Phase I clinical study of multiple epitope peptide vaccine combined with chemoradiation therapy in esophageal cancer patients

Hisae Iinuma, Ryoji Fukushima, Tsuyoshi Inaba, Junko Tamura, Taisuke Inoue, Etsushi Ogawa, Masahiro Horikawa, Yoshibumi Ikeda, Noriyuki Matsutani, Kazuyoshi Takeda, Koji Yoshida, Takuya Tsunoda, Tadashi Ikeda, Yusuke Nakamura, Kota Okinaga, Hisae Iinuma, Ryoji Fukushima, Tsuyoshi Inaba, Junko Tamura, Taisuke Inoue, Etsushi Ogawa, Masahiro Horikawa, Yoshibumi Ikeda, Noriyuki Matsutani, Kazuyoshi Takeda, Koji Yoshida, Takuya Tsunoda, Tadashi Ikeda, Yusuke Nakamura, Kota Okinaga

Abstract

Background: Chemoradiation therapy (CRT) has been widely used for unresectable esophageal squamous cell carcinoma (ESCC) patients. However, many patients develop local recurrence after CRT. In this study, we hypothesized that the immunotherapy by peptide vaccine may be effective for the eradication of minimal residual cancer cells after CRT. This study was conducted as a phase I clinical trial of multiple-peptide vaccine therapy combined with CRT on patients with unresectable ESCC.

Patients and methods: HLA-A*2402 positive 11 unresectable chemo-naïve ESCC patients were treated by HLA-A*2402-restricted multi-peptide vaccine combined with CRT. The peptide vaccine included the 5 peptides as follows; TTK protein kinase (TTK), up-regulated lung cancer 10 (URLC10), insulin-like growth factor-II mRNA binding protein 3 (KOC1), vascular endothelial growth factor receptor 1 (VEGFR1) and 2 (VEGFR2). CRT consisted of radiotherapy (60 Gy) with concurrent cisplatin (40 mg/m²) and 5-fluorouracil (400 mg/m²). Peptide vaccines mixed with incomplete Freund's adjuvant were injected subcutaneously once a week on at least 8 occasions combined with CRT.

Results: Vaccination with CRT therapy was well-tolerated, and no severe adverse effects were observed. In the case of grade 3 toxicities, leucopenia, neutropenia, anemia and thrombocutopenia occurred in 54.5%, 27.3%, 27.3% and 9.1% of patients, respectively. Grade 1 local skin reactions in the injection sites of vaccination were observed in 81.8% of patients. The expressions of HLA class I, URLC10, TTK, KOC1, VEGFR1 and VEGFR2 antigens were observed in the tumor tissues of all patients. All patients showed peptide-specific cytotoxic T lymphocytes responses in at least one of the 5 kinds of peptide antigens during the vaccination. Six cases of complete response (CR) and 5 cases of progressive disease (PD) were observed after the 8th vaccination. The 4 CR patients who continued the peptide vaccination experienced long consistent CR for 2.0, 2.9 4.5 and 4.6 years.

Conclusions: A combination therapy of multi-peptide vaccine with CRT can successfully be performed with satisfactory levels of safety, and application of this combination therapy may be an effective treatment for patients with unresectable ESCC.

Trial registration: ClinicalTrial.gov, number NCT00632333.

Figures

Figure 1
Figure 1
Immunohistochemical staining of HLA class I, URLC10, TTK, KOC1, VEGFR1 and VEGFR2 antigens in the ESCC tissues. Typical expression of HLA class I, URLC10, TTK, KOC1, VEGFR1 and VEGFR2 in the primary ESCC tissues obtained from case 6 are shown. The staining was undertaken using the serial sections of the biopsy sample.
Figure 2
Figure 2
Representative immunological monitoring assays detecting peptide-specific CTL response. PBLs obtained from CR patient (case 6) after the 8th vaccination were cultured with URLC10-peptide stimulation and subjected to the ELISPOT assay (a, b). The cultured lymphocytes were analyzed by flow cytometry, and proportion of URLC10 dextramer positive cells in CD8+CD3+CD4- cells were calculated (c).
Figure 3
Figure 3
Computed tomography (CT) and endoscopic images of CR patient (case 6). The CT and endoscopic images of the original site (a, b) and CT images of lymph nodes metastasis (c, d) in case 6, before (a, c) and after treatment by CRT with 8th vaccination (b, d) are shown.
Figure 4
Figure 4
Mechanism of peptide vaccine therapy combined with CRT.

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