Treatment Of NSCLC Research Articles

Potential of semen coicis in enhancing the anti-tumor effects of PD-1 inhibitor on A549 cell lines by blocking the PI3K-AKT-mTOR pathway.

The objective of this research was to investigate how the combination of semen coicis extract and PD-1 inhibitors can potentially work together to enhance the anti-tumor effects, with a focus on understanding the underlying mechanism. We obtained the active components and specific targets of semen coicis in the treatment of NSCLC from various databases, namely TCMSP, GeneCard, and OMIM. By utilizing the STRING database and Cytoscape software, we established a protein interaction network (PPI) for the active ingredient of semen coicis and the target genes related to NSCLC. To explore the potential pathways involved, we conducted gene ontology (GO) and biological pathway (KEGG) enrichment analyses, which were further supported by molecular docking technology. Additionally, we conducted cyto-inhibition experiments to verify the inhibitory effects of semen coicis alone or in combination with a PD-1 inhibitor on A549 cells, along with examining the associated pathways. Furthermore, we investigated the synergistic mechanism of these two drugs through cytokine release experiments and the PD-L1 expression study on A549 cells. Semen coicis contains two main active components, Omaine and (S)-4-Nonanolide. Its primary targets include PIK3R1, PIK3CD, PIK3CA, AKT2, and mTOR. Molecular docking experiments confirmed that these ingredients and targets form stable bonds. In vitro experiments showed that semen coicis demonstrates inhibitory effects against A549 cells, and this effect was further enhanced when combined with PD-1 inhibitors. PCR and WB analysis confirmed that the inhibition of the PI3K-AKT-mTOR pathway may contribute to this effect. Additionally, semen coicis was observed to decrease the levels of IFN-γ, IL-6, and TNF-α, promoting the recovery of the human anti-tumor immune response. And semen coicis could inhibit the induced expression of PD‑L1 of A549 cells stimulated by IFN‑γ as well. Semen coicis not only has the ability to kill tumor cells directly but also alleviates the immunosuppression found in the tumor microenvironment. Additionally, it collaboratively enhances the effectiveness of PD-1 inhibitors against tumors by blocking the activation of PI3K-AKT-mTOR.

Is it too early to recommend local treatment in oligometastatic non-small cell lung cancer: a plea for equipoise.

Oligometastatic non-small cell lung cancer (OMD NSCLC) has been proposed to bridge the spectrum between non-metastatic and widely metastatic states and is perceived as an opportunity for potential cure if removed. Twelve clinical trials on local treatment have been reported, yet none are conclusive. These trials informed the development of a joint clinical practice guideline by the American & European Societies for Radiation Oncology, which endorses local treatment for OMD NSCLC. However, the heterogeneity between prognostic factors within these trials likely influenced outcomes and can only support guidance at this time. Caution against an uncritical acceptance of the guideline is discussed, as strong recommendations are offered based on expert opinion and inconclusive evidence. The guideline is also examined by a patient's caregiver, who emphasizes that uncertain evidence impedes shared decision making.

PD-1 Blockade in Treatment of NSCLC: A Comprehensive Review of Clinical Efficacy, Safety and Cost-effective

Non-small cell lung cancer, also called NSCLC, makes up 85% incidence in lung cancer, presents a considerable challenge for its high mortality rate and poor prognosis. In recent years, the invention of programmed cell death 1 (PD-1) inhibitor nivolumab, which is a type of immune checkpoint inhibitors, emerging as a groundbreaking drug, has offered a potential solution to tackle the inescapable drawbacks of traditional cancer treatments. Several clinical trials assessing nivolumab's efficacy and safety in the form of monotherapy or combination therapy, demonstrated that nivolumab has remarkably increased the progression-free survival and overall survival for NSCC, in both first- and second-line. Notably, compared to chemotherapy drugs, nivolumab showed a favorable safety profile with fewer immune-related adverse events. Given the high cost and escalating use of nivolumab, finding more cost-effective treatment regimen is imperative. The purpose of this review is to delve into the clinical efficacy and safety of PD-1 inhibitor nivolumab as immunotherapy remedy against NSCLC, providing some therapeutic recommendations for cost-effective. While continued research and development are vital to further enhance nivolumab's potential as a more potent tool to combat cancer.

Abstract 3892: Systematic generation of a clinicogenomic harmonized oncologic real-world dataset (MSK-CHORD)

