Targeting therapeutic vulnerabilities associated with EWS fusion proteins in Ewing sarcoma

Ewing sarcoma (ES) is a are and aggressive bone tumour affecting children and young adults and that requires better therapeutic options to improve patient outcomes. ES is characterised by chromosomal rearrangements producing a fusion gene, the most predominant occurring between EWSR1 and FLI1 (85%). Recent evidence shows that the chimeric oncoprotein EWS-FLI recruits chromatin remodellers that epigenetically rewire transcription to establish its oncogenic programme. Additionally, transcriptional dysregulation is known to induce replication stress (RS) and genomic instability. To mitigate potential genotoxic damage, ES cells are particularly dependent on the replication stress response (RSR). Based on these EWS-FLI1-specific molecular effects, this thesis investigates two separate therapeutic strategies: (i) inhibition of the epigenetic modifier KDM1A, and (ii) exploiting the dependency on the RSR. Catalytic inhibition of histone demethylase KDM1A is demonstrated to be insufficient as a therapeutic strategy for ES, although roles beyond its demethylase function remain a possibility. To identify therapeutic combinations targeting the dependency on the RSR, clinically available drugs inhibiting the ATR-CHK1-WEE1 axis were tested in 3D spheroids of ES cell lines. Each drug candidate was combined at clinically relevant doses with SN-38, the active metabolite of topoisomerase I inhibitor irinotecan, currently used to treat relapsed ES. Combinations revealed cytotoxicity and decreased growth in ES spheroids following WEE1 and ATR inhibition, both concurrent with SN-38. Based on the strength of responses, further investigations prioritised the effects of the WEE1 inhibitor AZD1775 combined with SN-38 in additional ES cell lines and a model ectopically expressing EWS-FLI1. DNA damage, apoptosis, and cell cycle analysis uncovered two responses in ES cell lines, one characterised by cell death, the other resembling growth arrest. These may be dependent on the cell lines' mutational background and could act as a predictive biomarker. Taken together these findings identify a promising novel therapeutic strategy for ES

Targeting endothelial cells to improve cancer therapy.

PhD Medical.Antiangiogenic strategies have not provided the cancer control effects predicted from pre-clinical experiments. Obstacles include increasing hypoxia, reducing chemotherapy delivery and acquired resistance. Novel strategies to overcome these obstacles are thus warranted. In particular, exploiting endothelial cell-derived paracrine signalling (angiocrine signalling), manipulating blood vessel numbers and function whilst pursuing options of enhancing blood flow, chemo-delivery and reducing hypoxia become attractive possibilities. My PhD explores two such strategies: Targeting angiocrine signalling to enhance chemosensitivity Our laboratory's previously published work demonstrated that deletion of endothelial cell focal adhesion kinase (FAK) in mice with established subcutaneous tumour sensitised tumour cells to doxorubicin (Tavora, Reynolds, et al. 2014). This was through the alteration of endothelial cell signalling, known as angiocrine signalling. My experiments use pdgfb-iCreert;FAKfl/fl;R26K454R/K454R mice, where kinase-dead endothelial cell FAK is expressed in tumour blood vessels in vivo. My results demonstrate that loss of EC-FAK kinase activity had no impact on the tumour growth of PBS treated, established subcutaneous B16F0 melanomas but was sufficient to reduce tumour growth in doxorubicin treated animals. This change in tumour growth occurs with no apparent effect on blood vessel density or doxorubicin delivery but reduced perivascular tumour cell proliferation and increased DNA damage and apoptotic area relative to blood vessel number in vivo. Additionally, loss of EC-FAK kinase activity and pharmacological FAK inhibition were both sufficient to affect doxorubicin-induced cytokine production in vitro. The results suggest that EC-FAK kinase activity is involved in altering cytokine production and in modulating chemosensitisation of tumour cells. Testing the effect of a low dose of orally available vascular promotion agent 29P in enhancing chemosensitisation in pancreatic ductal adenocarcinoma. Our laboratory previously published that increasing blood vessel density by treating mice with intraperitoneal injections of low doses of the cyclic Arg-Gly-Asp (RGD)-mimetic, Cilengitide, was sufficient to increase gemcitabine delivery and decrease hypoxia, thus reducing tumour growth in mouse models of pancreatic ductal adenocarcinoma (Wong et al. 2015). Here I tested the utility of a similar RGD-peptide 29P that has been designed to be administered orally. I have carried out pilot in vivo experiments in various models 6 of PDAC, but due to technical difficulties, many of these data remain inconclusive. As part of dealing with these technical difficulties, I have also established a protocol for mouse pancreatic ductal adenocarcinom

Medical-Data-Models.org:A collection of freely available forms (September 2016)

MDM-Portal (Medical Data-Models) is a meta-data repository for creating, analysing, sharing and reusing medical forms, developed by the Institute of Medical Informatics, University of Muenster in Germany. Electronic forms for documentation of patient data are an integral part within the workflow of physicians. A huge amount of data is collected either through routine documentation forms (EHRs) for electronic health records or as case report forms (CRFs) for clinical trials. This raises major scientific challenges for health care, since different health information systems are not necessarily compatible with each other and thus information exchange of structured data is hampered. Software vendors provide a variety of individual documentation forms according to their standard contracts, which function as isolated applications. Furthermore, free availability of those forms is rarely the case. Currently less than 5 % of medical forms are freely accessible. Based on this lack of transparency harmonization of data models in health care is extremely cumbersome, thus work and know-how of completed clinical trials and routine documentation in hospitals are hard to be re-used. The MDM-Portal serves as an infrastructure for academic (non-commercial) medical research to contribute a solution to this problem. It already contains more than 4,000 system-independent forms (CDISC ODM Format, www.cdisc.org, Operational Data Model) with more than 380,000 dataelements. This enables researchers to view, discuss, download and export forms in most common technical formats such as PDF, CSV, Excel, SQL, SPSS, R, etc. A growing user community will lead to a growing database of medical forms. In this matter, we would like to encourage all medical researchers to register and add forms and discuss existing forms