Correlation Of PD-L1 Expression Research Articles

Correlation of PD-L1 expression with different clinico-pathological and immunohistochemical features of ovarian surface epithelial tumors.

Primary carcinoma of the ovary (OCs) are responsible for a significant number of deaths related to cancer, and have the highest rate of death related to cancers of the female reproductive organs. Programmed cell death 1 (PD1) protein, acts as an immune checkpoint, and has an important role in the down-regulation of the immune system by preventing the activation of T-cells, which will weaken the autoimmunity and increases self-tolerance. This study aimed at the evaluation of the immunohistochemical (IHC) expression of PD-L1 in various primary surface ovarian epithelial tumours and to test its correlation with different clinicopathological parameters together with the expression of a panel of P53, ER and PR. A set of 102 cases of primary ovarian surface epithelial neoplasms (benign, borderline and malignant) were collected to construct Tissue Microarray (TMA) using 3 tissue cores from each case. IHC for PD-L1, p53, PR and ER was performed. The expression of PD-L1 was evaluated in relation to some clinicopathological parameters and to the expression patterns of other markers. Expression of PD-L1 was detected in about 51% (n = 36) of malignant tumours. The malignant group significantly showed PD-L1 positivity compared to borderline and benign groups. The malignant tumours significantly showed PD-L1 and total p53 positivity in comparison to borderline group. Also, malignant tumours significantly showed higher combined positivity of PD-L1 and either PR or ER compared to borderline and benign lesions. No significant correlation was appreciated between PD-L1 expression and with any of the studied clinicopathological parameters. This study showed a significant PD-L1 expression in malignant primary surface epithelial tumours. Construction of a panel of IHC markers, including PD-L1, could have a potential value to define patients those would benefit from the addition of immunotherapy to the treatment plan.

Correlation of PDL-1 Expression with Tumour Budding and Tumour Infiltrating Lymphocytes in Colorectal Cancers

Objective: To study and correlate PDL-1 expression with Tumour budding and Tumour Infiltrating Lymphocytes in Colorectal Carcinoma. Background: Colorectal cancer (CRC) is third most common cancer with a high mortality. Many attempts have been made to raise overall survival of CRC patients. The immune system plays an important role in clearing the unhealthy cancer cells. Programmed death 1 (PD1) is a regulatory molecule which dampens the immune response when bound to one of its complementary ligands (PDL1). Its expression is related to the response of immunotherapy in CRC treatment which has been exploited in recent times. However, its prognostic value is still controversial, and the distribution of PD-L1 on tumour Cells or Immune Cells has not been comprehensively analysed. Method: A total of 30 patients diagnosed with CRCs were included who underwent surgical intervention. Cases who took preoperative neoadjuvant chemotherapy or radiotherapy were excluded. IHC analyses of PDL1 was done and was correlated with tumour budding and Tumour Infiltrating Lymphocytes (TILs) and statistical significance was assessed. Results: 11 cases showed low bud count at the invasive front out of which only 3 cases showed PDL1 positivity. The rest 19 cases had high bud count out of which 18 were PDL 1 positive. This difference was highly significant (p = 0.002). In Low Bud / High TILs, 75% cases showed no PDL1 expression in tumour cells, whereas 62.5% cases showed PDL1 positivity in TILs whereas in High Bud / Low TILs group, all the cases (100%) showed PDL1 expression in tumour cells whereas only 75% cases showed PDL1 positivity in TILs, again being statistically significant (p <0.001). Conclusion: This study showed an inverse correlation between PDL1 in tumour buds and immune cells, thus emphasising the role of tumour microenvironment. Our study reiterates the fact that high expression of PDL1 in tumour cells suppresses antitumor response whereas its high expression in TILS correlates with a better prognosis.

Durvalumab plus tremelimumab for the treatment of advanced neuroendocrine neoplasms of gastroenteropancreatic and lung origin

Single immune checkpoint blockade in advanced neuroendocrine neoplasms (NENs) shows limited efficacy; dual checkpoint blockade may improve treatment activity. Dune (NCT03095274) is a non-randomized controlled multicohort phase II clinical trial evaluating durvalumab plus tremelimumab activity and safety in advanced NENs. This study included 123 patients presenting between 2017 and 2019 with typical/atypical lung carcinoids (Cohort 1), G1/2 gastrointestinal (Cohort 2), G1/2 pancreatic (Cohort 3) and G3 gastroenteropancreatic (GEP) (Cohort 4) NENs; who progressed to standard therapies. Patients received 1500 mg durvalumab and 75 mg tremelimumab for up to 13 and 4 cycles (every 4 weeks), respectively. The primary objective was the 9-month clinical benefit rate (CBR) for cohorts 1-3 and 9-month overall survival (OS) rate for Cohort 4. Secondary endpoints included objective response rate, duration of response, progression-free survival according to irRECIST, overall survival, and safety. Correlation of PD-L1 expression with efficacy was exploratory. The 9-month CBR was 25.9%/35.5%/25% for Cohorts 1, 2, and 3 respectively. The 9-month OS rate for Cohort 4 was 36.1%, surpassing the futility threshold. Benefit in Cohort 4 was observed regardless of differentiation and Ki67 levels. PD-L1 combined scores did not correlate with treatment activity. Safety profile was consistent with that of prior studies. In conclusion, durvalumab plus tremelimumab is safe in NENs and shows modest survival benefit in G3 GEP-NENs; with one-third of these patients experiencing a prolonged OS.

