Subtype Of Non-small Cell Lung Cancer Research Articles

Neoadjuvant sintilimab plus chemotherapy in EGFR-mutant NSCLC: Phase 2 trial interim results (NEOTIDE/CTONG2104)

The clinical efficacy of neoadjuvant immunotherapy plus chemotherapy remains elusive in localized epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer (NSCLC). Here, we report interim results of a Simon's two-stage design, phase 2 trial using neoadjuvant sintilimab with carboplatin and nab-paclitaxel in resectable EGFR-mutant NSCLC. All 18 patients undergo radical surgery, with one patient experiencing surgery delay. Fourteen patients exhibit confirmed radiological response, with 44% achieving major pathological response (MPR) and no pathological complete response (pCR). Similar genomic alterations are observed before and after treatment without influencing the efficacy of subsequent EGFR-tyrosine kinase inhibitors (TKIs) invitro. Infiltration and Tcell receptor (TCR) clonal expansion of CCR8+ regulatory T (Treg)hi/CXCL13+ exhausted T (Tex)lo cells define a subtype of EGFR-mutant NSCLC highly resistant to immunotherapy, with the phenotype potentially serving as a promising signature to predict immunotherapy efficacy. Informed circulating tumor DNA (ctDNA) detection in EGFR-mutant NSCLC could help identify patients nonresponsive to neoadjuvant immunochemotherapy. These findings provide supportive data for the utilization of neoadjuvant immunochemotherapy and insight into immune resistance in EGFR-mutant NSCLC.

Unlocking therapeutic potential: IL-1β as a target in non-small cell lung cancer with oncogenic mutations—Prognostic and predictive insights.

8030 Background: Preclinical studies have demonstrated heterogeneity in interleukin-1β (IL-1β) expression and its potential implications in non-small cell lung cancer(NSCLC). The phase II CANTOS trial incidentally showed reduced lung cancer incidence and mortality with the IL-1β inhibitor canakinumab. However, the phase III CANOPY-1/2 trials failed to demonstrate survival benefit when combining canakinumab with chemo/immunotherapy. The impact of IL-1β inhibition in NSCLC with actionable mutations remains unknown. This study aims to evaluate the prognostic and predictive role of IL-1β in NSCLC and its potential as a target for combination therapy. Methods: 34,960 NSCLC tumors underwent next-gen sequencing of DNA (592-gene or whole exome) and RNA (whole transcriptome) at Caris Life Sciences (Phoenix, AZ). Tumors were stratified by IL-1β transcriptional expression quartiles (Q1: low expression and Q4: high expression). Quartiles were established within each subpopulation. Significance was calculated using chi-square, Fisher’s exact, or Mann-Whitney U test, with p-values adjusted for multiple comparisons (p < 0.05). Overall survival (OS) from either time of tissue collection to last contact or time on treatment (TOT) was estimated from insurance claims data using the Cox proportional hazards model to calculate hazard ratio (HR) and log-rank tests to calculate p-values. Results: In the entire cohort, low IL-1β expressors showed improved OS compared to high expressors (20.0 months (m) vs 17.3 m, HR 0.88, 95%CI 0.88-0.95, p < 0.0001). For NSCLC without driver mutations, IL-1β expression did not correlate with OS difference. IL-1β expression was positively associated with TP-53 mutations (Q4 76% vs Q1 57%), high TMB (Q4 44% vs Q1 37%) and PD-L1+ expression by IHC (Q4 67% vs Q1 44%), while negatively associated with mutations in KRAS(Q4 24% vs Q1 31%) , EGFR(Q4 8% vs Q1 14%) , ERBB2 (Q4 1% vs Q1 2%) , BRAF (Q4 4% vs Q1 5%) ,and STK11 (Q4 10% vs Q1 18%) (p < 0.05). In EGFR-mutant NSCLC, Q1 showed superior OS than Q4 (34.6 m vs 25.1 m, HR 0.79, 95% CI 0.68-0.92, p = 0.002). This survival benefit was amplified in adenocarcinoma (Q1 41.2 m vs Q4 27.1 m, HR 0.74, 95% CI 0.63-0.87, p < 0.001). For ALK fusion-positive NSCLC, low expressors demonstrated improved OS (Q1 63.4 m vs Q4 27.0 m, HR 0.53, 95% CI 0.37-0.76, p < 0.001). In KRAS-mutant adenocarcinoma, high expressors showed improved immune-checkpoint inhibitor (ICI) TOT (Q4 6.4 m vs Q1 5.4 m, HR 0.85, 95% CI 0.74-0.98, p = 0.026); however, this did not correlate with improved OS. Conclusions: Our study shows promising prognostic value of IL-1β expression in NSCLC, associating low expression with improved OS across NSCLC subtypes. IL-1β expression is closely linked to key driver mutations in NSCLC and may have predictive value for ICIs. The findings suggest the potential benefit of targeting IL-1β in high IL-1β expressing NSCLC with driver mutations.

