PURPOSE Universal cancer peptide–based vaccine (UCPVax) is a therapeutic vaccine composed of two highly selected helper peptides to induce CD4+ T helper-1 response directed against telomerase. This phase Ib/IIa trial was designed to test the safety, immunogenicity, and efficacy of a three-dose schedule in patients with metastatic non–small-cell lung cancer (NSCLC). PATIENTS AND METHODS Patients with refractory NSCLC were assigned to receive three vaccination doses of UCPVax (0.25 mg, 0.5 mg, and 1 mg) using a Bayesian-based phase Ib followed by phase IIa de-escalating design. The primary end points were dose-limiting toxicity and immune response after three first doses of vaccine. Secondary end points were overall survival (OS) and progression-free survival at 1 year. RESULTS A total of 59 patients received UCPVax; 95% had three prior lines of systemic therapy. No dose-limiting toxicity was observed in 15 patients treated in phase Ib. The maximum tolerated dose was 1 mg. Fifty-one patients were eligible for phase IIa. The third and sixth dose of UCPVax induced specific CD4+ T helper 1 response in 56% and 87.2% of patients, respectively, with no difference between three dose levels. Twenty-one (39%) patients achieved disease control (stable disease, n = 20; complete response, n = 1). The 1-year OS was 34.1% (95% CI, 23.1 to 50.4), and the median OS was 9.7 months, with no significant difference between dose levels. The 1-year progression-free survival and the median OS were 17.2% (95% CI, 7.8 to 38.3) and 11.6 months (95% CI, 9.7 to 16.7) in immune responders ( P = .015) and 4.5% (95% CI, 0.7 to 30.8) and 5.6 months (95% CI, 2.5 to 10) in nonresponders ( P = .005), respectively. CONCLUSION UCPVax was highly immunogenic and safe and provide interesting 1-year OS rate in heavily pretreated advanced NSCLC.

Background Several cancer immunotherapies that target the PD-L1/PD-1 pathway show promising clinical activity in patients with hepatocellular carcinoma (HCC). However, the standard of care in first-line treatment with atezolizumab (anti-PD-L1 therapy) in combination with bevacizumab is associated with a limited objective response rate. Telomerase reverse transcriptase (TERT) activation meets the criteria of oncogenic addiction in HCC and could be actionable therapeutic target and a relevant tumor antigen. Therefore we hypothesized that combining anti-PD-1/PD-L1 therapy with an anti-telomerase vaccine might be an attractive therapy in HCC. UCPVax is a therapeutic cancer vaccine composed of two separate peptides derived from telomerase (human TERT). UCPVax has been evaluated in a multicenter phase I/II study in non–small cell lung cancers and has demonstrated to be safe and immunogenic, and is under evaluation in combination with atezolizumab in a phase II clinical trial in tumors where telomerase reactivation contributes to an oncogene addiction (HPV + cancers). The aim of the TERTIO study is to determine the clinical interest and immunological efficacy of a treatment combining the CD4 helper T-inducer cancer anti-telomerase vaccine (UCPVax) with atezolizumab and bevacizumab in unresectable HCC in a multicenter randomized phase II study. Methods Patients with locally advanced, metastatic or unresectable HCC who have not previously received systemic anti-cancer treatment are eligible. The primary end point is the objective response rate at 6 months. Patients will be allocated to a treatment arm with a randomization 2:1. In both arms, patients will receive atezolizumab at fixed dose of 1200 mg IV infusion and bevacizumab at fixed dose of 15 mg/kg IV infusion, every 3 weeks, according to the standard of care. In the experimental arm, these treatments will be combined with the UCPVax vaccine at 0.5 mg subcutaneously. Discussion Combining anti-PD-1/PD-L1 therapy with an anti-telomerase vaccine gains serious consideration in HCC, in order to extend the clinical efficacy of anti-PD-1/PD-L1. Indeed, anti-cancer vaccines can induce tumor-specific T cell expansion and activation and therefore restore the cancer-immunity cycle in patients lacking pre-existing anti-tumor responses. Thus, there is a strong rational to combine immune checkpoint blockade therapy and anticancer vaccine (UCPVax) in order to activate antitumor T cell immunity and bypass the immunosuppression in the tumor microenvironment in HCC. This pivotal proof of concept study will evaluate the efficacy and safety of the combination of a CD4 Th1-inducer cancer vaccine derived from telomerase (UCPVax) and atezolizumab plus bevacizumab in unresectable HCC, as well as confirming their synergic mechanism, and settling the basis for a new combination for future clinical trials. Trial registration NCT05528952.