Abstract Clinical data storage in unstructured notes and siloed datasets present a major challenge for large-scale cancer informatics. Whether natural language processing (NLP) combined with multimodal integration across datasets can produce a mineable resource and improve discovery of relationships between tumor genomics and clinical phenotypes is unknown. We hypothesized that NLP could automatically annotate a pan-cancer corpus of 82,464 patients with tumor genomic sequencing. To develop algorithms to annotate free-text reports, we leveraged the AACR Project GENIE Biopharma Collaborative (BPC), a structured curation of EMR from five cancer types (non-small cell lung (NSCLC), breast, colorectal, prostate, and pancreatic cancer), to train and validate several Transformer and rule based-based NLP models. After automating the generation of NLP annotations alongside medication, demographic, tumor registry, survival, and tumor genomic sequencing data, we tested whether clinicogenomic relationships not apparent in the smaller BPC cohort might be discoverable in the larger cohort. In 5-fold cross-validation, NLP Transformers accurately annotated the presence of cancer (AUC=0.99), cancer progression (AUC=0.97), and sites of disease (AUC=0.99) from radiology reports, and presence of prior outside treatment (AUC=0.98) and hormone receptor (HR) and HER2 receptor status (AUC=0.98, 0.98) from clinician notes. In addition, rule-based models, trained on non-BPC data and validated on the whole BPC cohort, annotated smoking status from clinician notes (ACC=0.95), and Gleason score (ACC=1.0), PD-L1 status (ACC=0.98), and mismatch repair deficiency (ACC=0.98) from histopathology reports. NLP annotations were merged with genomic and other structured clinical data to create a Clinicogenomic, Harmonized Oncologic Real-world Dataset (MSK-CHORD). Finally, we tested if associations not apparent in the BPC might be discoverable in MSK-CHORD. We found positive associations between Gleason score and gene-level alterations in prostate cancer including TP53, PTEN and BRCA2 (q<0.1), none of which were adequately powered for detection in the BPC. We found PD-L1 status was associated with better survival following immunotherapy treatment in NSCLC, but only in the larger MSK-CHORD was this association statistically significant. In breast cancer, NF1 mutations were associated with prior therapy in both cohorts, but this association was only significant in MSK-CHORD. The infrastructure generating MSK-CHORD uses a combination of on-premise and cloud computing resources and open-source development operation applications to automate processes. Once annotations are created, data is imported into a local instance of cBioPortal, where researchers can visualize data and perform analyses. The system generating MSK-CHORD demonstrates how large-scale data delivery and integration can fuel cancer research. Citation Format: Christopher J. Fong, Karl Pichotta, Thinh Tran, Michele Waters, Tom Fu, Mono Pirun, Mirella Altoe, Brooke Mastrogiacomo, Anisha Luthra, Mehnaj Ahmed, Arfath Pasha, Armaan Kohli, Raymond Lim, Tom Pollard, Darin Moore, Benjamin Gross, Avery Wang, Calla Chennault, Ritika Kundra, Ramyasree Madupuri, Ino de Bruijn, Aaron Lisman, Walid K. Chatila, Subhiksha Nandakumar, Anika Begum, Doori Rose, Kenneth L. Kehl, Deborah Schrag, Michael Berger, Jian Carrot-Zhang, Pedram Razavi, Bob Li, Peter Stetson, Nikolaus Schultz, Justin Jee. Systematic generation of a clinicogenomic harmonized oncologic real-world dataset (MSK-CHORD) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3892.

Abstract 7162: Killing a fly with a sledgehammer: standard flat-dosing administration of Atezolizumab leads to marked overexposure in real-world lung cancer patients

Abstract Atezolizumab is an anti-PDL1 approved for treating small cell lung cancer and non-small cell lung cancers. A threshold of 6 µg/ml in plasma has been associated with target engagement with Atezolizumab. The extent to which patients could be overexposed with the standard 1200 mg Q3W flat-dosing remains unknown, but several in silico studies have suggested that flat-dosing with immune checkpoint inhibitors could lead to plasma concentrations far above the necessary levels to be efficacious. Here, we monitored Atezolizumab peak and trough levels in 27 real-world patients with lung cancer as part of routine therapeutic drug monitoring. Individual pharmacokinetic parameters were calculated using a population approach and optimal dosing-intervals were simulated with respect to the target trough levels of Atezolizumab. No patient displayed plasma levels below 6 µg/ml. The results showed that the mean trough level after the first course was 78.3 ± 17 µg/ml, i.e., 13 times above the target concentration associated with target engagement. The overall response rate was 55.5%. Low-grade IRAEs was observed in 37% of patients. No severe immune-related toxicities was observed. On univariate analysis, no relationship was found between exposure metrics of Atezolizumab (i.e., Cmin, Cmax, AUC at the first cycle or during the forthcoming courses) and pharmacodynamic endpoints (i.e., efficacy, toxicity). First-line Atezolizumab treatment and treatment for NSCLC were both significantly associated with efficacy on univariate analysis (p=0.0067 and p=0.0497, respectively), but further multivariate analysis did not confirm this relationship (p=0.2465 and p=0.9981). Smoking status was moderately associated with toxicities on univariate analysis (p=0.03), but this effect was not confirmed on further multivariate analysis (p=0.99). None of the other covariates (i.e., age, sex, NLR, number of metastatic sites, associated chemotherapy) were associated with efficacy or toxicity. Further simulations suggested that the dosing interval could be extended from the standard 21 days (i.e., Q3W) to 49 up to 136 days (mean: 85.7 days, i.e., Q12W dosing), while ensuring plasma levels always above the 6 µg/ml target threshold. This observational, real-world study confirms for the first time in real-world patients that the standard 1200 mg Q3W fixed-dose regimen of Atezolizumab results in significant overexposure in plasma. Of note, this overexposure was not associated with increased side-effects. As plasma levels always exceed pharmacologically active concentrations, the inter-individual variability we observed in PK parameters did not impact efficacy and could not explain differences in response. Our data suggest that dosing intervals could be markedly extended with respect to the target threshold associated with efficacy, thus lowering drug-cost in the future. Citation Format: Sophie Marolleau, Alice Mogenet, Mourad Hamimed, Clara Boeri, Laurent Greillier, Joseph Ciccolini. Killing a fly with a sledgehammer: standard flat-dosing administration of Atezolizumab leads to marked overexposure in real-world lung cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7162.

