Antigen-Specific Cancer Immunotherapeutic Research Articles

What future opportunities may immuno-oncology provide for improving the treatment of patients with lung cancer?

Lung cancer is the leading cause of cancer-related mortality worldwide, with non-small-cell lung cancer (NSCLC) accounting for approximately 85% of all cases. Most patients with NSCLC are diagnosed at an advanced stage and have a poor prognosis, with a 5-year survival rate of <5%. Despite the introduction of new chemotherapeutic agents and molecularly targeted drugs, outcomes remain poor, emphasising the need for new treatment approaches. Inducing or potentiating immune responses via immunotherapeutic manipulation is a viable treatment approach for lung cancer. Antigen-specific, tumour-cell, and dendritic cell-based vaccines have all been evaluated in lung cancer, and some have shown promising clinical activity in phase II trials. These include liposomal BLP25 vaccine (L-BLP25), which targets mucin 1, and melanoma-associated antigen 3 (MAGE-A3) antigen-specific cancer immunotherapeutic (ASCI), which targets MAGE-A3, a peptide expressed almost exclusively on tumour cells. MAGE-A3 ASCI is being evaluated in the adjuvant setting in a phase III trial of patients with early-stage NSCLC, while a phase III trial of L-BLP25 is enrolling patients with unresectable stage III NSCLC. T-cell modulating agents (e.g. antibodies against programmed death 1 and cytotoxic T-lymphocyte-associated antigen-4 [CTLA-4]) are also being investigated. For example, in patients with NSCLC treated with paclitaxel and carboplatin, the phased administration of ipilimumab (an antibody against CTLA-4) resulted in substantial improvements in immune-related progression-free survival compared with chemotherapy alone (5.7 versus 4.6 months; P=0.05). Immunotherapy in lung cancer is starting to deliver promising results in clinical trials. However, further research will be required to establish the optimal timing of therapy (i.e. in the adjuvant or metastatic settings). In addition, it will be important to determine if immunotherapies are most effective when used alone or in combination with other agents.

DERMA phase III trial of MAGE-A3 antigen-specific cancer immunotherapeutic (ASCI) as adjuvant therapy in patients with MAGE-A3-positive resected stage III melanoma.

TPS232 Background: Clinical activity of the MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (ASCI) was observed in a Phase II study in patients with metastatic melanoma (NCT00086866) and in a Phase II study in patients with completely resected non-small cell lung cancer (NSCLC) (NCT00290355). Based on these positive signals for the activity of MAGE-A3 ASCI, a Phase III trial is currently ongoing in patients with resectable regionally advanced melanoma (DERMA Phase III trial, NCT00796445). Methods: DERMA is a double-blind, placebo-controlled, 2:1 randomized trial of MAGE-A3 ASCI vs. Placebo. Eligible patients must have cutaneous melanoma expressing MAGE-A3 with histologically proven macroscopic nodal disease (AJCC Stage IIIB-C). Central randomization uses an internet-based algorithm accounting for the following factors: Stage of the disease (IIIB, IIIC and IIITx), TNM nodal category (N1, N2 and N3), primary category (Tx-0 or T1-2 or T3 or T4), extracapsular extension (Yes, No), study center, prior treatment with IFN or anti-CTLA-4 (Yes, No). Patients must have had complete surgical resection of all tumor. Disease-Free Survival (DFS) in the overall patient population is the primary endpoint, while overall survival, safety and health-related quality of life (EQ-5D) are secondary endpoints. A previously-defined gene signature associated with MAGE-A3 ASCI benefit is also being validated as a secondary endpoint of this trial (Louahed et al., EORTC-NCI-AACR 2009). The MAGE-A3 ASCI combines the tumor-specific antigen MAGE-A3, delivered as a recombinant protein, with the immunostimulant AS15. A maximum of 13 doses are administered by intramuscular injection over 27 months; the first 5 are given at 3-week intervals followed by 8 at 12-week intervals. This Phase III trial is designed to enroll 1300 patients from 250 centers worldwide. Since Dec 1, 2008, 2426 patients have been screened. From 1917 tumor samples valid for testing the expression of MAGE-A3, 1265 express MAGE-A3 (MAGE-A3 expression rate of 66%). 832 patients are currently enrolled to this study.

