Improved Recurrence Free Survival Research Articles

Gene signature to predict hypophysitis in patients with melanoma treated with anti-PD-1 in adjuvant setting.

e21562 Background: Adjuvant anti-PD-1 treatment has shown to improve recurrence free survival and distant metastases free survival in melanoma patients from stage II to IV. Until now ,we have no data about overall survival of these patients nor biomarkers to select them to the best therapy , avoiding in this way severe and some times durable toxicities.Hypophysitis is a rare immune-related adverse event (irAE) and in adjuvant setting may represent a chronic irAE that may affect quality of life , such as fertility in young people.There are no clinical factors able to predict hypophysitis due to adjuvant anti-PD-1 therapy. Methods: Gene profiling analysis was performed using NanoString IO360 panel from basal PBMCs of patients treated with anti-PD1 in adjuvant setting. Patient’s characteristics are reported in table1. ROC curves were used to determine the best cut off. Survival rates were evaluated and compared using the Kaplan–Meier method. Results: Eighty pts treated with adjuvant anti-PD-1were included in a retrospective analysis from December 2019 to September 2021. Among which 12 pts developed hyphosytis. Ten pts completed treatment regularly, while two were discontinued both at week 12 due to metastatic disease. A pts developed liver metastases, the other one liver, multiple lymph nodes and soft tissues metastases. Grade 2 hypophysitis was diagnosed as the only irAE in all pts (100%). Median time of onset was 3 months (9 pts ≤6 months and 7 pts ≤3 months). A specific gene signature for predicting the onset of hypophysitis has been identified and characterized with 4 genes: IFITM2,CTSS,DUSP1,HLA-B that are mainly involved in the activation of innate-like lymphocyte populations, such as natural killer (NK) cells and innate lymphoid cells (ILCs), and adaptive immune cells, such as T helper 1 (TH1) cells and CD8+ cytotoxic T lymphocytes (CTLs). High gene signature was mostly related to hypophysitis event than lower group (HR: 0.11 95%IC 0.03-0.39, p = 0,0007). Moreover, at data lock all pts who developed hypophysitis were censored with mean overall survival of 35.60 months, 95%CI 30,86-40, 41. Conclusions: A predictive gene signature of hypophysitis was found in melanoma patients treated with adjuvant anti-PD1.Moreover, hypophysitis seems to correlate to better outcome. [Table: see text]

Treatment package time < 14 weeks improves recurrence free and disease specific survival in HPV positive OPC with high-risk features

BackgroundGuidelines recommend treatment package time < 85 days and time from surgery to radiation initiation < 6 weeks in head and neck cancer patients. However, HPV positive primaries treated with TORS and adjuvant radiotherapy traditionally demonstrate favorable outcomes. MethodsSingle center retrospective chart review of patients diagnosed with HPV positive treatment naïve primary squamous cell carcinoma treated with TORS and postoperative radiation therapy with or without Chemotherapy from 2012 to 2022 with data collection from December 2022-April 2023. Kaplan-Meier survival analysis with log-rank testing assessed the impact of time intervalsbetween diagnosis, TORS, radiation initiation and radiation completion on recurrence free and disease specific survival. Univariate Cox proportional hazards regression analysis was performed to identify factors associated with recurrence free and disease specific survival. Subgroup analysis was done with high risk (positive lymph nodes > 5, >1mm extracapsular extension, positive margins) patients who underwent concurrent Chemotherapy. ResultsOf 255 patients (225 males [89 %], average age 58 years, 163 [64 %] high-risk, median follow-up 4.3 years), 22 (8.6 %) had recurrence and 14 died due after disease recurrence.Only radiation length of 5–7 weeks prolonged survival in the entire population. In the high-risk cohort, time from TORS to radiation initiation < 6 weeks improvedrecurrence free survival, while total package time < 14 weeks wasassociated with greater recurrence free and disease specific survival.

Contribution of neoadjuvant chemotherapy and radiation therapy (intensity-modulated radiation therapy with concurrent chemotherapy versus stereotactic body radiation therapy alone) on tumor volume regression in patients with borderline resectable or locally advanced pancreatic ductal adenocarcinoma: A single institution experience.

707 Background: Pancreatic ductal adenocarcinoma (PDAC) has a poor prognosis with a minority of patients (pts) eligible for curative resection. Pts with borderline resectable or locally advanced (BR/LA) PDAC frequently receive neoadjuvant chemotherapy (NAC) +/- radiation therapy (RT) to improve resectability. Here, we report the individual contribution of NAC and RT (either intensity modulated RT + concurrent chemotherapy (IMRT+CC) or stereotactic body RT (SBRT) alone) on tumor regression by measuring tumor volumes. Methods: We performed a single institution retrospective review of PDAC pts who underwent resection between 2011-2021 and were part of a tumor biobank registry. Tumor measurements were performed at baseline, post-NAC, and post-RT by five independent, blinded abdominal radiologists. Analyses were performed using paired t-tests, two-proportion z-tests, log-rank tests, and Mann-Whitney U test. Results: We identified 49 BR/LA pts who received NAC followed by RT who had complete pre- and post-treatment imaging. Fifteen pts received IMRT+CC and 28 received SBRT. The effect of NAC on tumor volumes (-29.2% vs -26.3%, p=0.66; mean percent change) and maximum diameter (-9.2% vs -12.0%, p=0.58) was similar within the IMRT+CC and SBRT groups. Both IMRT+CC (-7.12 cm3, p=0.003) and SBRT (-2.70 cm3, p=0.001) resulted in significant tumor regressions. IMRT+CC achieved a significantly greater overall mean percent change in tumor volume (-53.4% vs -28.8%, p=0.024) and maximum diameter (-30.4% vs -16.2%, p=0.020) compared to SBRT. Pts receiving IMRT+CC or SBRT had similar tumor volumes (10.88 vs 11.74 cm3, p=0.87, respectively) and maximum diameter (3.45 vs 3.17 cm, p=0.48) prior to RT. IMRT+CC resulted in improved tumor volume response (&gt;30% regression; 80.0% vs 46.4%; p=0.033) but not an improved response rate by RECIST criteria (46.7% vs 21.4%; p=0.085). Mean resected tumor volumes (by pathological review) for those receiving IMRT+CC (n=9) and SBRT (n=23) were 4.34 vs 13.1 cm3 (p=0.049). The R0 (86.7% vs 85.7%, p=0.93) and N0 (73.3% vs 53.6%, p=0.21) resection rates between the two groups were similar. Compared to SBRT, IMRT+CC resulted in an improved recurrence free survival (RFS; 2.36 vs 1.31 years; p=0.026), but did not meet statistical significance for an overall survival benefit (mOS; 2.85 vs 2.03 years, p=0.19). Conclusions: We found that IMRT+CC yielded superior radiographic volumetric regression and improved RFS but did not significantly impact mOS. Correspondingly, resected tumors from patients treated with IMRT+CC were smaller than those obtained from patients treated with SBRT. The role of IMRT+CC and SBRT in PDAC remains to be clearly defined.