Incidence trends and factors affecting survival in mucinous adenocarcinoma of the lung: Insights from SEER data.

e20090 Background: Lung cancer is the second most common cancer in terms of incidence and the leading cause of cancer-related deaths. Non-small cell lung cancer (NSCLC) accounts for 80 to 85% of cases. Adenocarcinoma, squamous cell carcinoma, and large-cell carcinoma are the main subtypes of NSCLC. Mucinous adenocarcinoma is the least common subtype of lung adenocarcinoma with poor prognosis. Our study aims to identify demographics, incidence trends, and factors impacting overall survival (OS) in mucinous adenocarcinoma of the lung. Methods: We conducted a retrospective cohort analysis utilizing the Surveillance, Epidemiology, and End Results (SEER) database to investigate patients diagnosed with mucinous adenocarcinoma of the lung between the years 2000 and 2020. Demographics, treatment data were characterized using descriptive statistics. The Cox proportional regression analysis was used to identify the factors that impact the OS. Results: Between 2000-2020, a total of 7,953 patients were diagnosed with mucinous adenocarcinoma of the lung. The highest incidence of mucinous adenocarcinoma was in older adults, accounting for 67% of all cases. Females had a higher incidence rate at 53.6%. Caucasians comprised the majority of patients at 83.1%, followed by blacks at 8.8%. The majority of patients, 89.3% resided in urban areas, and 43.6% had annual incomes exceeding $75k. Most of the patients were married at 58.2%. The incidence trends revealed a significant increase from 2000 to 2020. The incidence rates were: 15.76% (2000 -2004), 16.37% (2005 -2009), 31.08% (2010 - 2014), and 36.79% (2015-2020). However, the survival rates improved over time (See table). Among the demographic factors: adults, females, Asian/ Natives/ & other races combined, urban residence, higher annual income (> $75k), and married status had better OS outcomes than their counterparts. Among disease stages, localized disease had the best, while regional disease had a better OS than distant disease. In terms of treatment, surgery was associated with better OS; interestingly, receipt of radiation or chemotherapy was not associated with better OS. The HRs of all these factors are shown in Table. Conclusions: Our study, the largest to date on mucinous adenocarcinoma of the lung, demonstrated a rising incidence yet with improved OS rates from 2000-2020. Our findings shed light on the demographics and other factors influencing OS in this rare subtype of NSCLC. [Table: see text]

Evaluation of the genomic and immune profiles of patients with lung adenosquamous carcinoma (LUAS) and association of response to treatment.

8567 Background: LUAS is a rare subset of non-small cell lung cancer (NSCLC) comprising up to 4% of cases, characterized by at least 10% of both adenocarcinoma and squamous components. Limited studies exist on LUAS regarding the genomic/immune landscape and response to immunotherapy (IO). Methods: NSCLC samples (n = 35404) underwent NextGen Sequencing of DNA (592 genes or whole exome)/RNA (whole transcriptome) at Caris Life Sciences. PD-L1 expression was assessed using IHC (22c3; positive: TPS >1%). High tumor mutational burden (TMB-H) was defined as ≥10 mut/MB. Cell infiltration in the tumor microenvironment was estimated by quanTIseq. Gene expression profiles were analyzed for transcriptomic signatures (IFN-γ) predictive of IO response. Real-world overall survival (rwOS) was obtained from insurance claims data, calculated from time of biopsy to last contact. Time on treatment (TOT) was calculated from the first pembrolizumab (pembro) treatment to the last. Mann-Whitney U and X2/Fisher-Exact tests were applied where appropriate, with p-values adjusted ( p < .05). Results: Among NSCLC samples, 1.0% (n=374) were LUAS, 26.5% (n = 9365) were squamous (SCC) and 72.5% (n = 25665) were adenocarcinoma (AC). LUAS exhibited actionable alterations in 29% (n = 110/374) of cases, with MET exon 14 skipping alteration ( METex14) significantly higher in LUAS compared to SCC or AC (9.02% vs 1.21% vs 2.61%, p < 0.01). BRAF V600E (0.54% vs 0.13%, p = 0.041) and ALK-Fusion (1.13% vs 0.12%, p = 0.0145) were higher in LUAS compared to SCC, while PIK3CA mutation was higher in LUAS compared to AC (10.8% vs 4.1%, p <0.01). Notably, LUAS exhibited higher prevalence of KRAS, targetable EGFRand STK11 mutations compared to SCC but lower compared to AC ( KRAS G12C: 8.63% vs 1.37% vs 13.24%, p <0.01; EGFR: 13.1% vs 1.12% vs 17.53%, p<0.01; STK11: 6.97% vs 1.83% vs 17.22%, p<0.01). LUAS exhibited significantly higher PD-L1+ (65.5% vs 59.8% vs 53.7%, p <0.01), along with elevated expression of immune checkpoint genes and the IFN-γ signature compared to AC and SCC. Moreover, LUAS exhibited significantly higher neutrophils, CD8 T cells, M2 macrophages and Tregs compared to SCC. LUAS showed improved rwOS compared to SCC (HR 0.792, p<0.01), with no difference noted in the rwOS between LUAS and AC (HR = 0.98, p = 0.81). While PD-L1+ was not associated with TOT on pembro in LUAS, high TMB showed improved TOT on pembro in LUAS. Conclusions: This study provides a comprehensive genomic landscape of LUAS, highlighting actionable alterations in ~ 29% of cases. LUAS exhibits distinct molecular and clinical features compared to AC and SCC, with enrichment in actionable METex14, emphasizing the need for NGS testing. Although PD-L1 status may not significantly impact TOT in LUAS, it remains relevant in other NSCLC subtypes on IO and high TMB emerges as a potential biomarker for favorable TOT on pembro in LUAS, warranting further investigation.

