Hsp70 Inhibits Aminoglycoside-Induced Hair Cell Death and is Necessary for the Protective Effect of Heat Shock

Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC 29403, USA.
Journal of the Association for Research in Otolaryngology (Impact Factor: 2.6). 06/2008; 9(3):277-89. DOI: 10.1007/s10162-008-0122-2
Source: PubMed


Sensory hair cells of the inner ear are sensitive to death from aging, noise trauma, and ototoxic drugs. Ototoxic drugs include the aminoglycoside antibiotics and the antineoplastic agent cisplatin. Exposure to aminoglycosides results in hair cell death that is mediated by specific apoptotic proteins, including c-Jun N-terminal kinase (JNK) and caspases. Induction of heat shock proteins (Hsps) is a highly conserved stress response that can inhibit JNK- and caspase-dependent apoptosis in a variety of systems. We have previously shown that heat shock results in a robust upregulation of Hsps in the hair cells of the adult mouse utricle in vitro. In addition, heat shock results in significant inhibition of both cisplatin- and aminoglycoside-induced hair cell death. In our system, Hsp70 is the most strongly induced Hsp, which is upregulated over 250-fold at the level of mRNA 2 h after heat shock. Therefore, we have begun to examine the role of Hsp70 in mediating the protective effect of heat shock. To determine whether Hsp70 is necessary for the protective effect of heat shock against aminoglycoside-induced hair cell death, we utilized utricles from Hsp70.1/3 (-/-) mice. While heat shock inhibited gentamicin-induced hair cell death in wild-type utricles, utricles from Hsp70.1/3 (-/-) mice were not protected. In addition, we have examined the role of the major heat shock transcription factor, Hsf1, in mediating the protective effect of heat shock. Utricles from Hsf1 (-/-) mice and wild-type littermates were exposed to heat shock followed by gentamicin. The protective effect of heat shock on aminoglycoside-induced hair cell death was only observed in wild-type mice and not in Hsf1 (-/-) mice. To determine whether Hsp70 is sufficient to protect hair cells, we have utilized transgenic mice that constitutively overexpress Hsp70. Utricles from Hsp70-overexpressing mice and wild-type littermates were cultured in the presence of varying neomycin concentrations for 24 h. The Hsp70-overexpressing utricles were significantly protected against neomycin-induced hair cell death at moderate to high doses of neomycin. This protective effect was achieved without a heat shock. Taken together, these data indicate that Hsp70 and Hsf1 are each necessary for the protective effect of heat shock against aminoglycoside-induced death. Furthermore, overexpression of Hsp70 alone significantly inhibits aminoglycoside-induced hair cell death.