Real-World efficiency of pembrolizumab in metastatic melanoma patients following adjuvant anti-PD1 treatment

BackgroundPostoperative treatment of patients with either BRAF/MEK inhibitors or anti-PD1 antibodies in the adjuvant setting results in improved recurrence free survival and has therefore become a standard of care for most patients with resected stage III melanoma. For patients who need systemic treatment after failure of adjuvant immune checkpoint inhibitors, there is insufficient evidence regarding efficacy of a retreatment with anti-PD1 antibodies. MethodsFrom the European Melanoma Treatment Registry (EUMelaReg) we have identified 74 patients and evaluated the clinical characteristics and outcome of anti-PD1 retreatment with pembrolizumab in these patients. The primary objectives were overall response rate and progression-free survival stratified by type of recurrence, i.e. whether recurrence occurred while on adjuvant anti-PD1, or later during follow up. In addition, the analysis was stratified for patients, who terminated adjuvant treatment early due to side effects. ResultsThe ORR of pembrolizumab retreatment in 1st line after recurrence (n=51) was 37.3%, which did not differ significantly between type of recurrence (40.9% in early vs. 34.5% in late recurrence). Patients who discontinued adjuvant anti-PD1 for toxicity had a higher ORR (52.9%) compared to patients who completed the treatment (37.5%) or discontinued due to disease progression (23.1%). PFS in 1st line retreatment was comparable between the groups with a median PFS (95% CI) of 7.4 (2.3-NR) months and 6.1 (3.4-13.1) months in early and late recurrence, respectively. ConclusionRetreatment with pembrolizumab may be a valuable treatment option after failure of adjuvant immunotherapy, in particular in patients who have stopped adjuvant treatment for side effects.

Minimal residual disease by circulating tumor DNA as a biomarker of recurrence free survival in resected high-risk melanoma patients treated with mRNA-4157/V940, a personalized cancer vaccine, and pembrolizumab.

LBA9515 Background: The combination of mRNA-4157/V940 and pembrolizumab improved recurrence free survival (RFS) compared to pembrolizumab monotherapy in patients with resected high-risk stage III/IV cutaneous melanoma in the randomized Phase 2 mRNA-4157-P201/KEYNOTE-942 trial (RFS event rate of 22.4% (24/107) versus 40% (20/50); HR = 0.561; 95% CI: 0.309, 1.017). Across multiple cancer types and disease settings, detection of minimal residual disease (MRD) by circulating tumor DNA (ctDNA) assays in plasma identifies patients at higher risk of relapse and progression, and can monitor presence of disease throughout treatment. Herein, we report ctDNA analyses to explore the association between minimal residual disease and the probability of recurrence. Methods: In mRNA-4157-201, baseline tumor core biopsies and matched whole blood were subjected to whole exome sequencing (WES). The personalized amplicon-based NGS assay by Inivata (RaDaR) was used to identify and prioritize up to 48 patient-specific somatic variants to analyze ctDNA in longitudinal plasma samples for MRD detection. This method for ctDNA analysis is distinct from most prior studies in melanoma that report use of ddPCR for single mutations or fixed-gene panels. The association of MRD with RFS was evaluated with Kaplan Meier analyses and assessed with hazard ratio (95% CI) in ctDNApos and ctDNAneg subgroups and across study arms. Results: Of patients enrolled in this study with evaluable ctDNA, 88% (110/125) were ctDNAneg at start of treatment. Significantly longer RFS was observed in patients who were ctDNAneg compared to those with ctDNApos at baseline across study arms (RFS event rate of 20.9% (23/110) versus 80.0% (12/15); HR = 0.150; 95% CI: 0.073, 0.306). Within the mRNA-4157/V940 and pembrolizumab combination arm, significantly longer RFS was observed in patients with ctDNAneg compared to ctDNApos samples (RFS event rate of 10.4% (8/77) versus 76.9% (10/13); HR = 0.087; 95% CI: 0.034, 0.222). This trend was also observed in the pembrolizumab arm (RFS event rate of 45.5% (15/33) versus 100% (2/2); HR = 0.008; 95% CI: 0.001, 0.088) study arm, although the small sample size in pembrolizumab arm (n = 2) limits interpretation. Conclusions: MRD detection by plasma ctDNA assay at the start of adjuvant melanoma treatment is uncommon in mRNA4157-p201 but is associated with shorter RFS. Treatment with the combination of mRNA-4157/V940 and pembrolizumab was associated with prolonged RFS compared to pembrolizumab monotherapy in patients with high-risk resectable melanoma, irrespective of MRD status Additional analyses including assessment of longitudinal ctDNA patterns are ongoing. The association between MRD and mRNA-4157/V940 treatment effect will be further explored in upcoming planned studies. Clinical trial information: NCT03897881 .

Recurrence free survival in patients with HPV+ head and neck cancer treated with TORS and neck dissection.

e18076 Background: Prior research has demonstrated that delays to initiation of radiation therapy (RT) in the treatment of head and neck squamous cell carcinoma (HNSCC) after surgery result in poorer survival outcomes. However, there is limited data with little long-term follow up to evaluate whether human papilloma virus (HPV) positive primaries undergoing transoral robotic surgery (TORS) and neck dissection (ND) face a similar decrement in recurrence free survival (RFS) and disease specific survival (DSS). Methods: Single center retrospective chart review of patients diagnosed with HPV positive primary nonmetastatic HNSCC who underwent TORS with ND followed by RT with or without Chemotherapy and/or Immunotherapy from 2012-2022 with data collection from December 2022 to February 2023. 248 patients met inclusion criteria and were analyzed for DSS and RFS. Kaplan-Meier survival analysis with Log-rank testing was used to assess the impact of time intervals between diagnosis, TORS, radiation initiation and radiation completion on survival. Univariable and Multivariable Cox proportional hazards regression analysis was performed to identify factors associated with RFS and DSS and adjust for potential confounding variables. Subgroup analysis was completed with high risk (lymph nodes &gt; 4 with extracapsular extension) and low risk population. Results: Median follow-up time was 47.22 months (1452.07 days). Of 248 patients, 17 had a recurrence and 12 died due to disease. In patients who underwent radiation therapy for a time frame (TR1-R2) of 5-7 weeks compared with those who had a shortened or prolonged course of RT, RFS (140.116 months, p-value = .012) and DSS (142.641 months, p-value = .016) were improved. Although time from diagnosis to TORS did not impact RFS (p-value = .195), improved RFS persisted in patients who underwent time from diagnosis to radiation completion (TD-R2) or total treatment package time (TTPT) between 17-20 weeks (109.155 months, 95% CI: 105.276-113.034, p-value = .057) compared to patients with TTPT less than 17 or greater than 20 weeks. When the population is stratified into high risk and low risk groups, patients in the high risk group who underwent Chemotherapy had improved RFS compared to no chemotherapy (HR: 7.19, p-value = .09). On Univariate Analysis, TR1-R2 &lt; 5 weeks and &gt; 7 weeks, TD-R2 &lt; 17 weeks and &gt; 20 weeks, extracapsular extension, smoking history, and age &gt; 70 were associated with increased risk of recurrence. Conclusions: For HPV positive patients with OPSCC undergoing TORS and Neck dissection, time to radiation initiation may not be as critical in improving OS, DSS, and RFS as other patient populations. However, the length of radiation time between 5 to 7 weeks and overall treatment time from 17-20 weeks may improve recurrence free survival and disease specific survival.