Abstract 4666: HC-7366, a potent GCN2 kinase activator, augments osimertinib therapy to delay resistance in EGFR mutant NSCLC models

Abstract Despite the efficacy seen with EGFR tyrosine kinase inhibitors (TKIs) in the treatment of EGFR mutant NSCLC, the long-term prognosis remains unfavorable due to resistance. Osimertinib is used as the first- and second-line treatment for EGFR exon 19 deletion, L858R mutation, and acquired T790M mutation. Resistance is driven by both EGFR-dependent and independent events, such as gene amplification and upregulation of HIF1α. Metabolic events also drive resistance by increasing oxidative phosphorylation (OXPHOS) caused by osimertinib-induced glycolysis inhibition. HiberCell has developed HC-7366, a GCN2 activator currently being investigated in a Ph1 clinical trial in patients with solid tumors (NCT05121948). GCN2 activation by HC-7366 drives the integrated stress response (ISR) by inducing the expression of ATF4. This results in translation inhibition, cell cycle arrest, HIF1α suppression, and apoptosis in cancer cells. Our preclinical data suggest HC-7366 can reduce glycolysis, TCA cycle metabolites, and OXPHOS, thereby causing mitochondrial dysregulation. We hypothesized that combination of HC-7366 with osimertinib could reduce resistance by evading the adaptive resistance mechanisms. We treated the NCI-H1975 xenograft model with HC-7366, a clinically relevant dose of osimertinib, and the combination for 21 days and then followed for tumor regrowth. At day 21, the HC-7366 group showed efficacy with tumor growth inhibition (TGI) of 55%, while osimertinib resulted in tumor regression of 97% with 8/10 CRs and 2/10 PRs. The combination had tumor regression of 99% with 10/10 CRs. After treatment cessation for 74 days, tumors in the osimertinib group quickly regrew with 6/10 tumors progressing and 4/10 mice tumor-free. In contrast, the combination group had only 2/10 tumors regrow with 8/10 mice tumor-free. In an EGFR-dependent resistance model NCI-H1975 (EGFR C797S), combination achieved 63% TGI versus 31% with osimertinib. In an EGFR-independent resistant PDX with EGFR exon 19 deletion, combination benefit was seen at 70% TGI compared to 40% for osimertinib, while a PDX with EGFR L858R showed a trend for combination benefit. To understand the mechanism of action, we examined osimertinib-resistance pathways to determine how GCN2 activation could affect them. Combination with HC-7366 reduced the expression of short-lived proteins involved in osimertinib resistance, including HIF1α, MET, cyclin D1, and CDK4. Combination decreased TIM17A and TIM23, proteins essential for mitochondrial protein translocation, potentially causing mitochondrial dysfunction. We found a proliferation reduction by Ki67 and an increase in apoptotic proteins CHOP, PUMA, and cleaved caspase 3. Taken together, these data reveal that combination of HC-7366 and osimertinib, and potentially other EGFR TKIs, may be an effective treatment strategy at targeting adaptive resistance mechanisms. Citation Format: Crissy Dudgeon, Sho Fujisawa, Olivia Funk, Feven Tameire, Takashi O. Kangas, Weiyu Zhang, Eric S. Lightcap, David Surguladze, Nandita Bose. HC-7366, a potent GCN2 kinase activator, augments osimertinib therapy to delay resistance in EGFR mutant NSCLC models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4666.

Abstract 2492: Enhanced patient selection for anti-PD-L1 treatment in metastatic NSCLC with quantitative continuous scoring of PD-L1

Abstract Background: Anti-PD-L1 therapy has demonstrated clinical activity in patients with metastatic non-small cell lung cancer (mNSCLC). However, only subgroups of patients respond and their identification via PD-L1 as a biomarker remains imperfect. PD-L1 expression is commonly assessed by pathologist tumor cell (TC) scoring of immunohistochemically (IHC) stained tissue. We developed a system for digitally scoring PD-L1 in IHC (PD-L1 QCS), which demonstrated robust scoring across studies [1]. Here, we present a comparison of PD-L1 QCS against manual scoring of PD-L1 (SP263 assay, Ventana) in the MYSTIC clinical trial. Methods: PD-L1 QCS on digitized whole slide images (WSI) comprises two deep learning models, enabling segmentation of single TCs followed by PD-L1 expression quantification via their optical density (OD). Positive cells are classified based on an OD threshold, allowing robust digital calculation of the TC percentage [1]. The analysis included 502 WSI from the MYSTIC trial (NCT02453282), representing 256 patients treated with anti-PD-L1 therapy and 246 treated with chemotherapy as standard-of-care (SoC) [2]. First, an optimal cut-point was determined by optimization against outcome, classifying patients with ≥0.575% TC as biomarker positive (BM+). Next, the approach was compared against manual TC scoring at 1%, 25% and 50% cut-off. Results: In durvalumab treated patients, median overall survival (mOS) in the PD-L1 QCS BM+ subgroup (prevalence 54.3%) was 12.1 months longer than in the BM- subgroup (19.9m vs. 7.8m, HR=0.45, CI [0.33, 0.60]). Analogous comparison of subgroups based on manually scored TC proportion at 1% (prev. 75.0%), 25% (prev. 42.6%) or 50% (prev. 29.7%) cut-points yielded a mOS difference of 7.8m (HR=0.52, CI [0.38, 0.72]), 8.3m (HR=0.61, CI [0.45, 0.82]) and 11.0m (HR=0.55, CI [0.40, 0.77]) respectively. Comparing durvalumab treatment against SoC within the PD-L1 QCS BM+ subgroup yielded a HR of 0.62 (CI [0.46, 0.82], log rank p=0.0008, 7.4m mOS delta). In comparison, a HR of 0.69 (CI [0.46,1.02], p=0.0642, 8.4m mOS delta) was obtained for manual TC scoring at 50%. Conclusion: We compared a computational pathology approach for continuous PD-L1 scoring for the selection of mNSCLC patients for anti-PD-L1 treatment against established manual scoring. Our results suggest that PD-L1 QCS has the potential to identify a larger patient subgroup that retains benefit from anti-PD-L1 treatment and more precisely identifies non-responders.

