Geldanamycin Analog 17-DMAG Limits Apoptosis in Human Peripheral Blood Cells by Inhibition of p53 Activation and its Interaction with Heat-Shock Protein 90 kDa after Exposure to Ionizing Radiation

Radiation Combined Injury Program, Armed Forces Radiobiology Research Institute, Bethesda, MD 20889-5603, USA.
Radiation Research (Impact Factor: 2.91). 06/2011; 176(3):333-45. DOI: 10.2307/41318197
Source: PubMed


Exposure to ionizing radiation induces p53, and its inhibition improves mouse survival. We tested the effect of 17-dimethylamino-ethylamino-17-demethoxygeldanamycin (17-DMAG) on p53 expression and function after radiation exposure. 17-DMAG, a heat-shock protein 90 (Hsp90) inhibitor, protects human T cells from ionizing radiation-induced apoptosis by inhibiting inducible nitric oxide synthase (iNOS) and subsequent caspase-3 activation. Using ex vivo human peripheral blood mononuclear cells, we found that ionizing radiation increased p53 accumulation, acute p53 phosphorylation, Bax expression and caspase-3/7 activation in a radiation dose- and time postirradiation-dependent manner. 17-DMAG inhibited these increases in a concentration-dependent manner (IC(50)  =  0.93 ± 0.01 µM). Using in vitro models, we determined that inhibition of p53 by genetic knockout resulted in lower levels of caspase-3/7 activity 1 day after irradiation and enhanced survival at 10 days. Analysis of p53-Hsp90 interaction in ex vivo cell lysates indicated that the binding between the two molecules occurred after irradiation but 17-DMAG prevented the binding. Taken together, these results suggest the presence of p53 phosphorylation and Hsp90-dependent p53 stabilization after acute irradiation. Hsp90 inhibitors such as 17-DMAG may prove useful with radiation-based cancer therapy as well as for general radioprotection.