Correlation of PDL1 Expression with Genomic Alterations in Non-Small Cell Lung Cancer: a 7-month retrospective analysis

Abstract Background PD1/PDL1 inhibitors have significantly improved overall survival in non-small cell lung cancer (NSCLC). PDL1 is now a standard biomarker along with molecular testing for therapy selection. Checkpoint therapy is given front-line for patients with high PDL1 and no actionable mutations or fusions, or after progression with any PDL1 expression. Studies comparing PDL1 status in the context of actionable mutations or fusions in NSCLC are sparse. Design All patients undergoing molecular testing for a next-generation sequencing (NGS) lung cancer panel at our institution in a 7-month period were reviewed. This included an NGS DNA panel for actionable/driver genes: EGFR, ERBB2, BRAF, K/NRAS, PIK3CA, and IDH1/2; and a NGS RNA fusion panel for ALK, ROS1, RET, NTRK1/3, NRG1, and MET e14 skipping. PDL1 expression by IHC was scored for tumor proportion score (TPS). Statistical comparison was performed using Fisher’s Exact test. Results 186 patients with NSCLC were included (126 adenocarcinoma, 33 squamous/adeno-squamous, 19 poorly differentiated, and 8 NSCLC-NOS); 162 of those had concurrent PDL1 IHC. 121 (65%) patients had a genomic alteration, most commonly RAS [28%], EGFR [24%], ALK [8%], MET [7%], and BRAF [5%]. Cases with genomic alterations were classified into those with and without RAS mutation/s. The prevalence of high PDL1 expression was greater in RAS-driven tumors: 47% vs 22% with non-RAS bearing genomic alterations (p=0.0173). There was no significant difference in the PDL1 expression between the genomically negative tumors and those with non-RAS bearing mutations. Recently, the FDA granted approval to sotorasib, a KRAS G12C inhibitor, in patients with locally advanced or metastatic NSCLC. However, there was no significant difference in the PDL1 expression between the tumors with RAS G12C mutation and those with other RAS mutations. Conclusion Our data indicate that close to half of RAS-driven NSCLC have high PDL1 expression, an important consideration for front-line therapy selection. The majority of genomically-actionable tumors have at least low-positive expression of PDL1, highlighting the potential importance of immune checkpoint therapy in the resistant-progression setting. In comparison, only about one third of genomically-negative cases have high PDL1 expression, and a significant percent are completely negative. When the actionable KRAS G12C mutation bearing cases were compared to those with other RAS mutations, no significant difference was noted in the PDL1 expression between these two subsets; further investigations and a larger cohort of patients may be helpful in this case to determine whether PDL1 levels are affected by a specific RAS mutation.

Abstract 5200: The incidence and risk factors of hyperprogressive disease(HPD) in non-small cell lung cancer(NSCLC) treated with immune checkpoint inhibitors(ICI)

Abstract Background: HPD is an unexpected rapid acceleration of tumor growth reported for patients treated with ICI. The definition of HPD is not fully established and risk factors are uncertain. Our aim in this study was to find the incidence of HPD as well as its significant clinical and genetic risk factors. Method: A total of 203 patients with NSCLC who received ICI at Northwestern Memorial Hospital during 2008-2020 were included. Tumor growth kinetics (TGK) on immunotherapy and TGK pre-immunotherapy were collected and TGK ratio (TGKR) was calculated. HPD was defined as TGKR≥2. Result: HPD was observed in 16% (n=33) of patients. HPD was significantly associated with high ECOG (3-4 vs 1-2) [OR = 4.52, 95% CI = 1.31 to 16.29, P = 0.01], presence of bone metastasis [OR = 2.51, 95% CI = 1.10 to 5.28, P = 0.01], neutrophils/lymphocyte ratio(NLR) ≥5 [OR = 2.68, 95% CI = 1.24 to 5.81 P = 0.01], thrombocytosis (platelet count >450k) [OR = 5.31, 95% CI = 2.07 to 14.70, P=0.0003], and treatment with PD-L1 inhibitor vs PD-1 inhibitor [OR = 2.14, 95% CI= 0.97 to 4.68, P= 0.068]. HPD was inversely associated with positive PD-L1 expression [OR = 0.34, 95% CI = 0.13 to 0.86, P = 0.01], ICI combined with chemotherapy vs ICI monotherapy [OR = 0.24, 95% CI = 0.055 to 0.97, P=0.04], and positive TTF-1 expression [OR = 0.46, 95% CI = 0.21 to 1.02, P=0.057]. HPD was not associated with histology of NSCLC (adenocarcinoma, squamous cell carcinoma, large cell carcinoma, and large cell neuroendocrine carcinoma), age(<70, ≥70), sex, smoking history, number of metastatic lesions (<3, ≥3), brain metastasis, liver metastasis, brain metastasis before treatment initiation, EGFR mutation, TP53 mutation, BRAF mutation, and TNM staging. Conclusion: The incidence of HPD in NSCLC treated with ICI was consistent with current literature. High NLR and platelet count are suggestive of an immunosuppressive environment that could contribute to the rapid growth of tumors. To our knowledge, this is the first study to report an inverse correlation of PD-L1 expression and TTF-1 expression with HPD. The strength of our study lies in a large number of patients. Further studies on validating risk factors of HPD and exploring their associated mechanisms are warranted. Citation Format: Hyeonseon Kim, Yeun Ho Lee, Chiwoo Song, Leeseul Kim, Min Jeong Kim, Horyun Choi, Jinah Kim, Young Kwang Chae. The incidence and risk factors of hyperprogressive disease(HPD) in non-small cell lung cancer(NSCLC) treated with immune checkpoint inhibitors(ICI) [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 5200.

Correlation of PD-L1 Expression with Clinicopathological and Genomic Features in Chinese Non-Small-Cell Lung Cancer.

