Immune Checkpoint Inhibitors In Non-small Cell Lung Cancer Research Articles

Organ-specific efficacy in advanced non-small cell lung cancer patients treated with first-line single-agent immune checkpoint inhibitors.

Background:Response to immune checkpoint inhibitors (ICIs) is affected by multiple factors. This study aimed to explore whether sites of metastasis are associated with clinical outcomes of ICIs in advanced non-small-cell lung cancer (NSCLC) patients.Methods:The data of NSCLC patients with high programmed death-ligand 1 expression and good performance status receiving first-line ICIs monotherapy from Guangdong Provincial People's Hospital between May 2019 and July 2020 were retrospectively analyzed. Metastatic sites included liver, bone, brain, adrenal gland, pleura, and contralateral lung. Progression-free survival (PFS) and overall survival (OS) were compared between different metastatic sites and metastatic burden by the Kaplan-Meier method. Organ-specific disease control rate (OSDCR) of different individual metastatic sites was evaluated.Results:Forty NSCLC patients meeting the criteria were identified. The presence of liver metastasis was significantly associated with shorter PFS (3.1 vs. 15.5 months, P = 0.0005) and OS (11.1 months vs. not reached, P = 0.0016). Besides, patients with bone metastasis tend to get shorter PFS (4.2 vs. 15.5 months, P = 0.0532) rather than OS (P = 0.6086). Moreover, the application of local treatment could numerically prolong PFS in patients with brain metastasis (15.5 vs. 4.3 months, P = 0.1894). More metastatic organs involved were associated with inferior PFS (P = 0.0052) but not OS (P = 0.0791). The presence of liver metastasis or bone metastasis was associated with more metastatic organs (Phi[ϕ]: 0.516, P = 0.001). The highest OSDCR was observed in lung (15/17), and the lowest in the liver (1/4).Conclusions:Metastases in different anatomical locations may be associated with different clinical outcomes and local tumor response to ICIs in NSCLC. ICIs monotherapy shows limited efficacy in patients with liver and bone metastasis, thus patients with this type of metastasis might require more aggressive combination strategies.

Abstract 2027: Spatial omics and multiplexed imaging to discover new biomarkers of response or resistance to immune checkpoint inhibitors (ICI) in advanced non-small cell lung cancer (NSCLC)

Abstract Introduction: Despite the impressive outcomes with ICI in NSCLC, only a minority of the patients show long-term benefit from ICI. While PD-L1 immunohistochemistry is an approved companion diagnostic test, it is neither sensitive nor specific. Here, we describe the use of spatial transcriptomics using the GeoMx Digital Spatial Profiler (DSP) as a discovery platform to find biomarkers for ICI response or resistance. Methods: Pre-treatment samples from 56 patients with NSCLC treated with ICI were collected and analyzed in a tissue microarray (TMA) format in two-fold redundancy. The human whole transcriptome, represented by 18000 genes assessed by oligonucleotide-tagged in situ hybridization, was sequenced on the NovaSeq platform to quantify the mRNAs present in each region of interest. Three molecularly defined tissue compartments, defined by fluorescence co-localization (tumor [panCK+], leukocytes [CD45+/CD68-], macrophages [CD68+]) were generated to assess mRNA. All statistical testing was performed using a two-sided significance level of α=0.05. Result: 54000 gene variables were generated per case, from them about 27000 were analyzed after removing targets with expression lower than a prespecified frequency. Expression distribution of EPCAM in tumor, PTPRC in leukocytes and CD68 in macrophages confirmed that DSP successfully profiled the molecular compartments. Differential gene expression (DEG) analysis was performed for 6-month clinical benefit across two distinct tumor TMA spots per patient. This resulted in a DEG list of about 250, 125 and 122 genes in CK, CD45 and CD68 compartment respectively. When we used the intersection of significantly associated with benefit genes the candidate list was reduced dramatically to 5, 10 and 10 distinct genes in CK (CYBA, ESD, SH3BGRL3, TOP2B, UFM1), CD45 (ATRN, DGAT1, DNAAF9, GMPS, IFT20, MPHOSPH6, PKP4, SOCS2, TFAP4, TGFB3) and CD68 (ANKS3, ARFRP1, CTSD, DPYD, GTF3C5, HS2ST1, KIAA0895L, PIGR, SFTPA2, YEATS4) compartment respectively. From them, using the Cox Proportional-Hazards Model we further identified CYBA, ESD and UFM1 in CK, DGAT1 and IFT20 in CD45 and ANKS3 and DPYD in CD68 to predict survival as well. Conclusions: Using DSP technology allows rapid, patient-specific assessment of the transcriptome in TMAs by in situ hybridization in spatially defined molecular compartments. Here we show a small set of candidate genes that are associated with outcome in this ICI-treated cohort. By intersecting two non-adjacent cores of the same patient tissue sample, we have begun to dissect tumor heterogeneity and pilot biomarker candidate information with unique molecularly defined compartments for tumor cells, lymphocytes, and macrophages. Results from this cohort may lead to a novel, spatially defined, transcriptomic approach for developing new biomarkers for immunotherapy. Citation Format: Myrto Moutafi, Sandra Martinez-Morilla, Rolando Garcia-Milian, Thazin Nwe Aung, Ioannis Vathiotis, Niki Gavrielatou, Vasiliki Xirou, Leonidas Salichos, David L. Rimm. Spatial omics and multiplexed imaging to discover new biomarkers of response or resistance to immune checkpoint inhibitors (ICI) in advanced 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 2027.