Available from: Margaret I Lomax, May 20, 2015
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    • "Accumulating evidences indicated that hyperthermia induced HSP70 expression was related with chemotherapy resistance [25]. Moreover, Behnsawy et al had proved that the expression of HSP70 was correlated with cell survival when treated with chemotherapy [26]. "
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    ABSTRACT: Up-regulation of heat shock protein 70 (HSP70) could be elicited primarily by heat in former studies, and this was proved to be associated with cancer progression. Burkitt's lymphoma is one of highly aggressive B-cell non-Hodgkin's lymphoma and is one of the fastest growing human tumors. To investigate the effect of HSP70 expression on the sensitivity of human Burkitt lymphoma cells (Raji cells) to chemotherapy and its role in the involvement of PI3K/AKT pathway, we evaluated the effects of LY294002, a PI3K inhibitor, on the expression of HSP70 and cell sensitivity to adriamycin (ADM) or cisplatin (DDP). In present study, expressions of HSP70, AKT and phosphorylated AKT (p-AKT) in Raji cells were measured by Western-Blot. Apoptosis index of Raji cells was examined by flow cytometry. Cytotoxicities of adriamycin (ADM) and cisplatin (DDP) were determined by WST-8 assay. We found that hyperthermia (42 degrees for 1 hour) up-regulated the expression of HSP70 expression and blockade of PI3K/AKT pathway down-regulated HSP70 expression in Raji cells. Compared to cells treated with ADM or DDP alone, hyperthermia protected cells from chemotherapy while LY294002 enhanced sensitivity of Raji cells to chemotherapy. Our results suggested down-regulation of HSP70 expression by blockade of PI3K/AKT pathway maybe responsible for the increased sensitivity of Raji cells to chemotherapy. Targeting PI3K/AKT pathway or inhibiting HSP70 expression may be beneficial for chemotherapy treatment of Burkitt lymphoma patients.
    Cancer Cell International 05/2013; 13(1):48. DOI:10.1186/1475-2867-13-48 · 2.77 Impact Factor
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    • "Aside from this function, Hsps are also implicated in the general protection of stressed cells and organisms [4,5]. Many studies have reported that exposure of organisms to such diverse stressors as temperature extremes, pollutants, anoxia, parasitism, predation, or competition; all elicit reversible increases in Hsp70 expression that serve to protect the organism against cellular damage [6-9]. The involvement of Hsp70 in the acclimation of fish to salinity changes has also been well documented experimentally [10,11]. "
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    ABSTRACT: The relationship between environmental variation and induction of heat shock proteins (Hsps) has been much documented under experimental conditions. However, very little is known about such induction in natural populations acclimatised to prevailing environmental conditions. Furthermore, while induction of stress proteins has been well documented in response to environmental contaminants and thermal stressors, little is known about whether factors, such as extreme salinity, are also potential inductors. The black-chinned tilapia Sarotherodon melanotheron is unusual for its ability to colonise estuarine environments in West Africa that are characterised by extremely high salinities. The relationships between mRNA levels of the 70 kDa heat shock protein (Hsp70) and Na+, K+-ATPase1alpha (Naka) in the gills, environmental salinity, and a life-history trait (condition factor) were investigated in wild populations of this species sampled from three locations in the Saloum estuary, at salinities ranging from 40 to 100 psu. The highest Hsp70 and Naka mRNA levels, and the poorest condition factors were recorded in the most saline sampling site (100 psu). The Hsp70 and Naka mRNA were correlated amongst themselves and showed a direct positive correlation with environmental salinity, and a negative correlation with fish condition factor. Thus, the Hsp70 is constitutively overexpressed by S. melanotheron acclimatised to extreme hypersalinity. These results indicate that, although S. melanotheron can colonise extremely saline environments, the overexpression of Hsp70 combined with the higher Naka mRNA expression reveals that this represents a chronic stress. The induction of Hsp70 was, therefore, a biomarker of chronic hyper-osmotic stress which presumably can be linked to the impaired growth performance and precocious reproduction that have been demonstrated in the populations at the extremely saline sites.
    BMC Ecology 04/2010; 10(1):11. DOI:10.1186/1472-6785-10-11 · 2.36 Impact Factor
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    • "The otoprotective role of HSPs in the cochlea is well documented. Experiments with knock-out Hsf1-/- and Hsp70-/- mice, demonstrated that the expression of HSP70 is necessary to protect the cochlear hair cells from noise exposure and aminoglycoside ototoxicity [10,11]. "
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    ABSTRACT: Heat shock protein 70 (HSP70) protects inner ear cells from damage and death induced by e.g. heat or toxins. Benzoquinone ansamycin antibiotic geldanamycin (GA) was demonstrated to induce the expression of HSP70 in various animal cell types. The aim of our study was to investigate whether GA induces HSP70 in the organ of Corti (OC), which contains the auditory sensory cells, and whether GA can protect these cells from toxicity caused by a common aminoglycoside antibiotic gentamicin. To address these questions, we used the OC explants isolated from p3-p5 rats. As a read-out, we used RT-PCR, ELISA and immunofluorescence. We found that GA at the concentration of 2 microM efficiently induced HSP70 expression on mRNA and protein level in the OC explants. Confocal microscopy revealed that HSP70 induced by GA is expressed by hair cells and interdental cells of spiral limbus. Preincubation of explants with 2 muM GA prior to adding gentamicin (500 microM) significantly reduced the loss of outer but not inner hair cells, suggesting different mechanisms of otoprotection needed for these two cell types. GA induced HSP70 in the auditory sensory cells and partially protected them from toxicity of gentamicin. Understanding the molecular mechanisms of GA otoprotection may provide insights for preventative therapy of the hearing loss caused by aminoglycoside antibiotics.
    Journal of Biomedical Science 10/2009; 16(1):79. DOI:10.1186/1423-0127-16-79 · 2.76 Impact Factor
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