MHC-restricted phosphopeptide antigens: Preclinical validation and first-in-humans clinical trial in participants with high-risk melanoma

Victor H. Engelhard*, Rebecca C. Obeng, Kara L. Cummings, Gina R. Petroni, Angela L. Ambakhutwala, Kimberly A. Chianese-Bullock, Kelly T. Smith, Amanda Lulu, Nikole Varhegyi, Mark E. Smolkin, Paisley Myers, Keira E. Mahoney, Jeffrey Shabanowitz, Nico Buettner, Emily H. Hall, Kathleen Haden, Mark Cobbold, Donald F. Hunt, Geoffrey Weiss, Elizabeth GaughanCraig L. Slingluff

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Background Phosphorylated peptides presented by MHC molecules represent a new class of neoantigens expressed on cancer cells and recognized by CD8 T-cells. These peptides are promising targets for cancer immunotherapy. Previous work identified an HLA-A∗0201-restricted phosphopeptide from insulin receptor substrate 2 (pIRS2) as one such target. The purpose of this study was to characterize a second phosphopeptide, from breast cancer antiestrogen resistance 3 (BCAR3), and to evaluate safety and immunogenicity of a novel immunotherapic vaccine comprising either or both of these phosphorylated peptides. Methods Phosphorylated BCAR3 protein was evaluated in melanoma and breast cancer cell lines by Western blot, and recognition by T-cells specific for HLA-A∗0201-restricted phosphorylated BCAR3 peptide (pBCAR3 126-134) was determined by 51 Cr release assay and intracellular cytokine staining. Human tumor explants were also evaluated by mass spectrometry for presentation of pIRS2 and pBCAR3 peptides. For the clinical trial, participants with resected stage IIA-IV melanoma were vaccinated 6 times over 12 weeks with one or both peptides in incomplete Freund's adjuvant and Hiltonol (poly-ICLC). Adverse events (AEs) were coded based on National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) V.4.03, with provision for early study termination if dose-limiting toxicity (DLT) rates exceeded 33%. The enrollment target was 12 participants evaluable for immune response to each peptide. T-cell responses were assessed by interferon-γELISpot assay. Results pBCAR3 peptides were immunogenic in vivo in mice, and in vitro in normal human donors, and T-cells specific for pBCAR3 126-134 controlled outgrowth of a tumor xenograft. The pIRS2 1097-1105 peptide was identified by mass spectrometry from human hepatocellular carcinoma tumors. In the clinical trial, 15 participants were enrolled. All had grade 1 or 2 treatment-related AEs, but there were no grade 3-4 AEs, DLTs or deaths on study. T-cell responses were induced to the pIRS2 1097-1105 peptide in 5/12 patients (42%, 90% CI 18% to 68%) and to the pBCAR3 126-134 peptide in 2/12 patients (17%, 90% CI 3% to 44%). Conclusion This study supports the safety and immunogenicity of vaccines containing the cancer-associated phosphopeptides pBCAR3 126-134 and pIRS2 1097-1105, and the data support continued development of immune therapy targeting phosphopeptides. Future studies will define ways to further enhance the magnitude and durability of phosphopeptide-specific immune responses. Trial registration number NCT01846143

Original languageEnglish (US)
Article numbere000262
JournalJournal for immunotherapy of cancer
Volume8
Issue number1
DOIs
StatePublished - May 7 2020
Externally publishedYes

Keywords

  • antigens
  • immunogenicity
  • immunotherapy
  • melanoma
  • neoplasm
  • vaccination
  • vaccine

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Molecular Medicine
  • Oncology
  • Pharmacology
  • Cancer Research

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