Surgical Non-small Cell Lung Cancer Research Articles

Postoperative ctDNA in indicating the recurrence risk and monitoring the effect of adjuvant therapy in surgical non-small cell lung cancer.

Circulating tumor DNA (ctDNA) has emerged as a potential novel biomarker to predict molecular residual disease (MRD) in lung cancer after definitive treatment. Herein, we investigated the value of ctDNA in prognosing risk of relapse and monitoring the effect of adjuvant therapy in surgical non-small cell lung cancer (NSCLC). We enrolled 58 NSCLC patients in a real-world setting, and 58 tumor tissues and 325 plasma samples were analyzed. Tumor tissues and plasma samples were subjected to targeted next-generation sequencing (NGS) of 1021 cancer-related and ultra-deep targeted NGS covering 338 genes, respectively. ctDNA was detected in 31.0% of cases at the first postoperative time, which was associated with advanced tumor stage, T stage and KEAP1 or GRIN2A mutations in tissues. ctDNA positivity at landmark and longitudinal indicated the shorter disease-free survival. For patients with ctDNA positivity at the first postoperative time, regardless of adjuvant therapy, all patients who were persistently ctDNA positive during postoperative surveillance had disease recurrence. Among the patients who were ctDNA negative, only two patients (15.4%, 2/13) receiving adjuvant therapy relapsed, while one patient (50.0%, 1/2) without adjuvant therapy relapsed. For the first postoperative ctDNA negative patients, the recurrence rate of patients with adjuvant therapy was and higher than without adjuvant therapy (22.6% [7/31] vs. 11.1% [1/9]). The patients who became ctDNA positive may also benefit from intervention therapy. Postoperative ctDNA is a prognostic marker, and ctDNA-detection may facilitate personalized adjuvant therapy, and applying adjuvant therapy to the patients with detectable ctDNA could bring clinical benefits for them.

Optimum timing of lung resection surgery following SARS-CoV-2 infection for non-small cell lung cancer.

The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection on postoperative recovery of non-small cell lung cancer (NSCLC) is need to be understood, thereby informing the optimal timing of surgical decision-making during the COVID-19 pandemic for NSCLC patients. This study reports the postoperative outcomes of surgical NSCLC patients with preoperative SARS-CoV-2 infection. This single-center retrospective cohort study included 241 NSCLC patients who underwent lobectomy or sub-lobectomy between December 1, 2022 and February 14, 2023. Surgical outcomes of patients with preoperative SARS-CoV-2 infection (stratified by the time from diagnosis of SARS-CoV-2 infection to surgery) were compared with those without preoperative SARS-CoV-2 infection. The primary outcomes were total postoperative complications and postoperative pulmonary complications (PPCs), the secondary outcomes included operation time, total postoperative drainage and time, length of hospital stay (LOS), 30-day and 90-day postoperative symptoms. This study included 153 (63.5%) patients with preoperative SARS-CoV-2 infection and 88 (36.5%) patients without previous SARS-CoV-2 infection. In patients with a preoperative SARS-CoV-2 diagnosis, the incidence of total postoperative complications (OR, 3.00; 95% CI, 1.12-8.01; p = 0.028) and PPCs (OR, 4.20; 95% CI, 1.11-15.91; p = 0.035) both increased in patients infected having surgery within 2 weeks compared with non-infection before surgery. However, patients who underwent lung resection more than 2 weeks after SARS-CoV-2 diagnosis had a similar risk of postoperative complications and surgical outcomes with those non-infection before surgery. This is the first study to provide evidence regarding the optimum timing of lung resection surgery and assessing early outcomes after surgery in NSCLC patients with SARS-CoV-2 infection. Our study documents that the SARS-CoV-2 infection did not complicate surgical procedures for lung cancer, and suggest that lung surgery should be postponed at least 2 weeks after SARS-CoV-2 infection for NSCLC patients.

Blood regulator of G protein signalling 1 as a potential prognostic biomarker in surgical nonsmall cell lung cancer patients: Correlation with clinical features and survival.

Regulator of G protein signalling 1 (RGS1) closely regulates malignant phenotypes and tumour immunity in several cancers, while its clinical value in nonsmall cell lung cancer (NSCLC) is by far rarely reported. Consequently, this study aimed to explore the linkage of blood RGS1 with clinical features and prognosis in surgical NSCLC patients. Two-hundred and ten surgical NSCLC patients were consecutively enrolled in this study, whose RGS1 in peripheral blood mononuclear cells was determined before treatment via reverse transcriptional-quantitative polymerase chain reaction. Additionally, the blood RGS1 was also collected from 30 healthy controls (HCs). Blood RGS1 was increased in NSCLC patients compared with HCs (P < 0.001). Elevated blood RGS1 was related to lymph node (LYN) metastasis (P = 0.001), higher tumour-nodes-metastasis (TNM) stage (P = 0.004), neoadjuvant chemotherapy administration (P = 0.044), shortened accumulative disease-free survival (DFS) (P = 0.008) and overall survival (OS) (P = 0.013) in NSCLC patients. A multivariate Cox's regression analysis showed that blood RGS1 high expression could independently reflect shortened DFS (hazard ratio = 1.499, P = 0.023), whereas it could not independently predict OS (P > 0.050). Furthermore, blood RGS1 high expression was associated with shortened OS (P = 0.020) in patients with neoadjuvant therapy and with worse DFS (P = 0.028) and OS (P = 0.026) in patients with adjuvant therapy, while blood RGS1 was not linked with DFS or OS in patients without neoadjuvant or adjuvant therapy (all P > 0.050). Elevated blood RGS1 correlates with LYN metastasis, neoadjuvant chemotherapy administration, worse DFS and OS, which might serve as a useful prognostic biomarker for surgical NSCLC patients.