Triplet DCF (docetaxel, cisplatin and 5-flurouracil) and doublet CP/CF (carboplatin and paclitaxel/cisplatin and 5-fluorouracil) regimens were prospectively evaluated in advanced squamous anal cell carcinoma (SCCA), and validated as standard treatments. Even though the high efficacy and good tolerance of DCF regimen were confirmed in 3 independent prospective trials, doublet CP regimen is still recommended in several guidelines based in its better safety profile with similar efficacy compared to CF regimen. We performed a propensity score-adjusted method with inverse probability of treatment weighted (IPTW) and matched case control (MCC) comparison among patients with metastatic or non-resectable locally advanced recurrent SCCA, treated with chemotherapy as first line regimen. The primary endpoint was the overall survival (OS), and the secondary endpoint was the progression-free survival (PFS). 247 patients were included for analysis. 154 patients received DCF and 93 patients received a doublet regimen. The median OS was 32.3 months with DCF and 18.3 months with doublet regimens (HR 0.53, 95%CI 0.38–0.74; p = 0.0001), and the median PFS was 11.2 months with DCF versus 7.6 months with doublet regimens (HR 0.53, 95%CI 0.39–0.73; p < 0.0001). The hazard ratios by IPTW and MCC analyses were 0.411 (95% CI, 0.324–0.521; p < 0.0001) and 0.406 (95% CI, 0.261–0.632; p < 0.0001) for OS, and 0.466 (95% CI, 0.376–0.576; p < 0.0001) and 0.438 (95% CI, 0.298–0.644; P < 0.0001) for PFS. The triplet DCF regimen provides a high and significant benefit in OS and PFS over doublet regimens, and should be considered as upfront treatment for eligible patients with advanced SCCA.

Abstract BACKGROUND The Telomerase Reverse Transcriptase (TERT) is activated in 85%-90% of cancers, and glioblastomas (GBM) harbor the highest incidence of activating mutations within its promoter (over 85% of patients). This high expression of TERT in cancers and its role in oncogenesis make TERT a very promising tumor antigen for immunotherapy, especially in GBM. UCPvax is a therapeutic vaccine in which two CD4 helper peptides derived from TERT are mixed with the Montanide adjuvant. This phase IIa trial was designed to test the immunogenicity, safety and efficacy of UCPvax in patients with newly diagnosed GBM. PATIENTS AND METHODS In this first cohort of the UCPvax-glio trial (NCT04280848), patients with unmethylated MGMT non-mutated IDH1 GBM were included if they had a Karnofsky Performance status (KPS) ≥ 70%, a steroid treatment < 10mg/ day equivalent prednisone, and lymphocytes count ≥ 0.8 x 109/L. One month after completion of radiation/ temozolomide (TMZ), patients started UCPvax vaccinations on days 1, 8, 15, 29, 36 and 43, then every two months until tumor progression. Patients did not receive any additional cure of TMZ because of the unmethylated MGMT promoter status. Peripheral blood mononuclear cells were collected before treatment, at 1 and 2 months after treatment, and at each vaccination boost. The primary endpoint was the anti-TERT specific CD4 T-cell response at 2 months using IFN-gamma ELISPOT. Secondary endpoints included safety, progression-free survival (PFS) and overall survival (OS). RESULTS All the 31 patients (median age 60-yr old, median KPS 90%) included in this study received at least one vaccination. Vaccinations were given for 4.5 months on average (min 2- max 14). At baseline, only one patient had a pre-existing anti-TERT CD4 T-cell response (3%). After immunizations, de novo induction and/or amplification of an anti-TERT response were found in 29/30 patients (97%). An epitope spread response against other tumor-associated antigens was detected in 12/25 patients (48%). All patients developed local skin reactions (≤ grade 2), 16 patients complained of transient asthenia (≤grade 2), and 13 patients experienced local pain (≤grade 2). No severe (grade 3-4) toxicity was attributable to the vaccination. In the intent-to-treat population (n = 31), the PFS was 8.9 months (95% CI: 7.6-10.6) and the median OS was 17.9 months (95% CI: 16-23). Two years after diagnosis, 26% of the patients were still alive. OS was significantly improved in the patients developing an epitope spread response vs the others (20.8 versus 14.1 months, respectively, p = 0.03). CONCLUSION s: In this population of unmethylated MGMT GBM patients, UCPVax is highly immunogenic and provides an interesting OS rate. These data strongly support the completion of the second ongoing cohort, assessing UCPvax in combination with TMZ for GBM patients with methylated MGMT promoter status.