PREDICT phase II: A trial to prospectively validate a gene signature associated with clinical benefit from MAGE-A3 antigen-specific cancer immunotherapeutic (ASCI) in patients with unresected metastatic cutaneous melanoma.

TPS231 Background: Active immunization towards the tumor-specific antigen MAGE-A3 is a potential new option under investigation to treat cancer. Clinical activity of the MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (ASCI) has been observed in a Phase II trial (NCT00086866) in patients with unresected metastatic cutaneous melanoma. Current data showed that combination of MAGE-A3 recombinant protein with the AS15 immunostimulant induced a robust and long-lasting antibody and CD4 T-cell immune response as well as clinical responses. Retrospective gene expression profiling of the tumor taken prior immunization led to the characterization of a gene signature (GS) associated with clinical benefit (Louahed et al., EORTC-NCI-AACR 2009). Methods: The aim of this open-labeled, single-arm trial (PREDICT, NCT00942162) is to prospectively validate the GS as a predictive biomarker of the clinical outcome of the MAGE-A3 ASCI. Main eligibility criteria are patients with histologically proven measurable MAGE-A3+ metastatic cutaneous melanoma, (unresectable stage III or stage IV M1a melanoma - AJCC 2002), an ECOG performance status of 0-1. Patients will receive a maximum of 24 doses of MAGE-A3 ASCI (recMAGE-A3 + AS15) by i.m. injections over 48 months. The primary endpoint is 1-Year overall survival rate (OSR) in the GS+ population. Main secondary endpoints include 1-Year OSR in global and GS- populations but also the clinical response rate and other clinical activity indicators (PFS, median OS) in these 3 populations. The GS will be determined on the tumor collected for the MAGE-A3 screening. This trial is also an opportunity to validate the technology transfer to support large scale identification of the GS: from micro-array on fresh tissue (NCT00086866) to qRT-PCR on FFPE tumor samples (PREDICT). Data of Jan 10, 2011 showed that 280 patients were screened. Out of the 236 tumor samples valid for MAGE-A3 expression testing, 111 were found MAGE-A3+ with a MAGE-A3 expression rate of 47% and 82 patients were finally enrolled. The identification of the GS status is ongoing for the enrolled population.

MAGRIT (MAGE‐A3 as Adjuvant, Non‐small Cell Lung Cancer Immunotherapy)

PROTOCAL ID NCT00480025 CONTACT (877) 379-3718 GSKClinicalSupportHD@gsk.com , www.ascitrials.com/locator SUMARY The study will measure how well the MAGE-A3 antigen-specific cancer immunotherapeutic (ASCI) product works in preventing cancer from recurring when given to patients with non-small cell lung cancer (NSCLC) after their tumor is surgically removed. It also will evaluate the potential side effects of the MAGE-A3 ASCI. Patients' progress will be measured by periodic examinations (including x-rays or computed tomography scans, blood tests, and physical examinations). Approximately 400 hospitals in Europe, Asia, and the United States will participate. Two of 3 patients will receive the MAGE-A3 ASCI, while 1 patient will receive a placebo control. Participants will receive 13 injections over a 27-month period. After treatment patients will receive regular checkups every 6 months up until 5 years after treatment. They will then be checked annually by telephone for 10 years after treatment. ELIGIBILITY Participants must have NSCLC in stage 18, II, or IIIA and have had or will have the cancer surgically removed, agree to have a piece of their tumor tested for the presence of MAGE-A3 (this antigen must be found in the cancer), have no other significant health problems, not be pregnant or likely to become pregnant during the treatment period, and be at least aged 18 years.