Multimodal prehabilitation in patients with non-small cell lung cancer undergoing anatomical resection: protocol of a non-randomised feasibility study

BackgroundThe preoperative period can be used to enhance a patient’s functional capacity with multimodal prehabilitation and consequently improve and fasten postoperative recovery. Especially, non-small cell lung cancer (NSCLC) surgical patients may benefit from this intervention, since the affected and resected organ is an essential part of the cardiorespiratory fitness. Drafting a prehabilitation programme is challenging, since many disciplines are involved, and time between diagnosis of NSCLC and surgery is limited. We designed a multimodal prehabilitation programme prior to NSCLC surgery and aimed to conduct a study to assess feasibility and indicative evidence of efficacy of this programme. Publication of this protocol may help other healthcare facilities to implement such a programme.MethodsThe multimodal prehabilitation programme consists of an exercise programme, nutritional support, psychological support, smoking cessation, patient empowerment and respiratory optimisation. In two Dutch teaching hospitals, 40 adult patients with proven or suspected NSCLC will be included. In a non-randomised fashion, 20 patients follow the multimodal prehabilitation programme, and 20 will be assessed in the control group, according to patient preference. Assessments will take place at four time points: baseline, the week before surgery, 6 weeks postoperatively and 3 months postoperatively. Feasibility and indicative evidence of efficacy of the prehabilitation programme will be assessed as primary outcomes.DiscussionSince the time between diagnosis of NSCLC and surgery is limited, it is a challenge to implement a prehabilitation programme. This study will assess whether this is feasible, and evidence of efficacy can be found. The non-randomised fashion of the study might result in a selection and confounding bias. However, the control group may help putting the results of the prehabilitation group in perspective. By publishing this protocol, we aim to facilitate others to evaluate and implement a multimodal prehabilitation programme for surgical NSCLC patients.Trial registrationThe current study is registered as NL8080 in the Netherlands Trial Register on the 10th of October 2019, https://www.trialregister.nl/trial/8080. Secondary identifiers: CCMO (Central Committee on Research Involving Human Subjects) number NL70578.015.19, reference number of the Medical Ethical Review Committee of Máxima MC W19.045.

Quantitative Image Analysis of Fibrillar Collagens Reveals Novel Diagnostic and Prognostic Biomarkers and Histotype-Dependent Aberrant Mechanobiology in Lung Cancer

Fibrillar collagens are the most abundant extracellular matrix components in non–small cell lung cancer (NSCLC). However, the potential of collagen fiber descriptors as a source of clinically relevant biomarkers in NSCLC is largely unknown. Similarly, our understanding of the aberrant collagen organization and associated tumor-promoting effects is very scarce. To address these limitations, we identified a digital pathology approach that can be easily implemented in pathology units based on CT-FIRE software (version 2; https://loci.wisc.edu/software/ctfire) analysis of Picrosirius red (PSR) stains of fibrillar collagens imaged with polarized light (PL). CT-FIRE settings were pre-optimized to assess a panel of collagen fiber descriptors in PSR-PL images of tissue microarrays from surgical NSCLC patients (106 adenocarcinomas [ADC] and 89 squamous cell carcinomas [SCC]). Using this approach, we identified straightness as the single high-accuracy diagnostic collagen fiber descriptor (average area under the curve = 0.92) and fiber density as the single descriptor consistently associated with a poor prognosis in both ADC and SCC independently of the gold standard based on the TNM staging (hazard ratio, 2.69; 95% CI, 1.55-4.66; P < .001). Moreover, we found that collagen fibers were markedly straighter, longer, and more aligned in tumor samples compared to paired samples from uninvolved pulmonary tissue, particularly in ADC, which is indicative of increased tumor stiffening. Consistently, we observed an increase in a panel of stiffness-associated processes in the high collagen fiber density patient group selectively in ADC, including venous/lymphatic invasion, fibroblast activation (α-smooth muscle actin), and immune evasion (programmed death-ligand 1). Similarly, a transcriptional correlation analysis supported the potential involvement of the major YAP/TAZ pathway in ADC. Our results provide a proof-of-principle to use CT-FIRE analysis of PSR-PL images to assess new collagen fiber–based diagnostic and prognostic biomarkers in pathology units, which may improve the clinical management of patients with surgical NSCLC. Our findings also unveil an aberrant stiff microenvironment in lung ADC that may foster immune evasion and dissemination, encouraging future work to identify therapeutic opportunities.

Prognostic value of preoperative modified Glasgow prognostic score in surgical non-small cell lung cancer: A meta-analysis.