2005 Background: TERT, a subunit of the telomerase complex, is largely expressed in almost all cancers. Among those cancers, glioblastomas (GBM) harbor the highest incidence of activating mutations within the TERT promoter (over 85% of patients). This high incidence underlies the potential role of TERT in oncogenesis, and points out TERT as a highly relevant tumor target in GBM. UCPvax is a therapeutic vaccine composed of two CD4 helper peptides derived from TERT, combined with the montanide adjuvant. We conducted a phase IIa trial to test the immunogenicity, safety and efficacy of UCPvax in patients with newly diagnosed GBM. Methods: Key inclusion criteria were: histologically confirmed, non-mutated IDH1 glioblastoma, unmethylated MGMT promoter status, previous treatment with concomitant radiotherapy and temozolomide (TMZ), Karnofsky Performance status (KPS) ≥ 70%, steroids < 10mg/ day equivalent prednisone, and lymphocytes count ≥ 0.8 x 109/L. One month after completion of radiation/TMZ, patients started UCPvax vaccinations on days 1, 8, 15, 29, 36 and 43, then every two months until tumor progression, without additional cure of TMZ. Peripheral blood mononuclear cells were collected before treatment, at 1 and 2 months after treatment and, at each vaccination boost. The primary endpoint was an anti-TERT specific CD4 T-cell response measured ex vivo in peripheral blood using IFN-gamma ELISPOT at 2 months. Secondary endpoints included safety, overall survival (OS) and progression-free survival (PFS). Results: Thirty-one adult patients (median age 60-yr old, median KPS 90%) were included in this study. All patients received at least one vaccination, and vaccinations were given for 4.5 months on average (min 2– max 14). At baseline, only one patient had a pre-existing anti-TERT CD4 T-cell response (3%). After immunizations, de novo induction and/or amplification of an anti-TERT response were found in 29/30 pts (97%). An epitope spread response against other tumor-associated antigens was detected in 12/25 pts (48%). No severe (grade 3–4) toxicity was attributable to the vaccination. All patients developed local skin reactions (≤ grade 2), 16 patients complained of transient asthenia (≤grade 2), and 13 patients experienced local pain (≤grade 2). In the intent-to-treat population (n=31), the PFS was 8.9 months (95% CI: 7.6–10.6) and the median OS was 17.9 months (95% CI: 16–23). Two years after diagnosis, 26 % of the patients were still alive. OS was significantly improved in the patients developing an epitope spread response vs the others (19.3 vs 15.8 months, respectively, p=0.03). Conclusions: UCPVax is highly immunogenic and provides an interesting OS rate in this population of poor prognosis, unmethylated MGMT GBM patients. These data support further clinical studies of UCPvax in GBM patients. Clinical trial information: NCT04280848 .

Background: Squamous cell carcinoma of the anus (SCCA) is a rare disease often diagnosed at a localised stage. For locally advanced recurrence or metastatic disease, DCF (docetaxel, cisplatin, 5-fluorouracil) demonstrated high efficacy and became one of the standard regimens. However, there is no standard of care in the second line.Patients and methods: In the Epitopes-HPV01 and Epitopes-HPV02 prospective trials, 115 patients with advanced SCCA were treated with a DCF regimen in the first line. In these studies, second-line data were registered per protocol.Results: After a median follow-up of >40 months, at progression, 73 patients received a second-line (L2) treatment. In this L2 population, median overall survival (mOS) was 13.5 months (95%CI 9.4-19.8), and median progression-free survival (mPFS) was 5.7 months (3.4-7.3) in L2. Fourteen patients presented an oligometastatic progression and were treated with an ablative treatment (surgery or radiotherapy); mOS was 48.3 months (NE-NE), and mPFS was 31.3 months (23.2-NE). Fifty-nine patients received a systemic treatment (chemotherapy or immunotherapy); mOS was 11 months (8.4-15.4) and mPFS was 4.9 months (3.3-7). The most frequent chemotherapy regimens were the reintroduction of DCF, paclitaxel, FOLFIRI and mitomycin plus fluoropyrimidine. No significant difference was observed between regimens (p Z 0.26). Six patients received anti-PD1/L1-based immunotherapy.Conclusion: Second-line treatments are effective in patients with SCCA. Ablative treatment is feasible and is probably the best option for patients with oligometastatic progression. If this is not possible, systemic therapy by an anti-PD1/L1 immunotherapy or chemotherapy can be recommended. Reintroduction of DCF, paclitaxel, FOLFIRI or mitomycin-C plus fluoropyrimidine are possible options.(c) 2021 Elsevier Ltd. All rights reserved.

BackgroundThere is a strong rational of using anti–programmed cell death protein-1 and its ligand (anti–PD-1/L1) antibodies in human papillomavirus (HPV)–induced cancers. However, anti–PD-1/L1 as monotherapy induces a limited number of objective responses. The development of novel combinations in order to improve the clinical efficacy of an anti–PD-1/L1 is therefore of interest. Combining anti–PD-1/L1 therapy with an antitumor vaccine seems promising in HPV-positive (+) cancers. UCPVax is a therapeutic cancer vaccine composed of two separate peptides derived from telomerase (hTERT, human telomerase reverse transcriptase). UCPVax is being evaluated in a multicenter phase I/II study in NSCLC (non–small cell lung cancer) and has demonstrated to be safe and immunogenic. The aim of the VolATIL study is to evaluate the combination of atezolizumab (an anti-PD-L1) and UCPVax vaccine in a multicenter phase II study in patients with HPV+ cancers.MethodsPatients with HPV+ cancer (anal canal, head and neck, and cervical or vulvar), at locally advanced or metastatic stage, and refractory to at least one line of systemic chemotherapy are eligible. The primary end point is the objective response rate (ORR) at 4 months. Patients will receive atezolizumab every 3 weeks at a fixed dose of 1,200 mg in combination with the UCPVax vaccine at 1 mg subcutaneously.DiscussionAnti-cancer vaccines can restore cancer-immunity via the expansion and activation of tumor-specific T cells in patients lacking pre-existing anti-tumor responses. Moreover, preclinical data showed that specific TH1 CD4 T cells sustain the quality and homing of an antigen-specific CD8+ T-cell immunity. In previous clinical studies, the induction of anti-hTERT immunity was significantly correlated to survival in patients with advanced squamous anal cell carcinoma. Thus, there is a strong rational to combine an anti-cancer hTERT vaccine and an immune checkpoint inhibitor to activate and promote antitumor T-cell immunity. This pivotal proof of concept study will evaluate the efficacy and safety of the combination of a telomerase-based TH1 inducing vaccine (UCPVax) and an anti–PD-L1 (atezolizumab) immunotherapy in HPV+ cancers, as well as confirming their synergic mechanism, and settling the basis for a new combination for future clinical trials.Clinical Trial Registrationhttps://www.clinicaltrials.gov/, identifier NCT03946358.