Recent Advancements of Monotherapy, Combination, and Sequential Treatment of EGFR/ALK-TKIs and ICIs in Non-Small Cell Lung Cancer.

Lung cancer is the leading cause of cancer-related deaths with high morbidity and mortality. Non–small cell lung cancer (NSCLC) is the most common type of lung cancer, accounting for 85% of all cases. Fortunately, the development of molecular oncology provides a promising and effective therapeutic strategy for lung cancers, including specific gene mutations/translocations and immune checkpoints, with epidermal growth factor receptor (EGFR) common mutations first and anaplastic lymphoma kinase (ALK) translocations later as the targeted therapy and immune checkpoint inhibitors (ICIs) as immunotherapy. This review summarized the recent therapy advancements of TKIs and ICIs in NSCLC and focused on the clinical effect of combination or sequential treatment so as to provide the effective advice for the treatment of NSCLC.

Blood Biomarkers of Response to Immune Checkpoint Inhibitors in Non-Small Cell Lung Cancer.

Immune checkpoint inhibitors (ICIs) have revolutionized the treatment landscape of non-small cell lung cancer (NSCLC), either used in monotherapy or in combination with chemotherapy. While some patients achieve durable responses, some will not get benefit from this treatment. Early identification of non- responder patients could avoid unnecessary treatment, potentially serious immune-related adverse events and reduce treatment costs. PD-L1 expression using immunohistochemistry is the only approved biomarker for the selection of patients that can benefit from immunotherapy. However, application of PD-L1 as a biomarker of treatment efficacy shows many deficiencies probably due to the complexity of the tumor microenvironment and the technical limitations of the samples. Thus, there is an urgent need to find other biomarkers, ideally blood biomarkers to help us to identify different subgroups of patients in a minimal invasive way. In this review, we summarize the emerging blood-based markers that could help to predict the response to ICIs in NSCLC.

LungTS: A new gene expression signature for prediction of response to checkpoint inhibitors in non-small cell lung cancer.

e21143 Background: Immune checkpoint inhibitors (ICIs) are associated with durable antitumor activity in advanced non-small cell lung cancer (NSCLC), however, selection of the best candidates for this type of therapy is a challenge. PD-L1 expression, the only adequately validated predictive biomarker of response to ICIs, is imperfect, and not completely discriminatory between responders and non-responders. Gene expression profile (GEP) is a promising way to evolve in this field. The objective of this study was to identify a GEP of immune-oncology related genes, predictive of response to ICIs in NSCLC. Methods: This study analyzed two cohorts of advanced NSCLC patients treated with ICIs in any line: a discovery cohort composed of 66 Brazilian patients and a validation cohort composed of 54 Spanish patients. Clinical data were collected from medical records. Total RNA was extracted from FFPE tumor tissue. Gene expression profile (GEP) was assessed using the nCounter PanCancer IO360 panel (NanoString Technologies), which comprises 770 genes involved in the cancer immune response. LASSO regression with Cox proportional hazards approach was used to define the new gene expression signature. Results: In the discovery cohort, median age was 62.5 years (36 - 81 years). Most patients were males (57.6%), stage IV (91.9%), ECOG-PS≤1 (90.7%), smokers/former smokers (83.3%) and had adenocarcinoma histology (67.7%). The majority was treated with ICI in the first-line setting (42.4%) and nivolumab was the most frequently used drug (52.6%). At data cutoff, median follow-up for overall survival (OS - time from ICI initiation to death by any cause) was 27.7 months and for real-world progression-free survival (rwPFS - time from ICI initiation to disease progression as assessed by treating physician) was 28.2 months. The median OS and rwPFS were 17.7 months and 5.6 months, respectively. We identified a 6-gene signature (Lung Tumor Score – LungTS) that discriminate patients at low and high risk for death (median OS: 36.1 vs 15.5 months; p < 0.001) and for disease progression (median rwPFS: 10.1 vs 4.2 months; p < 0.001). Considering the expression of CD274 (PD-L1), the LungTS signature also discriminated two populations regarding OS and rwPFS (p < 0.0001). In the validation cohort, which had clinicopathological characteristics similar to the discovery cohort, the OS for high versus low LungTS was 17.0 months and 9.0 months, respectively (p = 0.004). Patients with high score also showed better rwPFS, although not statistically significant (median 10.0 versus 4.0 months; p = 0.11). Conclusions: Our study identified a novel 6-gene immune-signature predictive of response to ICI in NSCLC.