The predictive role of modified Glasgow prognostic score (mGPS) for long-term survival in several types of cancers has been well manifested. We supposed that preoperative mGPS might also be associated with long-term survival of operated non-small cell lung cancer (NSCLC) patients. The aim of this meta-analysis was to identify the prognostic value of preoperative mGPS in surgical NSCLC patients. The PubMed, Web of Science, EMBASE and CNKI databases were searched for relevant studies up to November 7, 2022. The primary and secondary outcomes were overall survival (OS) and disease-free survival (DFS), respectively. The hazard ratios (HRs) and 95% confidence intervals (CIs) were combined. A total of 3,803 patients from 11 studies were enrolled and analyzed. The combined results demonstrated elevated preoperative mGPS was significantly related to poorer OS (HR = 2.11, 95% CI: 1.83-2.44, P < 0.001) and DFS (HR = 1.70, 95% CI: 1.42-2.03, P < 0.001). Subgroup analysis for the OS further identified the predictive role of elevated preoperative mGPS for worse OS in NSCLC. Preoperative mGPS was significantly associated with prognosis in NSCLC and patients with elevated preoperative mGPS experienced poorer long-term survival.

The predictive value of the preoperative systemic immune‐inflammation index in the occurrence of postoperative pneumonia in non‐small cell lung cancer: A retrospective study based on 1486 cases

To investigate the correlation between the preoperative systemic immune-inflammation index (pSII) and postoperative pneumonia (POP) in surgical non-small cell lung cancer patients. Patients who underwent lung cancer surgery at West China Hospital of Sichuan University were retrospectively included. The indicators were collected, including basic information of patients, surgery-related variables and POP rate. The predictive value of the pSII in the occurrence of POP was analyzed. A total of 1486 patients (male: 748, 50.3%; female: 738, 49.7%; mean age: 58.2 ± 9.7 years; median age: 59 years old, interquartile range: 51-65 years old) were finally included in the study, of which 142 patients had POP with an incidence of 9.5% (142/1486), 9.2% (69/748) in males, and 9.9% (73/738) in females. The proportion of patients with diabetes in the pneumonia group was significantly higher than that in the nonpneumonia group (9.8%, 14/142 vs. 5.6%, 75/1344, p=0.041). Compared with the nonpneumonia group, the level of the preoperative body mass index (24.2 [21.9, 26.1] vs. 23.1 [21.1, 25.2], p=0.003) and SII (487 [350, 673] vs. 345 [230, 500], p < 0.001) in the pneumonia group were significantly higher. Multiple factor analysis showed that the pSII (odds ratio: 1.001, 95% confidence interval: 1.000-1.001, p < 0.001) was an independent risk factor for POP (487 [350, 673] vs. 345 [230, 500], p < 0.001); receiver operating characteristic curve analysis showed that the pSII was effective in predicting POP (area under curve: 0.751, p < 0.001). The pSII is closely related to and can effectively predict the occurrence of POP after lung cancer surgery.

Comparison of 22C3-PD-L1 Expression Between Paired Tumor Paraffin Blocks of Surgical Resection Specimens of Nonsmall Cell Lung Cancer.

In this study, our aim was to evaluate the discordance of programmed cell death ligand 1 (PD-L1) expression between 2 paired paraffin blocks from surgical nonsmall cell lung cancer (NSCLC) specimens, thus providing recommendations for choosing paraffin blocks for PD-L1 immunohistochemistry detection. A total of 460 paired paraffin blocks of surgical NSCLC specimens were analyzed using the 22C3-PD-L1 assay. PD-L1 expression between 2 paired paraffin blocks was calculated using 3 classification schemes, tumor proportion score (TPS)-based 3-level classification and TPS-based binary classification using TPS ≥1% or TPS ≥50% as the cutoff. Clinicopathological characteristics were analyzed for cases with discordant PD-L1 expression. PD-L1 expression in 2 paired paraffin blocks of surgical NSCLC specimens was highly correlated from a single tumor ( R2 =0.89) in the overall trend. The overall discordance rates were 10.9%, 7.4%, and 3.5% respectively when using TPS-based 3-level classification or binary classification. No statistical differences were observed in PD-L1 expression discordance rates when patients were stratified by age, sex, smoking status, histologic types, TNM stage, or years of paraffin blocks ( P >0.05). The presence of a solid histologic pattern was associated with a higher PD-L1 expression discordance rate in adenocarcinomas ( P <0.05). Twenty-five cases with discordant PD-L1 expression were divided into 2 categories: spatial heterogeneity without recognizable morphology difference (76%) and spatial heterogeneity with significantly different morphology (24%). The discordance rate in TPS scores was much higher in cases with different morphology than those without different morphology. One representative paraffin block containing adequate neoplastic tissue may be adequate to determine PD-L1 expression in most of the surgical specimens of NSCLC. In cases that harbored different morphology in different paraffin blocks, the dual-block immunohistochemistry assessment method is recommended due to the intratumoral heterogeneity.

SENP1 Aberrance and Its Linkage to Clinical Features, Adjuvant Regimen, and Prognosis in Patients With Surgical Non-small Cell Lung Cancer Receiving Adjuvant Chemotherapy.