BackgroundThe stimulation of antitumor CD4 T helper response represents a critical requirement for therapeutic cancer vaccine effectiveness. In this context, we developed a Th1-inducer anticancer peptide vaccine derived from telomerase called UCPVax (Dosset et al. Clin Can Res 2012). The safety and efficacy of UCPVax therapeutic vaccination were recently reported in metastatic non-small cell lung cancer (NSCLC) phase I/II trial (Adotévi et al., J Clin Oncol 2022, NCT02818426). Here, we described the immunological responses promoted by UCPVax.MethodsImmune monitoring was performed in 52 patients with refractory metastatic NSCLC previously vaccinated with three doses of UCPVax for six times (one per week) followed by boost every two months for one year. Blood samples were collected before and at different times after vaccination. Vaccine-specific CD4 T cell responses and epitope spreading were evaluated by IFN-γ ELISPOT assay. MHC class II pentamer staining and intracellular cytokine secretion assay were used for the assessment of phenotype function and polarization of antigen specific CD4 T cells. Vaccine induced antibody response was measured by ELISA.ResultsSeventy % of patients mounted strong UCP-specific CD4 T cells after vaccination regardless the dose level. UCPVax induced de novo and strong UCP-specific CD4 T-cell response and the intensity of the response increased according to the number of vaccinations. The UCP-specific CD4 T cells were effector memory phenotype, showed cytotoxic potential and (IFN-γ,TNF-a, IL-2)+ Th1 polarization which were maintained in long-term responders. Increased level of antibody response against UCP was found in 45% of patients which was correlated to the number of UCP-CD4 T cells. Furthermore, UCPVax induced epitope spread responses against various tumor antigens in most of patients analyzed. Finally, patients displayed polyfunctional anti-UCP CD4 response and epitope spreading had a better survival.ConclusionsUCPVax vaccination promotes highly functional and long-lasting tumor specific CD4 T cell responses associated with the improvement of patients' survival.Clinical trial identificationNCT02818426.Legal entity responsible for the studyO. Adotevi.FundingNational Cancer Institute INCa France PHRC program.DisclosureO. Adotevi: Financial Interests, Institutional, Funding: AstraZeneca; Financial Interests, Institutional, Research Grant: BMS; Non-Financial Interests, Personal, Principal Investigator: MSD. V. Westeel: Financial Interests, Personal, Advisory Role: Ipsen; Financial Interests, Personal, Speaker's Bureau: Amgen, Lilly, MSD, BMS. C. Borg: Financial Interests, Personal, Advisory Role: MSD Oncology, Roche; Financial Interests, Institutional, Sponsor/Funding: Bayer. All other authors have declared no conflicts of interest. BackgroundThe stimulation of antitumor CD4 T helper response represents a critical requirement for therapeutic cancer vaccine effectiveness. In this context, we developed a Th1-inducer anticancer peptide vaccine derived from telomerase called UCPVax (Dosset et al. Clin Can Res 2012). The safety and efficacy of UCPVax therapeutic vaccination were recently reported in metastatic non-small cell lung cancer (NSCLC) phase I/II trial (Adotévi et al., J Clin Oncol 2022, NCT02818426). Here, we described the immunological responses promoted by UCPVax. The stimulation of antitumor CD4 T helper response represents a critical requirement for therapeutic cancer vaccine effectiveness. In this context, we developed a Th1-inducer anticancer peptide vaccine derived from telomerase called UCPVax (Dosset et al. Clin Can Res 2012). The safety and efficacy of UCPVax therapeutic vaccination were recently reported in metastatic non-small cell lung cancer (NSCLC) phase I/II trial (Adotévi et al., J Clin Oncol 2022, NCT02818426). Here, we described the immunological responses promoted by UCPVax. MethodsImmune monitoring was performed in 52 patients with refractory metastatic NSCLC previously vaccinated with three doses of UCPVax for six times (one per week) followed by boost every two months for one year. Blood samples were collected before and at different times after vaccination. Vaccine-specific CD4 T cell responses and epitope spreading were evaluated by IFN-γ ELISPOT assay. MHC class II pentamer staining and intracellular cytokine secretion assay were used for the assessment of phenotype function and polarization of antigen specific CD4 T cells. Vaccine induced antibody response was measured by ELISA. Immune monitoring was performed in 52 patients with refractory metastatic NSCLC previously vaccinated with three doses of UCPVax for six times (one per week) followed by boost every two months for one year. Blood samples were collected before and at different times after vaccination. Vaccine-specific CD4 T cell responses and epitope spreading were evaluated by IFN-γ ELISPOT assay. MHC class II pentamer staining and intracellular cytokine secretion assay were used for the assessment of phenotype function and polarization of antigen specific CD4 T cells. Vaccine induced antibody response was measured by ELISA. ResultsSeventy % of patients mounted strong UCP-specific CD4 T cells after vaccination regardless the dose level. UCPVax induced de novo and strong UCP-specific CD4 T-cell response and the intensity of the response increased according to the number of vaccinations. The UCP-specific CD4 T cells were effector memory phenotype, showed cytotoxic potential and (IFN-γ,TNF-a, IL-2)+ Th1 polarization which were maintained in long-term responders. Increased level of antibody response against UCP was found in 45% of patients which was correlated to the number of UCP-CD4 T cells. Furthermore, UCPVax induced epitope spread responses against various tumor antigens in most of patients analyzed. Finally, patients displayed polyfunctional anti-UCP CD4 response and epitope spreading had a better survival. Seventy % of patients mounted strong UCP-specific CD4 T cells after vaccination regardless the dose level. UCPVax induced de novo and strong UCP-specific CD4 T-cell response and the intensity of the response increased according to the number of vaccinations. The UCP-specific CD4 T cells were effector memory phenotype, showed cytotoxic potential and (IFN-γ,TNF-a, IL-2)+ Th1 polarization which were maintained in long-term responders. Increased level of antibody response against UCP was found in 45% of patients which was correlated to the number of UCP-CD4 T cells. Furthermore, UCPVax induced epitope spread responses against various tumor antigens in most of patients analyzed. Finally, patients displayed polyfunctional anti-UCP CD4 response and epitope spreading had a better survival. ConclusionsUCPVax vaccination promotes highly functional and long-lasting tumor specific CD4 T cell responses associated with the improvement of patients' survival. UCPVax vaccination promotes highly functional and long-lasting tumor specific CD4 T cell responses associated with the improvement of patients' survival.

