Abstract P4-04-12: Enhancing the immunogenicity of breast cancer cells to stimulate innate immunity and augment the effects of cellular immunotherapy
Authors: AR Khaled, A Limaye, R Bassiouni, A Showalter, J Oyer, V Pandey, R Igarashi, DA Altomare, AJ Copik
Journal: Cancer Research (2016)

Abstract A promising strategy for treating metastatic breast cancer is engaging the immune system to destroy disseminated breast cancer cells. However, breast cancer is a challenge for immunotherapy because of its inherent genetic heterogeneity and decreased immunogenicity. There is an unmet need to develop new, targeted therapies for breast cancer that stimulate the immune system and can be used in combination with checkpoint blockade or adoptive cell transfer to fight metastatic disease. One potential approach is to generate dying breast cancer cells that operate like a vaccine to stimulate a tumor-specific immune response. This is termed immunogenic cell death (ICD) and is characterized by a unique molecular signature, involving the release of molecules that attract and stimulate phagocytes like dendritic cells (DCs) and that could sensitize tumor cells to killing by natural killer (NK) cells. With the discovery of new ICD-inducing agents, interest in this approach is increasing. However, the current doses of drugs used to induce ICD may be too high to translate into clinically relevant regimens. To address these problems, our group developed a novel cytotoxic peptide, CT20p, and a nanotechnology-based platform to deliver and concentrate CT20p in breast tumors. We found that treatment of a murine xenograft model with nanomolar amounts of CT20p, encapsulated in nanoparticles formed with a novel hyperbranched polyester polymer (HBPE-NPs), resulted in regression of breast cancer tumors, and that dying breast cancer cells expressed markers characteristic of ICD, such as the pre-mortem exposure of the "eat me" signal, calreticulin (Crt). The intracellular target of CT20p is a protein called chaperonin-containing T-complex (CCT), which is essential for the folding of actin and tubulin and other critical proteins into their native forms. Disruption of CCT could cause endoplasmic reticulum (ER) stress through the accumulation of misfolded proteins. ER stress in turn initiates intracellular pathways that generate the danger signals associate with ICD. To this end, we observed that cancer cells treated with CT20p displayed alterations in PERK, a key mediator of the unfolded protein response (UPR) and translocation of Crt. As a result, CT20p-treated cancer cells were more readily phagocytosed. More importantly, NK cells more effectively killed cancer cells pre-treated with CT20p. This suggests that CT20p may stimulate NK cell cytotoxicity by inducing activating signals on cancer cells, which would greatly augment the curative effects of adoptive transfer of NK cells. In our experiments normal breast epithelial cells, macrophages or NK cells, themselves, were unaffected by CT20p treatment due reduced CCT. These studies indicate that a single molecule, CT20p, can do both the killing of cancer cells and activation of immune cells and can be combined with cellular immunotherapy (e.g. expanded NK cells) to provide a more complete protection from breast cancer recurrence and metastasis. Citation Format: Khaled AR, Limaye A, Bassiouni R, Showalter A, Oyer J, Pandey V, Igarashi R, Altomare DA, Copik AJ. Enhancing the immunogenicity of breast cancer cells to stimulate innate immunity and augment the effects of cellular immunotherapy. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P4-04-12.