Implementing personalized lung cancer care with spatial profiling, patient derived organoids, and rationally designed drug testing.

e20521 Background: Lung cancer (LC) remains the leading cause of cancer-related death in 2024, with most LC diagnoses with non-small cell lung cancer (NSCLC) subtype. Tyrosine kinase inhibitors (TKI) and targeted therapy against oncogenic drivers (KRAS, EGFR, or ALK fusions, others) are effective initial therapies; however, nearly all treated patients eventually develop resistance to therapy. Key reasons are the focus on limited predictive biomarkers and lack of patient derived LC models that reflect the underlying heterogeneity and the various phenotypic cell states among subclones within and between patients. Methods: We report the establishment of a novel platform for personalized LC care by refining the resolution of molecular interrogation of diagnostic and resistance biopsies to the single cell level, and with spatial profiling using single cell spatial multiomic (scSpMO) and coupling these enhanced diagnostic tools with functional validation with rational designed drug screens in patient-derived organoids (PDOs). Results: For deriving LC PDOs in epithelial and tumor microenvironment (TME) conditions to faithfully maintain the histological and genetic features of their respective LC tissues, we developed enrichment conditions for LC PDOs guided by tumor mutation variants and activated pathways. PDOs are maintained under the same treatment condition in the clinic (e.g., Lorlatinib) and drug testing is tailored to identify potentially most effective next lines of therapy. With nine patients enrolled and others in ongoing studies, single cell PDOs (at > 80% establishment rate) were used to determine activated pathways, lineage plasticity and acquired resistance. Functional assays for acquired resistance such as PI3K/AKT signaling, Src kinase, BRAF fusion and MET hyperactivity allow the identification of potentially novel lines of therapy and treatment sequence and/or combinations for each patient, through the compassionate care program and/or trial participation. Conclusions: Our platform when empowered by combining datasets from scRNA-seq and scSpMO signatures, with validated functional drug responses in PDOs, offers the ability to perform high-content assays, predict and/or act on resistance to therapy for each patient, and provides a path for precision medicine-guided impact on patient care.

Stereotactic radiosurgery (SRS) for non-small cell lung cancer (NSCLC) in the oncogene-addicted era: A real-world UK tertiary center experience.

e14011 Background: 30-50% of NSCLC patients develop brain metastases (BM). Re-evaluation of the role of SRS is required with evolving understanding of molecular subtypes of NSCLC and increasing intracranial efficacy of systemic agents. Methods: We conducted a retrospective analysis of NSCLC patients receiving SRS at our centre from 05/16-11/23. Primary objective was median overall survival (mOS months). Analysis was performed on Prism 10.1.1. Multivariate analysis was conducted using Cox Proportional Hazards Regression, with age, gender, KPS, BM size, total BM volume and oncogene status as predictor variables. Extraction from medical records was undertaken by authors, with local R&D committee approval. Results: 237 patients were evaluable. 59% (N=139) were oncogene-addicted (oNSCLC) and 41% (N=98) non-oncogene-addicted (non-oNSCLC). Median duration of follow up was 43.3m. m OS was 11.9m overall. For oNSCLC mOS was 13.1m vs 8.8m for non-oNSCLC (HR 1.49, P=0.02). mOS was 20.7m for EGFR sensitizing NSCLC (N=48) vs 10.5m without (HR 1.40 P=0.12). mOS for ALK+ (N=22) was 29.3m vs 11.6m for non-ALK+ (HR 1.90, P=0.05). mOS for KRAS+ (N=32) was 11.6m vs 12.0m without (HR 0.91, P=0.68). Significantly inferior mOS was seen for single (N=186 [78%]) vs multiple (N=51[22%]) SRS courses (8.4m vs 38.8m, [HR 0.34 P=<0.0001]). SRS at diagnosis of BM (N=118) had a mOS of 12.3m vs at nadir (N=9, not reached [HR0.44, P=0.18]), progression on 1st line TKI (N=31, 22.1m [HR 1.1, P=0.79]) and at any other time (N=157, 7.8m [HR 1.33, P=0.11 ]). There was no significant difference in survival for patients diagnosed with brain metastases at the diagnosis of metastatic disease compared to those who developed brain metastases during their disease course (12.3m vs 10.6m [HR 1.18, P=0.33]). Survival of those with 1 site of metastatic disease (18.3m) was numerically improved compared to those with 2 (10.7m, HR 1.26, P=0.25) sites, and statistically significant compared to those with 3 (10.1m, HR 1.69, P<0.02) and ≥4 (4.4m, HR 4.41, P<0.0001) sites. Conclusions: Median OS in oNSCLC was superior to non-oNSCLC, driven by numerically superior survival in EGFR+ and ALK+ cohorts. Significantly improved OS was observed in patients with multiple vs single SRS treatments, however this may be confounded by biases. Patients treated with SRS at nadir had numerically superior OS to those treated at other timepoints, however this must be interpreted with caution due to small sample size. Further prospective research is needed to interrogate these findings and guide local therapies in NSCLC with BM. [Table: see text]

