Antibody-mediated delivery of LIGHT to the tumor boosts Natural Killer cells and delays tumor progression

Abstract

BackgroundLIGHT is a member of the TNF superfamily, which has been claimed to mediate anti-tumor activity on the basis of cancer cures observed in immunocompetent mice bearing transgenic LIGHT-expressing tumors. The preclinical development of a LIGHT-based therapeutic has been hindered by the lack of functional stability exhibited by this protein. MethodsHere, we describe the cloning, expression and characterization of five antibody-LIGHT fusion proteins, directed against the alternatively-spliced EDA domain of fibronectin, a conserved tumor-associated antigen. ResultsAmong the five tested formats, only the sequential fusion of the F8 antibody in single-chain diabody format, followed by the LIGHT homotrimer expressed as a single polypeptide, yielded a protein (termed "F8-LIGHT") which was not prone to aggregation. A quantitative biodistribution analysis in tumor bearing mice, using radioiodinated protein preparations, confirmed that F8-LIGHT was able to preferentially accumulate at the tumor site, with a tumor-to-blood ratio of ca. five to one twenty-four hours after intravenous administration. Tumor therapy experiments, performed in two murine tumor models (CT26 and WEHI-164), featuring different levels of lymphocyte infiltration into the neoplastic mass, revealed that F8-LiGHT could significantly reduce tumor cell growth and was more potent than a similar fusion protein (KSF-LIGHT), directed against hen egg lysozyme and serving as negative control of irrelevant specificity in the mouse. At a mechanistic level, the activity of F8-LiGHT was mainly due to an intratumoral expansion of Natural Killer cells, whereas there was no evidence of expansion of CD8+ T cells, neither in the tumor, nor in draining lymph nodes. ConclusionWe developed a novel recombinant LIGHT fusion protein, able to accumulate at the site of disease and which displayed anti-tumor activity in two mouse models of cancer.