Abstract CT224: mRNA-4157, a personalized cancer vaccine, in combination with pembrolizumab, demonstrates trend for improved recurrence free survival compared to pembrolizumab alone in adjuvant melanoma patients across tumor mutational burden subgroups

Abstract Background: The open-label randomized Phase 2 mRNA-4157-P201/Keynote-942 trial met its primary endpoint of prolonged recurrence free survival (RFS) in patients with resected high-risk stage III/IV melanoma. Tumor immunogenicity provides a favorable landscape for inflammatory processes associated with clinical benefit to checkpoint inhibitors (ICI) and tumor mutational burden (TMB) has been shown to be an independent predictor of treatment outcomes in patients treated with ICI therapy. mRNA-4157 is a novel mRNA-based personalized cancer vaccine which encodes up to 34 patient-specific tumor neoantigens. Here we report analyses of baseline biopsies from the trial to explore the novel mechanism of action hypothesized to augment endogenous anti-tumor responses and generate immunity to additional tumor neoantigens. Methods: Paraffin-embedded formalin-fixed baseline tumor core biopsies underwent whole exome sequencing (WES) and whole transcriptome sequencing. According to the established WES genomic score for pembrolizumab, the TMB high threshold utilized for analysis was 175/exome (10 mutations/megabase per F1CDx). The distribution of TMB expression in baseline tumor samples across study arms and their association with the primary RFS endpoint was evaluated. The association with RFS of other markers of inflamed tumors, including those established for pembrolizumab (e.g. gene expression profile (GEP) and PD-L1 expression) was also assessed. Results: The RFS benefit of mRNA-4157 and pembrolizumab combination compared to pembrolizumab monotherapy observed in the intention-to-treat population was maintained with a similar treatment effect magnitude across both high (HR = 0.65; 95% CI: 0.3, 1.5) and low (HR = 0.59; 95% CI: 0.3,1.4) TMB subpopulations. In line with observations from historical data for pembrolizumab, improved RFS was observed in high TMB compared to low TMB patient subgroups in the pembrolizumab monotherapy arm. The trend for increased RFS benefit in the high TMB subpopulation was maintained in the mRNA-4157 and pembrolizumab study arm. Additional subgroup analyses (e.g. GEP and PD-L1) were assessed and will be discussed. Conclusions: Our results indicate that mRNA-4157 demonstrates improvements in RFS irrespective of TMB status when administered in combination with pembrolizumab compared to pembrolizumab monotherapy in patients with resected high-risk cutaneous melanoma. The novel mechanism of action of mRNA-4157 may both deepen the activity of pembrolizumab and broaden the population of patients that can benefit from immune therapy. The association between TMB and mRNA-4157 treatment effect will be further explored in upcoming planned studies. Citation Format: Ryan J. Sullivan, Matteo Carlino, Jeffrey S. Weber, Tarek Meniawy, Matthew H. Taylor, Kevin Kim, Meredith McKean, Georgiana V. Long, Mark Faries, C. Lance Cowey, Andrew Pecora, Geoffrey T. Gibney, Jason Luke, Sajeve Thomas, Vasudha Sehgal, Igor Feldman, Praveen Aanur, Michelle Brown, Robert S. Meehan, Celine Robert-Tissot, Adnan Khattak. mRNA-4157, a personalized cancer vaccine, in combination with pembrolizumab, demonstrates trend for improved recurrence free survival compared to pembrolizumab alone in adjuvant melanoma patients across tumor mutational burden subgroups [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr CT224.

Abstract CT001: A personalized cancer vaccine, mRNA-4157, combined with pembrolizumab versus pembrolizumab in patients with resected high-risk melanoma: Efficacy and safety results from the randomized, open-label Phase 2 mRNA-4157-P201/Keynote-942 trial

Abstract Background: Targeting of mutation-derived epitopes (neoantigens) by T cells has been demonstrated to drive anti-tumor immune responses. mRNA-4157 is a novel mRNA-based personalized cancer vaccine, which encodes up to 34 patient-specific tumor neoantigens. It was hypothesized mRNA-4157 could synergize with adjuvant pembrolizumab to improve recurrence free survival (RFS) in patients with resected stages IIIB/IIIC/IIID and IV melanoma. Methods: Eligible patients with completely resected, high-risk cutaneous melanoma were randomly assigned 2:1 (stratified by stage) to receive mRNA-4157 in combination with pembrolizumab or pembrolizumab alone. mRNA-4157 (1mg) was administered intramuscularly every 3 weeks for a total of 9 doses and pembrolizumab (200mg) intravenously was given every 3 weeks for up to 18 cycles. Safety was evaluated as a secondary endpoint. RFS in the overall intention-to-treat population was the primary end point. The study was designed with 80% power to detect a hazard ratio (HR) of 0.5 with an overall 1-sided type I error of 0.1 when a total of 40 RFS events were observed. The primary analysis for RFS was specified to occur after all patients completed a minimum of 12 months on study and at least 40 RFS events were observed. Results: 107 patients received the combination of mRNA-4157 with pembrolizumab and 50 patients were treated with pembrolizumab monotherapy. Recurrence or death was reported in 24 of 107 patients (22.4%) in the combination arm and in 20 of 50 patients (40%) in the monotherapy arm, at a median follow-up of 101 and 105 weeks respectively. 18-month RFS rates (95% CI) were 78.6% (69.0%, 85.6%) vs 62.2% (46.9%, 74.3%) in the combination and monotherapy arm respectively. The combination showed protocol defined statistical significance and a clinically meaningful improvement in RFS compared to pembrolizumab, with a reduction in the risk of recurrence or death by 44% (HR = 0.561; 95% CI: (0.309, 1.017); stratified log-rank test 1-sided p-value of 0.0266. The majority of treatment related adverse events were Grade 1/2. The number of patients reporting treatment related Grade ≥ 3 adverse events was generally similar between the arms (25% vs 18%, respectively). The most common mRNA-4157 related Grade 3 event was fatigue. No Grade 4 or Grade 5 events related to mRNA-4157 were reported. No potentiation of immune-mediated adverse events were observed with the addition of mRNA-4157 to pembrolizumab. Conclusions: mRNA-4157 in combination with pembrolizumab as adjuvant therapy for resected high-risk melanoma significantly prolonged RFS compared to pembrolizumab without an increase in clinically meaningful adverse events. These results are the first to demonstrate improvement of RFS over adjuvant standard of care PD-1 blockade in resected high-risk melanoma and provide the first randomized evidence that a personalized neoantigen approach is potentially beneficial for cancer patients. A phase 3 study will be initiated in patients with melanoma. Citation Format: Adnan Khattak, Matteo Carlino, Tarek Meniawy, George Ansstas, Theresa Medina, Matthew H. Taylor, Kevin B. Kim, Meredith McKean, Georgina V. Long, Ryan J. Sullivan, Mark Faries, Thuy Tran, Charles Cowey, Andrew Pecora, Jennifer Segar, Victoria Atkinson, Geoffrey T. Gibney, Jason Luke, Sajeve Thomas, Elizabeth Buchbinder, Peijie Hou, Lili Zhu, Tal Zaks, Michelle Brown, Praveen Aanur, Robert S. Meehan, Jeffrey S. Weber. A personalized cancer vaccine, mRNA-4157, combined with pembrolizumab versus pembrolizumab in patients with resected high-risk melanoma: Efficacy and safety results from the randomized, open-label Phase 2 mRNA-4157-P201/Keynote-942 trial [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr CT001.