BackgroundSmall ubiquitin-like modifier-specific protease 1 (SENP1) plays vital roles in cancer progression and chemoresistance, but its prognostic value in non-small cell lung cancer (NSCLC) is vague. This study aimed to explore the correlation of SENP1 with clinical features, adjuvant chemotherapy regimen, and prognosis in patients with surgical NSCLC receiving adjuvant chemotherapy.MethodsTumor and adjacent tissues were collected from 157 patients with surgical NSCLC receiving adjuvant chemotherapy. Meanwhile, tumor tissue and paired adjacent tissue specimens were obtained to evaluate SENP1 protein expression by immunohistochemistry (IHC) assay; among which, 102 pairs were used to detect SENP1 messenger RNA (mRNA) by reverse transcription quantitative PCR.ResultsSENP1 IHC score and SENP1 mRNA expression were increased in tumor tissue than adjacent tissue (p < 0.001). Besides, elevated SENP1 IHC score was correlated with > 5 cm tumor size (p = 0.045), lymph node metastasis occurrence (p = 0.003), and advanced tumor-node-metastasis (TNM) stage (p = 0.012); meanwhile, increased SENP1 mRNA expression was associated with histopathological subtype (p = 0.011), lymph node metastasis occurrence (p = 0.008), and higher TNM stage (p = 0.015). Besides, no correlation was found in SENP1 IHC score (p = 0.424) or mRNA expression (p = 0.927) with specific adjuvant chemotherapy regimen. Additionally, both the SENP1 protein (high) (p = 0.003) and mRNA high (p = 0.028) were correlated with poor disease-free survival (DFS), while SENP1 protein high was also associated with shorter overall survival (OS) (p = 0.029). Furthermore, SENP1 protein (high vs. low) was independently associated with unsatisfying DFS [p = 0.009, hazard ratio (HR) = 1.798] and OS (p = 0.049, HR = 1.735).ConclusionSENP1 may serve as a potential biomarker to improve the management of patients with surgical NSCLC receiving adjuvant chemotherapy.

Abstract 5916: Tumor-informed patient-specific panel outperforms tumor-naïve and tumor-informed fixed panel for circulating tumor DNA (ctDNA)-based postoperative monitoring of non-small cell lung cancer (NSCLC)

Abstract Background: Patient-specific flexible gene panels designed based on whole-exome sequencing (WES) of resected tumor tissues is a promising strategy for ctDNA-based detection of molecular residual disease (MRD) in early-stage NSCLC. Flexible gene panels could potentially overcome the limitations of fixed panels by incorporating more unique genomic regions that might be absent in fixed panels; however, no study has reported a head-to-head comparison of these two approaches in postoperative disease monitoring. In this study, we investigated the clinical utility of a novel Patient-specific pROgnostic and Potential tHErapeutic marker Tracking (PROPHET) tumor-informed ctDNA assay. Using the same set of longitudinal blood samples, we further compared the performance of PROPHET assay with tumor-informed (TI) and tumor-naïve (TN) fixed panels for predicting MRD and prognosis in surgical NSCLC patients. Methods: Fifty-three patients with stage I-III resected NSCLC from the MEDAL study (NCT03634826) with adequate samples and median follow-up of 647 days were analyzed. Matched surgical tumor tissue and blood samples collected at various time points, including before surgery (baseline), 3-days (B) and 1-month (C) postoperative time points before any adjuvant therapy and subsequent follow-up time points (F) were analyzed. PROPHET assay involved four major steps: identify somatic mutations using WES, customized design of a patient-specific panel consisting of 50 single nucleotide variants, ultra-deep unique molecular-identifier-based next-generation sequencing (UMI-NGS) of serial blood samples using the patient-specific panel, and MRD risk prediction. Fixed panel assay of serial blood samples was performed using UMI-NGS with 168 gene panel spanning 273 kb of human genome. Results: At 1-month post-surgery, PROPHET assay accurately predicted MRD-positive cases among relapsed patients (50%, 13/26), whereas all disease-free patients were MRD-negative (100%, 21/21). Three-year prognostication with PROPHET assay at B+C yielded higher sensitivity (59% vs 26% vs 22%), negative predictive value (66% vs 51% vs 50%), and hazard ratio (7.15, 95%CI [3.2-15.9] vs 4.48 [1.9-10.9] vs 5.58 [2.1-14.7]) as compared with TI and TN fixed panel assays. Disease monitoring using PROPHET assay at B/C/F accurately predicted the MRD risk in 70% (21/30) of relapsed patients at a median lead time of 318 days (range: 20-751), whereas TI assay predicted 43% (13/30) at 282 days (range 20-716) and TN assay predicted 37% (11/30) at 282 days (range: 20-634). Conclusion: Patient-specific tumor-informed ctDNA-based postoperative monitoring enables risk stratification at early postoperative settings better than fixed panel, which paves an alternative strategy in the individualized management of surgical NSCLC patients. Citation Format: Kezhong Chen, Haifeng Shen, Shuailai Wu, Pengfei Zhu, Chenyang Wang, Analyn Lizaso, Guannan Kang, Yang Wang, Juan Lv, Shuai Fang, Wenjun Wu, Fujun Qiu, Yuan Sun, Qiang Lu, Heng Zhao, Shannon Chuai, Fan Yang, Zhihong Zhang. Tumor-informed patient-specific panel outperforms tumor-naïve and tumor-informed fixed panel for circulating tumor DNA (ctDNA)-based postoperative monitoring of non-small cell lung cancer (NSCLC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5916.

Postoperative ctDNA in indicating the recurrence risk and monitoring the effect of adjuvant therapy in surgical non–small cell lung cancers.