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    • "Injuries induced by ionizing radiation alone (RI) or in combination with trauma from blast and thermal energy exposure (CI) are expected after the detonation of radiation dispersal devices or nuclear weapons. In vivo [1] and in vitro [2] [3] studies indicate that RI induced DNA doublestrand breaks (DSBs), activated signal transduction pathways , elevated cytokine/chemokine concentrations in the peripheral blood, and increased systemic bacterial infection, thereby leading to cell death and multiple-organ dysfunction and failure [1, 4–6]. Traumatic injury followed by RI (i.e., CI) enhanced histopathological responses to RI, thereby increasing the mortality [1, 5–7]. "
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    ABSTRACT: Exposure to ionizing radiation alone (radiation injury, RI) or combined with traumatic tissue injury (radiation combined injury, CI) is a crucial life-threatening factor in nuclear and radiological accidents. As demonstrated in animal models, CI results in greater mortality than RI. In our laboratory, we found that B6D2F1/J female mice exposed to 60 Co- γ -photon radiation followed by 15% total-body-surface-area skin burns experienced an increment of 18% higher mortality over a 30-day observation period compared to irradiation alone; that was accompanied by severe cytopenia, thrombopenia, erythropenia, and anemia. At the 30th day after injury, neutrophils, lymphocytes, and platelets still remained very low in surviving RI and CI mice. In contrast, their RBC, hemoglobin, and hematocrit were similar to basal levels. Comparing CI and RI mice, only RI induced splenomegaly. Both RI and CI resulted in bone marrow cell depletion. It was observed that only the RI mice treated with pegylated G-CSF after RI resulted in 100% survival over the 30-day period, and pegylated G-CSF mitigated RI-induced body-weight loss and depletion of WBC and platelets. Peg-G-CSF treatment sustained RBC balance, hemoglobin levels, and hematocrits and inhibited splenomegaly after RI. The results suggest that pegylated G-CSF effectively sustained animal survival by mitigating radiation-induced cytopenia, thrombopenia, erythropenia, and anemia.
    Oxidative Medicine and Cellular Longevity 03/2014; 2014(12):481392. DOI:10.1155/2014/481392 · 3.36 Impact Factor
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    • "It is known that 17-DMAG inhibits activation of the iNOS pathway in vitro[37] and in vivo[20], and the p53 pathway ex vivo[13]. Further studies should explore the inter-relationship among iNOS, p53, and survivin pathways and the regulation of G-CSF. "
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    ABSTRACT: Our previous research demonstrated that one subcutaneous injection of 17-Dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG) 24 hours (h) before irradiation (8.75 Gy) increased mouse survival by 75%. However, the protective mechanism of 17-DMAG is currently unknown. The present study aimed to investigate whether oral administration of 17-DMAG was also radioprotective and the potential role it may play in radioprotection. A single dose of orally pre-administered (24, 48, or 72 h) 17-DMAG (10 mg/kg) increased irradiated mouse survival, reduced body weight loss, improved water consumption, and decreased facial dropsy, whereas orally post-administered 17-DMAG failed. Additional oral doses of pre-treatment did not improve 30-day survival. The protective effect of multiple pre-administrations (2-3 times) of 17-DMAG at 10 mg/kg was equal to the outcome of a single pre-treatment. In 17-DMAG-pretreated mice, attenuation of bone marrow aplasia in femurs 30 days after irradiation with recovered expressions of cluster of differentiation 34, 44 (CD34, CD44), and survivin in bone marrow cells were observed. 17-DMAG also elevated serum granulocyte-colony stimulating factor (G-CSF), decreased serum fms-related tyrosine kinase 3 ligand, and reduced white blood cell depletion. 17-DMAG ameliorated small intestinal histological damage, promoted recovery of villus heights and intestinal crypts including stem cells, where increased leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5) was found 30 days after irradiation. 17-DMAG is a potential radioprotectant for bone marrow and small intestine that results in survival improvement.
    Cell and Bioscience 09/2013; 3(1):36. DOI:10.1186/2045-3701-3-36 · 3.63 Impact Factor
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    • "Since many Hsp90 clients are important for disease development, 17-DMAG is extensively studied for possible treatments of various diseases [21-25]. In this study, we have determined the effects of 17-DMAG and TNF treatments on multiple cell lines. "
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    ABSTRACT: The tumor necrosis factor (TNF) and the cellular NF-κB pathway protein IKKβ play important roles in various cellular processes such as cell proliferation, survival, differentiation, and apoptosis. A heat shock protein 90 inhibitor, 17-DMAG, can induce apoptosis of some tumor cells. This study is to determine the combined effects of 17-DMAG and TNF on malignant cells and the related mechanisms. We have determined effects of 17-DMAG, an Hsp90 inhibitor, and TNF treatments on the small cell lung cancer cell line (MS-1), the adenocarcinoma cell line (A549), the squamous-cell carcinoma cell line (LK-2), and the normal human bronchial epithelium cell line (NuLi-1) by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrozolium bromide assay. To determine if 17-DMAG inhibit the expression of IKKβ in the normal human NuLi-1 cells, and the malignant MS-1, A549, and LK-2 cells, immunoblotting assays and luciferase assays were performed. It was found that the combined treatments resulted in synergistic killing of malignant cells, which was confirmed by the apoptosis determination using a fluorescence microscopic assay following staining of the drug-treated cells with Hoescht 33258. The immunoblotting results indicated that the synergistic killing due to 17-DMAG and TNF treatments may be related to the decreases in IKKβ levels in the presence of 17-DMAG. The results suggest that combination of 17-DMAG and TNF treatments might be useful for treating malignancies upon further study in the further. Virtual slides The virtual slide(s) for this article can be found here:
    Diagnostic Pathology 05/2013; 8(1):70. DOI:10.1186/1746-1596-8-70 · 2.60 Impact Factor
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