Abstract 4402: Survival impact of post-operative immunotherapy in resected stage III cutaneous melanomas in the checkpoint era

Abstract Introduction: Since 2015, immune checkpoint inhibitors have been approved as post-operative therapy for resected stage 3 cutaneous melanoma. This approval was a result of studies which demonstrated improved recurrence free survival. However, overall survival (OS) benefit has yet to be observed. We examine the OS of postoperative immunotherapy in stage 3 melanoma using real world data from the National Cancer Database (NCDB). Methods: Data was collected from the 2020 NCDB Participant Use File (October 2022 release). Patients were staged by both the 7th and 8th version of American Joint Committee on Cancers (AJCC). Those who underwent definitive surgery since 2016 and had sufficient pathologic data for staging were included. Patients who received systemic therapy prior to surgery were excluded. Patients were then sorted by the type of postoperative therapy started within 12 weeks from definitive surgery and classified into the no post-operative systemic therapy (NP) or postoperative immunotherapy (PI) groups. Survival time was calculated from the time of definitive surgery to last contact or death. OS was estimated by Kaplan-Meier method and compared by log-rank method. Results: 18,167 patients underwent surgery for stage 3 disease between 2016-2020. By AJCC v8 staging, 4,256 had Stage 3A disease, 3,752 stage 3B disease, 9,251 stage 3C disease, and 908 stage 3D disease. Median OS was not met in any stage 3 PI groups. The 36 - month landmark survival of all stage 3 subgroups is shown in Table 1. Only 13.3% of patient received PI in 2016. PI use increased to 24.8% in 2017, 43.8% in 2018, 46.8% in 2019, and 50.8% in 2020. Discussion: Using a large real-world database, we found that patients receiving post-operative immunotherapy in the checkpoint era have a significant survival benefit compared to those who did not. The benefit was significant in stage IIIB-C in AJCC v7 and Stage IIIC-D in AJCC v8. However, the benefit was not significant in stage 3A melanoma. Table 1. 36-month Landmark survival reported with 95% confidence interval for both AJCC versions No Post-Operative Immunotherapy Post-Operative Immunotherapy Given Stage Total Patients (n) n (%) 36- month survival % (95% CI) n (%) 36- month survival % (95% CI) p-value AJCC v 7 Stage 3 15272 Stage 3A 3818 2539 (66.5) 90 (89-91) 1141 (29.9) 91 (90-92) p=0.737 Stage 3B 4572 2990 (65.4) 78 (77-79) 1461 (32.0) 83 (82-94) p=0.001 Stage 3C 6882 4021 (58.4) 64 (63-65) 2637 (38.3) 73 (72-74) p&amp;lt;0.001 AJCC v 8 Stage 3 18167 Stage 3A 4256 2981 (70.0) 91 (90-92) 1145 (26.9) 93 (92-94) p=0.354 Stage 3B 3752 2255 (60.1) 84 (83-85) 1379 (36.8) 87 (86-88) p=0.057 Stage 3C 9251 5592 (60.4) 65 (64-66) 3372(36.5) 76 (75-77) p&amp;lt;0.001 Stage 3D 908 550 (60.6) 44 (42-46) 316 (34.8) 59 (56-62) p&amp;lt;0.001 Citation Format: Garo G. Hagopian, Christopher Grant, Danielle Brazel, Priyanka Kumar, Maki Yamamoto, James Jakowatz, Warren Chow, Thuy Tran, Justin Moyers. Survival impact of post-operative immunotherapy in resected stage III cutaneous melanomas in the checkpoint era. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4402.

LBA02-05 DEFINING INNATE IMMUNE MEMORY IN THE MECHANISM OF BCG IMMUNOTHERAPY FOR BLADDER CANCER

You have accessJournal of UrologyCME1 Apr 2023LBA02-05 DEFINING INNATE IMMUNE MEMORY IN THE MECHANISM OF BCG IMMUNOTHERAPY FOR BLADDER CANCER Song Jiang, Gil Redelman-Sidi, Eugene J. Pietzak, Bernard H. Bochner, and Michael Glickman Song JiangSong Jiang More articles by this author , Gil Redelman-SidiGil Redelman-Sidi More articles by this author , Eugene J. PietzakEugene J. Pietzak More articles by this author , Bernard H. BochnerBernard H. Bochner More articles by this author , and Michael GlickmanMichael Glickman More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003361.05AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Intravesical instillation of the live attenuated mycobacterium Bacillus Calmette-Guerin (BCG) remains the most effective therapy for the treatment of non-muscle invasive bladder cancer. An increasing body of evidence suggests that the innate immune system has characteristics that involve a heterologous memory of past insults through a process thought to involve epigenetic reprogramming, termed trained immunity. We seek to investigate the potential for innate immune training as a predictor of the anti-tumor effects of BCG. METHODS: A prospective biospecimen collection was performed on venous blood from non-muscle invasive bladder cancer patients prior to receiving intravesical BCG therapy. An ex vivo training protocol was performed on peripheral blood monocytes as previously described. Subsequently, cells were stimulated with 25 ng/ml lipopolysaccharide (a TLR4 agonist) for 24 hours. Cell free supernatant was measured for TNF-alpha production by enzyme-linked immunosorbent assay. RESULTS: In vitro training of primary monocytes induced an increase in proinflammatory cytokine production upon re-stimulation. In our in vitro training experiments, there was demonstrable interpatient variability seen pre-BCG exposure in a cohort of 30 NMIBC patients (Figure 1). Kaplan Meier analysis of this NMIBC patient cohort demonstrated improved recurrence free survival in individuals with a higher innate immune memory response. CONCLUSIONS: We present data on a trained innate immune phenotype on primary monocytes collected from patient samples. We found that innate immune training correlated improved recurrence free survival in NMIBC patients treated with BCG. Such insight will have potential clinical impact on the development of biomarkers and clinical assays that could predict immune responsiveness to BCG a priori, as well as identify possible immunological adjuvants that could enhance the effect of intravesical BCG. Source of Funding: ASCO YIANCI SPORE in Bladder Cancer © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e1188 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Song Jiang More articles by this author Gil Redelman-Sidi More articles by this author Eugene J. Pietzak More articles by this author Bernard H. Bochner More articles by this author Michael Glickman More articles by this author Expand All Advertisement PDF downloadLoading ...