Programmed cell death 1 ligand 1 (PD-L1) has been approved as predictive biomarker for non-small-cell lung cancer (NSCLC) patients treated with PD-(L)1 blockade therapy. The clinical/genomic features associated with PD-L1 are not well studied. Genomic profiling of tumor biopsies from 883 Chinese NSCLC patients was performed by targeted next-generation sequencing. Immunohistochemical analysis was conducted to evaluate PD-L1 expression levels using antibodies Dako 22C3 and 28-8, respectively. Our study showed distinct correlation between PD-L1 expression and clinical/genomic characteristics when using different PD-L1 antibodies and in different histological subtypes including adenocarcinoma (ADC) and squamous cell carcinoma (SCC), respectively. PD-L1 high expression (22C3) was associated with male and lymph node metastasis only in ADC patients. Furthermore, mutations of TP53 and KRAS, KIF5B-RET fusion, copy number gains of PD-L1 and PD-L2, and arm-level amplifications of chr.12p were significantly associated with PD-L1 positive status in ADC patients. For SCC patients, the gain of EGFR and MDM2 and loss of PTPRD were negatively associated with PD-L1 expression. We also compared our results with other studies and found conflicting results presumably because of the multiplicity of antibody clones and platforms, the difference of cutoffs for assigning PD-L1 expression levels, and the variation in study populations. Our study can help to understand the utility and validity of PD-L1 as biomarker of response to immune checkpoint inhibitors.

What Do We Have to Know about PD-L1 Expression in Prostate Cancer? A Systematic Literature Review (Part 6): Correlation of PD-L1 Expression with the Status of Mismatch Repair System, BRCA, PTEN, and Other Genes.

Pembrolizumab (anti-PD-1) is allowed in selected metastatic castration-resistant prostate cancer (PC) patients showing microsatellite instability/mismatch repair system deficiency (MSI-H/dMMR). BRCA1/2 loss-of-function is linked to hereditary PCs and homologous recombination DNA-repair system deficiency: poly-ADP-ribose-polymerase inhibitors can be administered to BRCA-mutated PC patients. Recently, docetaxel-refractory metastatic castration-resistant PC patients with BRCA1/2 or ATM somatic mutations had higher response rates to pembrolizumab. PTEN regulates cell cycle/proliferation/apoptosis through pathways including the AKT/mTOR, which upregulates PD-L1 expression in PC. Our systematic literature review (PRISMA guidelines) investigated the potential correlations between PD-L1 and MMR/MSI/BRCA/PTEN statuses in PC, discussing few other relevant genes. Excluding selection biases, 74/677 (11%) PCs showed dMMR/MSI; 8/67 (12%) of dMMR/MSI cases were PD-L1+. dMMR-PCs included ductal (3%) and acinar (14%) PCs (all cases tested for MSI were acinar-PCs). In total, 15/39 (39%) PCs harbored BRCA1/2 aberrations: limited data are available for PD-L1 expression in these patients. 13/137 (10%) PTEN- PCs were PD-L1+; 10/29 (35%) PD-L1+ PCs showed PTEN negativity. SPOP mutations may increase PD-L1 levels, while the potential correlation between PD-L1 and ERG expression in PC should be clarified. Further research should verify how the efficacy of PD-1 inhibitors in metastatic castration-resistant PCs is related to dMMR/MSI, DNA-damage repair genes defects, or PD-L1 expression.

Molecular and Clinical Characteristics of Primary Pulmonary Lymphoepithelioma-Like Carcinoma.

Objectives: Primary pulmonary lymphoepithelioma-like carcinoma (PPLELC) is an extremely rare subtype of non-small cell lung cancer (NSCLC). Currently, there are no established treatment protocols due to rarity of the cancer. Thus, this study aimed to explore the molecular and clinical characteristics of PPLELC. Study design and setting: Data from patients with PPLELC who were admitted to Zhejiang Cancer Hospital from August 2009 to September 2020 were retrospectively collected. Next-generation sequencing was performed to obtain a genomic profile and tumor mutation burden (TMB) value of patients with adequate tissue and divided them into two groups according to the expression level of PD-L1. The correlation of PD-L1 expression and the clinicopathological characteristics was evaluated by Pearson Chi-square test. Kaplan-Meier curves was applied to present the probability of survival between PD-L1 expression level and overall survival (OS). Moreover, the literature on the immunotherapy of advanced PPLELC published in PubMed between 2016 and 2020 were reviewed and the efficacy of immunotherapy were analyzed. Results: A total of 18 patients pathologically diagnosed as PPLELC were included. After a follow-up period of 8.8–138 months, 14 patients survived, three patients died and one patient lost, the median OS was 45.3 months Seven samples (tissue-available) tested by NGS and the median TMB was 2.5 mutations/Mb. 19 somatic mutated genes were recognized and TP53 (43%) and CYLD (43%) were the two most commonly mutated genes. Only seven patients who underwent NGS were tested for PD-L1. Three patients with high PD-L1 expression (PD-L1≥ 50%) and four patients with low PD-L1 expression (PD-L1 <50%) were included. No significant correlation was observed between PD-L1 expression and clinical characteristics (age, gender, smoking status, tumor stage, lymph node metastasis) (p > 0.05) and OS (p = 1). What’s more, 10 PPLELC patients involved in previous studies and one patient received nivolumab in the current study were collected retrospectively. 4/11 (36.4%) patients achieved PR, 6/11 (54.5%) patients achieved SD, and 1/11 (9.1%) patients achieved PD and the disease control rate (DCR) was 90.9%. Conclusions: The prognosis of PPLELC is better than that of other NSCLC, and immunotherapy may be a promising treatment to prolong the survival of advanced PPLELC patients. Whether the immunotherapy efficacy of PPLELC can be predicted by PD-L1 and TMB needs further clinical investigation. CYLD genetic alterations may participate in Epstein–Barr virus-mediated tumorigenesis in PPLELC, providing a novel therapeutic target.