New data on MAGE-A3 cancer immunotherapy may signal new options for patients with melanoma and non-small cell lung cancer

The MAGE-A3 gene, encodes a tumor-specific antigen expressed on tumor cells only and not on normal tissues. When combined with an immunological adjuvant designed to enhance the immune response to the MAGE-A3 antigen, immunization with the MAGE-A3 cancer immunotherapeutic was shown to induce specific T-cell responses and long-lasting clinical objective responses in metastatic melanoma. Data today highlights the encouraging potential of the MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (ASCI). In a Phase II study designed to evaluate the MAGE-A3 ASCI in patients with metastatic melanoma, objective long-term responses were recorded. These melanoma study results represent a second positive signal of clinical activity for the novel cancer immunotherapeutic, following encouraging clinical activity reported in a separate double-blind, placebo-controlled Phase II study of patients with Non-Small Cell Lung Cancer (NSCLC). In addition, we present compelling data regarding the use of the genetic profiling of melanoma in predicting clinical outcomes of treatment with MAGE-A3 ASCI. An independent characterization of biomarkers predictive to clinical response to the MAGE-A3 in the Phase II study in NSCLC provided similar observation resulting in 53% reduction in the relative risk of relapse. Phase II results led to phase 3 trials in both metastatic melanoma (DERMA trial) and stage IV NSCLC (MAGRIT trial) to show the efficacy of de MAGE-A3 ASCI. To date, more than 2,000 patients have been screened to enter in DERMA trial, with 67% MAGE-A3 expressing melanomas leading to 700 inclusions in that trial. Le gène MAGE-A3 code pour un antigène spécifique de la tumeur, exprimé sur les cellules tumorales uniquement et pas sur les cellules normales. L’immunothérapie anticancéreuse spécifique de MAGE-A3 contient l’antigène MAGE-A3 et un immunostimulant destiné à renforcer la réponse immunitaire envers l’antigène. Cette combinaison s’est montrée capable d’induire des réponses immunitaires et cliniques de longue durée chez des patients atteints de mélanome métastatique. Un second signal positif pour ce traitement potentiel a été obtenu avec ce même candidat MAGE-A3 dans une étude de Phase II indépendante réalisée chez des patients atteints de cancer du poumon non à petites cellules (CBNPC). En outre, nous présentons ici l’identification de biomarqueurs capables de prédire, avant traitement, le bénéfice des patients au traitement par MAGE-A3 ASCI. Ces biomarqueurs ont été identifiés dans l’étude de Phase II dans le mélanome, et leur valeur prédictive du bénéfice au traitement a également été montrée dans l’étude de phase II dans le cancer du poumon ; les patients traités par MAGE-A3 ASCI et dont la tumeur présente ces biomarqueurs dérivent un bénéfice plus important du traitement par MAGE-A3 ASCI, de l’ordre de 53 % de réduction relative du risque de rechute. Les résultats de ces études de Phase II ont mené à l’initiation de 2 études de Phase III visant à démontrer l’efficacité de MAGE-A3 ASCI dans le mélanome et le cancer du poumon : l’étude MAGRIT dans le cancer du poumon non à petites cellules, et l’étude DERMA. À ce jour, plus de 2 000 patients ont été screenés pour participer à l’étude DERMA : la tumeur de ∼ 67 % d’entre eux exprimait MAGE-A3 et plus de 700 patients sont entrés dans l’étude.

Abstract A37: Identification of a gene expression signature predictive of clinical activity following MAGE-A3 ASCI treatment