8533 Background: Circulating tumor DNA (ctDNA) has emerged as a potential novel biomarker to predict molecular residue disease in lung cancer after definitive treatment. Herein, we investigated the value of ctDNA in prognosing risk of relapse and monitoring the effect of adjuvant therapy in surgical non-small cell lung cancer (NSCLC) patients. Methods: Forty-one surgical NSCLC patients were enrolled. Tumor tissues were collected at surgery and subjected to targeted NGS of 1021 cancer-related genes. The serial peripheral blood samples were collected at postoperative one month and then at every third or sixth month and subjected to ultra-deep targeted NGS covering 338 genes. Results: From 41 eligible patients including 18 patients with stage Ⅰ disease, 2 with stage Ⅱ and 21 with stage Ⅲ, 41 tumor tissues and 137 plasma samples were enrolled and successfully tested. In tissue samples, 323 somatic variations were identified, with a median of 8 (range, 1-21) gene variations detected in each patient. TP53 was the most common mutation (63.41%), followed by EGFR (58.53%), LRP1B (17.07%) and KRAS (14.63%). The first-postoperative ctDNA positive was found in 13 of 41 patients (31.71%), 9 stage Ⅲ, 2 stage Ⅱ and 2 stage Ⅰ. During a median 9.47 months follow-up, 38.46% (5/13) patients with detectable ctDNA in the first postoperative blood sample experienced recurrence, while 3.57% (1/28) patients with undetectable ctDNA ultimately recurred. The DFS can be stratified by the first-postoperative ctDNA status, with ctDNA-positive groups having significantly reduced DFS (p &lt; 0.05). We detected ctDNA in at least one time point after surgery in 17 patients (41.46%), 5 of them (29.41%) experienced recurrence. Twenty-four patients without ctDNA detection during postoperative surveillance, one (4.17%) of them ultimately recurred. Serial ctDNA detection revealed disease recurrence ahead of radiologic imaging by a median of 5.25 months (range, 0.98-14.19). Among the 41 patients, 6 patients had surgery alone, 35 patients received adjuvant therapy. For these 35 patients, ctDNA analysis can stratify patients before and after adjuvant therapy. Recurrence ratio was 33.33% (4/12) in patients with detectable and 4.34% (1/23) in patients with undetectable ctDNA before adjuvant therapy. For ctDNA analysis after adjuvant therapy, no patients (11/11) with negative ctDNA had disease-recurrence while 33.33% (1/3) patients with positive ctDNA experienced recurrence. All four patients with clearance ctDNA by adjuvant therapy remained disease free. Conclusions: For NSCLC patients, postoperative ctDNA is a prognostic marker, which reveals disease recurrence ahead of radiographic examination. Importantly, ctDNA-detecting may facilitate personalized adjuvant therapy, and applying adjuvant therapy to the patients with detectable ctDNA could bring clinical benefits for them.

Prognostic value of pre-treatment advanced lung cancer inflammation index in non-small cell lung cancer: a meta-analysis

Purpose To identify the prognostic value of pre-treatment advanced lung cancer inflammation index (ALI) in non-small cell lung cancer (NSCLC) including surgical patients who were diagnosed with early stage. Methods The PubMed, EMBASE and Web of Science electronic databases were searched up to January 12, 2022 for relevant studies. The hazard ratios (HRs) with 95% confidence intervals (CIs) were combined to assess the association between pre-treatment ALI and overall survival (OS) or progression-free survival (PFS) of NSCLC patients. All statistical analyses were conducted by STATA 12.0 software. Results A total of 14 studies involving 3607 participants were included. The pooled results indicated that lower pre-treatment ALI was significantly related to poorer OS (HR = 2.20, 95% CI: 1.46–3.33, P < 0.001) and PFS (HR = 1.78, 95% CI: 1.49–2.13, P < 0.001). Besides, subgroup analysis also demonstrated that lower pre-treatment ALI was associated with worse OS in surgical (P < 0.001) and non-metastatic (P < 0.001) patients and worse PFS of surgical (P < 0.001) NSCLC patients. Conclusion Pre-treatment ALI was a novel and reliable prognostic indicator in NSCLC and lower pre-treatment ALI predicted worse survival including patients diagnosed with early stage. However, more prospective high-quality studies are still needed to verify the above findings.

Circulating tumor DNA predicts neoadjuvant immunotherapy efficacy and recurrence-free survival in surgical non-small cell lung cancer patients.

BackgroundThere is currently a lack of effective biomarkers to evaluate efficacy of neoadjuvant therapy (NAT) for resectable non-small cell lung cancer (NSCLC) patients. Circulating tumor DNA (ctDNA) has been investigated as a non-invasive tool for the assessment of tumor burden and minimal residual disease (MRD). The utility of ctDNA profiling in reflecting NAT efficacy, however, has not been confirmed. This study explored the association of ctDNA change with treatment response to NAT and recurrence-free survival (RFS) after surgery.MethodsEligible patients with stage IB–IIIA NSCLC were retrospectively included if they had received neoadjuvant immunotherapy combined with chemotherapy (IO+Chemo), dual immunotherapy (IO+IO), or chemotherapy alone (Chemo). We conducted ctDNA profiling before and after NAT, after surgery, and during follow-ups using an ultra-deep lung cancer-specific MRD (LC-MRD) sequencing panel.ResultsA total of 22 patients who received NAT followed by surgery between August 2018 and July 2019 were included in this study. The major pathological response (MPR) rates were 58.33% (7/12) in the IO+Chemo group, 25.00% (1/4) in the IO+IO group, and 16.67% (1/6) in the Chemo group. The ctDNA dynamics during NAT were highly concordant with pathologic response, demonstrating 100% sensitivity and 83.33% specificity, for an overall accuracy of 91.67%. Pre-surgery detectable ctDNA (after NAT) trended to correlate with inferior RFS [hazard ratio (HR), 7.41; 95% confidence interval (CI): 0.91–60.22, log-rank P=0.03]. At 3-8 days after surgery, ctDNA was detectable in 31.8% of patients and was an independent risk factor for recurrence (HR, 5.37; 95% CI: 1.27–22.67; log-rank P=0.01). The presence of ctDNA at 3 months after surgery showed 83% sensitivity and 90% specificity for predicting relapse (C-index, 0.79; 95% CI: 0.62–0.95). During disease monitoring after surgery, molecular recurrence by means of ctDNA preceded radiographic relapse, with a median time of 6.83 months.ConclusionsThis study investigated the potential of ctDNA in evaluating NAT efficacy in NSCLC, implying the high concordance between ctDNA and pathological response. We also set out the prognostic value of perioperative ctDNA in predicting recurrence.