Correlation of PD-L1 Expression with TILs CD8+ in Non-small Cell Carcinoma Lung Cancer

BackgroundNon-small cell lung cancer (NSCLC) is an epithelial carcinoma of the lung which consists of adenocarcinoma, squamous cellcarcinoma (SCC), and large-cell carcinoma (LCC). In a microtumor environment, PD-L1 and PD-1 bonds are immune checkpointsthat act in suppressing the activity of CD8+ TILs, thus affecting the prognosis of NSCLC. Aims of this study is to determine thecorrelation between PD-L1 expression with CD8+TILs in NSCLC.MethodsThis was a cross-sectional study of NSCLC registered in Department of Anatomical Pathology Faculty of Medicine University ofSriwijaya/Dr. Moh. Hoesin Palembang Hospital since 1 January 2017 to 31 January 2020 with a total of 24 samples. All sampleswere immunostained using PD-L1 and CD8 antibodies. PD-L1 expression was assessed in tumor mass and CD8 expression inintratumoral and peritumoral TILs.ResultsOnly two subtypes, adenocarcinoma and SCC, were found in this study. Adenocarcinoma was the most frequent subtype (54.1%),sample group is dominated by &lt;60 years old and was the subtype that expressed the PD-L1 most strongly (TPS ≥50%). There wasnot significant correlation between PD-L1 expression with CD8+ TILs in NSCLC (p=0.679 and p=0.826) but based onclinicopathological characteristics, there was a significant relationship between PD-L1 with disease stage and between intratumoralCD8+ TILs density with disease stage (p=0.048 and p=0.017).ConclusionAdenocarcinoma subtype had stronger expression of PD-L1 compared to SCC. There was no correlation between PD-L1 and CD8+TILs density in NSCLC.

Microsatellite instability (MSI-H) is associated with a high immunoscore but not with PD-L1 expression or increased survival in patients (pts.) with metastatic colorectal cancer (mCRC) treated with oxaliplatin (ox) and fluoropyrimidine (FP) with and without bevacizumab (bev): a pooled analysis of the AIO KRK 0207 and RO91 trials

IntroductionIn a retrospective analysis of two randomized phase III trials in mCRC patients treated first line with oxaliplatin, fluoropyrimidine with and without Bevacizumab (the AIO KRK 0207 and R091 trials) we evaluated the association of high microsatellite instability (MSI-H), immunoscore (IS) and PD-L1 expression in relation to overall survival (OS).MethodsIn total, 550 samples were analysed. Immunohistochemical analysis of the MMR proteins and additionally fragment length analysis was performed, molecular examinations via allele-discriminating PCR in combination with DNA sequencing. Furthermore PD-L1 and IS were assessed.ResultsMSI-H tumors were more frequent in right sided tumors (13.66% vs. 4.14%) and were correlated with mutant BRAF (p = 0.0032), but not with KRAS nor NRAS mutations (MT). 3.1% samples were found to be PD-L1 positive, there was no correlation of PDL1 expression with MSI-H status, but in a subgroup analysis of MSI-H tumors the percentage of PD-L1 positive tumors was higher than in MSS tumors (9.75% vs. 2.55%). 8.5% of samples showed a positive IS, MSI-H was associated with a high IS. The mean IS of the pooled population was 0.57 (SD 0.97), while the IS of MSI-H tumors was significantly higher (mean of 2.4; SD 1.4; p =< 0.0001).DiscussionRegarding OS in correlation with MSI-H, PD-L1 and IS status we did not find a significant difference. However, PD-L1 positive mCRC tended to exhibit a longer OS compared to PD-L1 negative cancers (28.9 vs. 22.1 months).

Abstract 5381: Elevated PD-L1 and HER2 expression on aneuploid carcinoma cells in malignant ascites accelerate stumor cell immune evasion in gastrointestinal cancer patients

Abstract Background Peritoneal dissemination (PD) is a type of metastasis frequently found in gastrointestinal (GI) cancer patients, resulting in the production of massive malignant ascites which associate with poor prognosis. However, the question of how the expression of tumor biomarkers on cancer cells in malignant ascites correlates with PD is yet to be answered. This study aims to investigate the clinical significance of PD-L1 and HER2 expression on malignant ascites disseminated tumor cells (maDTCs) in GI carcinoma patients. Patients and Methods Paired blood and malignant ascites specimens were respectively collected from 24 of metastatic GI subjects, including 15 gastric, 2 pancreatic, 1 esophageal, and 6 colorectal cancer patients. Non-hematologic CTCs and maDTCs were enriched by subtraction enrichment (SE), followed by immunostaining-FISH (iFISH) to co-examine PD-L1 and HER2 expression on aneuploid maDTCs and CTCs. In addition, CD4+, CD8+, Treg and naive T cells in the same cohort of samples were quantified by flow cytometry. Results In situ karyotypic and phenotypic characterization performed by iFISH showed that the average number of maDTCs in the 24 patients was 32755/ml ascites, which was significantly higher than the average number of CTCs detected (3.5/ml blood) (P&amp;lt;0.0001). The number of maDTCs and CTCs exhibited no correlation between each other (Pearson's product-moment correlation coefficient, R=-0.03, P=0.89). Further examination indicated that PD-L1 was expressed on 50% of maDTCs compared to only 11% for CTCs (P=0.0002). No correlation of PD-L1 expression between maDTCs and CTCs was observed (Pearson's product-moment correlation coefficient, R=0.25, P=0.24). Co-expression of PD-L1 and HER2 on CTCs and maDTCs was respectively examined in the recruited 15 gastric cancer (GA) patients. All 15 patients were histopathological HER2 negative (hHER2-). One subject (1/15=6.7%) showed 1 double HER2+/PD-L1+ CTC (1/63 CTCs in 15 GA patients=1.6%). In comparison, 6 patients (6/15=40%) had a total of 290377 HER2+/PD-L1+ maDTCs detected (290377/1306783 overall maDTCs=22%). In addition, flow cytometry analyses revealed that 5% of lymphocytes in malignant ascites were immunosuppression relevant regulatory T cells (Tregs), compared to only 2% of Tregs identified in the paired peripheral blood sample of the same patient. Conclusions Our study suggested that a higher proportion of Tregs in malignant ascites may contribute towards fostering an immunosuppressive microenvironment.PD-L1+/HER2+ maDTCs, in such an immunosuppressive microenvironment, might be able to evade immune system attack. Moreover, PD-L1 and HER2 identified on maDTCs could serve as therapy targets with respect to inhibition of peritoneal metastasis, potentially leading to an additional treatment strategy for GI cancer patients with malignant ascites. Citation Format: Yang Chen, Changsong Qi, JiaJia Yuan, Dan Liu, Zhi Peng, Xiaotian Zhang, Jian Li, Pin Peter Lin, Yilin Li, Lin Shen. Elevated PD-L1 and HER2 expression on aneuploid carcinoma cells in malignant ascites accelerate stumor cell immune evasion in gastrointestinal cancer patients [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5381.