Insights into the heterogeneity of the tumor microenvironment in lung adenocarcinoma and squamous carcinoma through single‐cell transcriptomic analysis: Implications for distinct immunotherapy outcomes

AbstractBackgroundImmune checkpoint blockade has emerged as a key strategy to the therapy landscape of non‐small cell lung cancer (NSCLC). However, notable differences in immunotherapeutic outcomes exist between the two primary NSCLC subtypes: lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). This disparity may stem from the tumor immune microenvironment's heterogeneity at the transcriptome level.MethodsBy integrative analysis of transcriptomic characterization of 38 NSCLC patients by single‐cell RNA sequencing, the present study revealed a distinct tumor microenvironment (TME) between LUAD and LUSC, with relevant results further confirmed in bulk transcriptomic and multiplex immunofluorescence (mIF) validation cohort of neoadjuvant immunotherapy patients.ResultsLUAD exhibited a more active immune microenvironment compared to LUSC. This included highly expression of HLA I/II in cancer cells, reinforced antigen presentation potential of dendritic cells and enhanced cytotoxic activity observed in T/NK cells. In LUSC, cancer cells highly expressed genes belonging to the aldo‐keto reductases, glutathione S‐transferases and aldehyde dehydrogenase family, negatively correlating with immunotherapy outcomes in the validation cohort of our center. Further analysis revealed elevated infiltrated cancer‐associated fibroblasts (CAFs) in LUSC, which was corroborated in The Cancer Genome Atlas cohort. Corresponding increased infiltration of ADH1B+ CAFs in major pathologic response (MPR) patients and the higher presence of FAP+ CAFs in non‐MPR patients were demonstrated by multiplex mIF. Moreover, upregulating immunosuppressive extracellular matrix remodeling was identified in LUSC.ConclusionsThese comprehensive analyses advance the understanding of the differences in TME between LUAD and LUSC, offering insights for patient selection and developing subtype‐specific treatment strategies.

Disparities in the prevalence of EGFR mutations and ALK rearrangements by racial-ethnic group in South Florida with a focus on Hispanic patients, 2011-2019.

e20528 Background: We examined the prevalence and survival outcomes of EGFR+, ALK+, and receipt of Tyrosine Kinase Inhibitors (TKIs), among all 1,654 patients diagnosed with de novo advanced non-small-cell lung cancer (NSCLC) during 2011-2019 in 2 predominantly Hispanic healthcare institutions in South Florida: Sylvester Cancer Center and University of Miami Hospital. Methods: Tumor registry data was linked with genomic databases. Electronic medical records were further reviewed. We employed Cox regression to assess survival outcomes among patients with documented EGFR+ and ALK+ results. Results: There were 748 Hispanics, 697 Non-Hispanic (NH)-Whites, 173 NH-Blacks, 32 Asians, and 4 other race patients in our study. The overall prevalence for EGFR+ and ALK+ was 11.1% and 1.3%, respectively. During the study period, as many as 16.7% and 4.1% of all patients received either EGFR- or ALK-targeting TKIs, respectively. EGFR+ prevalence varied among racial groups, with 11.3% in NH-Whites, 9.8% in NH-Blacks, 10.7% in Hispanics, and 25.0% in Asians. Among Florida Hispanics, there were significant variations in EGFR+ results from 7.0-8.1% in Caribbean Hispanics (Cuba, Puerto Rico, Dominican Republic) to 14.0% among Mexican and Central Americans, and 22.9% among South Americans (p < 0.05). Among all 184 patients with EGFR+ results, 43.5% exhibited a deletion 19 mutation, 26.1% had an L838R mutation, 17.4% had an unspecified targetable mutation, and 13.0% had rarer mutation types. Notably, the proportion of never-smokers varied significantly by studied group: 13.8% among NH-Whites, 36.4% among South Americans, 41.9% Mexicans and Central Americans, and 62.5% among Asians (p < 0.05). Much of the inter-racial and intra-ethnic variation in EGFR+ prevalence could be attributed to smoking status, with more similar proportions observed when comparing never-smokers and ever-smokers separately. The ALK+ prevalence ranged from 0.9% in NH-Whites, 1.3% among Hispanics, 2.3% among NH-Blacks, to 6.3% in Asians. Both EGFR+ and ALK+ were more prevalent among non-smokers and women. Still, smokers accounted for approximately half of all targetable mutations for both EGFR+ (90 never-smokers; 94 smokers) and ALK+ (12 never-smokers; 10 smokers). After adjusting for age group, sex, smoking status, and race-ethnicity, both EGFR+ and ALK+ NSCLC subtypes exhibited a lower risk of death over time compared to EGFR-/ALK- cases (EGFR+ aHR 0.68 95%CI 0.56-0.81; ALK+ aHR: 0.41 95%CI 0.23-0.73). Conclusions: Smoking status significantly impacts the documented varying prevalence of EGFR+ and ALK+ by race-ethnicity. TKIs were extensively used in 2011-2019 even without evidence of targetable mutations. Current/history of smoking should not prevent the study of EGFR+/ALK+ mutations/rearrangements. Patients with targetable mutations experienced significantly improved survival outcomes.