Obesity diabetes and dyslipidemia as determinants of immune checkpoint inhibitors response in non-small cell lung cancer.

e21104 Background: Immune checkpoint inhibitors (ICI) are standard of care in the treatment of non-small cell lung cancer (NSCLC). Predictive markers of response to ICI are being studied extensively. Obesity has been correlated with a better response to ICI in NSCLC and other types of cancer. This may in-part be related to high circulating leptin or impaired T-cell fatty acid oxidation within the tumor microenvironment. We aim to further explore the effects of obesity, dyslipidemia and other components of the metabolic syndrome such as diabetes on treatment outcomes on NSCLC patients receiving ICI. Methods: We conducted a retrospective cohort study in patients with NSCLC treated with pembrolizumab, nivolumab, durvalumab, cemiplimab or atezolizumab as monotherapy or combined with chemotherapy at Mount Auburn and Lahey Hospital between January 2016 to June 2021. Obesity was defined as body mass index (BMI) > 30 kg/m2, dyslipidemia and type 2 diabetes were defined by ICD-10 diagnosis codes. Overall survival was evaluated. Kaplan-Meier survival analysis and cox proportional hazards regression were used to assess correlation between overall survival and obesity, dyslipidemia, type 2 diabetes, metformin use and statin use. These were adjusted for age, biological sex, cancer stage, ECOG and smoking status. Patients with BMI < 18.5 were excluded. Results: We identified 408 patients with NSCLC who were treated with ICIs. Of these, 66.7% had adenocarcinoma, 29.1% had squamous and 4.2% had either adenosquamous, large cell or poorly differentiated carcinoma. Among these patients 23.3% were classified as obese, 55.6% had dyslipidemia and 23% had type 2 diabetes. Of these, 66% of patients received ICIs as monotherapy. Median survival was 840 days in obese patients compared to 483 days in non-obese patients (HR 0.66, CI 0.42-0.92, p = 0.014). Conversely, within this cohort, median survival in patients with dyslipidemia was 483 days when compared to 762 days in controls (HR 1.37, CI 1.07-1.81, p = 0.013). Overall survival was not significantly different between type 2 diabetes, metformin use or statin use. Conclusions: Our study shows a trend towards a better survival in obesity and poor survival associated with dyslipidemia. Obesity and dyslipidemia may be important markers of ICI response in patients with NSCLC regardless of treatment strategies. Further studies are needed to explore the molecular mediators within the metabolic syndrome that may provide a synergistic effect with immune checkpoint inhibitors.

PAPPA2 mutation as an indicator stratified patients benefit from immune checkpoint inhibitors in NSCLC and SKCM.

2617 Background: Pappalysin 2 ( PAPPA2) mutation, most occurring in skin cutaneous melanoma (SKCM) and non-small cell lung cancer (NSCLC), is found to be related to anti-tumor immune response. However, the association between PAPPA2 and efficacy of immune checkpoint inhibitors (ICIs) therapy remains unknown. Methods: Forty-one NSCLC patients receiving anti-PD-(L)1 (China cohort, n = 41) and seven public cohorts with whole-exome sequencing (WES) data were included to analyze the performance of PAPPA2 mutation as an indicator stratifying patients benefit from ICIs, consolidating a discovery set (n = 34) and two validation sets (NSCLC validation set, n = 172 ; SKCM validation set, n = 210). The mechanism was analyzed in the Cancer Genome Atlas (TCGA) database (n = 1467). Results: In the discovery set, patients with PAPPA2 mutation exhibited a significantly predominant PFS (HR, 0.11 [95% CI, 0.01-0.83]; P = 0.01), ORR (100.0% vs 21.4%; P < 0.001) and durable clinical benefit (DCB, 83.3% vs 32.1%; P = 0.012) compared to those with wide-type PAPPA2, consistent in NSCLC validation set (PFS, HR, 0.3 [95% CI: 0.16-0.56], P < 0.001; ORR, 70.8% vs. 20.9%, P < 0.001; DCB, 83.3% vs. 36.5%, P < 0.001) and SKCM validation set (OS, HR, 0.49 [95% CI: 0.31-0.78]; P = 0.002; ORR, 34.1% vs. 16.9%, P = 0.039; DCB, 50.0% vs. 30.7%; P = 0.036). Similar results were observed in multivariable models. In contrast, no association between PAPPA2 mutation and OS or PFS was observed in TCGA datasets, suggesting that PAPPA2 mutation may be a predictive but not a prognostic factor in ICI treatment for NSCLC patients. Based on CIBERSORT-inferred tumor infiltrating lymphocytes from TCGA, we found that PAPPA2 mutation was significantly associated with higher activated CD4 memory T cells and lower Treg cells in tumor immune microenvironment, related with higher TMB ( P < 0.001) and neoantigen load ( P < 0.001). In addition, gene set enrichment analysis (GSEA) analysis revealed that PAPPA2 mutation was correlated with upregulated DNA damage repair (DDR) signaling pathway. Conclusions: Our findings indicated that PAPPA2 mutation could serve as a novel indicator to stratify beneficiaries from ICIs therapy in NSCLC and SKCM, warranting further prospective studies.