Krüppel-Like Factor 6 Downregulation Is Connected with a Poor Prognosis and Tumor Growth in Non-Small-Cell Lung Cancer.

Purpose Research in this article was performed to explore the biological role and clinical significance of Krüppel-like transcription factor 6 (KLF6) in non-small-cell lung cancer (NSCLC). Methods KLF6 expression in NSCLC cell lines was analyzed using reverse transcription PCR and Western blot. The expressed KLF6 protein was examined in 50 surgical NSCLC tissues using immunohistochemistry. Statistical analyses were employed for clinical association examinations. CCK8 assay and Annexin V/PI analysis were used to execute cell proliferation and apoptosis in KLF6-overexpression cell lines and the control groups. Cleaved caspase-3 expression was also detected in KLF6-overexpression cells and NSCLC tissues. KLF6 expression correlation with cleaved caspase-3 was also examined. Results It was discovered that downregulation of KLF6 was seen in human NSCLC cell lines. Low KLF6 expression in NSCLC tissues was correlated with poor patient prognosis (P < 0.005); patients with less KLF6 expression possessed a lower cumulative 5-year survival rate. Multivariate analysis showed KLF6 expression as an independent prognostic indicator for NSCLC individuals. Expression levels of KLF6 were associated with NSCLC tumor size (P = 0.041). Overexpression of KLF6 inhibited cell proliferation and stimulated A549 and H322 cell line apoptosis. Cleaved caspase-3 protein had higher expression levels in KLF6-overexpressed cells than in the control group. The KLF6 expression levels were positively related to the cleaved caspase-3 protein expression in NSCLC tissues (r = 0.689, P = 0.001). Conclusions The results indicate that downregulation of KLF6 is a significant NSCLC progression marker. KLF6 prevents cell growth and promotes cell apoptosis, possibly caspase-3 activations.

NaPi2b expression in a large surgical Non-Small Cell Lung Cancer (NSCLC) cohort

BackgroundNaPi2b is a multi-transmembrane sodium-dependent phosphate transporter expressed at normal levels in several organs, including lung. High expression levels have been reported in various tumors including breast, thyroid, ovarian and non-small cell lung cancer. To date evaluation of NaPi2b expression has mostly been restricted to smaller lung cancer cohorts. MethodsAnalyses were performed on archival formalin fixed paraffin embedded primary tumor specimens from patients who had undergone curative intent resection at an Australian tertiary hospital. Tissue microarrays were constructed and stained with the chimeric anti-NaPi2b antibody, MERS67. Semi-quantitative H-scores (range 0 – 300) were calculated for each core tissue sample (H-score = % tumor cells staining for NaPi2b multiplied by staining intensity). An overall average H-score was reported for each specimen, with a cut-off score of 50 considered positive. ResultsOf 438 cases, high NaPi2b expression was observed in 151 (34.5%) overall, high expression in 137 of 208 (65.9%) adenocarcinoma cases, and 5 of 179 (2.8%) squamous cases (P < .0001). High NaPi2b expression was associated with female sex, EGFR or KRAS mutation, and TTF1 positivity (adenocarcinoma cases only). High NaPi2b expression was associated with improved overall survival (median 54 vs. 35 months, P = .029). ConclusionHigh NaPi2b expression was noted in a significant subset of adenocarcinoma cases, and in particular amongst those who were TTF1+, or exhibited EGFR or KRAS mutations. This agrees with earlier reports and highlights the significance that NaPi2b may have a role as a possible target for delivery of cytotoxic agents via antibody-drug conjugate models for some patients with lung adenocarcinoma.

Abstract 1710: A T-SIGn cancer gene therapy virus expressing an EpCAM-targeted bispecific T-cell activator stimulates T-cells and induces tumor cell death in patient-derived organotypic non-small cell lung cancer (NSCLC) spheroids ex vivo

