Insights into immuno-oncology drug development landscape with focus on bone metastasis

Bone is among the main sites of metastasis in breast, prostate and other major cancers. Bone metastases remain incurable causing high mortality, severe skeletal-related effects and decreased quality of life. Despite the success of immunotherapies in oncology, no immunotherapies are approved for bone metastasis and no clear benefit has been observed with approved immunotherapies in treatment of bone metastatic disease. Therefore, it is crucial to consider unique features of tumor microenvironment in bone metastasis when developing novel therapies. The vicious cycle of bone metastasis, referring to crosstalk between tumor and bone cells that enables the tumor cells to grow in the bone microenvironment, is a well-established concept. Very recently, a novel osteoimmuno-oncology (OIO) concept was introduced to the scientific community. OIO emphasizes the significance of interactions between tumor, immune and bone cells in promoting tumor growth in bone metastasis, and it can be used to reveal the most promising targets for bone metastasis. In order to provide an insight into the current immuno-oncology drug development landscape, we used 1stOncology database, a cancer drug development resource to identify novel immunotherapies in preclinical or clinical development for breast and prostate cancer bone metastasis. Based on the database search, 24 immunotherapies were identified in preclinical or clinical development that included evaluation of effects on bone metastasis. This review provides an insight to novel immuno-oncology drug development in the context of bone metastasis. Bone metastases can be approached using different modalities, and tumor microenvironment in bone provides many potential targets for bone metastasis. Noting current increasing interest in the field of OIO, more therapeutic opportunities that primarily target bone metastasis are expected in the future

Imaging studies of cell physiology with particular reference to Na,K-ATPase function

The membrane protein Na,K-AT'Pase is well known for its critical function of transporting sodium out of the cell and potassium into the cell, thereby creating a fundamental electrochemical gradient upon which several other important cell functions are dependent. In the current study we have investigated novel aspects of Na,K-AT'Pase function in cell physiology. In order to study this, a considerable part of the present thesis has involved methodological development of different microscopic techniques. It is well established that NaX-AT'Pase is important for maintaining cell volume, membrane potential and reabsorption of electrolytes in the kidney. The new findings in this study are: 1) A role for Na,K-AT'Pase in the regulation of cell adhesion. Partial inhibition of Na,K-ATPase activity significantly reduced cell attachment to fibronectin. The results suggest that this effect is mediated by perturbation of normal Ca2+ signaling and a reduction of focal adhesion kinase activity. These findings indicate the importance of Na,K-ATPase during development and differentiation. 2) A major role of Na,K-ATPase activity in regulatory volume decrease (RVD). A direct link between Na,K-ATPase activity and the ability of COS-7 cells to perform RVD was demonstrated. With site directed mutagenesis of the a subunit of Na,K-ATPase it was demonstrated that Na,K-ATPase may have both negative and positive effects on the rate of RVD. Further knowledge about the interaction between ouabain and NaK-ATPase was achieved by demonstrating that a Leu-799 to Cys substitution in the alpha subunit of rat Na,K-ATPase produced complete ouabain resistance. This residue belongs to the extracellular loop between transmembrane segment 5 and 6 of the enzyme which is of importance for ion occlusion and ion transport. The same mutant has been demonstrated to have a positive effect on the rate of RVD. This thesis was also concerned with the rigorous use of fluorescence lifetime imaging to investigate intracellular pH. We made theoretical predictions concerning sensitivity and noise which were supported by experimental results. Our study on the influence of probe binding on pH based fluorescent lifetime imaging indicates that the method is not a straightforward approach to measure pH in the absence of correction for the effect of probe binding. We find it likely that other fluorescent ion probes have similar probe-binding-sensitive fluorescence lifetimes. However, the overall effect is difficult to predict. In conclusion, this thesis demonstrates that Na,K-ATPase is a key enzyme in a variety of important cell functions beyond those that were previously known. In addition to being the dynamic modulator of ion transport, Na,K-ATPase also serves a primary role in regulatory cell volume decrease and cell attachment

Experimental and numerical investigations of jet mixing in a multifunctional channel reactor: Passive and reactive systems

Mixing of two liquids in a new multifunctional channel reactor developed by AlfaLaval has been studied both experimentally and through computational fluid dynamics (CFD). As the channels are quite narrow the Reynolds numbers are low and the bulk of the channel is within the turbulent boundary layer. This makes accurate a priori predictions of the flowfield difficult and experimental validation necessary. Particle image velocimetry (PIV) was used to measure the flowfield, whereas planar laser-induced fluorescence was used for a scalar concentration field. CFD simulations were performed with the commercial software Fluent 5.5. Different turbulence models were tested and compared with PIV. The best predictions were obtained with a low Reynolds boundary layer k-ε turbulence model. Mixing of a passive tracer, including mean concentration and concentration variance, was calculated with the turbulent mixer model of Baldyga. A reactive system with diazo coupling between 1-naphthols, 2-naphthols and diazotized sulphanilic acid was studied both experimentally and theoretically due to its sensitivity to mixing conditions. The interpolation model of Baldyga was used to predict the evolution of the species. Good agreement was found between simulations and experiments for both the flow field and the reactive system

Experimental and numerical investigations of jet mixing in a multifunctional channel reactor: Passive and reactive systems

Mixing of two liquids in a new multifunctional channel reactor developed by AlfaLaval has been studied both experimentally and through computational fluid dynamics (CFD). As the channels are quite narrow the Reynolds numbers are low and the bulk of the channel is within the turbulent boundary layer. This makes accurate a priori predictions of the flowfield difficult and experimental validation necessary. Particle image velocimetry (PIV) was used to measure the flowfield, whereas planar laser-induced fluorescence was used for a scalar concentration field. CFD simulations were performed with the commercial software Fluent 5.5. Different turbulence models were tested and compared with PIV. The best predictions were obtained with a low Reynolds boundary layer k-ε turbulence model. Mixing of a passive tracer, including mean concentration and concentration variance, was calculated with the turbulent mixer model of Baldyga. A reactive system with diazo coupling between 1-naphthols, 2-naphthols and diazotized sulphanilic acid was studied both experimentally and theoretically due to its sensitivity to mixing conditions. The interpolation model of Baldyga was used to predict the evolution of the species. Good agreement was found between simulations and experiments for both the flow field and the reactive system