Durvalumab following multimodality therapy for locally advanced esophageal and GEJ adenocarcinoma: Updated survival and early translational results from Big Ten Cancer Research Consortium Study.

4572 Background: Concurrent chemoradiation(CRT) followed by esophagectomy is a standard of care for locally advanced esophageal(LA-EAC) and GEJ adenocarcinoma. Approximately 50% of patients(pts) experience disease relapse within the 1st yr after treatment(tx) completion. Immune checkpoint inhibitors have activity in metastatic PD-L1 positive EAC. Preclinical studies have shown radiation +/- chemotherapy upregulates PD-1/PD-L1 pathway. Methods: We conducted a phase II trial evaluating safety and efficacy of PD-L1 inhibitor durvalumab(durva) in pts with LA-EAC and GEJ adenocarcinoma who had residual disease in surgical specimen after neoadjuvant CRT and R0 resection. Pts received durva 1500mg IV every 4 weeks for up to 1yr. Results: Initially 24 pts were enrolled, study was expanded to enroll additional 13 pts. Median age: 61yrs (range, 43-73). 31 received carbo/paclitaxel and 6 received cis/5-FU concurrently with RT. 24(64.9%) pts had positive lymph nodes(LN) at the time of surgery following CRT: N3(n = 3,8.1%), N2(n = 10, 27%), N1(n = 11,29.7%).17 pts relapsed: 11 on tx, 6 had late relapses. 3/5 late relapses were locoregional and were re-treated with chemo-RT. Remaining relapses were systemic with lung and LN being the most common sites. 2 of 3 pts who developed grade 3 irAEs are alive and disease free at 17 and 23 mo. RFS/OS:1 yr- 79.2%/95.5%, 2yr-55.5%/67.4%. 20/37 pts have HER-2 status available: 5/6 HER2 positive pts had disease relapse, 1 is undergoing tx. Molecular profiling is available on 8 relapsed pts: all were microsatellite stable with low TMB and PD-L1 &lt; 10% CPS. Mutations in DNA repair genes ( ARID1A, ATM, ATR, CHEK2), and PIK3CA E542K were more prevalent among late relapsing pts. Circulating tumor cells (CTCs) analysis is available for 10/37 pts. 4/5 pts where CTCs increased from C1 to C4 had disease relapse. Molecular profiling of the remaining pts and correlation of PD-L1 expression, TMB, specific genes mutations, CTCs, and Immunoscore with outcomes with durva is being evaluated will be presented at the meeting. Conclusions: Adjuvant durva following trimodality therapy for LA-EAC and GEJ adenocarcinoma improved 1-yr RFS to 79.2% compared to historical rate of 50%.2-yr RFS and OS data are encouraging in this high risk pt population. HER-2 positivity may be associated with lack of benefit from durva. Mutations in DNA repair genes are prevalent in pts with delayed relapse. Rise in CTCs during durva tx may be an early marker of disease relapse. Clinical trial information: NCT02639065 .

Abstract B57: MUC3A induces PD-L1 and reduces tyrosine kinase inhibitor effects functions in EGFR-mutant non-small cell lung cancer