Abstract Tumor-Specific Immuno-Gene (T-SIGn) cancer gene therapy vectors, which are based on the oncolytic adenovirus enadenotucirev, carry a gene cassette encoding combinations of human transgenes under control of the virus major late promoter (MLP) which restricts expression to tumor cells. These vectors can be dosed intravenously for delivery to tumors and selective expression of therapeutic proteins locally within the tumor microenvironment. Because these viruses only infect human tumor cells, in vivo studies are limited to human tumor xenografts in immunodeficient mice. Recently, we have optimized a 3D microfluidic system for evaluating response to immunotherapies using patient-derived organotypic tumor spheroids (PDOTS). PDOTS contain stromal as well as autologous myeloid and lymphoid cell populations and respond to PD-1 blockade in short-term ex vivo culture. As an alternative preclinical approach for evaluating activity and mechanistic underpinnings of T-SIGn viruses expressing human immunomodulatory transgenes, we have been developing the PDOTS ex vivo culture model. Here we report on the visualization of oncolytic virus uptake, replication in tumor cells and activation of T cells with a T-SIGn virus encoding an EpCAM-targeted bispecific T-cell activator (NG-611) within NSCLC PDOTS. Surgical NSCLC cases from St. Elizabeth's Medical Center, collected under an IRB approved protocol, were processed in PDOTS and assessed for EpCAM expression and number of T cells. Five different samples were treated with NG-611 (mixed with a GFP expressing vector) at concentrations ranging from 2x109 to 2x106 vp/mL final volume of culture. Virus infection was shown after 3 and 5 days of culture by dose-dependent expression of GFP reporter transgene driven by a CMV promoter, and virus replication in tumor cells was confirmed by using GFP expressed under the virus MLP promoter. Upregulation of IFNγ and Granzyme B in culture media indicated a strong activation of T-cells via expression of the bispecific T-cell activator. Cytotoxic T-cell activation also led to dose-dependent cell killing in NSCLC spheroids as shown by life/death analysis. Having established this proof of concept for functionality of T-SIGn viruses in PDOTS, ongoing studies are evaluating the immunomodulatory activities of a series of T-SIGn viruses expressing human IL-12 with or without other cytokine/chemokines. Citation Format: Moataz Noureddine, Katy West, Elizabeth Isaacson, Sylvia Alarcon, Christopher S. Lathan, John Wain, David A. Barbie, Cloud P. Paweletz, Elena Ivanova, Brian R. Champion. A T-SIGn cancer gene therapy virus expressing an EpCAM-targeted bispecific T-cell activator stimulates T-cells and induces tumor cell death in patient-derived organotypic non-small cell lung cancer (NSCLC) spheroids ex vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1710.

Monitoring HOTTIP levels on extracellular vesicles for predicting recurrence in surgical non-small cell lung cancer patients

In resected non-small cell lung cancer (NSCLC), postsurgical recurrence is the major factor affecting long-term survival. The identification of biomarkers in extracellular vesicles (EV) obtained from serial blood samples after surgery could enhance early detection of relapse and improve NSCLC outcome. Since EV cargo contains long non-coding RNAs (lncRNAs), we aimed to analyze whether the oncogenic lncRNA HOTTIP, which higher expression in tumor tissue was related to worse outcome in NSCLC, could be detected in EV from NSCLC patients and serve as recurrence biomarker. After purification of EVs by ultracentrifugation in 52 serial samples from 18 NSCLC patients, RNA was isolated and HOTTIP was quantified by Real time PCR. We observed that patients that relapsed after surgery displayed increased postsurgical EV HOTTIP levels in comparison with presurgical levels. In the relapsed patients with several samples available between surgery and relapse, we observed an increment in the EV HOTTIP levels when approaching to relapse, which indicated its potential utility for monitoring disease recurrence. When we focused in EV HOTTIP levels in the first post-surgical sample, we observed that the detection of an increment of the expression levels in comparison to presurgical sample, predicted recurrence with high sensitivity (85.7%) and specificity (90.9%) and that patients had shorter time to relapse and shorter overall survival. In conclusion, our pilot study showed that EV HOTTIP is a potential biomarker for monitoring disease recurrence after surgery in NSCLC.

Katanin P60: a potential biomarker for lymph node metastasis and prognosis for non-small cell lung cancer

BackgroundThis study aimed to assess the correlation of katanin P60 expression with clinical characteristics and survival profiles of surgical non-small cell lung cancer (NSCLC) patients.MethodsTwo hundred and sixty-five primary NSCLC patients treated by surgical resection were retrospectively viewed. The expression of katanin P60 in the tumor specimen was detected by the immunohistochemical (IHC) staining assay. Preoperative clinical data were collected from patients’ medical records, and survival data were extracted from follow-up records.ResultsThere were 127 (47.9%) and 138 (52.1%) patients with katanin P60-low expression and -high expression, respectively; in addition, patients presenting katanin P60-high+, -high++, and -high+++ expression were 62 (23.4%), 63 (23.8%), and 13 (4.9%), respectively. Katanin P60 expression was correlated with lymph node (LYN) metastasis and advanced TNM stage but not pathological grade, tumor size, carcinoembryonic antigen (CEA) level or other non-tumor features in NSCLC patients. Regarding survival profiles, disease-free survival (DFS) and overall survival (OS) were both the lowest in katanin P60-high+++ expression patients, followed with katanin P60-high++ patients, katanin P60-high+ patients, and the highest in katanin P60-low expression patients. Further analysis illustrated that katanin P60-high expression was an independent predictive factor for unfavorable DFS and OS in NSCLC patients.ConclusionsKatanin P60 presents potential as a biomarker for lymph node metastasis and prognosis in NSCLC patients.