Abstract Clinical data today highlight the encouraging potential of the MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (ASCI) for cancer treatment. The MAGE-A3 gene encodes a tumor-specific antigen expressed on tumor cells only. When used as a recombinant protein and combined with an immunological Adjuvant System designed to enhance the immune response to the MAGE-A3 antigen, it was shown to induce specific T-cell responses and long-lasting clinical objective responses in metastatic melanoma (Phase II NCT00086866 trial) (Kruit et al. JCO 2008 26:9065), and demonstrated proof-of-concept in Non-Small Cell Lung Cancer (NSCLC; Phase II NCT00290355 trial) (Vansteenkiste et al. JCO 2007 25:7554). In these 2 trials, gene expression profiling by microarrays was used to identify biomarkers predictive of the clinical activity of the MAGE-A3 ASCI. A first analysis carried out on samples from melanoma patients using supervised hierarchical clustering of 2 objective responders and 2 non-responders identified 2 gene clusters based on differential gene expression. This immune gene expression signature was then confirmed in an analysis of 22-patient samples, and independently validated on an additional 30-patient testing set; furthermore these analyses confirmed the association of clinical benefit and the molecular signature. Most of the identified genes are immune-related, defining a particular biological context present in the tumor environment before immunization. This was confirmed by selecting genes using all patients eligible for gene expression profiling. Upon crossvalidation, median overall survival was improved significantly in the population of patients whose tumor presented the gene signature (GS): 28 months in the GS+ population, 16.2 months in the GS−. The predictive value of the melanoma signature was tested in NSCLC. A subset of 49 genes discovered in the melanoma Phase II trial was assayed by qPCR on biopsies taken prior to any ASCI treatment from 137 patients of the lung Phase II trial. Applied to NSCLC patients, this biomarker showed that the relative reduction in the risk of recurrence upon MAGE-A3 ASCI treatment is increased by about 2 fold in the patients with the predictive gene signature as compared to the overall population: from 25% relative improvement in the overall population to 53% in patients whose tumor presents the predictive gene signature. In conclusion, two clinical proof-of-concepts have been obtained with the MAGE-A3 ASCI in 2 different types of tumors, NSCLC and melanoma. More importantly, we have identified a gene expression signature predictive of clinical activity of the MAGE-A3 ASCI treatment. The initiation of Phase III studies in NSCLC (MAGRIT) and melanoma (DERMA) is a unique opportunity to validate prospectively these biomarkers with the ultimate goal to select patients that are the most likely to benefit from the MAGE-A3 ASCI therapy. Citation Information: Mol Cancer Ther 2009;8(12 Suppl):A37.

9021 Preliminary results of MAGE-A3 expression and baseline demographic data from MAGRIT, a large phase III trial of MAGE-A3 ASCI (Antigen-Specific Cancer Immunotherapeutic) in adjuvant NSCLC

In this study, we examined whether epidermal growth factor receptor (EGFR) mutations were detectable using a polymerase chain reaction-based assay and wash fluid of computed tomography (CT)-guided lung biopsy needles.DNA was extracted from wash fluid of CT-guided biopsy needles of 53 lung tumors (as diagnosed according to the results of the CT-guided biopsies). EGFR mutations, specifically exon19 deletions and exon21 L858R mutations, were examined using a mutant-enriched polymerase chain reaction assay. We also examined the presence of EGFR mutations in 26 surgically resected tumor specimens and compared the results with those obtained for the corresponding wash fluid samples.The amount of DNA obtained for the wash fluid of the CT-guided biopsy needles ranged from 35 to 2360 ng. There were no significant differences in the amount of extracted DNA according to the tumor characteristics, including tumor size and the percentage of ground glass opacity. Thirty-four of the 53 lung tumor samples were histologically diagnosed as non-small cell lung cancer (NSCLC). Exon19 deletions and exon21 L858R mutations in EGFR were detected in 4 (12%) and 13 (38%) of 34 NSCLC cases, respectively. No EGFR mutations were found in the non-NSCLC cases. The EGFR mutation status in the wash fluid samples was consistent with those obtained for all 26 corresponding surgical specimens.Our results indicate that EGFR mutations can be detected using wash fluid of CT-guided biopsy needles. In this manner, the DNA genotype can be determined even in extremely small clinical specimens using highly sensitive assays.

MAGRIT: The Largest-Ever Phase III Lung Cancer Trial Aims to Establish a Novel Tumor-Specific Approach to Therapy

This clinical trial summary provides the background and rationale for a randomized trial, MAGRIT, to investigate the efficacy of MAGE-A3 antigen-specific cancer immunotherapeutic (ASCI) agents in preventing cancer relapse, when administered after tumor resection, in patients with MAGE-A3—positive stages IB, II, and IIIA Non–Small-cell lung cancer. The study will also evaluate potential side effects of MAGE-A3 ASCIs. The primary endpoint is disease-free survival. The secondary endpoint is prospective validation of the gene signature predictive of benefit from MAGE-A3 ASCI therapy.