Abstract 5857: Trastuzumab deruxtecan resistance can be mediated by payload resistance or secondary extracellular ERBB2 mutations but sensitivity to HER2 tyrosine kinase inhibitors is maintained

Abstract Among NSCLC patients, the most common HER2 mutation is the exon 20 insertion mutation, Y772dupYVMA, which accounts for over 40% of all HER2 mutations in lung cancer. Trastuzumab deruxtecan (T-DXd), a HER2 antibody-drug conjugate (ADC), received FDA approval for the treatment of HER2 mutant NSCLC yielding a confirmed objective response of 52% and a median duration of response of greater than 9 months. Unfortunately, patients that initially respond to T-DXd will eventually acquire resistance, and the mechanisms of resistance as well as effective targeting strategies to overcome resistance are not yet elucidated. We generated Ba/F3 cells expressing the HER2 YVMA insertion mutation with acquired resistance to T-DXd by culturing cells in T-DXd until resistance occurred. T-DXd resistant cells retained expression of HER2, but were resistant to the payload, deruxtecan, as well as other topoisomerase inhibitors. T-DXd resistant cells exhibited loss of topoisomerase I, a previously reported mechanism of topoisomerase inhibitor resistance. In parallel, we generated a PDX model of acquired T-DXd resistance and observed sustained expression of HER2 but loss of TOP1 in resistant tumors. T-DXd resistant cell lines were sensitive to payloads with alternate mechanisms of action including maytansine and likewise retained sensitivity to the HER2 ADC trastuzumab emtansine (T-DM1) which utilizes DM1 as a payload. Given that T-DXd resistant cells retained HER2 expression, we assessed whether they were sensitive to HER2 tyrosine kinase inhibitors (TKI). T-DXd resistant cells remained highly sensitive to HER2 TKIs including poziotinib, afatinib, BI-4142, and BI-1622. To investigate whether genomic alterations of HER2 could facilitate HER2 ADC resistance, we utilized the LentiMutate approach which employs an error-prone HIV-1 reverse transcriptase to produce a high frequency of mutations to identify resistance-associated mutations in cells treated with a HER2 ADC. Sequencing analysis revealed that resistant cells had an enrichment in point mutations within HER2 domain IV (D582N, F595C/S, E580K, C623Y), which includes the binding site of trastuzumab. To validate the impact of these mutations on HER2 ADC resistance, we generated cells expressing a HER2 exon 20 insertion in combination with the observed domain IV mutations. D582N and F695C/S co-mutations conferred resistance to T-DXd and T-DM1 but not to HER2 TKIs. These data demonstrate that resistance to T-DXd can be mediated by loss of sensitivity to the ADC payload in which case ADCs bearing payloads with alternate mechanisms of action as well as HER2 TKIs retain anti-tumor cell activity. Moreover, resistance to trastuzumab-based ADCs can also be mediated by secondary mutations within domain IV of HER2. However, these resistance mutations do not diminish HER2 TKI activity. Citation Format: Monique B. Nilsson, Xiuning Le, Junqin He, Xiaoxing Yu, Xiaofang Huo, Ashwani Kumar, Alissa Poteete, Qian Huang, Ralf Kittler, John Heymach. Trastuzumab deruxtecan resistance can be mediated by payload resistance or secondary extracellular ERBB2 mutations but sensitivity to HER2 tyrosine kinase inhibitors is maintained [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5857.

