this study aims at exploring the possible role and underlying mechanisms of PDCD5 in the pathogenesis of cardiac fibrosis.METHODS AND RESULTS:PDCD5 levels were found to be elevated in the serum obtained from patients with cardiac fibrosis, in fibrotic mice heart tissues after myocardial infarction, and in cardiac fibroblasts stimulated by Ang II (angiotensin II)- or TGF-β1 (transforming growth factor-β1). Overexpression of PDCD5 in cardiac fibroblasts or treatment with PDCD5 protein reduced the expression of profibrogenic proteins in response to TGF-β1 stimulation, while knockdown of PDCD5 increased fibrotic responses. It has been demonstrated that SMAD3, a protein that is also known as mothers against decapentaplegic homolog 3, directly upregulated PDCD5 during cardiac fibrosis. Subsequently, the increased PDCD5 promoted HDAC3 (histone deacetylase 3) ubiquitination, thus, inhibiting HDAC3 to reduce fibrotic responses. Fibroblast-specific knock-in of PDCD5 in mice ameliorated cardiac fibrosis after myocardial infarction and enhanced cardiac function, and these protective effects were eliminated by AAV9-mediated HDAC3 overexpression.CONCLUSIONS:The findings of this study demonstrated that PDCD5 is upregulated by SMAD3 during cardiac fibrosis, which subsequently ameliorated progressive fibrosis and cardiac dysfunction through HDAC3 inhibition. Thus, this study suggests that PDCD5 functions as a negative feedback factor on fibrotic signaling pathways and might serve as a potential therapeutic target to suppress the progression of fibrotic responses.