MP47-17 SINGLE CENTER COMPARISON OF RADIOGRAPHIC AND MORPHOMETRIC CHARACTERISTICS AND OUTCOMES IN T3A PATHOLOGICALLY UPSTAGED AND NON-UPSTAGED RENAL CELL CARCINOMA

You have accessJournal of UrologyCME1 Apr 2023MP47-17 SINGLE CENTER COMPARISON OF RADIOGRAPHIC AND MORPHOMETRIC CHARACTERISTICS AND OUTCOMES IN T3A PATHOLOGICALLY UPSTAGED AND NON-UPSTAGED RENAL CELL CARCINOMA Saeed Ghassemzadeh, Aastha Shah, Dhruv Puri, Luke Wang, Franklin Liu, Sohail Dhanji, Kevin Hakimi, Mimi Nguyen, Ryan Nasseri, Margaret Meagher, Clara Cerrato, Juan Javier-Desloges, and Ithaar Derweesh Saeed GhassemzadehSaeed Ghassemzadeh More articles by this author , Aastha ShahAastha Shah More articles by this author , Dhruv PuriDhruv Puri More articles by this author , Luke WangLuke Wang More articles by this author , Franklin LiuFranklin Liu More articles by this author , Sohail DhanjiSohail Dhanji More articles by this author , Kevin HakimiKevin Hakimi More articles by this author , Mimi NguyenMimi Nguyen More articles by this author , Ryan NasseriRyan Nasseri More articles by this author , Margaret MeagherMargaret Meagher More articles by this author , Clara CerratoClara Cerrato More articles by this author , Juan Javier-DeslogesJuan Javier-Desloges More articles by this author , and Ithaar DerweeshIthaar Derweesh More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003293.17AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: A significant portion of patients presenting with Stage 1-2 Renal Cell Carcinoma (RCC) are pathologically upstaged to Stage 3 following surgery. This is due to previously undetected extension into the renal venous system, perirenal or renal sinus fat, or collecting system. Improved detection of potential T3 upstaging may prompt changes in disease management and may impact patient survival. We sought to compare pathologically upstaged and non-upstaged T3a RCC cases to identify characteristics of upstaged masses, predictors of T3a disease, and impact on oncological outcomes. METHODS: We conducted a single center retrospective analysis of patients with pathologic T3a RCC who underwent surgical intervention. The cohort was divided into a group of patients with masses which were upstaged, (cT1-cT2/pT3a) and a group with masses preoperatively identified as cT3a (non-upstaged, cT3a/pT3a) for descriptive and outcomes analyses. We sought to delineate proportion of underdiagnosed pT3a RCC, location of upstaged disease, and predictors of upstaging. Outcomes were overall survival (OS) and recurrence-free survival (RFS). Multivariate analyses (MVA) were performed to identify predictors of T3a invasion site and outcomes. Kaplan Meier survival analyses (KMA) were performed to compare survival outcomes. RESULTS: We analyzed 185 patients, of which 120 (64.9%) were upstaged and 65 (35.1%) were non-upstaged. When compared to non-upstaged masses, upstaged masses were significant for smaller size (6.8 cm vs 8.2 cm, p=0.008), lower RENAL score (8.7 vs 9.9, p<0.001), less hilar involvement (29.2% vs 86.2%, p<0.001), and increased exophyticity (41.7% vs 23.1%, p=0.011). On pathology, upstaged masses had greater proportions of perirenal fat invasion (53.3% vs 33.8%, p=0.011), but less venous system (44.2% vs 78.5%, p<0.001) and sinus fat invasion (35.8% vs 63.1%, p<0.001) compared to non-upstaged masses. RENAL domains R, E, and L were independent predictors for T3a disease foci (OR=0.053-7.70, p=0.001-0.042). MVA demonstrated that non upstaged cT3a RCC had a higher risk of recurrence (HR=2.01, p=0.043). KMA noted better RFS in upstaged patients compared to non-upstaged patients (80.8% vs 75.4%, p=0.002). CONCLUSIONS: Pathologically upstaged T3a RCC is associated with distinct morphology and invasion patterns, and improved recurrence free survival when compared to non-upstaged T3a RCC. RENAL domains can aid in identifying masses with upstaging potential. Source of Funding: UC San Diego Summer Research Fellowship © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e651 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Saeed Ghassemzadeh More articles by this author Aastha Shah More articles by this author Dhruv Puri More articles by this author Luke Wang More articles by this author Franklin Liu More articles by this author Sohail Dhanji More articles by this author Kevin Hakimi More articles by this author Mimi Nguyen More articles by this author Ryan Nasseri More articles by this author Margaret Meagher More articles by this author Clara Cerrato More articles by this author Juan Javier-Desloges More articles by this author Ithaar Derweesh More articles by this author Expand All Advertisement PDF downloadLoading ...