Abstract 1871: Discovery and characterization of NXV01c, an EGFR × cMET bispecific nanobody drug conjugate with potent anti-tumor activity

Abstract EGFR and cMET are proven cancer targets co-expressed in diverse tumor types. EGFR × cMET bispecific antibody has been approved for the treatment of NSCLC, supporting a simultaneous targeting strategy. In addition to this dual targeting benefit, bispecific antibody drug conjugates (ADCs) targeting EGFR and cMET have also been developed to further improve anti-tumor activity and tissue selectivity. Sufficient target affinity, good cellular internalization and high tumor infiltration are critical for an ADC to mediate therapeutic activity. To this end, we developed the first EGFR × cMET bispecific nanobody conjugated with monomethyl auristatin E (MMAE) as payload (NXV01c). Lead nanobodies against EGFR and cMET were respectively selected from immune libraries by phage display, followed by highly efficient humanization and optimization by neoX’s computation platform. The bispecific nanobody with Fc (NXV01) was constructed by “knobs-into-holes” heterodimerization and then homogeneously conjugated with MMAE via a lysosomal cleavable valine-citrulline dipeptide linker. The resulting bispecific nanobody drug conjugate (NDC), NXV01c, was evaluated in multiple tumor cell lines and tumor xenograft models. NXV01, the pre-conjugate bispecific nanobody, bound EGFR and cMET with nanomolar potency (BLI) and inhibited the phosphorylation of cellular EGFR and cMET with nanomolar IC50 (ELISA). Moreover, it did not activate cellular cMET. Importantly, the NDC NXV01c is highly homogeneous: it has an average DAR of 3.87 and >95% of NXV01c has a DAR of 4. To examine stability of conjugation, NXV01C was incubated in human plasma (37°C) for 14 days, the maximum free drug release rate (by LC/MS/MS) is 0.68%, which is much lower than that of DS8201. In vitro, NXV01c was rapidly internalized into H1975 cells which co-expressed EGFR and cMET. It inhibited the growth of H1975 (lung) and SNU5 (gastric) cancer cells with picomolar activity but spared normal keratinocytes. NXV01c also inhibited the proliferation of additional cell lines derived from lung, gastric, esophageal, and liver cancer, and the level of inhibition positively associated with the expression density of both targets. In an H1975 cell line-derived xenograft model, NXV01C exhibited potent and dose-dependent anti-tumor activity. Treatments at 3 and 10 mg/kg once per week for 2 weeks led to shrinkage and complete regression of tumor, respectively. In patient-derived xenograft models of NSCLC and esophageal cancer, NXV01C led to tumor regression and elimination without noticeable toxic effects. EGFR × cMET bispecific nanobody drug conjugate NXV01c has favorable drug-like properties and demonstrated superior anti-tumor effect in vitro and in vivo. The results suggest NXV01C can be an effective solution for EGFR/cMET bearing tumors commonly found in diverse malignancies. Citation Format: Ran Wu, Puwei Yuan, Yang Xie, Jianxiu Guo, Fei Zhang, Fan Liu, Taylor B. Guo. Discovery and characterization of NXV01c, an EGFR × cMET bispecific nanobody drug conjugate with potent anti-tumor activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1871.

Abstract 6360: A novel CAR-NK cell therapy to target lung adenocarcinoma cells

Abstract Cell Adhesion Molecule (CADM1) is a Bonafede tumor suppressor with 40% of NSCLC tumors lose CADM1 expression either by loss of heterozygosity or promoter hypermethylation. The tumor suppressive ability of CADM1 is attributed to its cell adhesion function. However, we previously demonstrated that CADM1 also mediates its tumor suppression through regulation of NK-mediated immunosurveillance. In tumors that do not lose CADM1, it is expressed on cell surface and up regulated during TGF-β-induced EMT making it a biomarker for potential therapeutic targeting. Here we report the design and development of T and NK specific CAR that would specifically target lung cancer cells in CADM1 dependent manner. Anti-CADM1 single chain variable fragment (CADM1-scFv) was fused to NK or T cell specific transmembrane and co-stimulatory domains. The resulting CAR constructs were cloned into a retroviral vector, from which viral stocks were generated to transduce human and mouse cells. Next, we introduced anti-CADM1-CAR-NK construct into NK92 cell line and tested it in a cytotoxicity assay using A549-GFP-ffluc cell line as a target. We observed a significant increase in the killing of target cells with anti-CADM1-CAR-NK cells compared with non-transduced NK92 cells. This effect was significantly reduced when A549- GFP-ffluc-CADM1 KO cells were used as target. Next, we generated anti-CADM1-CAR-T cells by transducing human CD3+ T-cells purified from human PBMCs. Similar to anti -CADM1-CAR-NK, anti-CADM1-CAR-T cells showed increased T-cell specific cytotoxicity in a CADM1-dependent manner. Next, we evaluated the efficacy of anti-CADM1-CAR-T cells through adoptive transfer into NSG mice xenografted with human lung adenocarcinoma cells into mouse lungs. The administration of anti-CADM1-CAR-T cells markedly reduced tumor size and enhanced the survival rate of the mice. Importantly this effect was dependent on CADM1 expression, as mice xenografted with human CADM1 KO cells showed neither tumor size reduction nor increased survival. Finally, using IHC staining, we demonstrated that anti-CADM1-CAR-T cells directly target xenografted tumors in the mouse lungs. In conclusion, our findings indicate that CAR-NK and CAR-T cells targeting CADM1-expressing lung tumors have a significant therapeutic potential for the treatment of NSCLC. Citation Format: Sergey Zolov, Sergei Chuikov, Shiva Krishna Katkam, Delaney Shield, Venkateshwar G. Keshamouni. A novel CAR-NK cell therapy to target lung adenocarcinoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6360.