CD4 T cells play a key role in anticancer immunity. In this study, we investigate the clinical relevance of circulating CD4 T helper type 1 (Th1) response against telomerase (anti-TERT Th1 response) in patients with melanoma. The spontaneous anti-TERT Th1 response was detected in 54.5% (85/156) of patients with melanoma before treatment. The prevalence of this systemic response was inversely related to Breslow thickness >1 mm and American Joint Committee on Cancer stage >= II (P = 0.001 and 0.032, respectively). In contrast to patients treated with targeted therapies, the anti-TERT Th1 immunity was associated with an objective response after immune checkpoint inhibitors treatment. Hence, 86% (18/21) of responder patients exhibited pre-existing anti-TERT Th1 versus 35% (6/19) in nonresponders (P = 0.001). This response was also associated with increased progression-free survival and overall survival in patients with melanoma treated with immune checkpoint inhibitors (P = 0.0008 and 0.012, respectively). Collectively, the presence of circulating anti-TERT Th1 response is inversely related to melanoma evolution and appears to be a predictive factor of response to immunotherapy. Our results highlight the interest in telomerase-specific CD4 Th1 response as a promising blood-based biomarker of immune checkpoint inhibitors therapy in melanoma.

Aim Raltitrexed (RTX) is a thymidylate synthase inhibitor with large pharmacokinetics (PK) variability that can be administered in case of 5-fluorouracil (5FU) intolerance or dihydropyrimidine dehydrogenase deficiency. While it is a more potent thymidylate synthase inhibitor than 5FU, RTX failed to replace this drug for colorectal cancer patients, mainly due to its toxicity at the recommended dose of 3 mg/m(2)every 3 weeks. However, every 2 weeks administration at 2 mg/m(2)demonstrated a favourable toxicity profile. Method We performed a randomized crossover comparative population PK study between every 2 weeks TOMOX (RTX 2 mg/m(2)) and every 3 weeks TOMOX (RTX 3 mg/m(2)). Results A three-compartment model and a proportional error model best describe the data. Creatinine clearance and sex, but not body surface area (BSA), were covariates of RTX clearance leading to decrease of its interindividual variability of 28%. Weight and body surface area were covariates of central and peripheral volumes of distribution, respectively, leading to decreases of interindividual variability of 34.6% and 100%, respectively. In contrast to the dose, AUC was a good predictor of liver toxicity (P= 0.006, OR = 3.91, 95%CI = [1.48-10.34]). Using covariates to compute individual clearance and a threshold AUC (1.639, determined in this study), a covariates-based dose was calculated, leading to less variability in AUC than observed with the actual BSA-based or fixed doses. Conclusion These results advocate for the use of creatinine clearance and sex to determine the RTX dose instead of BSA.

