Introduction: Endothelial progenitor cells (EPCs) are a subtype of stem cells capable of differentiating into mature endothelial cells. Tumor-derived paracrine signals activate the bone marrow-residing EPCs to home to the tumor to promote angiogenesis. The aim of this study was to utilize the EPCs and their specific pathophysiologic homing to tumors as a biologically targeted drug-delivery system, as a novel “Trojan horse” cellular immunotherapy in cancer.

Methods: To test the specificity of EPCs’ homing to cancer, chemotaxis of EPCs towards conditioned media derived from a series of cancer cell lines or towards nonconditioned media was performed in vitro using Boyden Chamber. In vivo biodistribution of EPCs was studied in diverse tumor models (localized and disseminated) of different cancers (myeloma, lymphoma, leukemia, breast, lung, pancreatic and glioma) after injecting EPCs intravenously, by analysis of the accumulation of EPCs in the tumors and other organs. To enhance the ability of EPCs to home to tumors, EPCs were “primed” in vitro by exposing them to conditioned media from hypoxic (1% O2) tumor cells, followed by testing the homing of “primed” EPCs to tumors, both in vitro and in vivo. Next, EPCs were loaded with LS-542 (NIR imaging dye), and with Titanocene (Tc) (a nontoxic compound used in phototherapy that can be activated by positrons to become toxic) and tested for the effect of the loading on viability and functionality of EPCs, using MTT and migration assays, respectively. Also, we tested the effect of Tc-loaded EPCs on killing of cancer cells when activated by 18FDG/PET.

Results: We found that EPCs specifically homed to tumors in vitro and in vivo, in tumor size-dependent and time-dependent manner, and that EPCs selectively accumulated in tumor tissues, with significantly negligible amounts in other organs. Moreover, EPCs homed to primary tumors and followed the metastatic pattern of the tumor. Hypoxia “priming” of EPCs significantly improved their homing to cancer in vitro and in vivo. Loading EPCs with LS-542 or with Tc did not affect their viability or functionality. Additionally, Tc-loaded EPCs, when activated by 18FDG, induced a dose-dependent killing of cancer cells.

Conclusions: EPCs have a highly specific and efficient biologic machinery to target tumors in as a pan-cancer phenomenon, especially when “primed” with hypoxic tumor media. Loading EPCs with LS-542 or Tc did not change their viability and functionality; thus, both are promising molecules for imaging and therapy of cancer. These results provide a proof of concept that EPCs can be used as a novel cellular immunotherapy for specific and efficacious delivery of theranostic agents in cancer. Also, unlike engineered/targeted immunotherapies such as cellular (CAR-T cells), bispecific T cell engagers, or monoclonal antibodies that target specific molecules on a specific tumor type, EPCs can be used as a pan-cancer “Trojan horse” drug-delivery platform due to the universality of (angiogenesis) in all cancer types.

Citation Format: Barbara Muz, Pilar de la Puente, Shruti Shah, Cinzia Federico, Feda Azab, Justin King, Mark Fiala, Rui Tang, Partha Karmakar, Ravi Vij, Samuel Achilefu, Abdel Kareem Azab. Endothelial progenitor cells as drug-delivery Trojan horses for treatment and imaging of cancer [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr A27.