Cervical cancer: ESMO Clinical Recommendations for diagnosis, treatment and follow-up

Objectives Gene expression profiling by microarrays is used to identify biomarkers predictive of the observed clinical activity of the MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (ASCI) recorded in a Phase II study in metastatic melanoma (EORTC 16032-18031). Clinical activity to MAGE-A3 ASCI treatment in metastatic melanoma patients and a gene signature discriminating between clinical benefit and non clinical benefit patients were previously demonstrated (ASCO 2008). The predictive signature improved significantly the median time to treatment failure (2.3 months in the GS (-) and 10.3 months in the GS (+) population. Here we report on a new improved classifier to select patients with a higher likelihood of clinical response to treatment. Methods 75 patients with progressive, unresectable stage III or stage IV M1a MAGEA3 (+) melanomas, were randomized as 1st line therapy between immunization with MAGE-A3 recombinant protein combined with GSK Adjuvant Systems AS15 or AS02B. Gene expression profiling was performed on tumor biopsies taken prior to any immunization. Results Supervised classification experiments were conducted on a subset of 62 patients. A multivariate gene selection process was embedded in the estimation of a mathematical predictive model (linear support vector machine). Using bootstrap resamplings, gene selection was repeated on 30 independent sample sets. A final gene signature of 33 probes was derived from genes appearing most frequently after the various resamplings. The vast majority of the identified genes are immune-related. The final classifier correctly predicts clinical response with 91% sensitivity, 95% specificity and 91% positive predictive value. These performances were estimated on new independent resamplings of the same data. Conclusions The gene signatures found in metastatic melanoma patients are strongly correlated with the response to the MAGE-A3 ASCI treatment, reflecting an immune microenvironment in the tumor present prior to any therapeutic intervention. Such signatures and associated classifier could be used to select patients with a higher likelihood of response to MAGE-A3 ASCI treatment. This abstract was published in Cancer Immunity, a Cancer Research Institute journal that ceased publication in 2013 and is now provided online in association with Cancer Immunology Research.

MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (ASCI) as adjuvant therapy in resected stage IB/II non-small cell lung cancer (NSCLC): from proof-of-concept to phase III trial (MAGRIT)

MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (ASCI) as adjuvant therapy in resected stage IB/II non-small cell lung cancer (NSCLC): from proof-of-concept to phase III trial (MAGRIT)

Cancer immunotherapy targeting tumour-specific antigens: towards a new therapy for minimal residual disease

Background: Clinical investigation of cancer immunotherapy has been very active and several approaches have been evaluated in Phase III trials. In particular, the characterisation at the molecular level of tumour-specific antigens, together with expert knowledge from GSK Biologicals in recombinant protein manufacturing and immunological Adjuvant Systems, has led the company to develop Antigen-Specific Cancer Immunotherapeutic (ASCI). Objective/methods: This paper reviews the different cancer immunotherapy approaches that have reached Phase III clinical development. A special attention is given to GSK's ASCI approach. Conclusion: Based on encouraging data in a double-blind Phase II trial in non-small-cell lung cancer, the selection of the most suitable adjuvant system in melanoma and the choice of the adequate clinical setting for the clinical development of cancer immunotherapy, the ASCI approach offers the perspective that the long quest towards a new cancer treatment approach is about to succeed.

Association of gene expression signature and clinical efficacy of MAGE-A3 antigen-specific cancer immunotherapeutic (ASCI) as adjuvant therapy in resected stage IB/II non-small cell lung cancer (NSCLC)