Background Modified docetaxel, cisplatin, and 5-fluorouracil (mDCF) regimen has become a new standard for the treatment of metastatic or unresectable locally advanced recurrent squamous cell carcinoma of the anus (SCCA) after demonstrating improved efficacy (12-month PFS of 47%) in the Epitopes-HPV02 trial. Antibodies targeting the checkpoint inhibitor (CKI) programmed cell death protein-1 (PD1) have demonstrated the efficacy as monotherapies in second-line treatment of SCCA. The aim of this study is to evaluate the combination of atezolizumab and mDCF as first-line chemotherapy in a non-comparative multicentre randomized phase II study of advanced SCCA patients. Methods Patients with chemo-naive advanced histologically proven SCCA, metastatic or unresectable locally advanced recurrence, and Eastern Cooperative Oncology Group-performance status (ECOG-PS) < 2 will be eligible. The primary endpoint is a 12-month PFS rate. Using one-arm non-parametric survival with unilateral alpha type I error of 5% and a statistical power of 80%, the upper critical value for the 12-month PFS rate is 47% to reject H0. Assuming 5% lost to follow-up, 99 patients will be randomized on a 2:1 basis, 66 to the experimental arm (arm A, mDCF plus atezolizumab) and 33 to the standard arm (arm B, mDCF). In both arms, 8 cycles of mDCF will be administered. In arm A, patients receive mDCF with a fixed dose of atezolizumab (800 mg every 2 weeks) and are followed up to 1 year. Secondary endpoints are overall survival, PFS, response rate, safety, health-related quality of life, and an extensive biomarker programme and its correlation with the treatment efficacy. Discussion Although the Epitopes-HPV02 trial has changed long-lasting prognosis of patients with SCCA in advanced stage disease, more than 50% of patients will progress at 12 months. The purpose of the SCARCE trial to establish the addition of atezolizumab to mDCF as a new standard in this rare disease. Associated biomarker studies and the control arm could contribute to better understanding of the potential synergic and tumour resistance mechanisms in SCCA. Trial registration NCT03519295 .

Aims: The addition of docetaxel to cisplatin and 5-fluorouracil (DCF) has shown promising efficacy in advanced squamous cell carcinoma of the anus (SCCA). Preliminary results of Epitopes-HPV01 study showed a high rate of long-lasting complete response to DCF. The prospective, multicenter, Epitopes-HPV02 trial then confirmed the high efficacy of the modified DCF (mDCF) regimen in terms of complete response rate and long-term survival in metastatic or non-resectable locally advanced recurrent SCCA. Here, we present updated results of the Epitopes-HPV01 and Epitopes-HPV02 studies. Patients & methods: Epitopes-HPV01 is a prospective study performed by the regional cancer network of Franche-Comté, France. Epitopes-HPV02 is a phase II study supported by two French collaborative oncological groups, performed in 25 centers. Both studies included patients with metastatic, or with unresectable local recurrent SCCA, treated with DCF regimen. Results: In Epitopes-HPV01, 51 patients were enrolled between September 2012 and January 2019, and 49 patients were included for analysis; while 69 patients were included between September 2014 and December 2016 in Epitopes-HPV02, and 66 patients for analysis. Pooled analysis of 115 patients showed a median progression-free survival of 12.2 months [95% confidence interval (CI) 10.6–16.1] [11.0 months (9.3–16.0) in -HPV02, and 15.6 months (11.2–34.5) in -HPV01, ( p = 0.06)]. The median overall survival was 39.2 months (26.0–109.1) [36.3 in -HPV02 (25.2–NR), and 61.1 months (21.4–120.0) in -HPV01 ( p = 0.62)]. Objective response rate was 87.7% (90.9% in -HPV02 and 83.3% in -HPV01) with 40.3% of complete response (45.5% in -HPV02 and 33.3% in -HPV01). No differences were observed between standard DCF ( n = 54) and mDCF ( n = 58) in terms of OS ( p = 0.57) and PFS ( p = 0.99). 5-years PFS and OS rates were 24.5% and 44.4%, respectively, in the whole population. No treatment-related death was observed. Conclusion: Updated results of Epitopes-HPV01 and 02 studies, as well as the pooled analysis, confirm mDCF as a standard treatment in patients with advanced SCCA.