Abstract 5785: Tumors with higher heterogeneity were associated with poorer survival outcome in early stage lung adenocarcinoma cancer patients with low tumor mutational burden

Abstract Background: Response to immune checkpoint inhibitors is associated with tumor mutational burden (TMB). Pembrolizumab has recently been approved by the FDA for use in patients with high TMB. However low TMB patients are not considered good responders and could benefit from stratification. The predictive value of other biomarkers such as intratumor heterogeneity (ITH) has been studied across a range of cancer indications. The utility of ITH in Lung Adenocarcinoma (LUAD) has not been established. Here we evaluate the relationship between ITH and outcome in early stage low TMB patients and explore differences in immune cell enrichment and predicted driver genes. Stratifying early stage patients may yield an opportunity to improve Recurrence Free Survival (RFS) and lower the risk of progression to late stage disease. Methods: All molecular data was obtained from the Tumor Cancer Genome Atlas via the GDC Data Portal. Clinical data was from the PanCancer Atlas dataset through cBioPortal. We estimated ITH using PyClone, an algorithm that estimates the number of tumor clones. We generated our ITH values in triplicate across 3 separate days, yielding an average PyClone clone number. Clones with &amp;gt;2 mutations were counted. We used p-value minimization to choose 1.4 for the TMB cutoff and 2.6 for the PyClone cutoff. We generated 4 groups: HTHI, HTLI, LTHI, and LTLI, where H - high, L- low, T-TMB, and I-ITH. Immune cell enrichment analysis was obtained from the xCell webtool. Driver gene prediction was performed with OncoDriveClust via the R package maftools. Results: We found a statistically significant difference in RFS between early stage (Stage I and Stage II) LTHI (N = 18) and LTLI (N = 40) patients (16.69 months vs. 33.27 months, p-value = 0.0056). Interestingly the predicted driver genes did not significantly differ between these two groups. One driver (EGFR) and two drivers (EGFR and TP53) genes were predicted for LTHI and LTLI, respectively. Gene set enrichment analysis showed statistically significant differences in key immune subtypes. B-cells were depressed and Th1 cells were elevated in LTHI, both of which are associated with poorer survival outcomes. Conclusion: We found an association between high ITH and poor survival in low TMB early stage LUAD patients. Early stage patients can be stratified using heterogeneity metrics like PyClone. Given the presence of immune subtypes known to associate with poorer outcomes, early stage LTHI patients may benefit from additional adjuvant therapies. Citation Format: Stanislav Fridland, Young Kwang Chae. Tumors with higher heterogeneity were associated with poorer survival outcome in early stage lung adenocarcinoma cancer patients with low tumor mutational burden [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 5785.

Tumour infiltrating lymphocyte density differs by meningioma type and is associated with prognosis in atypical meningioma

Tumour infiltrating lymphocyte (TIL) density is prognostically significant in various tumours, but few studies have investigated its significance in meningioma. This study aimed to investigate how TIL density differs by meningioma histology and whether it is a predictor of meningioma recurrence. We studied CD3, CD8, CD4, FOXP3 and PD-1 positive (+) TIL density in a continuous cohort of 476 meningiomas resected at Auckland Hospital between 2002 and 2011 using tissue microarrays and computer assisted image analysis. TILs were identified in all meningiomas except one (median CD3+ TIL density across entire cohort 53.0 cells/mm2). Most TILs were CD8+ (median 33.6 cells/mm2) with smaller numbers of CD4+ TILs (median 2.9 cells/mm2). PD-1+ (median 0.32 cells/mm2) and FOXP3+ (median 0.0 cells/mm2) TILs were scarce. Reduced CD3+ (p=0.0066), CD8+ (p=0.0029) and PD-1+ (p=0.0375) TIL density was seen in WHO grade II/III meningioma compared with WHO grade I. Pairwise comparison confirmed statistically significant differences in TIL density existed between meningioma types (CD3, CD8, CD4, p<0.0001; FOXP3, p=0.0096; PD-1, p=0.0090) with chordoid meningioma having the lowest overall CD3+ TIL density (median 12.5 cells/mm2). Despite its low TIL density, chordoid meningioma had a higher FOXP3:CD8 ratio than several meningioma types. Atypical meningioma had a higher FOXP3:CD8 ratio than transitional meningioma (p=0.0045). No association between TIL density and recurrence was seen across the entire cohort or by WHO grade. However, CD3+ and CD8+ TIL density was associated with recurrence in atypical meningioma on multivariable analysis (CD3, p=0.0012; CD8, p=0.0071). A higher CD3+ and CD8+ TIL density was associated with improved recurrence free survival. Our findings suggest CD3+ and CD8+ TIL density is prognostically significant in atypical meningioma. Further investigation of this observation and its biological basis is warranted. The differences in TIL density by meningioma histology may be of relevance in studies of therapeutic immune checkpoint inhibition.

Consider Routine Administration of Neoadjuvant Treatment for Resectable Pancreatic Ductal Adenocarcinoma – A Single Center Experience

Introduction: Major postoperative complications (MPC) after pancreatoduodenectomy limit adjuvant chemotherapy (AC) administration, which may factor in increasing use of neoadjuvant chemotherapy (NC) for resectable pancreatic ductal adenocarcinoma (PDAC). Here we examined effects of NC on outcomes and MPC on AC administration. Methods: We performed a retrospective chart review of patients who underwent pancreatoduodenectomy for PDAC at our institution, 2015-2020. Demographic, clinicopathologic, and postoperative variables were analyzed. Results: We identified 146 patients, 68 (47%) received NC (Table). NC patients had smaller tumors [median 2.4cm vs. 2.8cm, p<0.002]; higher R0 resection rates (94% vs. 85%, p=0.048); lower vascular invasion rates (35% vs. 71%, p<0.001); lower LODDS [log of positive to negative lymph nodes; -1.2 (-1.5-(-0.9) vs. (-0.8 (-1.3-(-0.4), p<0.001]. NC was not associated with MPC or AC administration. With 21 months median follow-up, there was a trend towards improved recurrence free survival (RFS) (median 40 vs. 24 months, p=0.327) in NC patients. Of 78 (53%) patients who did not receive NC, 13 (17%) experienced MPC, and AC receipt was lower in this group (80% vs 39% p=0.006), with a trend toward lower RFS (median 25 vs 16 months p=0.456). On univariate regression analysis, MPC were associated with lower AC administration rates, both in patients that received and did not receive NC (OR 0.091, p=0.002; and OR 0.156, p=0.006, respectively). Conclusions: NC before pancreatoduodenectomy for PDAC was associated with favorable pathology and a trend toward improved RFS. As MPC limits AC administration, NC should be considered for all patients with resectable PDAC.

Optimal Management for Residual Disease Following Neoadjuvant Systemic Therapy.