Cancer cachexia as a predictor of adverse outcomes in patients with non-small cell lung cancer: A meta-analysis

IntroductionCancer cachexia is a complex problem characterized by weight loss due to skeletal muscle and adipose tissue reduction. The purpose of this meta-analysis is to examine the association between cancer cachexia and adverse outcomes in patients with non-small cell lung cancer (NSCLC). MethodsA comprehensive search was conducted in the PubMed, Web of Science, and Embase databases from their inception to January 15, 2024. Retrospective or prospective studies that investigated the cancer cachexia as a predictor of overall survival (OS), progression-free survival (PFS), overall response rate (ORR), or disease control rate (DCR) in NSCLC patients were included in this analysis. ResultsSixteen studies, comprising 5,919 NSCLC patients, were identified. The pooled prevalence of cachexia in NSCLC patients was 39%, with individual studies reporting rates ranging from 19% to 63.8%. A meta-analysis using a random effects model showed that cachexia was associated with reduced OS (hazard ratio [HR] 1.84; 95% confidence interval [CI] 1.54-2.21) and PFS (HR 1.49; 95% CI 1.27-1.73).Subgroup analysis indicated that cancer cachexia significantly predicted OS, regardless of study design, NSCLC subtypes, cancer stage, definitions of cachexia, or follow-up duration. However, there was no clear association between cancer cachexia and ORR or DCR. ConclusionsCancer cachexia emerges is a negative prognostic factor for OS and PFS in NSCLC patients. Assessing cancer cachexia can improve risk classification for survival outcomes in this patient population.

Focal adhesion kinase-YAP signaling axis drives drug-tolerant persister cells and residual disease in lung cancer

Targeted therapy is effective in many tumor types including lung cancer, the leading cause of cancer mortality. Paradigm defining examples are targeted therapies directed against non-small cell lung cancer (NSCLC) subtypes with oncogenic alterations in EGFR, ALK and KRAS. The success of targeted therapy is limited by drug-tolerant persister cells (DTPs) which withstand and adapt to treatment and comprise the residual disease state that is typical during treatment with clinical targeted therapies. Here, we integrate studies in patient-derived and immunocompetent lung cancer models and clinical specimens obtained from patients on targeted therapy to uncover a focal adhesion kinase (FAK)-YAP signaling axis that promotes residual disease during oncogenic EGFR-, ALK-, and KRAS-targeted therapies. FAK-YAP signaling inhibition combined with the primary targeted therapy suppressed residual drug-tolerant cells and enhanced tumor responses. This study unveils a FAK-YAP signaling module that promotes residual disease in lung cancer and mechanism-based therapeutic strategies to improve tumor response.

First insight about the ability of specific glycerophospholipids to discriminate non-small cell lung cancer subtypes.

Introduction: Discrimination between adenocarcinoma (ADC) and squamous cell carcinoma (SCC) subtypes in non-small cell lung cancer (NSCLC) patients is a significant challenge in oncology. Lipidomics analysis provides a promising approach for this differentiation. Methods: In an accompanying paper, we explored oxPCs levels in a cohort of 200 NSCLC patients. In this research, we utilized liquid chromatography coupled with mass spectrometry (LC-MS) to analyze the lipidomics profile of matching tissue and plasma samples from 25 NSCLC patients, comprising 11 ADC and 14 SCC cases. This study builds upon our previous findings, which highlighted the elevation of oxidised phosphatidylcholines (oxPCs) in NSCLC patients. Results: We identified eight lipid biomarkers that effectively differentiate between ADC and SCC subtypes using an untargeted approach. Notably, we observed a significant increase in plasma LPA 20:4, LPA 18:1, and LPA 18:2 levels in the ADC group compared to the SCC group. Conversely, tumour PC 16:0/18:2, PC 16:0/4:0; CHO, and plasma PC 16:0/18:2; OH, PC 18:0/20:4; OH, PC 16:0/20:4; OOH levels were significantly higher in the ADC group. Discussion: Our study is the first to report that plasma LPA levels can distinguish between ADC and SCC patients in NSCLC, suggesting a potential role for LPAs in NSCLC subtyping. This finding warrants further investigation into the mechanisms underlying these differences and their clinical implications.

Comprehensive genomic profiling of pulmonary spindle cell carcinoma using tissue and plasma samples: insights from a real-world cohort analysis.