3044 Background: Increasing evidence highlights the crucial roles played by CD4+ Th1 cells in cancer immunity and immunotherapy (Spitzer et al., Cell 2017, Borst et al., Nat rev Immunol 2018). Here, we investigate the relevance of circulating CD4 Th1 response against shared tumor-associated antigens (TAA) in cancer patients treated by anti-PD-1 immunotherapy. Methods: A total of 46 advanced cancer patients (pts) including 32 pts with non-small cell lung cancer (NSCLC), 14 pts with melanoma, were enrolled (ITHER trial NCT02840058). Patients were treated with anti-PD-1 therapy as standard of care (26 pts with nivolumab and 20 pts with pembrolizumab). Peripheral blood mononuclear cells were collected before and after treatment at 1 and 3 months. The presence of circulating TAA-specific Th1 response was measured by IFNy ELISPOT assay using a mixture of 15mer peptides derived from telomerase (TERT) (Laheurte et al., Oncoimmunology 2016 and Br J C 2019). Results: At the baseline, the anti-TERT Th1 response was observed in 37% of pts. After anti-PD-1 therapy, de novo induction and/or amplification of pre-existing anti-TERT Th1 response was found in 26 % of pts (12/46). Whereas, a decrease of this response was documented in 15% of pts (7/46).The presence of anti-TERT Th1 response in peripheral blood during anti-PD-1 treatment was associated with a prolonged progression free-survival (PFS) as compared to the immune non responder pts (14.4 vs 2.6 months respectively, p = 0.006, HR 0.39 [0.2;0.76]). Similar observation was made for the overall survival (OS) (22.3 vs 12.3 months respectively, p = 0.04 HR 0.45 [0.21;0.96]). Notably, de novo reinvigoration of peripheral anti-TERT Th1 response after anti-PD-1 therapy was associated with a better clinical outcome as compared to the group of pts with decreased immune response after treatment (Median OS not reached vs 5.8 months). In contrast, no association with anti-PD-1 response was observed neither with circulating anti-NY-ESO-1 or with anti-viral Th1 response, concurrently measured in these patients. Conclusions: The reinvigoration of circulating CD4 Th1 against telomerase in patients treated by anti-PD-1 is associated with a better clinical outcome. These results underline the potential interest of monitoring circulating antitumor CD4 Th1 response for immune checkpoint inhibitors management.

Biochemical recurrence (BCR) occurs in up to 40% of prostate cancer patients after prostatectomy. In our study, we performed an immune monitoring study in 20 prostate cancer patients with BCR previously treated with metronomic cyclophosphamide (mCTX). We observed a decrease of regulatory T cells (Tregs) from 2 months and this was more pronounced after 6 months of mCTX treatment. This drop of Tregs was associated with increased level of activated HLADR+ CD45R0+ T cells in peripheral blood. Furthermore, a reactivation of Th1 polarized anti-PSA T-cell response was detected in BCR patients treated with mCTX. However, dendritic cell subsets counts and activation were not influenced by the treatment. In the clinical setting, we found that PSA level control was observed in 82% (9/11) of patients with a significant diminution of Tregs after mCTX compared to 33% (3/9) in patients without Tregs decrease. In addition, 30% (6/20) of patients previously treated with mCTX remained free for androgen deprivation therapy. In conclusion, Tregs diminution and immune activation associated with PSA level control occurred after mCTX in prostate cancer patients with BCR.

Background The addition of docetaxel to cisplatin and 5FU (DCF) has shown a promising efficacy in Epitopes-HPV01 study with 4 of first 8 consecutive patients presenting a long-lasting complete response. Then, the prospective, multicenter, Epitopes-HPV02 trial, settled the modified DCF regimen as a new standard of care in metastatic or non-resectable locally advanced recurrent anal squamous cell carcinoma (ASCC). Here we present updated results of Epitopes-HPV02 study, as well as final results of Epitopes-HPV01 study. Methods Epitopes-HPV02 was a phase 2 study supported by the GERCOR and FFCD collaborative oncological groups, performed in 25 academic and community hospitals in France. Epitopes-HPV01 was a real-life based cohort study performed by the regional cancer network of Franche-Comte, France, and including one university hospital, 5 community hospitals, and 1 private center. Both studies included patients with histologically confirmed ASCC, with metastatic disease, or with unresectable local recurrence after chemoradiotherapy, and treated with DCF regimen. Results In Epitopes-HPV02, 69 patients were enrolled between Sept 2014-Dec 2016, and 66 patients were included for analysis; while 51 patients were included between Sept 2012- January 2019 in Epitopes-HPV01, and 49 patients for analysis. Pooled analysis of 115 patients showed a median PFS of 12 months (95% CI 10.6-16.0) [11.0 months (9.3-16.0) in -HPV02, and 12.7 months (11.2-34.5) in -HPV01, (p = 0.14)]. The median OS was 50.2 months (26.0-120.0) [not reached in -HPV02, and 50.2 months (21.4-120.0) in -HPV01 (p = 0.73)]. ORR was 87.7% (89% in -HPV02 and 85.1% in -HPV01) with 40.7% of CR (45% in -HPV02 and 34% in -HPV01). No difference was observed between standard DCF (n = 54) and modified DCF (n = 58) in OS (p = 0.93) and PFS (p = 0.52).No treatment-related death was observed in both studies. The median PFS in second-line was 5.9 months (3.3-7.1). Conclusions Pooled analysis of Epitopes-HPV01 and 2 results, confirm mDCF as the regimen of choice in fit patients with metastatic or locally advanced recurrent ASCC. Clinical trial identification NCT01845779, NCT02402842. Legal entity responsible for the study CHU Jean Minjoz, Besancon. Funding Besancon University Hospital and Ligue Contre le Cancer Grand-Est. Disclosure All authors have declared no conflicts of interest.