Abstract Objective: Non-small cell lung cancer (NSCLC) patients with specific EGFR mutations benefited from the emergence of EGFR tyrosine kinase inhibitors (EGFR-TKIs). Unfortunately, almost all cases eventually recrudesced after a median of 10 months. Therapeutic antibodies blocking PD-L1 strikingly regressed NSCLC development. However, only 20% patients responded to the αPD-L1 therapy. EGFR activation was reported to induce PD-L1 expression in NSCLC. MUC3A is a transmembrane mucin with 2 EGF-like domains in the extracellular segment. This study aimed to investigate the effects of MUC3A on PD-L1 expression and EGFR-TKIs sensitivity in EFGR-mutant NSCLC. Methods: Expression levels of MUC3A and PD-L1 in a tissue microarray of 92 patients’ lung tumor samples were measured by immunohistochemistry. Data of lung cancer were downloaded from the TCGA database, and the correlation of PD-L1 expression between EGFR wild type and mutation in lung cancer patients was analyzed. H1975 and PC-9 cells were infected with lentiviral vectors to downregulate MUC3A. The expression pattern of PD-L1 in NSCLC cells was detected by qRT-PCR and flow cytometry. Cell proliferation was assessed with CCK8. Cell apoptosis was examined by flow cytometry. Immunoblotting was used to identify molecular markers of signaling pathways. A xenograft nude mouse transplantation model was established to examine the effects of MUC3A and EGFR-TKIs on tumorigenesis in vivo. The expression levels of MUC3A, EGFR and PD-L1 in harvested tumors were measured by immunohistochemistry. Results: The tissue microarray of 92 patients’ lung tumor samples indicated that high levels of MUC3A were positively correlated with poor prognosis and PD-L1 expression. In the TCGA database, the levels of PD-L1 were independent of EGFR protein levels but correlated with EGFR mutation. Our results showed that induction of PD-L1 by EGF stimulation was only detected in EGFR-mutated NSCLC cell lines, such as H1975 and PC-9, but not in H1299 and H460. Knockdown of MUC3A in H1975 and PC-9 reduced PD-L1 expression, as well as PI3K/Akt and MAPK pathways. In addition, MUC3A deficiency potentiated gefitinib and AZD-9291-induced growth inhibition in PC-9 and H1975 cells by downregulating proliferation and inducing apoptosis. More interestingly, knockdown of MUC3A decreased EGFR protein levels, but not mRNA levels. Our findings suggested that MUC3A deficiency improved EGFR-mutated NSCLC sensitivity to tyrosine kinase inhibitors via increasing EGFR protein stability and decreasing PD-L1 expression through PI3K/Akt and MAPK pathways. Mice inoculated with MUC3A deficiency tumor cells treated with AZD-9291 had the fastest tumor regression and delayed relapse. Conclusions: The transmembrane mucin MUC3A increased EGFR stability. MUC3A induced PD-L1 and reduced tyrosine kinase inhibitor effects via EGFR modulation in NSCLC, suggesting potential implications for a novel immunotherapeutic approach. Citation Format: Yan Gong, Yuan Luo, Shan Peng, Shijing Ma, Yingming Sun, Jiangbo Ren, Conghua Xie. MUC3A induces PD-L1 and reduces tyrosine kinase inhibitor effects functions in EGFR-mutant non-small cell lung cancer [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2019 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(3 Suppl):Abstract nr B57.

Abstract CT121: Neoadjuvant combination targeted and immunotherapy for high-risk resectable stage III melanoma (NeoACTIVATE), NCT03554083

Abstract Background: Standard treatment for stage III melanoma is surgery followed by consideration of adjuvant therapy. This strategy is inadequate for high-risk stage III patients who have a substantial risk of subsequent relapse and distant metastasis. The goals of this trial are to improve oncologic outcomes, assess treatment response and spare patients with poorly responsive, rapidly metastatic disease from a futile operation. This protocol based on the hypothesis that treatment before surgery will immunize patients against his or her own unique tumor. While new drugs are now FDA-approved for adjuvant treatment of stage III melanoma, outcomes are still suboptimal in high-risk patients. The neoadjuvant approach is largely untested and preclinical data suggest neoadjuvant therapy is more effective than adjuvant immunotherapy. While targeted therapies can be rapidly effective, immunotherapy responses, when they occur, tend to be much more durable, prompting this study. Our specific strategy is based on the fact that BRAFm melanoma patients benefit both from BRAF/MEK inhibition (vemurafenib/cobimetinib) and PD-1 axis inhibition (atezolizumab) while patients with BRAFwt melanoma benefit from PD-1 axis inhibition (atezolizumab) and intermittent MEKi (cobimetinib) may increase effector function in tumor-infiltrating T cells. A second component of the study is robust evaluation of biomarkers of response, toxicity and outcome. Specific correlatives include assessment of the predictive value of Bim levels in tumor-related T cells, correlation of sPD-L1 with efficacy and toxicity, correlation of PD-L1 expression with efficacy, and determination of key relationships between tumor and immune cells via mIHC spatial analysis, gene expression profiles and microbiome associations with clinical response. Methods: The NeoACTIVATE trial enrolls melanoma patients with resectable clinically detectable nodal metastases (by physical examination or imaging), dual nodal basin disease or nodal recurrence of melanoma. Neoadjuvant systemic therapy is based on BRAF mutation status; participants receive either vemurafenib, cobimetinib and atezolizumab (vem/cobi/atezo), or cobimetinib and atezolizumab (cobi/atezo) before surgery. Following surgery, all patients receive atezo immunotherapy. The primary aim of the study is RFS estimates for each cohort. Secondary aims are safety/toxicity, pathologic complete response rate and prospective evaluation of biomarkers of treatment response and toxicity. This study is significant in that it is the first to test 1) combined of targeted and immunotherapy in the neoadjuvant setting for patients with resectable Stage III melanoma and 2) the novel combination of MEK-inhibition with immunotherapy in the neoadjuvant setting for resectable Stage III melanoma. As of January 1, 2019, 10% of planned patients have been enrolled and enrollment continues. Citation Format: Tina J. Hieken, Vera J. Suman, Shernan G. Holtan, Thomas J. Flotte, Lisa A. Kottschade, Matthew S. Block. Neoadjuvant combination targeted and immunotherapy for high-risk resectable stage III melanoma (NeoACTIVATE), NCT03554083 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr CT121.

Correlation between PD-L1 level and immunotherapy outcome in NSCLC in Lebanon, 2012-2018: A multicenter observational review.

