NCALD as a potential predictive biomarker for the efficacy of platinum-based chemotherapy in ovarian cancer.

Aim: Since reliable response predictors to platinum-based chemotherapy in ovarian cancer (OC) are scarce, we characterize NCALD as a predictive biomarker. Materials & methods: NCALD mRNA (n = 100) and protein (n = 102) expression was analyzed in OC samples and associated with patient outcome. A stable OC cell line knockdown was generated and cellular response to platinum was explored. Results: High NCALD mRNA and protein expression was significantly associated with longer overall patient survival (p = 0.037/0.002). Knockdown experiments revealed a significant association between cisplatin sensitivity and NCALD expression. Conclusion: Low NCALD expression was associated with reduced sensitivity to platinum-based chemotherapy. NCALD may be a new biomarker candidate to identify patients who might benefit from platinum-based chemotherapy.

NZ28-induced inhibition of HSF1, SP1 and NF-κB triggers the loss of the natural killer cell-activating ligands MICA/B on human tumor cells.

The activity of natural killer (NK) cells is regulated by activating and inhibiting receptors, whereby the C-type lectin natural killer group 2D (NKG2D) receptor serves as the major activating receptor on NK cells which recognizes major histocompatibility class I chain-related proteins A and B (MICA/B). The MICA/B expression has been described to be regulated by the transcription factor heat shock factor 1 (HSF1). Inhibition of heat shock protein 90 (Hsp90) is known to induce the heat shock response via activation of HSF1 which is associated with tumor development, metastasis and therapy resistance and also with an increased susceptibility to NK cell-mediated lysis. Therefore, we compared the effects of Hsp90 inhibitor NVP-AUY922, HSF1 inhibitor NZ28 and HSF1 knockdown on the sensitivity of lung (H1339) and breast (MDA-MB-231, T47D) cancer cells to NK cell-mediated cytotoxicity and the expression of the NKG2D ligands MICA/B. Although NVP-AUY922 activates HSF1, neither the MICA/B surface density on tumor cells nor their susceptibility to NK cell-mediated lysis was affected. A single knockdown of HSF1 by shRNA decreased the surface expression of MICB but not that of MICA, and thereby, the NK cell-mediated lysis was only partially blocked. In contrast, NZ28 completely blocked the MICA/B membrane expression on tumor cells and thereby strongly inhibited the NK cell-mediated cytotoxicity. This effect might be explained by a simultaneous inhibition of the transcription factors HSF1, Sp1 and NF-κB by NZ28. These findings suggest that new anticancer therapeutics should be investigated with respect to their effects on the innate immune system.

Sensitizing tumor cells to radiation by targeting the heat shock response.

Elevated levels of heat shock proteins (HSPs) contribute to tumor cell survival and mediate protection against radiation-induced cell death. Hsp90 inhibitors are promising radiosensitizers but also activate heat shock factor 1 (HSF1) and thereby induce the synthesis of cytoprotective Hsp70. In this study the heat shock response inhibitor NZ28 either alone or in combination with the Hsp90 inhibitor NVP-AUY922 was investigated for radiosensitizing effects, alterations in cell cycle distribution and effects on migratory/invasive capacity of radioresistant tumor cells. NZ28 reduced the constitutive and NVP-AUY922-induced Hsp70 expression by inhibition of the HSF1 activity and inhibited migration and invasion in human lung and breast tumor cells. Treatment of tumor cells with NZ28 significantly increased their radiation response. One possible mechanism might be a decrease of the radioresistant S-phase. When combined with the Hsp90 inhibitor NVP-AUY922 the concentration of NZ28 could be significantly reduced (1/10th-1/20th) to achieve the same radiosensitization. Our results demonstrate that a dual targeting of Hsp70 and Hsp90 with NZ28 and NVP-AUY922 potentiates the radiation response of tumor cells that are otherwise resistant to ionizing radiation.

A hypoxia-induced decrease of either MICA/B or Hsp70 on the membrane of tumor cells mediates immune escape from NK cells.

