Abstract B123: Local treatment with PeptiCRAd-1, a novel cancer immunotherapy approach, mediates a systemic antitumour CD8+ T-cell response and infiltration of CD8+ and CD4+ T-cells into distant untreated tumors in a clinically relevant humanized mouse melanoma model
Authors: Ylosmaki Erkko , Ranki Tuuli , Priha Petri , Backman Charlotta , Vaughn Matthew , Cerullo Vincenzo , Pesonen Sari 
Published: February 2019

Abstract
Multiple oncolytic viruses (OV) are currently in clinical development as cancer immunotherapy candidates, due to their high immunogenicity and immune activation capacity. However, only anecdotal clinical evidence for OV-mediated tumor-specific immune responses exists. Peptide vaccines are highly tumor-specific, but the low immunogenicity and lack of CD8+ T-cell responses limit the clinical utility of this class of cancer immunotherapy. We have developed a unique cancer immunotherapy platform, PeptiCRAd™, which utilizes a highly immunogenic genetically engineered oncolytic adenovirus to deliver tumor-specific peptides to the immune system. This creates robust Th1 biased cellular immune responses against multiple tumor-specific targets simultaneously. This study compares the immune activation characteristics of intratumorally administered PeptiCRAd-1 to intratumoral OV treatment and standard intradermal peptide vaccination in a humanized mouse melanoma model. PeptiCRAd™ is an immunotherapy platform where multiple tumor-specific peptides are adsorbed onto the negatively charged capsid of genetically engineered oncolytic adenovirus via electrostatic interactions. Our lead clinical candidate PeptiCRAd-1 is based on a state-of-the-art oncolytic adenovirus coding for human CD40L and OX40L transgenes, coated with poly-lysine extended NY-ESO-1 and MAGE-A3 peptides. NOD/Shi-scid/IL-2Rγnull immunodeficient mice were humanized using hematopoietic stem cells (CD34+, HLA-B35+) isolated from human cord blood. A375 melanoma tumors were implanted subcutaneously and treated either with PeptiCRAd-1 or the naked virus. Peptide vaccines were given intradermally with Poly-IC as an adjuvant. Secondary tumors were implanted into the contralateral flank two days after the treatments were stopped. No treatments for secondary tumors were given. Peripheral blood mononuclear cells (PBMCs) and tumor infiltrating CD8+ lymphocytes (TILs) were analyzed for NY-ESO-1 and MAGE-A3 specific CD8+ T-cells by flow cytometry with dextramer analysis. Different immune cell subsets among PBMCs and TILs were assessed. All active treatments increased the number of immune cells in primary tumors in comparison to mock treated animals. Both OV and PeptiCRAd-1 treated animals showed more T-cells (CD3, CD4, CD8) in primary tumors in comparison to peptide vaccine or mock treated animals post treatment. Furthermore, the number of T regulatory cells (CD3+/CD4+/FoxP3+) was smaller in OV and PeptiCRAd-1 treated primary tumors in comparison to primary tumors from peptide vaccine or mock treated animals. This suggests that intratumorally administered immunogenic adenovirus (either naked virus or PeptiCRAd-1) modulates the tumor microenvironment by reducing local immune-suppression. Unlike OV treated animals, PeptiCRAd-1 treated animals showed more CD4+ and CD8+ T-cells in untreated secondary tumors than in treated primary tumors, suggesting that tumor-targeting via peptide-coating of the virus was critically important for the induction of an effect in distant untreated tumors. Furthermore, PeptiCRAd-1 treated animals had more NYESO-specific CD8+ T-cells in blood post priming (mean= 4.3% of total CD8+ cells) in comparison to OV treated (mean= 0.6%) or peptide vaccine treated (mean= 0.6%) animals. PeptiCRAd™ is superior to naked oncolytic adenovirus or standard peptide vaccination in triggering systemic tumor-targeted CD8+ T-cell responses and infiltration of CD8+ TILs into untreated distant tumors. The data suggest that PeptiCRAd improves the tumor targeting specificity of a standard OV. This easily adaptable technology could potentially be used to improve the tumor-focused immune responses generated by any oncolytic adenovirus currently in development. Importantly, PeptiCRAd™ could potentially be used for personalized tumor vaccination by delivering patient-specific neoantigens. A phase I clinical trial with PeptiCRAd-1 in several solid tumor indications is under preparation.
Citation Format: Erkko Ylosmaki, Tuuli Ranki, Petri Priha, Charlotta Backman, Matthew Vaughn, Vincenzo Cerullo, Sari Pesonen. Local treatment with PeptiCRAd-1, a novel cancer immunotherapy approach, mediates a systemic antitumour CD8+ T-cell response and infiltration of CD8+ and CD4+ T-cells into distant untreated tumors in a clinically relevant humanized mouse melanoma model [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B123.