Abstract 1709: Targeting immune checkpoint therapy to the lung tumor microenvironment
Authors: Cohen Allison S. , Doligalski Michael L. , Zheng Hong , Tafreshi Narges K. , Estrella Veronica , Delva Nella , Nguyen Jonathan , Beg Amer , McLaughlin Mark L. , Morse David L. 
Published: July 2017

Abstract
Lung cancer is the leading cause of cancer deaths worldwide. Many lung cancer patients are diagnosed with advanced disease. These patients have a low 5-year survival rate and limited treatment options. Novel therapies, which target biomarkers that are overexpressed in lung cancer but have basal expression in benign tissue, are desperately needed. The goal of this work was to develop a targeted agent for immune checkpoint therapy of lung cancer. Recently, immune checkpoint inhibitors have been approved for use in lung cancer and many more are being tested in clinical trials. The current immune checkpoint inhibitor agents are not tumor-targeted. Targeting the immune checkpoint inhibitor to tumor cell-surface markers should concentrate the conjugate in the tumor microenvironment and enhance the immune response in the tumor while reducing the systemic dosages needed, resulting in lower systemic toxicity.
The delta opioid receptor (DOR) is expressed in some lung cancers, but is not expressed or is expressed only at basal levels in normal tissues outside the brain. We have previously synthesized fluorescently-labeled DOR-targeted imaging agents based on a synthetic peptide antagonist (DORL). These targeted fluorescent agents have high affinity and selectivity for DOR, and exhibit good pharmacokinetic (PK) and biodistribution (BD) profiles, i.e. specific tumor uptake with rapid systemic clearance and no uptake in tissues of concern, e.g. brain. We are now developing lung cancer-specific immunotherapy agents that target the DOR by conjugating DORL to immunomodulatory molecules. In the current work, we synthesized a fluorescently-labeled DOR targeting ligand and conjugated it to an anti-PD1 antibody (DORL-PD1). We synthesized immunoconjugates with several targeting ligand-to-antibody ratios (TARs). We engineered murine lung cancer cells to constitutively express the DOR. By lanthanide time-resolved fluorescence (LTRF) competitive binding assays, we have shown that the agents have high avidity for the DOR in vitro with higher TARs resulting in higher binding avidity. We characterized the uptake of DORL-PD1 in vitro using live-cell fluorescence microscopy. Using syngeneic engraftment tumor models in immunocompetent mice, we performed longitudinal fluorescence imaging studies to determine the agent circulation time (PK), tumor selectivity and tissue distribution (BD). Immune checkpoint efficacy studies were performed using the DOR negative mouse tumor models.
In conclusion, we have synthesized fluorescent DOR-targeted immune checkpoint therapy agents, DORL-PD1; demonstrated avidity and selectivity for the DOR in vitro and in vivo; and immune checkpoint therapy efficacy in vivo. Future studies will evaluate the efficacy of DORL-PD1 in immune competent mice bearing DOR positive tumors. These agents could be useful for increasing the efficacy and reducing systemic toxicity of immune-checkpoint therapy of lung cancer.
Citation Format: Allison S. Cohen, Michael L. Doligalski, Hong Zheng, Narges K. Tafreshi, Veronica Estrella, Nella Delva, Jonathan Nguyen, Amer Beg, Mark L. McLaughlin, David L. Morse. Targeting immune checkpoint therapy to the lung tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1709. doi:10.1158/1538-7445.AM2017-1709