7501 Background: A phase II randomized trial in 182 patients (pts) with completely resected stage IB or II MAGE-A3 (+) NSCLC comparing postoperative MAGE-A3 recombinant protein combined with an Adjuvant System to placebo (q3w x 5, followed by q3m x 8) showed a strong positive signal for activity (J Clin Oncol 25 Suppl 18:398S, 2007). We report here the analysis of gene expression profiling of tumors prior to treatment to identify a predictive signature that correlates with clinical activity of MAGE-A3 ASCI treatment. Methods: Microarray analysis (Affymetrix) was performed on 159 fresh-frozen tumor biopsies (MAGE-A3: 108; placebo: 51; stage IB: 109; stage II: 50). Clinical data on relapse are based on a median follow-up of 44 months. Results: Study of tumor samples from pts in the placebo arm led to identification of a gene signature associated with a high risk of postoperative relapse. The absence of this signature correlated with a very low relapse rate in stage IB pts: placebo 0%; MAGE-A3: 4%. In the MAGE-A3 treated pts with the high risk of relapse signature, we compared 10 biopsies from stage IB pts with recurrence to 10 pts without recurrence. A 25 regulated gene probe predicted benefit from MAGE-A3 treatment. The relevance of this signature was confirmed on the remaining 139 stage IB and II biopsies. The predictive signature improved clinical efficacy (disease-free interval) (HR in GP+: 0.57; HR in GP-: 0.78) with no impact on relapse rate in the placebo arm. The signature consisted of immune- related genes associated to the pre-therapeutic tumor microenvironment. Conclusions: We identified a gene expression signature associated with high risk of relapse. Pts with tumors not presenting this signature have a very low risk of relapse after surgery (<3%). We also described another signature that is predictive to clinical activity of the MAGE-A3 ASCI treatment. Selection of pts with this predictive signature increases clinical efficiency by a factor 2. Interestingly, this signature is also associated to clinical activity to MAGE-A3 in a study in metastatic melanoma. Data will be validated in the ongoing NSCLC adjuvant phase III study (MAGRIT). Author Disclosure Employment or Leadership Consultant or Advisory Role Stock Ownership Honoraria Research Expert Testimony Other Remuneration GlaxoSmithKline

Expression of defined genes identified by pretreatment tumor profiling: Association with clinical responses to the GSK MAGE- A3 immunotherapeutic in metastatic melanoma patients (EORTC 16032–18031)

9045 Background: Gene expression profiling by microarrays was used to identify markers predictive of the clinical activity of the MAGE-A3 Antigen-Specific Cancer Immunotherapeutic (ASCI) recorded in a phase II study in metastatic melanoma. Methods: 75 patients with progressive, unresectable stage III or stage IV M1a MAGE-A3 (+) melanomas, were randomized as 1st-line therapy between immunization with MAGE-A3 protein combined with GSK proprietary Adjuvant Systems AS15 or AS02B. Gene expression profiling was performed on tumor biopsies taken prior to any immunization. Results: Initial analysis using supervised hierarchical clustering of 2 objective responders (OR) with 7 non-responders identified 2 gene clusters based on differential expression. The correlation of this gene expression signature (GS) and objective response was further confirmed on 22 patients. Interestingly, mixed response (MR) and stable disease > 6 months (SD) clustered with OR, suggesting a strong association between the identified signature and the MAGE-A3 induced clinical benefit. Independent validation on additional 30 patients confirmed the association of clinical benefit and the molecular signature. Most of the identified genes are immune-related, defining a particular biological context present in the tumor environment before immunization. The signature was randomly distributed in the 2 study arms and identified all patients with clinical benefit (OR, MR, SD). The AS15 arm has shown to induce a higher MAGE-A3 immune response and more frequent clinical activity. The predictive signature improves significantly the median time to treatment failure (2.3 months in the GS (-) and 10.3 months in the GS (+) population (HR = 0.31; 95% CI: 0.13–0.76). Conclusions: The gene expression in metastatic melanoma reported here is strongly correlated with a clinical response to the MAGE-A3 ASCI treatment. This signature reflects an immune microenvironment in the tumor present prior to any therapeutic intervention following MAGE-A3 ASCI administration. Such molecular signature could be used to select patients with a higher likelihood of clinical response the MAGE-A3 ASCI. Author Disclosure Employment or Leadership Consultant or Advisory Role Stock Ownership Honoraria Research Expert Testimony Other Remuneration GlaxoSmithKline Biologicals GlaxoSmithKline Biologicals