Abstract 7442: Exploring the role of chinese herbal medicine in adjunctive therapy for non-small cell lung cancer: A systematic review and meta-analysis

Abstract Non-small cell lung cancer (NSCLC) treatment traditionally relies on Platinum-based chemotherapy along with targeted therapy and immunotherapy. However, the emergence of drug resistance due to genetic factors and the progression of late-stage metastasis pose significant challenges. In Asian medical practices, the integration of Chinese herbal medicine (CHM) alongside conventional therapy has shown promise in improving treatment efficacy, ensuring safety, and mitigating adverse effects. This study aims to comprehensively summarize the prevalence and efficacy of CHM when combined with platinum-based chemotherapy in managing advanced NSCLC. Our analysis encompasses data from 14 randomized controlled trials involving 1451 patients, of which 739 received combined chemotherapy with CHMs while 712 received chemotherapy alone. Notably, the adjunctive use of traditional CHM demonstrated a marked increase in overall survival rates. Patients with NSCLC who received CHMs in combination with chemotherapy exhibited significantly higher Objective Response Rate (ORR) (RR = 1.37, 95% CI [1.20-1.58], p = .000) and Disease Control Rate (DCR) (RR = 1.13, 95% CI [1.07-1.21], p = .000) compared to those solely undergoing chemotherapy. The Karnofsky Performance Status (KPS) (SMD = 1.47, 95% CI [0.30-2.64], p = 0.014) also indicated substantial improvements in the quality of life assessment, involving a total of 277 cases. The systematic review identified 88 herbs and formulas, classified into two primary categories. The first category involves tonifying qi and nourishing yin, thereby enhancing the body's immune function and boost immune therapy, fortifying its ability to combat pathogens, and inhibiting cancer cell proliferation. Herbs such as Rx. Astragali (Huangqi), Rz. Atractylodis Macrocephalae (Baizhu), Rx. Glehniae (Shashen), and Rx. Ophiopogonis (Maidong) were among the most frequently prescribed CHMs. The second category consists of heat-clearing and toxin-removing medicines, effectively regulating the inflammatory response and infection around tumors while exhibiting potent anti-tumor activity, such as Herba Salviae Chinensis (ShiJianChuan) and Selaginella Doederleinii (ShiShangBai). The evidence derived from the meta-analysis strongly suggests that adjunctive CHM treatment can significantly enhance chemotherapy's effectiveness and improve tumor response. Subsequent research focusing on the development and application of CHM is warranted for the advancement of NSCLC treatment methodologies. Citation Format: Li-Chi Wang, Tai-Lin Chen, Li Wei Chen, Ming-Yen Tsai. Exploring the role of chinese herbal medicine in adjunctive therapy for non-small cell lung cancer: A systematic review and meta-analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7442.

The Tumor Immune Microenvironment Is Associated With Recurrence in Early-Stage Lung Adenocarcinoma

IntroductionImmune checkpoint inhibitors have recently been approved for the treatment of early-stage NSCLC in the perioperative setting on the basis of phase 3 trials. However, the characteristics of such patients who are susceptible to recurrence after adjuvant chemotherapy or who are likely to benefit from postoperative immunotherapy have remained unclear. MethodsThis biomarker study (WJOG12219LTR) was designed to evaluate cancer stem cell markers (CD44 and CD133), programmed death-ligand 1 (PD-L1) expression on tumor cells, CD8 expression on tumor-infiltrating lymphocytes, and tumor mutation burden in completely resected stage II to IIIA NSCLC with the use of archived DNA and tissue samples from the prospective WJOG4107 trial. Tumors were classified as inflamed or noninflamed on the basis of the PD-L1 tumor proportion score and CD8+ tumor-infiltrating lymphocyte density. The association between each potential biomarker and relapse-free survival (RFS) during adjuvant chemotherapy was assessed by Kaplan-Meier analysis. ResultsA total of 117 patients were included in this study. The median RFS was not reached (95% confidence intervals [CI]: 22.4 mo–not reached; n = 39) and 23.7 months (95% CI: 14.5–43.6; n = 41) in patients with inflamed or noninflamed adenocarcinoma, respectively (log-rank p = 0.02, hazard ratio of 0.52 [95% CI: 0.29–0.93]). Analysis of the combination of tumor inflammation category and TP53 mutation status revealed that inflamed tumors without TP53 mutations were associated with the longest RFS. ConclusionsPD-L1 expression on tumor cells, CD8+ T cell infiltration, and TP53 mutation status may help identify patients with early-stage NSCLC susceptible to recurrence after adjuvant chemotherapy.