Treatment sequencing in early-stage breast cancer has significantly evolved in recent years, particularly in the triple negative (TNBC) and human epidermal growth factor receptor 2 (HER2)-positive subsets. Instead of surgery first followed by chemotherapy, several clinical trials showed benefits to administering systemic chemotherapy (and HER2-targeted therapies) prior to surgery. These benefits include more accurate prognostic estimates based on the extent of residual cancer that can also guide adjuvant treatment, and frequent tumor downstaging that can lead to smaller surgeries in patients with large tumors at diagnosis. Patients with extensive invasive residual cancer after neoadjuvant therapy are at high risk for disease recurrence, and two pivotal clinical trials, CREATE-X and KATHERINE, demonstrated improved recurrence free survival with adjuvant capecitabine and ado-trastuzumab-emtansine (T-DM1) in TNBC and HER2-positive residual cancers, respectively. Patients who achieve pathologic complete response (pCR) have excellent long-term disease-free survival regardless of what chemotherapy regimen induced this favorable response. This allows escalation or de-escalation of adjuvant therapy: patients who achieved pCR could be spared further chemotherapy, while those with residual cancer could receive additional chemotherapy postoperatively. Ongoing clinical trials are testing this strategy (CompassHER2-pCR: NCT04266249). pCR also provides an opportunity to assess de-escalation of locoregional therapies. Currently, for patients with residual disease in the lymph nodes (ypN+), radiation therapy entails coverage of the undissected axilla, and may include supra/infraclavicular/internal mammary nodes in addition to the whole breast or chest wall, depending on the type of surgery. Ongoing trials are testing the safety of omitting post-mastectomy breast and post-lumpectomy nodal irradiation (NCT01872975) as well as omitting axillary lymph node dissection (NCT01901094) in the setting of pCR. Additionally, evolving technologies such as minimal residual disease (MRD) monitoring in the blood during follow-up may allow early intervention with "second-line systemic adjuvant therapy" for patients with molecular relapse which might prevent impending clinical relapse.

Propensity weighted indirect treatment comparison of nivolumab (NIVO) versus placebo (PBO) as adjuvant therapy for resected melanoma: A number needed to treat and overall survival analysis.

9572 Background: The CheckMate 238 trial demonstrated that NIVO improved recurrence free survival (RFS) vs. ipilimumab (IPI). The EORTC 18071 trial demonstrated that IPI improved RFS and overall survival (OS) vs. PBO. The current study pooled data from these two trials to indirectly assess the RFS and OS of NIVO vs. PBO and the numbers needed to treat (NNTs) for one additional recurrence-free survivor and survivor over 4 years. Methods: Patients with resected AJCC 7th edition stage IIIB/C cutaneous melanoma from CheckMate 238 (NIVO vs. IPI) and EORTC 18071 (IPI vs. PBO) were pooled together with inverse probability weighting to balance between-trial differences in baseline characteristics. NNTs were calculated for RFS and OS comparing NIVO vs. IPI and PBO over 4 years. To account for improved post-recurrence survival over time, a sensitivity analysis that adjusted for post-recurrence survival in the PBO arm of EORTC 18071 was performed. Results: A total of 278, 643, and 365 patients treated with NIVO, IPI, and PBO, respectively, were included. In the weighted samples, patients treated with NIVO had consistently higher RFS rates than those treated with IPI (HR [95% CI]: 0.69 [0.56, 0.85]) and PBO (HR: 0.49 [0.39, 0.61]). NIVO was associated with similar OS as IPI (HR: 0.80 [0.60, 1.08]) and superior OS compared to PBO (HR: 0.45 [0.33, 0.60]). At 4 years, the weighted RFS rate was 53.1% for NIVO, 41.8% for IPI, and 29.1% for PBO. The NNT to achieve one additional recurrence-free survivor was 4.2 for NIVO vs. PBO and 8.9 for NIVO vs. IPI. The NNT to obtain one additional survivor was 4.8 for NIVO vs. PBO and 22.2 for NIVO vs. IPI. The OS rate for PBO after adjusting for differences in post-recurrence treatments at 4 years was 64.1%, and the corresponding NNT of OS comparing NIVO vs. adjusted PBO was 8.5. Conclusions: In patients with resected AJCC 7th edition stage IIIB/C cutaneous melanoma, this indirect comparison showed that NIVO improved RFS and OS vs placebo, with OS improvement maintained after adjustment for post-recurrence therapy.[Table: see text]

The relationship between post-diagnostic statin usage and breast cancer prognosis varies by hormone receptor phenotype: a systemic review and meta-analysis.

Preclinical studies and epidemiologic data had indicated statins had antineoplastic properties in breast cancer patients. Since breast cancer treatment is based on its phenotype, it is important to explore influence of post-diagnosis statin usage on breast cancer patients with different phenotypes. We searched the related studies between inception and August, 2019 from MEDLINE and EMBASE. A total of 7 studies with 24,541 patients were identified. Stata/SE 15.0 and Review Manager 5.3 were used to analyze data. Inconsistency index was used to estimate heterogeneity. Begg's and Egger's regression test was used to examine publication bias. Overall post-diagnostic statin use was associated with improved recurrence free survival (recurrence free survival (RFS); hazard ratio (HR) 0.74; 95% confidential interval (95% CI) 0.57-0.98), overall survival (overall survival (OS); HR 0.53; 95% CI 0.31-0.91) and cancer-specific survival (cancer-specific survival (CSS); and HR 0.61; 95% CI 0.41-0.91). In hormone receptor positive patients, statin use was associated with improved CSS (HR 0.74, 95% CI 0.65-0.84). No protective effect was found in either OS or RFS. In hormone receptor negative patients, statin was associated with reduced OS (HR 2.19, 95% CI 1.34-3.59) and reduced RFS, but without statistical significance. Post-diagnostic statin use was associated with improved RFS, OS and CSS in breast cancer patients. Subgroup analysis indicted that the benefits of statin usage varied from hormone receptor phenotype type. Prospective randomized trial with patients of different hormone receptor types might be needed to help identify which subtype of breast cancer patients would benefit from post-diagnostic statin usage.

Characterizing the Tumor Immune Microenvironment (TIME) in Hepatocellular Carcinoma (HCC) And An Exploration Of The Prognostic Implications Of TIME Phenotypes On Treatment Response And Survival