XIRP2 mutation as an indicator stratified patients benefit from immune checkpoint inhibitors in NSCLC.

e14573 Background: Immune checkpoint inhibitors (ICIs), targeting the programmed cell death 1 (PD-1), programmed cell death ligand 1 (PD-L1) or cytotoxic T lymphocyte-associated protein 4 (CTLA-4) have demonstrated impressive anti-tumor efficacy in multiple tumors, particularly non-small cell lung cancer (NSCLC). However, only a minority of patients could clinically benefit from the ICIs. Therefore, it is of vital clinical significance to explore underlying predictive biomarkers to identify to identify these most potentially ICIs-benefitable patients. Xin-actin binding repeat containing 2 (XIRP2) encodes an actin-cross-linking protein that was previously reported to be significantly mutated in breast cancer metastases. However, its role in lung cancer immunotherapy remains unknown. Here we aimed to explore the association between XIRP2 and ICIs. Methods: 165 NSCLC patients from three public immunotherapy cohorts (Hellmann 2018, Miao 2018, and Rizvi 2015 cohort) were included to analyze the association between XIRP2 gene mutation and efficacy of ICI therapy. Genomic, survival and mRNA data of NSCLC patients from the Cancer Genome Atlas (TCGA) database was used to explore the potential mechanisms of anti-tumor immunity. Results: NSCLC patients with XIRP2 mutation were significantly associated with better PFS (HR = 0.4; 95% CI, 0.21-0.75; P = 0.003) after ICIs therapy, compared with those with wide-type XIRP2. XIRP2 mutation were associated with increased TMB ( P < 0.001). In the multivariable Cox proportional hazards regression model adjusted by smoking status, PD-L1 expression and TMB, the association between XIRP2 mutation and PFS remained significant (HR = 0.4; 95% CI, 0.20-0.81; P = 0.011). In contrast, no association between XIRP2 mutation and OS (LUAD: P = 0.43 ; LUSC: P = 0.17) or PFS (LUAD: P = 0.54; LUSC: P = 0.19) was observed in TCGA datasets, suggesting that XIRP2 mutation may be a predictive but not a prognostic factor in ICI treatment for NSCLC patients. Based on CIBERSORT-inferred tumor infiltrating lymphocytes from TCGA, XIRP2 mutation was significantly associated with higher M1 macrophage, CD8+ T cell, as well as lower Treg cell. Moreover, XIRP2 mutation was associated with increased TMB ( P < 0.001) and neoantigen load ( P < 0.001) in TCGA. Conclusions: Our results demonstrated that XIRP2 mutation is an underlying classifier that could stratify patients with NSCLC for ICIs. The role of XIRP2 in immunotherapy is needed to be further studied.

ANK2 mutation as an indicator stratified patients benefit from immune checkpoint inhibitors in NSCLC.

e21080 Background: Immune checkpoint inhibitors (ICIs), targeting the programmed cell death 1 (PD-1), programmed cell death ligand 1 (PD-L1) or cytotoxic T lymphocyte-associated protein 4 (CTLA-4) have demonstrated impressive anti-tumor efficacy in multiple cancers, in particular with non-small cell lung cancer (NSCLC). However, only a minority of patients could benefit from ICIs. Therefore, investigating predictive biomarkers of ICIs is important to identify the target population that derives the best therapeutic benefit. ANK2 encodes ankyrin-2, which plays a crucial role in cell development. Studies have shown a significant correlation between ANK2 expression levels and clinical outcomes in gastrointestinal cancers and pancreatic cancer. However, its role in lung cancer immunotherapy remains unknown. Here we aimed to demonstrate the association between ANK2 and ICIs. Methods: 165 patients from three public immunotherapy cohorts (Hellmann 2018, Miao 2018, and Rizvi 2015 cohort) were included to analyze the association between ANK2 gene mutation and efficacy of ICI therapy. Genomic, survival and mRNA data of NSCLC patients from the Cancer Genome Atlas (TCGA) database was used to explore the potential mechanisms of anti-tumor immunity. Results: Patients with ANK2 mutation were significantly associated with better PFS (HR = 0.28; 95% CI, 0.15-0.53; P < 0.001), objective response rate (ORR, 69.0% vs 24.3%; P < 0.001) and durable clinical benefit (DCB, 72.4% vs 41.9%; P = 0.003) compared with those with wide-type in NSCLC. ANK2 mutation were associated with increased TMB (P < 0.001). In the multivariable Cox proportional hazards regression model adjusted by the smoking status, PD-L1 expression and TMB, the association between ANK2 mutation and PFS remained significant (HR = 0.28; 95% CI, 0.20-0.82; P = 0.012).In TCGA, no association between ANK2 mutation and OS (LUAD: P =0.27 ; LUSC: P = 0.39) or PFS (LUAD: P = 0.21; LUSC: P = 0.63) was observed, suggesting that ANK2 mutation may be a predictive but not a prognostic factor in ICI treatment for NSCLC patients. Based on CIBERSORT-inferred tumor infiltrating lymphocytes from TCGA, ANK2mutation was significantly associated with higher CD8+ T cells, higher M1 macrophage, and lower M2 macrophage. Moreover, ANK2 mutation was associated with increased TMB (P < 0.001) and neoantigen load (P < 0.001) in TCGA. In addition, GSEA analysis revealed that ANK2 mutation was correlated with upregulated DNA damage repair (DDR) signaling pathway in LUAD. Conclusions: Our results demonstrated that ANK2 mutation is an independent classifier that could stratify patients with NSCLC for ICIs. The role of ANK2 in immunotherapy is needed to be further studied.