Real-World Treatment and Outcomes in ALK-Rearranged NSCLC: Results From a Large U.S.-Based Database

IntroductionALK–rearranged advanced NSCLC (aNSCLC) represents 4% of all NSCLCs, and multiple ALK-targeted therapies (ALK-inhibitors) are now available for use. Little is known about changes in treatment patterns, or how prognostic factors and sequence of therapy may impact overall survival in the real-world setting. We aim to describe initial and subsequent treatments used, survival outcomes, prognostic factors, and the impact of treatment on overall survival in the largest (N = 739) real-world cohort of patients with ALK+ aNSCLC reported in the literature. MethodsRetrospective observational cohort study with data drawn from a U.S.-based electronic health record–derived, deidentified database. Eligible patients were diagnosed with ALK+ aNSCLC between 2011-2020 and were treated in multiple different cancer clinics and across multiple geographic regions throughout the United States. ResultsFrom a cohort of 63,667 patients with aNSCLC, 739 patients with ALK+ NSCLC were eligible for analysis, median age was 63 years, 54% patients were female, and 85% were managed in community setting. More than 168 different treatment sequences were observed, and treatment utilization changed over time. Cohort median overall survival was 37 months (95% confidence interval: 33–45). Positive prognostic factors were as follows: never-smoking history, younger age, treatment in an academic setting, and initial early stage at diagnosis. Initial treatment with a second-generation ALK-inhibitor was associated with improved survival compared with chemotherapy. ConclusionsFor people with ALK+ aNSCLC, this study has identified several important clinical prognostic factors and is practice affirming; first-line treatment with a second-generation ALK-inhibitor improves survival compared with chemotherapy.

128TiP A phase II, two parallel group study of neoadjuvant and adjuvant targeted treatment in NSCLC with BRAF V600 or MET exon 14 mutations

128TiP A phase II, two parallel group study of neoadjuvant and adjuvant targeted treatment in NSCLC with BRAF V600 or MET exon 14 mutations

79P Real-world overall survival after planned alternative dosing for pembrolizumab in the treatment of NSCLC: A nationwide retrospective non-inferiority analysis

79P Real-world overall survival after planned alternative dosing for pembrolizumab in the treatment of NSCLC: A nationwide retrospective non-inferiority analysis

244MO Primary results from IMscin002: A study to evaluate patient (pt)- and healthcare professional (HCP)-reported preferences for atezolizumab (atezo) subcutaneous (SC) vs intravenous (IV) for the treatment of NSCLC

244MO Primary results from IMscin002: A study to evaluate patient (pt)- and healthcare professional (HCP)-reported preferences for atezolizumab (atezo) subcutaneous (SC) vs intravenous (IV) for the treatment of NSCLC

Hybridization-based discovery of novel quinazoline-2-indolinone derivatives as potent and selective PI3Kα inhibitors

IntroductionPhosphatidylinositol 3-kinases (PI3Ks) overexpression can elicit cellular homeostatic dysregulation, which further contributes to tumorigenesis, with PI3Kα emerging as the most prevalent mutant isoform kinase among PI3Ks. Therefore, selective inhibitors targeting PI3Kα have attracted considerable interest in recent years. Molecular hybridization, with the advantage of simplified pharmacokinetics and drug-drug interactions, emerged as one of the important avenues for discovering potential drugs. ObjectivesThis study aimed to construct PI3Kα inhibitors by hybridization and investigate their antitumor activity and mechanism. Methods26 quinazoline-2-indolinone derivatives were obtained by molecular hybridization, and their structure–activity relationship was analyzed by MTT, in vitro kinase activity and molecular docking. The biological evaluation of compound 8 was performed by transwell, flow cytometry, laser scanning confocal microscopy, Western blot, CTESA and immunohistochemistry. ResultsHere, we employed molecular hybridization methods to construct a series of quinazoline-2-indolinone derivatives as PI3Kα selective inhibitors. Encouragingly, representative compound 8 exhibited a PI3Kα enzymatic IC50 value of 9.11 nM and 10.41/16.99/37.53-fold relative to the biochemical selectivity for PI3Kβ/γ/δ, respectively. Moreover, compound 8 effectively suppressed the viability of B16, HCT116, MCF-7, H22, PC-3, and A549 cells (IC50 values: 0.2 μM ∼ 0.98 μM), and dramatically inhibited the proliferation and migration of NSCLC cells, as well as induced mitochondrial apoptosis through the PI3K/Akt/mTOR pathway. Importantly, compound 8 demonstrated potent in vivo anti-tumor activity in non-small cell lung cancer mouse models without visible toxicity. ConclusionsThis study presented a new avenue for the development of PI3Kα inhibitors and provided a solid foundation for novel QHIDs as potential future therapies for the treatment of NSCLC.

Network pharmacology analysis, molecular docking integrated experimental verification reveal β-sitosterol as the active anti-NSCLC ingredient of Polygonatum cyrtonema Hua by suppression of PI3K/Akt/HIF-1α signaling pathway