Recent findings suggest that hypoxia of the tumor microenvironment contributes to immune escape from natural killer (NK) cell-mediated cytotoxicity. Heat shock protein 70 (Hsp70) and the stress-regulated major histocompatibility class I chain-related protein A and B (MICA/B) both serve as ligands for activated NK cells when expressed on the cell surface of tumor cells. Herein, we studied the effects of hypoxia and hypoxia-inducible factor-1α (HIF-1α) on the membrane expression of these NK cell ligands in H1339 with high and MDA-MB-231 tumor cells with low basal HIF-1α levels and its consequences on NK cell-mediated cytotoxicity. We could show that a hypoxia-induced decrease in the membrane expression of MICA/B and Hsp70 on H1339 and MDA-MB-231 cells, respectively, is associated with a reduced sensitivity to NK cell-mediated lysis. A knockdown of HIF-1α revealed that the decreased surface expression of MICA/B under hypoxia is dependent on HIF-1α in H1339 cells with high basal HIF-1α levels. Hypoxia and HIF-1α did not affect the MICA/B expression in MDA-MB-231 cells but reduced the Hsp70 membrane expression which in turn also impaired NK cell recognition. Furthermore, we could show that the hypoxia-induced decrease in membrane Hsp70 is independent of HIF-1α in MDA-MB-231. Our data indicate that hypoxia-induced downregulation of both NK cell ligands MICA/B and Hsp70 impairs NK cell-mediated cytotoxicity, whereby only MICA/B appears to be regulated by HIF-1α.

Application of a degree-day model of West Nile virus transmission risk to the East Coast of the United States of America.

A geographical information systems model that identifies regions of the United States of America (USA) susceptible to West Nile virus (WNV) transmission risk is presented. This system has previously been calibrated and tested in the western USA; in this paper we use datasets of WNV-killed birds from South Carolina and Connecticut to test the model in the eastern USA. Because their response to WNV infection is highly predictable, American crows were chosen as the primary source for model calibration and testing. Where crow data are absent, other birds are shown to be an effective substitute. Model results show that the same calibrated model demonstrated to work in the western USA has the same predictive ability in the eastern USA, allowing for a continental-scale evaluation of the transmission risk of WNV at a daily time step. The calibrated model is independent of mosquito species and requires inputs of only local maximum and minimum temperatures. Of benefit to the general public and vector control districts, the model predicts the onset of seasonal transmission risk, although it is less effective at identifying the end of the transmission risk season.

A geographical information system-based web model of arbovirus transmission risk in the continental United States of America.

A degree-day (DD) model of West Nile virus capable of forecasting real-time transmission risk in the continental United States of America up to one week in advance using a 50-km grid is available online at https://sites. google.com/site/arbovirusmap/. Daily averages of historical risk based on temperatures for 1994-2003 are available at 10km resolution. Transmission risk maps can be downloaded from 2010 to the present. The model can be adapted to work with any arbovirus for which the temperature-related parameters are known, e.g. Rift Valley fever virus. To more effectively assess virus establishment and transmission, the model incorporates "compound risk" maps and forecasts, which includes livestock density as a parameter.

A temperature-limited assessment of the risk of Rift Valley fever transmission and establishment in the continental United States of America.

The rapid spread of West Nile virus across North America after its introduction in 1999 highlights the potential for foreign arboviruses to become established in the United States of America. Of particular concern is Rift Valley fever virus (RVFV), which has been responsible for multiple African epidemics resulting in death of both humans and livestock, as well as major economic disruption due to livestock loss and trade restrictions. Modern globalization, travel, and commerce allow viruses to easily jump from one continent to another; and it is likely only a matter of time before RVFV reaches North American shores. We used a degree-day model in combination with livestock population data and a pathways analysis to identify regions and times where RVFV is most likely to enter and become established in the United States of America. Transmission risk of the disease varies across the country from 325 annual risk days in parts of Florida to zero risk days in the far North and in high mountain regions. Areas of particular concern are where there are a high number of possible tranmission days, a large livestock population, and proximity to likely locations for the disease to enter the country via mosquito vector or human host. These areas should be monitored closely during transmission "risk seasons" so that if the virus does enter the country and begins to become established, it can be quickly controlled and eliminated before spreading further. Areas most at risk include the Baltimore and New York City metro areas as well as much of the region between these urban centers; most of Texas, especially around Houston; Florida; Atlanta; southwest Nebraska; southern California and Arizona; and the central valley of California.

Spatially explicit West Nile virus risk modeling in Santa Clara County, California.

A geographic information system model designed to identify regions at risk for West Nile virus (WNV) transmission was calibrated and tested with data collected in Santa Clara County, California. American Crows that died from WNV infection in 2005 provided spatial and temporal ground truth. When the model was run with parameters based on Culex tarsalis infected with the NY99 genotype of the virus, it underestimated WNV occurrence in Santa Clara Co. The parameters were calibrated to fit the field data by reducing the number of degree-days necessary to reach the mosquito's extrinsic incubation period from 109 to 76. The calibration raised model efficiency from 61% to 92% accuracy, and the model performed well the following year in Santa Clara Co.