Slit mutation as an indicator stratified patients benefit from immune checkpoint inhibitors in NSCLC.

e14572 Background: Immune checkpoint inhibitors (ICIs), targeting the programmed cell death 1 (PD-1), programmed cell death ligand 1 (PD-L1) or cytotoxic T lymphocyte-associated protein 4 (CTLA-4) have demonstrated impressive anti-tumor efficacy in multiple tumors, particularly non-small cell lung cancer (NSCLC). However, only a minority of patients could clinically benefit from the ICIs. Therefore, it is of vital clinical significance to explore underlyingpredictive biomarkers to identify to identify these most potentially ICIs-benefitable patients. Slits (Slit1, 2 and 3) are highly conserved secreted glycoproteins that play important roles in tumorigenesis, including cancer cell proliferation, motility and angiogenesis. However, its role in lung cancer immunotherapy remains unknown. Here we aimed to explore the association between SLIT and ICIs. Methods: 165 NSCLC patients from three public immunotherapy cohorts (Hellmann 2018, Miao 2018, and Rizvi 2015 cohort) were included to analyze the association between SLIT gene mutation and efficacy of ICI therapy. Genomic, survival and mRNA data of NSCLC patients from the Cancer Genome Atlas (TCGA) database was used to explore the potential mechanisms of anti-tumor immunity. Results: NSCLC patients with SLIT mutation were significantly associated with better PFS (HR = 0.27; 95% CI, 0.14-0.52; P < 0.001), objective response rate (ORR, 63.3% vs 25.2%; P < 0.001) and durable clinical benefit (DCB, 86.7% vs 63.0%; P = 0.048) after ICIs therapy, compared with those with wide-type SLIT. SLIT mutation were associated with increased TMB ( P < 0.001). In the multivariable Cox proportional hazards regression model adjusted by smoking status, PD-L1 expression and TMB, the association between SLIT mutation and PFS remained significant (HR = 0.38; 95% CI, 0.19-0.77; P = 0.007). In contrast, no association between SLIT mutation and OS (LUAD: P = 0.57 ; LUSC: P= 0.1) or PFS (LUAD: P = 0.71; LUSC: P = 0.55) was observed in TCGA datasets, suggesting that SLIT mutation may be a predictive but not a prognostic factor in ICI treatment for NSCLC patients. Based on CIBERSORT-inferred tumor infiltrating lymphocytes from TCGA, SLIT mutation was significantly associated with higher naive B cell, plasma B cell, activated memory CD4+ T cell, CD8+ T cell, as well as lower M2 macrophage and Treg cell. Moreover, SLIT mutation was associated with increased TMB ( P < 0.001) and neoantigen load ( P < 0.001) in TCGA. In addition, GSEA analysis revealed that SLIT mutation was correlated with upregulated DNA damage repair (DDR) signaling pathway and activation of NF-κB in B-cell. Conclusions: Our results demonstrated that SLIT mutation is an underlying classifier that could stratify patients with NSCLC for ICIs. The role of SLIT in immunotherapy is needed to be further studied.

NPAP1 mutation as an indicator stratified patients benefit from immune checkpoint inhibitors in NSCLC.

e14574 Background: Immune checkpoint inhibitors (ICIs), targeting the programmed cell death 1 (PD-1), programmed cell death ligand 1 (PD-L1) or cytotoxic T lymphocyte-associated protein 4 (CTLA-4) have demonstrated impressive anti-tumor efficacy in multiple tumors, particularly non-small cell lung cancer (NSCLC). However, only a minority of patients could clinically benefit from the ICIs. Therefore, it is of vital clinical significance to explore underlying predictive biomarkers to identify to identify these most potentially ICIs-benefitable patients. Nuclear pore associated protein 1 ( NPAP1) encodes an protein is associated with the nuclear pore complex and has been previously reported as a potential prognostic biomarker for laryngeal, colorectal and lung cancers. However, its role in lung cancer immunotherapy remains unknown. Here we aimed to explore the association between NPAP1 and ICIs. Methods: 165 NSCLC patients from three public immunotherapy cohorts (Hellmann 2018, Miao 2018, and Rizvi 2015 cohort) were included to analyze the association between NPAP1 gene mutation and efficacy of ICI therapy. Genomic, survival and mRNA data of NSCLC patients from the Cancer Genome Atlas (TCGA) database was used to explore the potential mechanisms of anti-tumor immunity. Results: NSCLC patients with NPAP1 mutation were significantly associated with better PFS (HR = 0.37; 95% CI, 0.19-0.71; P = 0.002), objective response rate (ORR, 54.2% vs 28.4%; P = 0.048) and durable clinical benefit (DCB, 75.0% vs 42.6%; P= 0.003) after ICIs therapy, compared with those with wide-type NPAP1. NPAP1 mutation were associated with increased TMB ( P < 0.001). In the multivariable Cox proportional hazards regression model adjusted by smoking status, PD-L1 expression and TMB, the association between NPAP1 mutation and PFS remained significant (HR = 0.46; 95% CI, 0.23-0.92; P = 0.028). In contrast, no association between NPAP1 mutation and OS (LUAD: P = 0.24 ; LUSC: P = 0.94) or PFS (LUAD: P = 0.3; LUSC: P = 0.72) was observed in TCGA datasets, suggesting that NPAP1 mutation may be a predictive but not a prognostic factor in ICI treatment for NSCLC patients. Based on CIBERSORT-inferred tumor infiltrating lymphocytes from TCGA, NPAP1 mutation was significantly associated with higher activated memory CD4+ T cell and CD8+ T cell. Moreover, NPAP1 mutation was associated with increased TMB ( P < 0.001) and neoantigen load ( P < 0.001) in TCGA. Conclusions: Our results demonstrated that NPAP1 mutation is an underlying classifier that could stratify patients with NSCLC for ICIs. The role of NPAP1 in immunotherapy is needed to be further studied.