e20524 Background: Immunotherapy targeting the PD1/PDL1 checkpoint pathway represents an important innovation in lung cancer treatment. In our research, we evaluate the correlation of PD-L1 expression with clinical outcomes (progression free survival, overall survival, objective response rate) in lung cancer patients in 4 centers in Lebanon. Methods: The 57 patients enrolled in this observational study were stage IIIB/IV NSCLC and SCLC who received immunotherapy with nivolumab, pembrolizumab or atezolizumab at any line of treatment, and were recruited between February 2012 and august 2018. Immunotherapy was continued until progression, death or unacceptable toxicity. PD-L1 expression was assessed on tumor samples either on diagnosis or at progression. Patients were divided into 3 categories according to PD-L1 level (&lt; 1, 1-49, &gt; 50%). Progression free survival defines the time from randomization to disease progression after each line of therapy. Overall survival defines time from randomization to death from any cause. Results: 70.18% had adenocarcinoma, 26.32% had squamous cell carcinoma, 1.75% had small cell lung cancer and one pathology was missing from the file. 66.66% were stage IV at diagnosis, 26.31% were stage 3. PD-L1 expression was &gt; 50% in 40.35% of patients, 24.56% had a level between 1 and 50% and the remaining had a value of less than 1%. PD-L1 was missing or not done in 15.79%. Median PFS was 13.9 months. Median OS was 14.2 months. 22.81% received immunotherapy as first line and 75% of the remaining patients upon first progression. PFS1 was 4.2 months. Mean time to first progression on immunotherapy was 3.6 months in patients with PD-L1 &gt; 50%, 10.5 months in PD-L1 between 1 and 50% and 14.3 months in those who had PD-L1 &lt; 1%. At the date of data cutoff, 49.12 % of patients were still receiving immunotherapy. No grade 3 or 4 immune related adverse events were seen during follow-up. Conclusions: The efficacy and safety of anti-PD-1 medications for advanced NSCLC were comparable in real-world and clinical settings. Prediction of treatment response from tumor PD-L1 expression seemed less practical than that was expected

Correlation Between PD-L1 Expression and Clinicopathologic Features in 404 Patients with Lung Adenocarcinoma.

PD-1/PD-L1 inhibitors is the important drugs of immunotherapy for malignant tumors. PD-L1 expression is an important biomarker of selecting patients for ICIs therapy. However, the correlation of PD-L1 expression with clinicopathologic features in lung adenocarcinoma remains controversial. PD-L1 expression was tested using clone SP263 by immunohistochemistry (IHC) on Ventana automated Benchmark in tissue micro-arrays (TMA) in lung adenocarcinoma. The association of PD-L1 expression with clinicopathologic characteristics, including gender, age, histological subtype, smoking history, stage, and genotype were analyzed. 404 patients were available for analyzing. The incidence of PD-L1 expression was 22.5% (using a cutoff of ≥ 25%). Statistical analysis showed PD-L1 expression was associated with advanced stage, lymph node (LN) metastasis, solid predominant subtype and wild-type epidermal growth factor receptor (EGFR) gene. In subgroup analysis, PD-L1 expression in patients with EGFR exon 19 deletions was higher than that of with EGFR L858R mutation at exon 21 (21.6% vs. 10.2%, P = 0.046). In multivariate analysis for overall survival (OS) by Cox hazard proportion model, patients with EGFR gene mutations (HR 1.635, 95% CI 1.310-2.040, P < 0.001) and early stage (HR 2.495, 95% CI 2.003-3.106, P < 0.001) had a longer survival, while PD-L1 expression was not associated with overall survival (HR 0.847, 95% CI 0.655-1.094, P = 0.203). For patients with lung adenocarcinoma, LN metastasis, wild-type EGFR, advanced stage, solid predominant subtype were independent predictors of PD-L1 expression by multivariate analysis. PD-L1 expression may be not a predictor of OS for patients with lung adenocarcinoma.

PD-L1 expression in carcinoma of the esophagogastric junction is positively correlated with T-cell infiltration and overall survival

IntroductionIn recent years antitumor immunity and inhibition of checkpoint molecules, such as PD-1 and PD-L1, have emerged as potential therapeutic strategies in advanced stages of various malignancies. We investigated PD-L1 expression in adenocarcinomas of the esophagogastric junction and correlated the results with densitiy of intratumoral T-lymphocytes. MethodsImmunohistochemical staining for PD-L1 was carried out on 135 samples using a tissue microarray. Scoring was done according to the combined positivity score. Results48.1% of tumors (65 cases) showed PD-L1 positivity with a score ≥ 1 while 51.9% were PD-L1 negative (70 cases). A positive correlation between PD-L1 negativity and mucinous and poorly cohesive carcinoma could be shown (p = 0.043), while no association existed for either gender, T-stage, N-stage, grading, surgical resection status, neoadjuvant therapy, distant metastases, lymphovascular or perineural invasion. No correlation of PD-L1 expression and overall survival could be detected (p = 0.497). Again, when stratified according to presence or absence of neoadjuvant therapy, no survival differences could be shown for either group (p = 0.540 and p = 0.736). When PD-L1 expression was correlated with density of tumor-infiltrating T-lymphocytes a positive correlation between PD-L1 positivity and denser T-cell infiltration could be shown (p = 0.001). Concerning overall survival, in PD-L1 negative cases, denser CD8-positive T-cell infiltrates were associated with prolonged survival times (p = 0.045). No differences could be shown for PD-L1 positive cases or CD103-positive T-cells. ConclusionPD-L1 expression is frequent in esophagogastric adenocarcinoma and - when combined with dense CD8 infiltration – PD-L1 negativity correlates with prolonged overall survival.

