Proteotoxic stress targeted therapy (PSTT): induction of protein misfolding enhances the antitumor effect of the proteasome inhibitor bortezomib

Nickolay Neznanov,Lubov Neznanova,Patricia Stanhope-Baker,Andrei P Komarov,Andrei V Gudkov

doi:10.18632/oncotarget.246

Abstract

Proteotoxic stress (PS) is generated in cells under a variety of conditions involving accumulation of misfolded proteins. To avoid the toxicity of unmitigated PS, cells activate the heat shock response (HSR). HSR involves upregulation of factors such as ubiquitin and the non-housekeeping chaperone Hsp70 which assist with metabolism of aberrant proteins. The PS-HSR axis is a potential anticancer treatment target since many tumor cells display constitutive PS and dependence on HSR due to their rapid rates of proliferation and translation. In fact, induction of PS via stimulation of protein misfolding (hyperthermia), inhibition of proteasomes (bortezomib) or inhibition of Hsp90 (geldanamycin) have all been considered or used for cancer treatment. We found that combination of bortezomib with an inducer of protein misfolding (hyperthermia or puromycin) resulted in enhanced PS. HSR was also induced, but could not mitigate the elevated PS and the cells died via largely p53-independent apoptosis. Thus, combination treatments were more cytotoxic in vitro than the component single treatments. Consistent with this, combination of non-toxic doses of puromycin with bortezomib significantly increased the antitumor activity of bortezomib in a mouse model of multiple myeloma. These results provide support for using combination treatments that disrupt the balance of PS and HSR to increase the therapeutic index of anticancer therapies.

Highlights

Proteotoxic stress (PS) is generated by accumulation of misfolded proteins in cells under a variety of conditions including hyperthermia, hypoxia, and exposure to denaturing agents or drugs that inhibit proteasome or chaperone activities [1,2,3]
Using inducible Hsp70 expression as a marker of PS, we found that combined treatment with hyperthermia and bortezomib resulted in a much higher level of PS than either agent alone (Fig. 1A, B)
As expected based on the well-established relationship between Hsp70 expression and PS, we observed much higher levels of ubiquitinated proteins in cells treated with hyperthermia and bortezomib than in cells treated with either agent alone (Fig. 1C)

Summary

Introduction

Proteotoxic stress (PS) is generated by accumulation of misfolded proteins in cells under a variety of conditions including hyperthermia, hypoxia, and exposure to denaturing agents or drugs that inhibit proteasome or chaperone activities [1,2,3]. PS can be toxic if misfolded proteins accumulate and aggregate in the cells. To avoid the toxicity of PS, cells activate an adaptive response, known as the heat shock response (HSR)[4]. Additional chaperones (e.g., Hsp70) are synthesized to assist in metabolism of misfolded proteins, ubiquitin is upregulated to allow for more proteasomemediated degradation, and general translation is attenuated [5,6]. The adaptive HSR depends upon activation of the transcription factor HSF1 [7]. HSF1 is required for expression of the inducible form of Hsp as well as other chaperones and some stressrelated transcription factors [1]

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Conclusion