DYNC2H1 Mutation as an indicator stratified patients benefit from Immune Checkpoint Inhibitors in NSCLC.

e21052 Background: Immune checkpoint inhibitors (ICIs), targeting the programmed cell death 1 (PD-1), programmed cell death ligand 1 (PD-L1) or cytotoxic T lymphocyte-associated protein 4 (CTLA-4) have demonstrated impressive anti-tumor efficacy in multiple tumors, particularly non-small cell lung cancer (NSCLC). However, only a minority of patients could clinically benefit from the ICIs. Therefore, it is of vital clinical significance to explore underlying predictive biomarkers to identify to identify these most potentially ICIs-benefitable patients. Dynein cytoplasmic 2 heavy chain 1 (DYNC2H1) encodes a large cytoplasmic dynein protein that involved in intraflagellar transport. However, its role in lung cancer immunotherapy remains unknown. Here we aimed to explore the association between DYNC2H1 and ICIs. Methods: 165 NSCLC patients from three public immunotherapy cohorts (Hellmann 2018, Miao 2018, and Rizvi 2015 cohort) were included to analyze the association between DYNC2H1 gene mutation and efficacy of ICI therapy. Genomic, survival and mRNA data of NSCLC patients from the Cancer Genome Atlas (TCGA) database was used to explore the potential mechanisms of anti-tumor immunity. Results: NSCLC patients with DYNC2H1 mutation were significantly associated with better PFS (HR = 0.3; 95% CI, 0.14-0.64; P= 0.001), objective response rate (ORR, 65.0% vs 27.6%; P = 0.003) and durable clinical benefit (DCB, 75.0% vs 43.4%; P = 0.031) after ICIs therapy, compared with those with wide-type DYNC2H1. DYNC2H1 mutation were associated with increased TMB ( P < 0.001). In the multivariable Cox proportional hazards regression model adjusted by smoking status, PD-L1 expression and TMB, the association between DYNC2H1 mutation and PFS remained significant (HR = 0.32; 95% CI, 0.14-0.73; P = 0.007). In contrast, no association between DYNC2H1 mutation and OS (LUAD: P = 0.97 ; LUSC: P = 0.34) or PFS (LUAD: P = 0.54; LUSC: P = 0.39) was observed in TCGA datasets, suggesting that DYNC2H1 mutation may be a predictive but not a prognostic factor in ICI treatment for NSCLC patients. Based on CIBERSORT-inferred tumor infiltrating lymphocytes from TCGA, DYNC2H1 mutation was significantly associated with higher CD8+ T cell. Moreover, DYNC2H1 mutation was associated with increased TMB ( P < 0.001) and neoantigen load ( P < 0.001) in TCGA. Conclusions: Our results demonstrated that DYNC2H1 mutation is an underlying classifier that could stratify patients with NSCLC for ICIs. The role of DYNC2H1 in immunotherapy is needed to be further studied.

MER4 endogenous retrovirus correlated with better efficacy of anti-PD1/PD-L1 therapy in non-small cell lung cancer

BackgroundEndogenous retroviruses (ERVs) are highly expressed in various cancer types and are associated with increased innate immune response and better efficacy of antiprogrammed death-1/ligand-1 (anti-PD1/PD-L1)-directed immune checkpoint inhibitors (ICI) in...

Influence of DNA Mismatch Repair (MMR) System in Survival and Response to Immune Checkpoint Inhibitors (ICIs) in Non-Small Cell Lung Cancer (NSCLC): Retrospective Analysis.

Mutations in the mismatch repair (MMR) system predict the response to immune checkpoint inhibitors (ICIs) like colon or gastric cancer. However, the MMR system’s involvement in non-small cell lung cancer (NSCLC) remains unknown. Addressing this issue will improve clinical guidelines in the case of mutations in the main genes of the MMR system (MLH1, MSH2, MSH6, and PMS2). This work retrospectively assessed the role that these gene mutations play in the response to and survival of ICIs in NSCLC. Patients with NSCLC treated with nivolumab as the second-line treatment in the University Hospital of Salamanca were enrolled in this study. Survival and response analyses were performed according to groups of MMR system gene expression (MMR expression present or deficiency) and other subgroups, such as toxicity. There was a statistically significant relationship between the best response obtained and the expression of the MMR system (p = 0.045). The presence of toxicity grade ≥ 3 was associated with the deficiency expression of MMR (dMMR/MSI-H) group (p = 0.022; odds ratio = 10.167, 95% confidence interval (CI) 1.669–61.919). A trend towards greater survival and response to ICIs was observed in NSCLC and dMMR. Assessing the genes in the MMR system involved in NSCLC is key to obtaining personalized immunotherapy treatments.