Pulmonary spindle cell carcinoma (PSCC) is a rare and aggressive non-small cell lung cancer (NSCLC) subtype with a dismal prognosis. The molecular characteristics of PSCC are largely unknown due to its rarity, which limits the diagnosis and treatment of this historically poorly characterized malignancy. We present comprehensive genomic profiling results of baseline tumor samples from 22 patients histologically diagnosed with PSCC, representing the largest cohort to date. Somatic genetic variant detection was compared between paired plasma samples and primary tumors from 13 patients within our cohort. The associations among genomic features, treatment, and prognosis were also analyzed in representative patient cases. TP53 (54.5%), TERT (36.4%), CDKN2A (27.3%), and MET (22.7%) were most frequently mutated. Notably, 81.8% of patients had actionable targets in their baseline tumors, including MET (22.7%), ERBB2 (13.6%), EGFR (9.1%), KRAS (9.1%), ALK (9.1%), and ROS1 (4.5%). The median tumor mutation burden (TMB) for PSCC tumors was 5.5 mutations per megabase (muts/Mb). TMB-high tumors (>10 muts/Mb) exhibited a significantly higher mutation frequency in genes such as KRAS, ARID2, FOXL2, and LRP1B, as well as within the DNA mismatch repair pathway. The detection rates for single nucleotide variants and structural variants were comparable between matched tumor and plasma samples, with 48.6% of genetic variants being mutually identified in both sample types. Additionally, a patient with a high mutation load and positive PD-L1 expression demonstrated a 7-month survival benefit from chemoimmunotherapy. Furthermore, a patient with an ALK-rearranged tumor achieved a remarkable 3-year progression-free survival following crizotinib treatment. Overall, our findings deepen the understanding of the complex genomic landscape of PSCC, revealing actionable targets amenable to tailored treatment of this poorly characterized malignancy.

Advances of Molecular Targeted Therapy in EGFR-mutated Squamous Cell Lung Cancer

Non-small cell lung cancer (NSCLC) is a prevalent tumour type in our country, with lung squamous carcinoma being a commonly observed NSCLC subtype besides lung adenocarcinoma. Epidermal growth factor receptor (EGFR) is a significant driver gene in lung cancer, and EGFR mutation frequency is considerably lower in lung squamous carcinoma in comparison to lung adenocarcinoma. Although targeted therapy against EGFR has demonstrated significant advancements in lung adenocarcinoma, while progress in lung squamous carcinoma has been relatively sluggish. This paper reviews recent studies on molecular targeted therapy for EGFR-mutated lung squamous carcinoma and summarises the efficacy of EGFR-tyrosine kinase inhibitors (TKIs) in treating squamous carcinoma of the lung, in order to provide a reference for treating patients with EGFR-mutated squamous carcinoma of the lung. .

Endobronchial Ultrasound Guided Transbronchial Needle Aspiration and PD-L1 Yields

PurposeImmunotherapy is a leading approach for treating advanced non-small cell lung cancer (NSCLC) by targeting the PD-1/PD-L1 checkpoint signaling pathway, particularly in tumors expressing high levels of PD-L1 (Jug et al. in J Am Soc Cytopathol 9:485–493, 2020; Perrotta et al. in Chest 158: 1230–1239, 2020). Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a minimally invasive method to obtain tissue for molecular studies, including PD-L1 analysis, in unresectable tumors (Genova et al. in Front Immunol 12: 799455, 2021; Wang et al. in Ann Oncol 29: 1417–1422, 2018). This study aimed to assess the adequacy of PD-L1 assessment in EBUS-TBNA cytology specimens.MethodsData was collected retrospectively from patients who underwent EBUS-TBNA between 2017 and 2021 for suspected lung cancer biopsy. Samples positive for NSCLC were examined for PD-L1 expression. EBUS was performed by experienced practitioners, following institutional guidelines of a minimum of five aspirations from positively identified lesions. Sample adequacy for molecular testing was determined by the pathology department.ResultsThe analysis involved 387 NSCLC cases (149 squamous cell, 191 adenocarcinoma, 47 unspecified). Of the 263 EBUS-TBNA specimens tested for PD-L1, 237 (90.1%) were deemed adequate. While 84% adhered to the protocol, adherence did not yield better results. Significantly higher PD-L1 adequacy was observed in squamous cell carcinomas (93.2%) compared to adenocarcinoma (87.6%). The number of aspirations and sedation type did not correlate with PD-L1 adequacy in either cancer type, but lesion size and location had a significant impact in adenocarcinomas. Adenocarcinoma exhibited higher PD-L1 expression (68%) compared to squamous cell carcinoma (48%).ConclusionEBUS-TBNA offers high yields for assessing immunotherapy markers like PD-L1, with satisfactory adequacy regardless of NSCLC subtype, lesion size, or location.

NCI 159456 PERK Inhibitor as a Targeted Therapy for Lung Cancer: An In Vitro Study.

Non-small cell lung cancer (NSCLC) represents the most common histological type of lung cancer, characterized by a five-year survival rate of 15% and poor prognosis. Accumulating evidence indicates a prominent role of endoplasmic reticulum (ER) stress and the protein kinase RNA-like ER kinase (PERK)-dependent pathway of the unfolded protein response (UPR) in the pathogenesis of NSCLC. Increased expression of downstream targets of PERK was observed in various subtypes of NSCLC, and it was associated with a more aggressive phenotype, high risk of recurrence, and poor prognosis. Therefore, the present study aimed to investigate the biological effect of the selective PERK inhibitor NCI 159456 on A549 NSCLC cells and Human Pulmonary Fibroblasts (HPF) in vitro. Treatment of both normal and ER-stressed A549 cells with NCI 159456 resulted in a significant increase in the mRNA expression level of pro-apoptotic genes like activating transcription factor 4 (ATF4), DNA damage inducible transcript 3 (DDIT3), and BCL2 Associated X, Apoptosis Regulator (BAX) as well as a decreased level of the anti-apoptotic gene B-cell lymphoma 2 (Bcl-2). Cytotoxicity and genotoxicity analyses revealed that NCI 159456 significantly decreased viability and increased DNA damage in A549 cells under normal and ER stress conditions. Caspase-3 and reactive oxygen species (ROS) detection assays demonstrated that NCI 159456 significantly induced apoptosis and increased the ROS level in normal and ER-stressed A549 cells. Importantly, treatment with the inhibitor did not affect substantially normal HPF cells at any used concentration. The results indicate that PERK inhibitors could potentially be applied as a targeted therapy for NSCLC.