The poor five-year survival in HCC is due in large part to frequent recurrence after primary resection and a lack of highly effective adjuvant treatments. While adjuvant sorafenib has not been shown to improve recurrence free survival in comparison with placebo, both external beam radiation therapy (EBRT) and stereotactic body radiation therapy (SBRT) have shown promise in the adjuvant setting. Specifically, EBRT was superior to RFA and TACE in patients with microvascular invasion, and 2-year survival in patients treated with SBRT was comparable to that in patients treated with RFA and TACE. Genomic sequencing advances have enabled the TIME in HCC to be characterized by a shift from effector CD8+ T cells to suppressive FoxP3+ Tregs. A higher Treg fraction confers a poor prognosis, and strategies targeting Tregs are gaining traction. Using data from the cancer genome atlas (TCGA), we characterize the HCC TIME and assess the relationship between TIME phenotype and response to therapies currently employed in recurrent HCC. RNA Seq by expectation maximization (RSEM) values for markers of interest were collected from the CBio portal and normalized. Matched clinical, pathological, and treatment data was downloaded from the Broad Institute’s Firebrowse tool. CIBERSORT “in silico flow cytometry” was used to quantify the relative fraction of immune cells in TCGA samples. Kaplan-Meier curves were constructed and compared using the log-rank test. Mean fractions of immune cell subsets and Treg marker expression between cohorts were compared by ANOVA. Clinical (ALBI, AFP, CP, ECOG) and pathologic (T stage) features were similar between the TCGA HCC cohort (n = 347) and the subset with available treatment data (n = 77). Mean post-treatment survival in the subset was 25.9 months. When divided by PD-1/PD-L1 expression and treatment, patients with high intratumoral co-expression of PD-1/PD-L1 had significantly increased median survival compared to other cohorts (45.5 mos, Xˆ2 = 10.56, p = 0.01). The PD-1/PD-L1 H/H group had a significantly higher relative Treg concentration (p = 0.001), and expression of Treg specific markers including CD25 (p = 1.4x10ˆ-5), FoxP3 (p = 2.7x10ˆ-4), and CTLA4 (p = 9.8x10ˆ-11). In this study, we demonstrate that the cohort with high PD-1/PD-L1 co-expression and an increased prevalence of active Tregs in the TIME had a significant survival advantage compared to other cohorts when treated with sorafenib, chemotherapy, or radiation - all of which have been shown to target Tregs as part of their mechanism of action. This survival difference was not observed in the same cohort treated with local therapies (TACE and RFA) that haven't been shown to have the same immune modulatory effects. The potential for targeting the immune suppressive microenvironment with radiation and systemic drug therapy, and stratification for treatment selection by TIME characteristics described here warrants further study as a strategy to approach recurrent HCC.

The prognostic significance of GRB7 protein expression and localization in human breast and ovarian cancers.

Objective: To study GRB7 protein expression in normal human tissues and breast and ovarian cancers, and determine its clinical significance. Results: GRB7 protein was expressed in multiple tissues, including myoepithelial cells of normal breast and fibroadenoma. Cytoplasmic GRB7 expression was seen predominantly in HER-2 positive and, to a lesser extent, triple negative breast cancer. Membrane localization of GRB7 was present in a subset of breast cancers with high cytoplasmic GRB7 expression. Univariate and multivariate analysis found that cytoplasmic GRB7 expression was associated with a negative progesterone receptor status, while membrane GRB7 expression was associated with a negative axillary nodal status. Membrane associated GRB7 expression was present in a subset of ovarian cancers with high cytoplasmic GRB7 expression. Membrane GRB7 expression displayed a trend towards improved recurrence free survival (RFS). Landmark analysis suggested an RFS advantage for ovarian cancers that had GRB7 membrane expression and survived beyond 27 months; GRB7 membrane expression in two or more cores (out of three) predicted an improved RFS. Membrane expression of GRB7 protein was observed in breast cancer cell lines with high GRB7 protein expression in vitro. Conclusion: GRB7 protein membrane expression may be associated with a better prognosis in breast and ovarian cancers. The favorable prognostic value of GRB7 protein membrane expression and its underlying mechanism is worthy of further investigation. Methods: Immunohistochemistry of normal human tissues, breast tissues of various pathologies, and clinically annotated ovarian cancers was performed to correlate the patterns of GRB7 expression with biomarkers or clinical outcome.

Abstract PD5-07: The immune landscape of residual triple-negative breast cancers after neoadjuvant chemotherapy

Abstract Background: Although neoadjuvant chemotherapy (NAC) induces complete response in 30-40% of triple-negative breast cancers (TNBC), patients with residual disease at surgery have poor prognosis and limited treatment options until recurrence. Tumor-infiltrating lymphocytes (TILs) in the residual disease are a positive prognostic factor, but how specific immune composition of the tumor guides outcome is unclear, resulting in a lack of understanding of how to employ immunotherapies in the adjuvant setting. Methods: We assessed multiple immunologic biomarkers in a series of 99 residual TNBCs after NAC. Immune markers were assessed by multiple methods, including H&amp;E-based TILs analysis, standard immunohistochemistry (IHC; CD8, CD20, CD56, FOXP3, LAG-3, B7-H4, HLA-A) and multiplexed immunofluorescence (mIF; HLA-DR, GZMB, CD4, CD8, PD-L1, pan-CK, PD1, CD3). Association among parameters were assessed (up to n=98) including clinical outcome after surgery (n=94). Results: As previously demonstrated, TILs in residual tumors after NAC predicted both RFS and OS (p=0.0093 and p=0.0376, respectively). H&amp;E-scored TILs were highly correlated to CD3, CD4 and CD8 positive T cells (Spearman r range 0.34-0.4850; all p&amp;lt;0.001), while CD20 and CD56 positive cells only make up a small and non-significant proportion of the TILs composition. Likewise, infiltration by overall T cell populations were associated with significantly improved recurrence free survival (RFS; HR range 0.34-0.54) and overall survival (OS; HR range 0.38-0.43), whereas CD20+ B cells were not. Additionally, infiltration by cells expressing LAG3 (HR 0.49) and CD56 (HR 0.44), were each associated with significantly improved OS. Interestingly, colocalizing GZMB+ and CD8+ T cells by mIF yielded a paradoxical finding that cytotoxic CD8+ T cells (CD8+GZMB+ T cells) were associated with worse RFS and OS (HR 1.8 p=0.0338 and HR=2.8 p=0.0013, respectively). In a multivariate model, only CD8+GZMB+ T cell infiltration remained significant for OS (p=0.0224), suggesting independence. CD8+GZMB+ T cell infiltration inversely correlated with TILs (p=0.0293) and CD8/mm2 counts in the tumor core (p=0.0020) and showed a positive correlation with B7H4 tumor expression (p=0.0183). Additionally, high CD8+GZMB+ cases showed low HLA-A (p=0.0189) expression in tumor cells. In accordance, high CD8+GZMB+ tumors mainly correspond to stroma-restricted, margin-restricted and immune-desert tumor immune microenvironment landscapes. Conclusions: We have found a paradoxical association of GZMB+ CD8+ T cells with a negative prognosis in NAC-treated TNBC. We hypothesize that while these T cells are poised for cytotoxic activity, they remain restricted through sub-localization outside the tumor core, downregulation of HLA-A on tumor cells preventing interaction with the T cell receptor, and upregulation of B7H4 expression, which has been shown to inhibit cytotoxic T cell activity. Patients with high CD8+GZMB+ tumors in the post-NAC setting may benefit from adjuvant immunotherapy, particularly in combination with therapies that enhance MHC-I (e.g. HLA-A) antigen presentation. Citation Format: Paula I Gonzalez-Ericsson, Violeta Sanchez, Roberto Salgado, Jennifer Bordeaux, Ju Y Kim, Christine Vaupel, Henry Gomez, Melinda E Sanders, Justin M Balko. The immune landscape of residual triple-negative breast cancers after neoadjuvant chemotherapy [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr PD5-07.