Crosstalk between developmental and tumour-specific signalling pathways : kallikrein-related serine peptidases and nodal in prostrate cancer

Prostate cancer is an important male health issue. The strategies used to diagnose and treat prostate cancer underscore the cell and molecular interactions that promote disease progression. Prostate cancer is histologically defined by increasingly undifferentiated tumour cells and therapeutically targeted by androgen ablation. Even as the normal glandular architecture of the adult prostate is lost, prostate cancer cells remain dependent on the androgen receptor (AR) for growth and survival. This project focused on androgen-regulated gene expression, altered cellular differentiation, and the nexus between these two concepts. The AR controls prostate development, homeostasis and cancer progression by regulating the expression of downstream genes. Kallikrein-related serine peptidases are prominent transcriptional targets of AR in the adult prostate. Kallikrein 3 (KLK3), which is commonly referred to as prostate-specific antigen, is the current serum biomarker for prostate cancer. Other kallikreins are potential adjunct biomarkers. As secreted proteases, kallikreins act through enzyme cascades that may modulate the prostate cancer microenvironment. Both as a panel of biomarkers and cascade of proteases, the roles of kallikreins are interconnected. Yet the expression and regulation of different kallikreins in prostate cancer has not been compared. In this study, a spectrum of prostate cell lines was used to evaluate the expression profile of all 15 members of the kallikrein family. A cluster of genes was co-ordinately expressed in androgenresponsive cell lines. This group of kallikreins included KLK2, 3, 4 and 15, which are located adjacent to one another at the centromeric end of the kallikrein locus. KLK14 was also of interest, because it was ubiquitously expressed among the prostate cell lines. Immunohistochemistry showed that these 5 kallikreins are co-expressed in benign and malignant prostate tissue. The androgen-regulated expression of KLK2 and KLK3 is well-characterised, but has not been compared with other kallikreins. Therefore, KLK2, 3, 4, 14 and 15 expression were all measured in time course and dose response experiments with androgens, AR-antagonist treatments, hormone deprivation experiments and cells transfected with AR siRNA. Collectively, these experiments demonstrated that prostatic kallikreins are specifically and directly regulated by the AR. The data also revealed that kallikrein genes are differentially regulated by androgens; KLK2 and KLK3 were strongly up-regulated, KLK4 and KLK15 were modestly up-regulated, and KLK14 was repressed. Notably, KLK14 is located at the telomeric end of the kallikrein locus, far away from the centromeric cluster of kallikreins that are stimulated by androgens. These results show that the expression of KLK2, 3, 4, 14 and 15 is maintained in prostate cancer, but that these genes exhibit different responses to androgens. This makes the kallikrein locus an ideal model to investigate AR signalling. The increasingly dedifferentiated phenotype of aggressive prostate cancer cells is accompanied by the re-expression of signalling molecules that are usually expressed during embryogenesis and foetal tissue development. The Wnt pathway is one developmental cascade that is reactivated in prostate cancer. The canonical Wnt cascade regulates the intracellular levels of β-catenin, a potent transcriptional co-activator of T-cell factor (TCF) transcription factors. Notably, β-catenin can also bind to the AR and synergistically stimulate androgen-mediated gene expression. This is at the expense of typical Wnt/TCF target genes, because the AR:β-catenin and TCF:β-catenin interactions are mutually exclusive. The effect of β-catenin on kallikrein expression was examined to further investigate the role of β-catenin in prostate cancer. Stable knockdown of β-catenin in LNCaP prostate cancer cells attenuated the androgen-regulated expression of KLK2, 3, 4 and 15, but not KLK14. To test whether KLK14 is instead a TCF:β-catenin target gene, the endogenous levels of β-catenin were increased by inhibiting its degradation. Although KLK14 expression was up-regulated by these treatments, siRNA knockdown of β-catenin demonstrated that this effect was independent of β-catenin. These results show that β-catenin is required for maximal expression of KLK2, 3, 4 and 15, but not KLK14. Developmental cells and tumour cells express a similar repertoire of signalling molecules, which means that these different cell types are responsive to one another. Previous reports have shown that stem cells and foetal tissues can reprogram aggressive cancer cells to less aggressive phenotypes by restoring the balance to developmental signalling pathways that are highly dysregulated in cancer. To investigate this phenomenon in prostate cancer, DU145 and PC-3 prostate cancer cells were cultured on matrices pre-conditioned with human embryonic stem cells (hESCs). Soft agar assays showed that prostate cancer cells exposed to hESC conditioned matrices had reduced clonogenicity compared with cells harvested from control matrices. A recent study demonstrated that this effect was partially due to hESC-derived Lefty, an antagonist of Nodal. A member of the transforming growth factor β (TGFβ) superfamily, Nodal regulates embryogenesis and is re-expressed in cancer. The role of Nodal in prostate cancer has not previously been reported. Therefore, the expression and function of the Nodal signalling pathway in prostate cancer was investigated. Western blots confirmed that Nodal is expressed in DU145 and PC-3 cells. Immunohistochemistry revealed greater expression of Nodal in malignant versus benign glands. Notably, the Nodal inhibitor, Lefty, was not expressed at the mRNA level in any prostate cell lines tested. The Nodal signalling pathway is functionally active in prostate cancer cells. Recombinant Nodal treatments triggered downstream phosphorylation of Smad2 in DU145 and LNCaP cells, and stably-transfected Nodal increased the clonogencity of LNCaP cells. Nodal was also found to modulate AR signalling. Nodal reduced the activity of an androgen-regulated KLK3 promoter construct in luciferase assays and attenuated the endogenous expression of AR target genes including prostatic kallikreins. These results demonstrate that Nodal is a novel example of a developmental signalling molecule that is reexpressed in prostate cancer and may have a functional role in prostate cancer progression. In summary, this project clarifies the role of androgens and changing cellular differentiation in prostate cancer by characterising the expression and function of the downstream genes encoding kallikrein-related serine proteases and Nodal. Furthermore, this study emphasises the similarities between prostate cancer and early development, and the crosstalk between developmental signalling pathways and the AR axis. The outcomes of this project also affirm the utility of the kallikrein locus as a model system to monitor tumour progression and the phenotype of prostate cancer cells.