Despite the critical roles played by IFN-γ+ CD4 Th1 response in tumor immunity, the translation of their potential in clinic remains challenging. Here, we evaluate the clinical significance of circulating anti-tumor CD4 Th1 response in non-small cell lung cancer (NSCLC) patients. 170 naïve-treatment patients were enrolled in this immune monitoring study. The antitumor adaptive Th1 response was assessed by IFN-γ ELISPOT assay in blood lymphocytes using a mixture of eight highly promiscuous and Th1-polarized HLA class II-restricted epitopes from telomerase (TERT). The presence of anti-TERT Th1 response was detected in 59/170 patients (35%). We found an opposite link between anti-TERT Th1 and hyper exhausted T cells co-expressing PD-1+ and TIM-3+ in NSCLC patients. In contrast to hyper exhausted CD8 T cells, the presence of anti-TERT Th1 response was associated with a low rate of hyper exhausted CD4 T cells. We showed that NSCLC stage dissemination is associated with a decrease of anti-TERT Th1 response but an increase of circulating hyper exhausted PD-1+/TIM-3+ CD4 T cells. Notably, anti-telomerase Th1 response and hyper exhausted CD4 T-cells displayed opposite prognosis value in NSCLC. While high level of anti-TERT Th1 cells play a protective role, hyper-exhausted PD-1+TIM-3+ CD4 T cells negatively affect patients’ survival. By using these two circulating immune factors, we stratified patients in distinct prognostic group. Patients with anti-TERT Th1high/CD4 PD-1 TIM-3low immune profile had better overall survival than anti-TERT Th1low/CD4 PD-1 TIM-3high group (median OS : not reached versus 4 months respectively p<0.0001). In conclusion, high circulating anti-TERT Th1 response plays a strong protective role in NSCLC by counteracting the deleterious effect associated with hyper exhausted T cells in blood. Our study provides a dynamic blood-based immunomonitoring tool allow patients stratification. Citation Format: Caroline Laheurte, Magalie Dosset, Dewi Vernerey, Elodie Lauret Marie Joseph, Laura Boullerot, Vincent Kaulec, Marion Jacquin, Laurie Cuche, Guillaume Eberst, Pascale Jacoulet, Virginie Westeel, Olivier Adotevi. Adaptive CD4 Th1 response against telomerase in blood counteracts T-cell exhaustion in non-small cell lung cancer [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 575.

Natural killer (NK) cells are innate effector lymphocytes widely involved in cancer immunosurveillance. In this study, we described three circulating NK cell subsets in patients with non-small cell lung cancer (NSCLC). Compared to healthy donors (HD), lower rate of the cytotoxic CD56dim CD16+ NK cells was found in NSCLC patients (76.1% vs 82.4%, P = 0.0041). In contrast, the rate of CD56bright NK cells was similar between patients and HD. We showed in NSCLC patients a higher rate of a NK cell subset with CD56dim CD16− phenotype (16.7% vs 9.9% P = 0.0001). The degranulation property and cytokines production were mainly drive by CD56dim CD16− NK cell subset in patients. Analysis of natural cytotoxicity receptors (NCRs) expression identified four distinct clusters of patients with distinct NK cell subset profiles as compared to one major cluster in HD. Notably the cluster characterized by a low circulating level of NKp46+ NK cell subsets was absent in HD. We showed that the rate of circulating NKp46+ CD56dim CD16+ NK cells influenced the patients’ survival. Indeed, the median overall survival in patients exhibiting high versus low level of this NK cell subset was 16 and 27 months respectively (P = 0.02). Finally, we demonstrated that blocking NKp46 receptor in vitro was able to restore spontaneous tumor specific T cell responses in NSCLC patients. In conclusion, this study showed a distinct distribution and phenotype of circulating NK cell subsets in NSCLC. It also supports the regulatory role of NKp46+ NK cell subset in NSCLC patients.

Background Despite the critical roles of Th1-polarised CD4 + T cells in cancer immunosurveillance, the translation of their potential to clinical use remains challenging. Here, we investigate the clinical relevance of circulating antitumor Th1 immunity in non-small cell lung cancer (NSCLC). Methods The circulating antitumor Th1 response was assessed by the ELISpot assay in 170 NSCLC patients using a mixture of HLA class II-restricted peptides from telomerase (TERT). Phenotyping of blood immune cells was performed by flow cytometry. Results TERT-reactive CD4 T-cell response was detected in 35% of NSCLC patients before any treatment. Functional analysis showed that these cells were effector memory and Th1 polarised capable to produce effector cytokines, such as IFN-γ, TNF-α and IL-2. The presence of anti-TERT Th1 response was inversely correlated with the level of exhausted PD-1 + /TIM-3 + CD4 T cells. The level of these two immune parameters differentially affected the survival, so that increased level of anti-TERT Th1 response and low rate of exhausted PD-1 + TIM-3 + CD4 + T cells were associated with a better prognosis. Conclusions Systemic anti-TERT Th1 response plays a strong antitumor protective role in NSCLC. This study underlines the potential interest of monitoring circulating antitumor Th1 response for patients’ stratification and therapy decision.