Cancer-associated fibroblasts expressing fibroblast activation protein and podoplanin in non-small cell lung cancer predict poor clinical outcome.

Cancer-associated fibroblasts (CAFs) are a dominant cell type in the stroma of non-small cell lung cancer (NSCLC). Fibroblast heterogeneity reflects subpopulations of CAFs, which can influence prognosis and treatment efficacy. We describe the subtypes of CAFs in NSCLC. Primary human NSCLC resections were assessed by flow cytometry and multiplex immunofluorescence for markers of fibroblast activation which allowed identification of CAF subsets. Survival data were analysed for our NSCLC cohort consisting of 163 patients to understand prognostic significance of CAF subsets. We identified five CAF populations, termed CAF S1-S5. CAF-S5 represents a previously undescribed population, and express FAP and PDPN but lack the myofibroblast marker αSMA, whereas CAF-S1 populations express all three. CAF-S5 are spatially further from tumour regions then CAF-S1 and scRNA data demonstrate an inflammatory phenotype. The presence of CAF-S1 or CAF-S5 is correlated to worse survival outcome in NSCLC, despite curative resection, highlighting the prognostic importance of CAF subtypes in NSCLC. TCGA data suggest the predominance of CAF-S5 has a poor prognosis across several cancer types. This study describes the fibroblast heterogeneity in NSCLC and the prognostic importance of the novel CAF-S5 subset where its presence correlates to worse survival outcome.

Histology-driven hypofractionated radiation therapy schemes for early-stage lung adenocarcinoma and squamous cell carcinoma

Background and PurposeHistology was found to be an important prognostic factor for local tumor control probability (TCP) after stereotactic body radiotherapy (SBRT) of early-stage non-small-cell lung cancer (NSCLC). A histology-driven SBRT approach has not been explored in routine clinical practice and histology-dependent fractionation schemes remain unknown. Here, we analyzed pooled histologic TCP data as a function of biologically effective dose (BED) to determine histology-driven fractionation schemes for SBRT and hypofractionated radiotherapy of two predominant early-stage NSCLC histologic subtypes adenocarcinoma (ADC) and squamous cell carcinoma (SCC). Material and MethodsThe least-χ2 method was used to fit the collected histologic TCP data of 8510 early-stage NSCLC patients to determine parameters for a well-developed radiobiological model per the Hypofractionated Treatment Effects in the Clinic (HyTEC) initiative. ResultsA fit to the histologic TCP data yielded independent radiobiological parameter sets for radiotherapy of early-stage lung ADC and SCC. TCP increases steeply with BED and reaches an asymptotic maximal plateau, allowing us to determine model-independent optimal fractionation schemes of least doses in 1–30 fractions to achieve maximal tumor control for early-stage lung ADC and SCC, e.g., 30, 44, 48, and 51 Gy for ADC, and 32, 48, 54, and 58 Gy for SCC in 1, 3, 4, and 5 fractions, respectively. ConclusionWe presented the first determination of histology-dependent radiobiological parameters and model-independent histology-driven optimal SBRT and hypofractionated radiation therapy schemes for early-stage lung ADC and SCC. SCC requires substantially higher radiation doses to maximize tumor control than ADC, plausibly attributed to tumor genetic diversity and microenvironment. The determined optimal SBRT schemes agree well with clinical practice for early-stage lung ADC. These proposed optimal fractionation schemes provide first insights for histology-based personalized radiotherapy of two predominant early-stage NSCLC subtypes ADC and SCC, which require further validation with large-scale histologic TCP data.

“Crosstalk between non-coding RNAs and transcription factor LRF in non-small cell lung cancer”

Epigenetic approaches in direct correlation with assessment of critical genetic mutations in non-small cell lung cancer (NSCLC) are currently very intensive, as the epigenetic components underlying NSCLC development and progression have attained high recognition. In this level of research, established human NSCLC cell lines as well as experimental animals are widely used to detect novel biomarkers and pharmacological targets to treat NSCLC. The epigenetic background holds a great potential for the identification of epi-biomarkers for treatment response however, it is highly complex and requires precise definition as these phenomena are variable between NSCLC subtypes and systems origin.We engaged an in-depth characterization of non-coding (nc)RNAs prevalent in human KRAS-mutant NSCLC cell lines A549 and H460 and mouse KRAS-mutant NSCLC tissue by Next Generation Sequencing (NGS) and quantitative Real Time PCRs (qPCRs). Also, the transcription factor (TF) LRF, a known epigenetic silencer, was examined as a modulator of non-coding RNAs expression. Finally, interacting networks underlying epigenetic variations in NSCLC subtypes were created. Data derived from our study highlights the divergent epigenetic profiles of NSCLC of human and mouse origin, as well as the significant contribution of 12qf1: 109,709,060–109,747,960 mouse chromosomal region to micro-RNA upregulated species. Furthermore, the novel epigenetic miR-148b-3p/lncPVT1/ZBTB7A axis was identified, which differentiates human cell line of lung adenocarcinoma from large cell lung carcinoma, two characteristic NSCLC subtypes.The detailed recording of epigenetic events in NSCLC and combinational studies including networking between ncRNAs and TFs validate the identification of significant epigenetic features, prevailing in NSCLC subtypes and among experimental models. Our results enrich knowledge in the field and empower research on the epigenetic prognostic biomarkers of the disease progression, NSCLC subtypes discrimination and advancement to patient-tailored treatments.