Immunotherapy in Early Stage Non-Small Cell Lung Cancer

Immune-checkpoint inhibitors are extensively used in cancer treatment and have transformed the therapeutic landscape by inducing durable responses. Immunotherapy with checkpoint inhibitors targeting programmed death 1 (PD-1) receptor and programmed death ligand-1 (PDL-1) are used alone or with chemotherapy for treatment of metastatic non-small cell lung cancer (NSCLC). There is a great need for improving outcomes of patients with early stage NSCLC after surgical resection and with recent F. D. A. approval, immune checkpoint inhibitors are used as neoadjuvant or adjuvant therapy to enable curative resection and prevent or delay disease progression. In this article, we review the clinical studies evaluating the role of adjuvant and neoadjuvant immune checkpoint inhibitors in NSCLC and discuss the role of immunotherapy with radiation therapy in locally advanced non-metastatic NSCLC.

Genomic data from NSCLC tumors reveals correlation between SHP-2 activity and PD-L1 expression and suggests synergy in combining SHP-2 and PD-1/PD-L1 inhibitors.

The identification of novel therapies, new strategies for combination of therapies, and repurposing of drugs approved for other indications are all important for continued progress in the fight against lung cancers. Antibodies that target immune checkpoints can unmask an immunologically hot tumor from the immune system of a patient. However, despite accounts of significant tumor regression resulting from these medications, most patients do not respond. In this study, we sought to use protein expression and RNA sequencing data from The Cancer Genome Atlas and two smaller studies deposited onto the Gene Expression Omnibus (GEO) to advance our hypothesis that inhibition of SHP-2, a tyrosine phosphatase, will improve the activity of immune checkpoint inhibitors (ICI) that target PD-1 or PD-L1 in lung cancers. We first collected protein expression data from The Cancer Proteome Atlas (TCPA) to study the association of SHP-2 and PD-L1 expression in lung adenocarcinomas. RNA sequencing data was collected from the same subjects through the NCI Genetic Data Commons and evaluated for expression of the PTPN11 (SHP-2) and CD274 (PD-L1) genes. We then analyzed RNA sequencing data from a series of melanoma patients who were either treatment naïve or resistant to ICI therapy. PTPN11 and CD274 expression was compared between groups. Finally, we analyzed gene expression and drug response data collected from 21 non-small cell lung cancer (NSCLC) patients for PTPN11 and CD274 expression. From the three studies, we hypothesize that the activity of SHP-2, rather than the expression, likely controls the expression of PD-L1 as only a weak relationship between PTPN11 and CD274 expression in either lung adenocarcinomas or melanomas was observed. Lastly, the expression of CD274, not PTPN11, correlates with response to ICI in NSCLC.

Prognostic significance of pre-treatment ALBI grade in advanced non-small cell lung cancer receiving immune checkpoint therapy

The liver is an essential organ for regulating innate and acquired immunity. We hypothesized that the pre-treatment hepatic function affects the clinical outcome of immune checkpoint inhibitors (ICIs) in non-small cell lung cancer (NSCLC). We analyzed 140 patients with NSCLC who received ICIs. We investigated the association between pre-treatment liver function, assessed using the albumin–bilirubin (ALBI) grade, and clinical outcomes in univariate, multivariate, and propensity score matching analyses. Patients were divided into four grades according to pre-treatment liver function. Eighty-eight patients had good hepatic reserve (ALBI grade 1 or 2a), whereas 52 patients had poor hepatic reserve (ALBI grade 2b or 3). In the univariate Kaplan–Meier analysis, the ALBI grade 1, 2a group had a significantly prolonged progression-free survival (PFS, 5.3 versus 2.5 months, p = 0.0019) and overall survival (OS, 19.6 vs. 6.2 months, p = 0.0002). These results were consistent, regardless of whether the analysis was performed in patients with a performance status of 0 or 1 at pre-treatment (N = 124) or in those selected using propensity score matching (N = 76). In the multivariate analysis, pre-treatment ALBI grade was an independent prognostic factor for both PFS (hazard ratio [HR] 0.57, 95% confidence interval [95% CI] 0.38–0.86, p = 0.007) and OS (HR 0.45, 95% CI 0.29–0.72, p = 0.001). Our results suggest that pre-treatment hepatic function assessed by ALBI grade could be an essential biomarker for predicting the efficacy of treatment with ICIs in NSCLC.

The Impact of NOTCH Pathway Alteration on Tumor Microenvironment and Clinical Survival of Immune Checkpoint Inhibitors in NSCLC.

