Abstract 4268: Protein folding pathway modulation upon Hsp70 inhibition in cancer cells

Cancer cells experience acute stress conditions such as low oxygen and energy, and exposure to toxic agents. To survive proliferate without accumulating toxic misfolded proteins, cancer cells constantly modulate protein homeostasis. Thus, it is not surprising that molecular chaperones, like Hsp70, as well as protein degradation pathways are upregulated in cancer cells compared to their normal counterparts. These data suggest that chaperones are potential targets for cancer therapy. We previously demonstrated the dependence of patient-driven rhabdomyosarcoma cell survival on cytoplasmic Hsp70 activity, thanks to the use of a specific Hsp70 inhibitor, MAL3-101. In particular, we discovered that MAL3-101-mediated Hsp70 inhibition activates the PERK arm of the unfolded protein response (UPR) that results in CHOP-dependent cell death (Sabinis et al., 2016). Moreover, by taking advantage of a MAL3-101-resistant cell line (RMS13-R), we recently determined which compensatory mechanism alters MAL3-101-driven cell death. We found that both endoplasmic reticulum-associated degradation (ERAD) and autophagy are upregulated in RMS13-R cells, underlying increased demand on two protein degradation pathways upon inhibition of Hsp70. However, only autophagy inhibition—but not inhibition of ERAD—re-sensitized RMS13-R cells to Hsp70 inhibition, suggesting that autophagy was the key compensatory mechanism for Hsp70 inhibition. Autophagy was further induced by MAL3-101 treatment in RMS13-R cells, as evidenced by an increase in the messages and proteins corresponding to key autophagy components as well as to the accumulation of autophagic-like structures detected by electron microscopy (Sannino et al., 2018). These data highlight a pro-survival role for autophagy induction upon exposure to an Hsp70 inhibitor in cancer, and provide a link between Hsp70, proteasomal degradation, UPR, and autophagy in rhabdomyosarcoma. We next asked if other cancer types might be sensitive to Hsp70 inhibition, and we investigated the potential benefit of combined treatment with autophagy and/or proteasome inhibitors together with MAL3-101. Specifically, we are investigating the effects of Hsp70 inhibition in breast cancer cells, a cancer type in which higher levels of Hsp70 correlate to increased metastasis and poor prognosis in patients. Our preliminary data suggest that HER2-expressing cells are less sensitive to MAL3-101-mediated Hsp70 inhibition and combinatory treatments including, MAL3-101 and autophagy inhibitors promoted HER2-breast cancer cell death. Further investigations will reveal the potential carcinogenic role of Hsp70 inhibitors in breast cancer treatment and highlight which pathways reduce proteotoxicity in different breast cancer subtypes. Citation Format: Sara Sannino, Christopher J. Guerriero, Amit J. Sabnis, Jeffrey J. Bridsky. Protein folding pathway modulation upon Hsp70 inhibition in cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4268.