Six-Gene Signature for Differential Diagnosis and Therapeutic Decisions in Non-Small-Cell Lung Cancer-A Validation Study.

Non-small-cell lung cancer (NSCLC) poses a challenge due to its heterogeneity, necessitating precise histopathological subtyping and prognostication for optimal treatment decision-making. Molecular markers emerge as a potential solution, overcoming the limitations of conventional methods and supporting the diagnostic-therapeutic interventions. In this study, we validated the expression of six genes (MIR205HG, KRT5, KRT6A, KRT6C, SERPINB5, and DSG3), previously identified within a 53-gene signature developed by our team, utilizing gene expression microarray technology. Real-time PCR on 140 thoroughly characterized early-stage NSCLC samples revealed substantial upregulation of all six genes in squamous cell carcinoma (SCC) compared to adenocarcinoma (ADC), regardless of clinical factors. The decision boundaries of the logistic regression model demonstrated effective separation of the relative expression levels between SCC and ADC for most genes, excluding KRT6C. Logistic regression and gradient boosting decision tree classifiers, incorporating all six validated genes, exhibited notable performance (AUC: 0.8930 and 0.8909, respectively) in distinguishing NSCLC subtypes. Nevertheless, our investigation revealed that the gene expression profiles failed to yield predictive value regarding the progression of early-stage NSCLC. Our molecular diagnostic models manifest the potential for an exhaustive molecular characterization of NSCLC, subsequently informing personalized treatment decisions and elevating the standards of clinical management and prognosis for patients.

Abstract 4286: Proteolytic degradation of BIGH3 can be quantified non-invasively in serum with biomarker potential for patients with non-small cell lung cancer

Abstract Introduction: Transforming growth factor beta induced protein ig-h3 (BIGH3/TGFBI) is expressed widely by multiple cell types, including macrophages and fibroblasts, and is participating in various biological processes including adhesion, migration, angiogenesis and wound healing. BIGH3 is known to bind multiple collagens and are often embedded in the matrix where it seems to function as a linker between ECM and cell surfaces. The increased proteolytic activity found in non-small cell lung cancer (NSCLC) may degrade BIGH3 that affect this interaction as well as generate small peptide fragments that can serve as novel non-invasive biomarker targets if released to circulation. The aim of this study is to develop a tool to quantify degraded BIGH3 non-invasively and explore its potential as a biomarker in NSCLC. Methods: An ELISA, named nordicBIGH3M-N targeting a cleaved fragment of BIGH3 was developed to reflect BIGH3 degradation and enable serological quantification. We incubated recombinant BIGH3 with and without collagenase to confirm that nordicBIGH3M-N only measured degraded BIGH3. In addition, we generated matrixes from normal fibroblasts and cancer associated fibroblasts (CAFs) grown with or without tgf-b. These matrixes were cleaved with collagenase to confirm that NordicBIGH3M-N could be generated from degradation of matrix. nordicBIGH3M-N was also measured in serum from 39 patients with NSCLC (18 patients with adenocarcinoma and 21 patients with squamous cell carcinoma) and 35 healthy controls. BIGH3M-N levels in serum from patients with the two subtypes of NSCLC and healthy controls was compared using Dunn’s multiple comparison test and area under receiver operation characteristic curves (AUROC). Results: BIGH3M-N was only detectable after incubating recombinant BIGH3 and fibroblasts matrices with collagenase. A 7-fold increase of BIGH3M-N levels was seen between the collagenase degraded matrix from tgf-b treated fibroblasts compared to the degraded matrix from untreated fibroblasts. No difference in BIGH3M-N levels was observed between degraded matrix from tgf-b treated or untreated CAFs. BIGH3M-N was significantly elevated in patients with squamous cell carcinoma compared to healthy controls (p = 0.006, AUROC = 0.73) and patients with adenocarcinoma (p = 0.022, AUROC = 0.78). However, there was no significant difference between BIGH3M-N levels in serum from patients with adenocarcinoma and healthy controls (p>0.99, AUROC = 0.51). Conclusion: Degradation of BIGH3 can be reflected by non-invasive quantification of the cleaved fragment of BIGH3, BIGH3M-N, in serum. BIGH3M-N is a promising biomarker in NSCLC with potential for discriminating between subtypes. Moreover, BIGH3M-N is connected to fibroblast matrix biology so the optimal use for this biomarker might be in combination with other ECM biomarkers. Citation Format: Rasmus S. Pedersen, Morten Karsdal, Nicholas Willumsen. Proteolytic degradation of BIGH3 can be quantified non-invasively in serum with biomarker potential for patients with non-small cell lung cancer [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 4286.