The treatment of non-small cell lung cancer (NSCLC) with immune checkpoint inhibitors (ICIs) has been proven to induce lasting tumor remission. Screening suitable populations for immunotherapy through predictive markers is an important approach to improving the clinical benefits of patients. Evidence has shown that there may be a close connection between NOTCH signaling and the tumor microenvironment (TME). Hence, we explored the impact of the mutation status of NOTCH signaling on the prognosis of NSCLC patients treated with immunotherapy with the aim to apply NSCLC immunotherapy to the greatest extent possible. We examined two clinical cohorts of NSCLC patients receiving ICIs (MSKCC and NG cohorts). The mutation and prognostic data of the ICI-treated cohort were used to evaluate the association between the mutation status of NOTCH signaling and prognosis following immunotherapy. The expression and mutation data of The Cancer Genome Atlas (TCGA)-NSCLC cohort were used to analyze the differences in the immune microenvironment under different NOTCH signaling mutation states. In the ICI-treated cohorts, the univariate and multivariate Cox regression analyses indicated that high-mutated NOTCH signaling could serve as an independent predictor of NSCLC patients receiving ICIs. Patients with high-mutated NOTCH signaling had significantly improved progression-free survival (PFS) (P = 0.03, HR = 0.69; MSKCC cohort) and prolonged overall survival (OS) (P = 0.004, HR = 0.34; NG cohort). Additionally, high-mutated NOTCH signaling was related to the inflammatory immune microenvironment, inflammatory expression profile, and enhanced immunogenicity. According to this study, high-mutated NOTCH signaling may serve as a biomarker for the prediction of the prognosis of NSCLC patients treated with ICIs. A series of prospective clinical studies and molecular mechanism explorations are still needed in the future.

Abstract 166: Real world genomic data supports combined use of SHP-2 and PD-1/PD-L1 inhibitors in solid tumors

Abstract The identification of novel therapies, new strategies for combination of therapies, and repurposing of drugs approved for other indications are all important for continued progress in the fight against lung cancers. Antibodies that target immune checkpoints can unmask an immunologically hot tumor from the immune system of a patient. However, despite accounts of significant tumor regression resulting from these medications, most patients do not respond. In this study, we sought to use protein expression and RNA sequencing data from The Cancer Genome Atlas and two smaller studies deposited onto the Gene Expression Omnibus (GEO) to advance our hypothesis that inhibition of SHP-2, a tyrosine phosphatase, will improve the activity of immune checkpoint inhibitors (ICI) that target PD-1 or PD-L1 in lung cancers. We first collected protein expression data from the The Cancer Proteome Atlas (TCPA) to study the association of SHP-2 and PD-L1 expression in lung adenocarcinomas. RNA sequencing data was collected from the same subjects through the NCI Genetic Data Commons and evaluated for expression of the PTPN11 (SHP-2) and CD274 (PD-L1) genes. We then analyzed RNA sequencing data from a series of melanoma patients who were either treatment naïve or resistant to ICI therapy. PTPN11 and CD274 expression was compared between groups. Finally, we analyzed gene expression and drug response data collected from 21 non-small cell lung cancer (NSCLC) patients for PTPN11 and CD274 expression. From the three studies, we hypothesize that the activity of SHP-2, rather than the expression, likely controls the expression of PD-L1 as only a weak relationship between PTPN11 and CD274 expression in either lung adenocarcinomas or melanomas was observed. Lastly, the expression of CD274, not PTPN11, correlates with response to ICI in NSCLC. Citation Format: Keller Toral. Real world genomic data supports combined use of SHP-2 and PD-1/PD-L1 inhibitors in solid tumors [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 166.

Potential role of serum proteome in predicting immune-related adverse events from immune checkpoint inhibitors in non-small cell lung cancer.

e21218 Background: Early recognition of immune-related adverse events (irAEs) of immune checkpoint inhibitors(ICI) is important. Circulating proteome reflects host response to diseases and is being explored as a marker for response to immunotherapy. We previously have reported that a serum-based proteomics test, Primary Immune Response (PIR) demonstrated a trend that PIR-sensitive patients are more likely to tolerate ICI treatment longer without developing irAEs in non-small cell lung cancer (NSCLC) patients. The VeriStrat test is another serum-based proteomic assay, which was reported to be predictive of survival outcomes for all treatment regimens and lines of therapy including ICI in NSCLC. We explored the associations between the VeriStrat test and developing irAEs in NSCLC patients treated with ICI. Methods: Data of 70 consented NSCLC patients treated with any regimens and lines of therapy including ICI were collected. Samples were grouped into either VeriStrat ‘Good’(VS-G) or VeriStrat ‘Poor’(VS-P). We analyzed the durations from the immunotherapy initiation to each episode of irAE and each irAE above grade 2 using log-rank test. IrAEs were graded according to Common Terminology Criteria for Adverse Events (CTCAE) v5.0. Results: Among the 70 patients, 18 patients (25%) experienced one or more irAEs. There was no significant difference in ‘Time to first irAE’ between VS-G and VS-P (p = 0.72, HR = 0.82, 95% CI = 0.29-2.32). Among 48 VS-G patients, 12(25%) had one or more irAE and 5(10%)had irAE graded over 2. Among 22 VS-P patients, 6(27%) had one or more irAE and 2(9%) had irAE graded over 2. There was no significant difference between VS-G and VS-P groups in the development of irAE and irAE graded over 2. Conclusions: There was no statistically significant association between the VeriStrat test and the development of irAEs. Further studies are warranted to investigate proper serum based proteomic assay to predict the development of irAE.