Abstract A043: NaPi2b expression in a large surgical non-small cell lung cancer cohort

Abstract Background: NaPi2b (SLC34A2) is a sodium-dependent phosphate transporter expressed in a high proportion of non-small cell lung cancer (NSCLC), especially adenocarcinoma, and ovarian carcinoma cases, as well as other tumour types. A clinical trial of the NaPi2b-targeting antibody drug conjugate XMT-1536 is currently underway. This study investigates NaPi2b expression in NSCLC emphasising correlations to histology. Methods: Tissue Microarrays (TMAs) were constructed using triplicate 1mm cores from FFPE primary NSCLC tumours resected by lobectomy or pneumonectomy with curative intent at Austin Health, Melbourne, Australia. Pathology review identified the best site for coring of tumour blocks. For IHC, TMAs were stained with an anti-human NaPi2b primary antibody (MERS67) on the Leica Bond RX auto-stainer and the H-score (0 – 300) calculated based on the percentage of tumour cells stained multiplied by the staining intensity (0 - 3+). The mean score of multiple cores was calculated. A high H-score was defined as ≥50. Results: High NaPi2b expression was seen in 153/439 (35%) of cases. High NaPi2b expression was determined in 132/215 (61%) of adenocarcinoma cases, while only 12/178 (7%) of squamous cell and 9/46 (20%) of other histology cases (p &amp;lt; 0.001, chi-square) had high expression. Considering adenocarcinoma subtypes, high NaPi2b was seen with 18/21 (86%) micropapillary, 35/61 (57%) solid, 3/13 (23%) mucinous, 54/84 (64%) acinar, 8/13 (62%) papillary, 9/9 (100%) lepidic and 3/9 (33%) adenosquamous. Overall high NaPi2b was seen in 56/95 (59%) with poor risk adenocarcinoma subtypes (micropapillary, solid, mucinous) and 71/106 (67%) with good risk subtypes (papillary, acinar, lepidic). High scores were more frequent for females (52% vs 27%; p &amp;lt; 0.001), never smokers vs smokers (63% vs 33%; p &amp;lt; 0.001), EGFR mutation vs wt (75% vs 22%; p &amp;lt; 0.001) and KRAS mutation vs wt (71% vs 22%; p &amp;lt; 0.001). There were no significant differences in expression considering stage, primary tumour diameter, FEV1 (% predicted) or level of PD-L1 expression. For the whole cohort, a high NaPi2b score was associated with improved overall survival on univariate analysis (median 58 months vs 36 months; HR 0.77 [95% CI 0.61-0.97], p=0.03), while for adenocarcinoma there was a trend for improved survival (median 54 months vs 35 months; HR 0.76 [95% CI 0.55-1.04], p=0.09), and for both high risk adenocarcinoma subtypes (micropapillary, solid) and good risk subtype (acinar), high NaPi2b expression was associated with a weak trend to better survival. On multivariable analysis, NaPi2b remained the strongest predictor of improved survival (HR 0.77, 95% CI 0.58 - 1.02, p=0.07), though stage remained the only statistically significant variable. Conclusion: In this large early stage surgical NSCLC cohort, a high level of NaPi2b expression was seen with adenocarcinoma histology and also across the range of adenocarcinoma subtypes. Female gender, never smokers, and EGFR mutations were independently predictive of high level expression. For the whole cohort, high level expression was associated with improved overall survival. There was a similar, but non-significant, trend for high level expression in adenocarcinoma cases to be associated with improved survival, which appeared to be the case for both high risk and good risk adenocarcinoma subtypes. Citation Format: Paul L Mitchell, Hui Yu, Gareth Rivalland, Khashayar Asadi, Rebecca Mosher, Fred Hirsch, Christopher Rivard. NaPi2b expression in a large surgical non-small cell lung cancer cohort [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A043. doi:10.1158/1535-7163.TARG-19-A043

T cell immunoglobulin and mucin-domain containing-3 in non-small cell lung cancer.

Immunotherapy has shown promising effect for non-small cell lung cancer (NSCLC) patients. Yet the biomarkers for predicting immunotherapy efficiency are still lacking. We tested 139 surgical resected NSCLC primary tumor samples from Medical University of Gdansk, Poland, for T cell immunoglobulin and mucin-domain containing-3 (TIM-3) level by immunohistochemistry (IHC), analyzed the expression of TIM-3 protein on NSCLC tumor cells and tumor infiltrating lymphocytes (TILs). TIM-3 on TILs was correlated with programmed cell death protein-1 (PD-1) on TILs (correlation coefficient =0.346, P<0.001) and programmed cell death protein-ligand 1 (PD-L1) on TILs (correlation coefficient =0.313, P<0.001), PD-L1 level on tumor cells (correlation coefficient =0.255, P=0.002), TIM-3 level on tumor cells (correlation coefficient =0.262, P=0.002) and TIL percentage (correlation coefficient =0.172, P=0.043). TIM-3 level on tumor cells only had correlation with PD-L1 level (correlation coefficient =0.170, P=0.045). High level of TIM-3 on TILs indicated shorter recurrence-free survival (RFS) and overall survival (OS) (RFS 1.800 years, 95% CI, 1.230-2.370 vs. 0.870 years, 95% CI, 0.212-1.528, P=0.048) (OS 2.960 years, 95% CI, 2.268-3.652 vs. 1.080 years, 95% CI, 0.228-1.932, P=0.034). TIM-3 is expressed on NSCLC tumor cells and TILs in all NSCLC pathological type. TIM-3 level on TILs had correlation with PD-1 and PD-L1 level. NSCLC patients with high TIM-3 level on TILs were more likely to have poor prognosis.