Ethnopharmacological relevancePolygonatum cyrtonema Hua (Huangjing) is a Chinese herb that is considered by ancient Chinese healers to have the effect of nourishing yin and moisturizing the lungs. It is clinically used to treat diseases of the pulmonary system, including non-small cell lung cancer. However, the precise active components and underlying mechanisms of Huangjing in the context of treating NSCLC remain uncertain. Aim of the studyThis study aimed to explore the active components and mechanisms of Huangjing for the treatment of NSCLC by means of data mining, network pharmacology, and in vitro and vivo experiments. Materials and methodsFirst, the main active compounds and key targets of Huangjing were predicted by network pharmacology. The potential key targets of Huangjing were molecularly docked with the main active compounds using Pymol. In vivo, we verified whether Huangjing and its main active compound have anti-lung cancer effects. Key targets were verified by PCR and immunohistochemistry. In vitro, we verified the effects of Huangjing's main active compound on the proliferation, apoptosis, and migration of A549 cells by CCK-8, colony formation, wound healing assay, and flow cytometry. Key targets and signaling pathway were validated by PCR and Western blot. ResultsThe network pharmacology results suggested that β-sitosterol was the main active substance. TP53, JUN, AKT1, MAPK14, ESR1, RELA, HIF1A, and RXRA were potential targets of Huangjing. Molecular docking results suggested that MAPK14, HIF-1α, and RXRA docked well with β-sitosterol. In vivo tests also confirmed that Huangjing could significantly inhibit the growth of lung cancer tumors, while PCR and immunohistochemistry results suggested that the expression of HIF-1α was significantly decreased. Critically, KEGG analysis indicated that the PI3K/Akt/HIF-1α signaling pathway was recommended as one of the main pathways related to the anti-NSCLC effect of Huangjing. We conducted in vitro experiments to confirm the significant impact of β-sitosterol on the proliferation, apoptosis, migration, and colony formation of A549 cells. Furthermore, our findings indicate that a high dosage of β-sitosterol may effectively decrease the expression of HIF-1α, AKT1, JUN and RELA in A549 cells. Similarly, in vitro experiments also revealed that high doses of β-sitosterol could inhibit the PI3K/Akt/HIF-1α signaling pathway. ConclusionsWe discovered Huangjing and its main active ingredient, β-sitosterol, can reduce HIF-1α, AKT1, JUN and RELA expression and decrease non-small cell lung cancer growth through the PI3K/Akt/HIF-1α signaling pathway.

The role of leptin in regulation of the soluble amyloid precursor protein α (sAPPα) levels in lung cancer cell media

Previously, we found that the levels of soluble amyloid precursor protein α (sAPPα) are regulated, in part, by acetylcholinesterase (AChE) in human A549 (p53 wild-type) and H1299 (p53-null) NSCLC cell lines. In this study, we found regulation of sAPPα levels in the media by leptin, a widely recognized obesity-associated adipokine that has recently been shown to play a possible role in cancer signaling. Increased levels of sAPPα, that were accompanied by lower Aβ40/42 levels in the media of A549 and H1299 cells, were detected upon cell incubation with leptin. Conversely, knockdown of leptin or its receptor led to reduced levels of sAPPα and increased levels of Aβ40/42 in the media of A549 and H1299 cells, suggesting that leptin likely shifts APP processing toward the non-amyloidogenic pathway. A549 cell treatment with leptin increased acetylcholine levels and blocked the activities of AChE and p53. Treatment with leptin resulted in increased activation of PKC, ERK1/2, PI3K, and the levels of sAPPα, effects that were reversed by treatment with kinase inhibitors and/or upon addition of AChE to A549 and H1299 cell media. Cell viability increased by treatment of A549 and H1299 cells with leptin and decreased upon co-treatment with AChE and/or inhibitors targeting PKC, ERK1/2, and PI3K. This study is significant as it provides evidence for a likely carcinogenic role of leptin in NSCLC cells via upregulation of sAPPα levels in the media, and highlights the importance of targeting leptin as a potential therapeutic strategy for NSCLC treatment.

LINAC-based SBRT in treating early-stage NSCLC patients-single institution experience and survival data analysis.

Aim: This single institute prospective study aimed to evaluate the feasibility of LINAC-based stereotactic body radiotherapy (SBRT) in treating patients with early-stage non-small cell lung cancer (NSLSC). We focused on the survival data with the local and distant control profiles and the cancer- and non-cancer-specific survival. Treatment-related side effects were also collected and analyzed. Methods: Patients with early-stage NSCLC between January 2018 and October 2021 were included in our prospective study; a total of 77 patients receiving LINAC-based SBRT were analyzed. All patients had pretreatment multidisciplinary tumor board decisions on SBRT. The average patient age was 68.8years (median: 70years, range: 52-82); 70 patients were in ECOG 0 status (91%), while seven patients were in ECOG 1-2 status (9%). 52% of the patients (40) had histologically verified NSCLC, and the other 48% were verified based on PETCT results. We applied the SBRT scheme 8 x 7.5 Gy for central tumors (74%) or 4 x 12 Gy for peripheral tumors (26%). Results: The mean follow-up time was 25.4months (median 23, range 18-50). The Kaplan-Meier estimation for overall survival in patients receiving LINAC-based SBRT was 41.67months. Of the 77 patients treated by SBRT, death was reported for 17 patients (9 cases cancer-specific, 8 cases non-cancer specific reason). The mean local tumor control was 34.25months (range 8.4-41), and the mean systemic control was 24.24months (range 7-25). During the treatments, no Grade I-II were reported; in 30 cases, Grade I non-symptomatic treatment-related lung fibrosis and two asymptomatic rib fractures were reported. Conclusion: In the treatment of early-stage NSCLC, LINAC-based SBRT can be a feasible alternative to surgery. Although we reported worse OS data in our patient cohort compared to the literature, the higher older average age and the initial worse general condition (ECOG1-2) in our patient cohort appear to be the reason for this difference. With the comparable local control and survival data and the favorable side effect profile, SBRT might be preferable over surgery in selected cases.