Phase I trial of a multi-epitope-pulsed dendritic cell vaccine for patients with newly diagnosed glioblastoma

Abstract

Background: This study evaluated the safety and immune responses to an autologous dendritic cell vaccine pulsed with class I peptides from tumor-associated antigens (TAA) expressed on gliomas and overexpressed in their cancer stem cell population (ICT-107).

Methods: TAA epitopes included HER2, TRP-2, gp100, MAGE-1, IL13Rα2, and AIM-2. HLA-A1- and/or HLA-A2-positive patients with glioblastoma (GBM) were eligible. Mononuclear cells from leukapheresis were differentiated into dendritic cells, pulsed with TAA peptides, and administered intradermally three times at two-week intervals.

Results: Twenty-one patients were enrolled with 17 newly diagnosed (ND-GBM) and three recurrent GBM patients and one brainstem glioma. Immune response data on 15 newly diagnosed patients showed 33 % responders. TAA expression by qRT-PCR from fresh-frozen tumor samples showed all patient tumors expressed at least three TAA, with 75 % expressing all six. Correlations of increased PFS and OS with quantitative expression of MAGE1 and AIM-2 were observed, and a trend for longer survival was observed with gp100 and HER2 antigens. Target antigens gp100, HER1, and IL13Rα2 were downregulated in recurrent tumors from 4 HLA-A2+ patients. A decrease in or absence of CD133 expression was seen in five patients who underwent a second resection. At a median follow-up of 40.1 months, six of 16 ND-GBM patients showed no evidence of tumor recurrence. Median PFS in newly diagnosed patients was 16.9 months, and median OS was 38.4 months.

Conclusions: Expression of four ICT-107 targeted antigens in the pre-vaccine tumors correlated with prolonged overall survival and PFS in ND-GBM patients. The goal of targeting tumor antigens highly expressed on glioblastoma cancer stem cells is supported by the observation of decreased or absent CD133 expression in the recurrent areas of gadolinium-enhanced tumors.

Fig. 1

Diagram of treatment schedule and events

Fig. 2

a Correlation of quantitative antigen expression on primary tumor from ND patients with progression-free survival (n = 13). Logarithmic plots of antigen expression determined by qRT-PCR (see “Patients and methods”) showed correlations of increasing antigen expression with longer progression-free survival times (PFS). Antigen expression was measured from fresh-frozen samples and calculated relative to GAPDH. b Correlation of quantitative antigen expression on primary tumor from ND patients with overall survival (n = 13). Logarithmic plots of antigen expression determined by qRTPCR (see “Patients and methods”) showed correlations of increasing antigen expression with longer overall survival time (OS)

Fig. 3

Downregulation of target antigens in recurrent tumors from HLA-A2+ patients. Antigen expression was from FFPE samples and calculated relative to GADPH. Significant downregulation post-vaccine of A2 epitopes gp100, HER2, and IL13Rα2 relative to upregulations was observed (p = 0.023, Fisher’s exact test). Downregulation of the HLA-A1 antigen, AIM2, was not significant (p = 0.21) in these patients

Fig. 4

CD133 expression by RT-PCR in primary tumor and samples from subsequent surgeries from newly diagnosed and recurrent patients. Expression is from FFPE samples and calculated relative to GADPH as described in “Patients and methods”. The sample for Patient 19 was negative for tumor

Fig. 5

a Kaplan–Meier estimates of progression-free survival for newly diagnosed patients (n = 16). Dotted lines illustrate the 95 % confidence intervals, and hash marks denote censored patients. b Kaplan–Meier estimates of overall survival for newly diagnosed patients (n = 16). Dotted lines illustrate the 95 % confidence intervals, and hash marks denote censored patients

Fig. 6

Gating strategy for intracellular cytokine analysis in CD8⁺ cells. Identical gain and gating was employed on all stimulated, unstimulated, pre-, and post-vaccine plots prior to analysis. Representative antigen-stimulated, post-vaccine plots from a IFNγ-responsive patient (#4) are shown