Learning to win process-control games watching game-masters

The present paper focuses on some interesting classes of process-control games, where winning essentially means successfully controlling the process. A master for one of these games is an agent who plays a winning strategy. In this paper we investigate situations in which even a complete model (given by a program) of a particular game does not provide enough information to synthesize—even incrementally—a winning strategy. However, if in addition to getting a program, a machine may also watch masters play winning strategies, then the machine is able to incrementally learn a winning strategy for the given game. Studied are successful learning from arbitrary masters and from pedagogically useful selected masters. It is shown that selected masters are strictly more helpful for learning than are arbitrary masters. Both for learning from arbitrary masters and for learning from selected masters, though, there are cases where one can learn programs for winning strategies from masters but not if one is required to learn a program for the master's strategy itself. Both for learning from arbitrary masters and for learning from selected masters, one can learn strictly more by watching m+1 masters than one can learn by watching only m. Last, a simulation result is presented where the presence of a selected master reduces the complexity from infinitely many semantic mind changes to finitely many syntactic ones.

Cerebellar augmented joint control for a humanoid robot

The joints of a humanoid robot experience disturbances of markedly different magnitudes during the course of a walking gait. Consequently, simple feedback control techniques poorly track desired joint trajectories. This paper explores the addition of a control system inspired by the architecture of the cerebellum to improve system response. This system learns to compensate the changes in load that occur during a cycle of motion. The joint compensation scheme, called Trajectory Error Learning, augments the existing feedback control loop on a humanoid robot. The results from tests on the GuRoo platform show an improvement in system response for the system when augmented with the cerebellar compensator.

Multi-robot control in highly dynamic, competitive environments

The control and coordination of multiple mobile robots is a challenging task; particularly in environments with multiple, rapidly moving obstacles and agents. This paper describes a robust approach to multi-robot control, where robustness is gained from competency at every layer of robot control. The layers are: (i) a central coordination system (MAPS), (ii) an action system (AES), (iii) a navigation module, and (iv) a low level dynamic motion control system. The multi-robot coordination system assigns each robot a role and a sub-goal. Each robots action execution system then assumes the assigned role and attempts to achieve the specified sub-goal. The robots navigation system directs the robot to specific goal locations while ensuring that the robot avoids any obstacles. The motion system maps the heading and speed information from the navigation system to force-constrained motion. This multi-robot system has been extensively tested and applied in the robot soccer domain using both centralized and distributed coordination.

Distributed control of gait for a humanoid robot

This paper describes a walking gait for a humanoid robot with a distributed control system. The motion for the robot is calculated in real time on a central controller, and sent over CAN bus to the distributed control system. The distributed control system loosely follows the motion patterns from the central controller, while also acting to maintain stability and balance. There is no global feedback control system; the system maintains its balance by the interaction between central gait and soft control of the actuators. The paper illustrates a straight line walking gait and shows the interaction between gait generation and the control system. The analysis of the data shows that successful walking can be achieved without maintaining strict local joint control, and without explicit global balance coordination.

Operation and control of a DVR in the presence of interharmonics

The paper discusses the operating principles and control characteristics of a dynamic voltage restorer (DVR). It is assumed that the source voltages contain interharmonic components in addition to fundamental components. The main aim of the DVR is to produce a set of clean balanced sinusoidal voltages across the load terminals irrespective of unbalance, distortion and voltage sag/swell in the supply voltage. An algorithm has been discussed for extracting fundamental phasor sequence components from the samples of three-phase voltages or current waveforms having integer harmonics and interharmonics. The DVR operation based on extracted components is demonstrated. The switching signal is generated using a deadbeat controller. It has been shown that the DVR is able to compensate these interharmonic components such that the load voltages are perfectly regulated. The DVR operation under deep voltage sag is also discussed. The proposed DVR operation is verified through the computer simulation studies using the MATLAB software package.