Abstract C222: Diverting Hsp70 trafficking as a tool for stimulating anticancer immunity.

Abstract

Abstract The stress-inducible heat shock protein 70 (Hsp70) is normally destined for cytosolic localization. In tumors, Hsp70 is frequently localized to the cell surface and released from cells, unusual features that stimulate the immune system against cancer cells. Interestingly, it has been shown that overexpression of Hsp70 enhances the immunogenicity of tumor cells in animal models. Here we investigated potential mechanisms by which overexpression of Hsp70 in B16 melanoma cells may increase surface expression and release. Flow cytometry measurements revealed an increased cell membrane concentration upon elevated level of intracellular Hsp70. Hsp70 was detectable in subcellular fractions enriched in endosomes/ lysosomes. An accumulation of excess Hsp70 was seen in high Hsp70 expressing cells, in particular in the purified lyosomal fractions. In low expressing cells apparent trafficking of Hsp70 was confined to the endosomal system. In contrast, high intracellular Hsp70 concentration was associated with a predominant lysosomal routing. Remarkable changes were observed in Hsp70 trafficking upon heat stress, a common approach used to facilitate Hsp70 release. In control cells a dramatic depletion of surface-exposed Hsp70 was paralleled with increased endosomal recycling of internalized protein. In contrast, excess intracellular Hsp70 prevented the membrane transport from heat-induced depletion, and allowed transport of surface-exposed Hsp70 to lysosomes secreting their cargo in a soluble form. Therefore excess Hsp70 appears to be decisive in terms of both trafficking to the plasma membrane and stress-induced active release. Altogether, our findings demonstrate a vesicular trafficking system bringing excess intracellular Hsp70 to fulfill its immunostimulatory role. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr C222.