ATM Signaling to TSC2: Mechanisms and Implications for Cancer Therapy

Ataxia telangiectasia mutated (ATM) is a critical component of the cellular response to DNA damage, where it acts as a damage sensor, and signals to a large network of proteins which execute the important tasks involved in responding to the damage, namely inducing cell cycle checkpoints, inducing DNA repair, modulating transcriptional responses, and regulating cell death pathways if the damage cannot be repaired faithfully. We have now discovered that an additional novel component of this ATM-dependent damage response involves induction of autophagy in response to oxidative stress. In contrast to DNA damage-induced ATM activation however, oxidative stress induced ATM, occurs in the cytoplasm, and does not require nuclear-to-cytoplasmic shuttling of ATM. Using several cell culture systems including MCF7 breast carcinoma cells, SKOV3 ovarian cancer cells, and various lineages of mouse embryonic fibroblasts, we showed that once activated by reactive oxygen species (ROS), ATM signals to mTORC1 to induce autophagy via the LKB1-AMPK-TSC2 pathway. Targeting dysregulation of mTORC1 in Atm-deficient mice, which succumb to lymphomagenesis within 3-4 months of age with daily administration of rapamycin, could significantly extend survival and cause regression of tumors, suggesting that pharmacologically targeting this pathway has therapeutic implications in cancer. We also identified a second contrasting pathway for DNA damage-induced mTORC1 repression which does not require AMPK activation, but does require ATM and TSC2. Several potential mechanisms including mTOR localization and p53-mediated pathways were ruled out however we identified that TSC2 may be an additional cytoplasmic direct ATM substrate that is engaged in response to DNA damage specifically. Lastly, a study was performed to examine whether autophagy induced by ovarian cancer therapeutics (focusing on cisplatin, since paclitaxel does not induce autophagy in the SKOV3 cell line model we used) plays a role in resistance to therapy since autophagy can play both pro-survival mechanisms or be a mechanism of cell death. Using a genetic approach to knock-down Atg5 expression with shRNA in SKOV3 ovarian carcinoma cells, we compared the cytotoxicity of cisplatin in vector or Atg5 knock-down cells, and demonstrated that autophagy does not play any significant role in the response to cisplatin in this cell line.

Culture and the social context of health inequalities

There has been a great deal of interest and debate recently concerning the linkages between inequality and health cross-nationally. Exposures to social and health inequalities likely vary as a consequence of different cultural contexts. It is important to guide research by a theoretical perspective that includes cultural and social contexts cross-nationally. If inequality affects health only under specific cultural conditions, this could explain why some of the literature that compares different societies finds no evidence of a relationship between inequality and health in certain countries. A theoretical framework is presented that combines sociological theory with constructs from cultural psychology in order to identify pathways that might lead from cultural dimensions to health inequalities. Three analyses are carried out. The first analysis explores whether there is a relationship between cultural dimensions at the societal level and self-rated health at the individual level. The findings suggest that different cultural norms at the societal level can produce both social and health inequalities, but the effects on health may differ depending on the socio-cultural context. The second analysis tests the hypothesis that health is affected by the density of social networks in a society, levels of societal trust, and inequality. The results suggest that commonly used measures of social cohesion and inequality may have both contextual and compositional effects on health in a large number of countries, and that societal measures of social cohesion and inequality interact with individual measures of social participation, trust, and income, moderating their effects on health. The third analysis explores whether value systems associated with vertical individualist societies may lead to health disparities because of their stigmatizing effects. I test the hypothesis that, within vertical individualist societies, subjective well-being will be affected by a social context where competition and the Protestant work ethic are valued, mediated by inequality. The hypothesis was not supported by the available cross-national data, most likely because of inadequate measures, missing data, and the small sample of vertical individualist countries. The overall findings demonstrate that cultural differences are important contextual factors that should not be overlooked when examining the causes of health inequalities. ^

Corporate decision-making: A case study of decision support systems and financial performance in health care organizations

The purpose of this study was twofold: (1) To describe the relation of the intensity of DSS implementation to financial performance as an empirical exploration of improved performance at the organizational level. (2) To describe the relation of the intensity of DSS implementation to the type of organizational decision culture. A multiple case study design was utilized to compare three groups of paired cases. A pattern matching strategy was applied in this study. Four predictions were specified and compared to the empirical data. A progressively upward trend in the scores was predicted for the following theoretical relationships. (1) The greater the number of DSSs, the higher the sophistication index. (2) The greater the number of DSSs, the higher the financial ratios. (3) The greater the number of DSSs, the higher the culture score. (4) The higher the culture score, the higher the financial ratios. The data did not support any of the predicted trends except the relation between the number of DSSs and the financial ratios. The Income/Revenue ratio indicates the efficiency of a company's operations. One would expect that this ratio would be most affected by the operational and financial decision support systems. The majority of the systems measured in the study supported decisions tangential to the patient service areas. The evidence suggested that the type and number of decision support systems affects the bottom line. ^

The p53 tumor suppressor pathway in culture and {\it in vivo\/}

p53 functions as a tumor suppressor through its ability to initiate either growth arrest or apoptosis in cells which have sustained DNA damage. p53 elicits these cellular phenotypes through its biochemical function as a transcriptional activator. By inducing the expression of a battery of target genes, p53 is able to prevent the propagation of cells with damaged DNA. However, the genes transcriptionally induced by p53 which have been identified to date do not fully explain p53 function. p53 has been demonstrated to activate genes involved in cell cycle inhibition, apoptosis and cell proliferation. The reasons for simultaneous activation of p53 targets with disparate, opposing functions are not clear, but may be due to the use of transformed cell lines in previous experiments. In the studies presented in this thesis, the pathway of p53 tumor suppression has been studied in detail in two systems chosen for their relevance to the natural cell environment. One utilizes a normal, unaltered cultured cell system; the other the whole mouse. In order to better understand the role of the known p53 targets in effecting p53 function in normal cells, early rat embryo fibroblasts were irradiated with ultraviolet light to induce DNA damage. It was discovered that p53 protein levels increased in response to irradiation. The known targets of p53, namely, $p21\sp{WAF1/CIP1},\ mdm2,\ cyclin\ G,$ and bax, were shown for the first time to have a differential temporal induction. The growth suppressor $p21\sp{WAF1/CIP1}$ was induced first, followed by cyclin G then mdm2, which is involved in proliferation through its inactivation of p53, and finally, the apoptosis promoter, bax. These findings indicated that p53 activates its target genes in a manner to allow maximum effectiveness of target function. The rat embryo fibroblasts were shown to undergo apoptosis 24 h after irradiation. Additionally, investigation of these cells for cell cycle alterations demonstrated a brief arrest in G1. In the second study, thymocytes from mice with wild type p53 were shown to undergo apoptosis and activate p53 target genes upon ionizing radiation treatment, while thymocytes from mice deficient in p53 could not. The p53 target genes mdm2 and fas were tested in vivo for their ability to mediate p53-regulated apoptosis, and were found dispensible for that cellular function. Therefore, the p53 targets identified to date do not fully explain the ability of p53 to function as a tumor suppressor. Potentially, functional redundancy between the known targets would account for the data seen in these experiments. Additionally, identification of additional target genes should add further understanding of the p53 pathway of tumor suppression. ^