Abstract A008: Neoadjuvant nivolumab, gemcitabine and cisplatin in muscle-invasive bladder cancer: Study update

Abstract Background: Cisplatin-based neoadjuvant chemotherapy in muscle-invasive bladder cancer (MIBC) improves survival which correlates with pathologic response (PaR) at radical cystectomy (RC). Immunotherapy with checkpoint inhibitors has improved outcomes in advanced bladder cancer and immunotherapy is synergistic with chemotherapy. Our ongoing trial is aimed at studying the efficacy and safety of nivolumab (N)with gemcitabine-cisplatin (GC) as neoadjuvant therapy for MIBC (NCT03294304). Methods: This is a single-arm, multicenter phase II study for patients with MIBC eligible for neoadjuvant GC and planned for RC. Forty-one patients will be treated with N+ GC every 21 days for 4 treatment cycles over 12 weeks followed by RC. The primary endpoint is PaR. Key inclusion criteria are presence of MIBC (predominantly urothelial carcinoma) with clinical stage T2-T4a and N≤1 disease (solitary lymph node measuring &amp;lt; 2 cm) and M0 and deemed eligible for radical cystectomy, ECOG PS of 0 or 1, Cr Cl ≥ 50 ml/min, adequate organ function, eligible to receive GC and no contraindications to receiving N. Patients will receive C (70mg/m2) IV on D1, G (1000mg/m2) on D1,D8 and N (360 mg) IV on D8 every 21 days for 4 cycles followed by radical cystectomy within 6-8 weeks. Secondary objectives are to determine safety of combination of N+GC in pts with MIBC prior to RC and PFS. The correlative objectives include: 1) correlation of whole-genome sequencing (WGS) of the pre-treatment bladder cancer biopsy with PaR; 2) correlation of molecular subtypes of bladder cancer using Genomedx to evaluate whether addition of N alters the response to neoadjuvant GC; 3) correlation of PD-L1 expression in tumor tissue at baseline with PaR; 4) Nanostring pancan immune panel gene expression will be studied at baseline and at RC. At the time of submission of this abstract, 12/41 pts. have been enrolled and the combination has not resulted in any added toxicities or immune-related AEs so far. There were no delays in RC. PaR and correlative analysis is ongoing in enrolled pts. and further enrollment is ongoing. Citation Format: Shilpa Gupta, Christopher J. Weight, Neeraj Agarwal, Sumati Gupta, Badrinath Konety, Ewan Gibb, Elai Davicioni, Bharat Thyagarajan, Guru Sonpavde. Neoadjuvant nivolumab, gemcitabine and cisplatin in muscle-invasive bladder cancer: Study update [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A008.

Discordance of high PD-L1 expression in primary and metastatic urothelial carcinoma lesions

Immune checkpoint inhibitors (ICI) targeting PD-(L)1 are effective in select patients with advanced urothelial carcinoma (UC). High PD-L1 expression enriches for response to ICIs; however, the predictive value of PD-L1 expression is limited, which may be due in part to dynamic expression of PD-L1 in the tumor environment. We sought to characterize PD-L1 expression in primary UC and paired metastatic lesions to gain insight into the potential discordance of tumor PD-L1 expression during the metastatic process. Materials and methods: Immunohistochemical staining for PD-L1 using the SP-142 antibody was performed on primary tumors and matched metastatic specimens in 77 evaluable subjects with advanced UC. Immunohistochemical staining was scored for the percentage of cells positive (<5%, ≥5%) in tumor cell (TC) and immune cell (IC) compartments. Correlation of PD-L1 expression in TCs and ICs was estimated using Spearman's correlation coefficients (rho, ρ). Cohen's kappa statistics (κ) were utilized to assess the agreement in PD-L1 expression between groups. Results: High (≥5%) PD-L1 expression in primary and metastatic biopsies, respectively, was observed in 6.0% and 7.7% of TCs and in 14.5% and 11.5% of ICs. IC PD-L1 expression in primary tumors was not correlated with IC PD-L1 expression in paired metastatic lesions (ρ = 0.05, P = 0.67) and there was poor agreement in high expression rates between primary and metastatic lesions in the IC compartment (κ= 0.086). Conclusion: High PD-L1 IC expression is temporally and spatially discordant between primary and metastatic UC lesions. Future studies of PD-(L)1 targeted therapies in patients with metastatic UC may benefit from use of fresh biopsies of metastatic lesions to define PD-L1 expression when feasible.

A nomogram-based immunoprofile predicts overall survival for previously untreated patients with esophageal squamous cell carcinoma after esophagectomy

BackgroundImmunoscore, as a prognostic tool defined to quantify in situ immune cell infiltrates, appears to be superior to the TNM staging system. In esophageal squamous cell carcinoma (ESCC), no immunoscore has been established; however, in situ tumor immunology is recognized as highly important. Our study aimed to construct a comprehensive immunoprofile for ESCC.MethodsThe infiltration of four immune cell types (CD8+/CD4+/Foxp3+/CD33+ cells), the expression of both inhibitory (PD-1/PD-L1/Tim-3/LAG-3) and stimulatory checkpoints (OX-40/ICOS), and IDO1 were evaluated by IHC staining and multi-color immunofluorescence in two independent cohorts (95 patients in the primary cohort and 55 patients in the validation cohort). The association with patients’ overall survival was analyzed by the Kaplan-Meier method and the Cox model. Nomogram-based immunoprofile was established using the independent prognostic variables. To determine its predictive accuracy and discriminatory capacity, the C-index and calibration curve were calculated.ResultsSignificant correlation of PD-L1 expression in tumor cells with PD-1+ T cell infiltration was found (P = 0.035), indicating the activation of the inhibitory PD-1/PD-L1 pathway in ESCC cases. More PD-L1+ ICs, Tim-3+ ICs and LAG-3+ ICs were found in the CD8-rich tumor microenvironment, which is in accordance with the feedback nature of immune system. After adjustment by TNM stage, four immune variables including the infiltration of CD8+/Foxp3+/CD33+ cells and the PD-L1 expression by tumor cells were selected to construct a prognostic nomogram. The calibration curves showed good accuracy of the nomogram for survival prediction. To overcome the complexity of applying a nomogram in a clinical setting, a simple immunoprofile was then established according to the points of each factor from the nomogram. Our immunoprofile model could separate same-stage patients into different risk subgroups, and showed superior accuracy for survival prediction than the TNM staging system based on the C-index calculation and ROC analysis.ConclusionsOur nomogram-based immunoprofile can provide more accurate prognosis prediction and is an important complement to the TNM staging system for operable ESCC patients.