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Project Title:

Pre-clinical evaluation of Oct-1002 to control prostate cancer malignant progression
Supervisors:
Jenny Worthington, Paul Thompson
Contact Details:
Jenny Worthington
Paul Thompson
Transcriptional Regulation & Epigenetics Group
School of Biomedical Sciences
University of Ulster, Coleraine

Level:
PhD

Background to the project:
Androgen deprivation therapy (ADT) is commonly used to treat locally advanced prostate cancer
and although this treatment is initially successful, after a median of around 18 months a castrate resistant, more aggressive form of the disease emerges (CRPC). We and others have identified increased tumour hypoxia in LNCaP prostate cancer xenografts following treatment with ADT (Ming et al., 2012). Hypoxia is a common feature of solid tumours and drives malignant progression. Prostate tumours show very low median oxygen levels i.e.0.3 – 0.8% pO2 (2.4-6.8mmHg) (Movsas et al., 2001); this is extremely important since increased levels of hypoxia correlate with clinical staging and biochemical failure after radiotherapy. Additionally there is growing clinical evidence that hypoxia is a key driver of malignant progression in prostate cancer (Milosevic et al., 2012; Turkada et al., 2012). Hypoxia is thought to cause malignant progression through selection for hypoxia-tolerant stress-resistant cells; in addition to showing a more malignant phenotype, these cells frequently have an enhanced ability to metastasise. It therefore makes sense to target tumour hypoxia as a strategy to increase the efficacy of ADT. We have previously used the bioreductive drug AQ4N in combination with ADT and identified increased primary tumour control and a reduction in lung metastases using LNCaP tumours (Ming et al., 2012). These experiments provided ‘proof-of-principle’ that bioreductive drugs can play a role in prostate cancer treatment when used in combination with other chemotherapeutics. For this study we will use the novel bio-reductive drug Oct-1002 in combination with ADT for treatment of androgen-sensitive tumours and with docetaxel for treatment of castrate-resistant prostate cancer. We will also use vitamin D analogues to increase cytochrome levels within the tumours (Dr Thompsons group have identified increases in CYP 3A4 and 3A5 using this approach in vitro (Maguire et al., 2012). As these cytochromes are known to metabolise Oct-1002, this will increase metabolism of the bioreductive drug and increase associated cytotoxicity. Proof-of-principle of increasing bioreductive drug efficacy by increasing cytochrome expression has already been established (McCarthy et al., 2003). We will assess any therapeutic gain from this approach in vitro Methods to be used:
Cell culture

Initially the efficacy of Oct-1002 will be assessed in vitro using the spheroid model which contains a hypoxia core – this will give us a biologically relevant model to work with. Cell viability, clonogenicity invasion and migration will be measured following treatment with Oct-1002 and in combination (with bicalutamide for LNCaP and docetaxel for PC3). In vivo analysis
Tumour growth and metastases will be measured following in vivo treatment of animals bearing either LNCaP or PC3 tumours. Combination studies detailed above will also be carried out. Molecular techniques
RNA and protein will be extracted from Vitamin D analogue treated cells/tumours and the expression of CYP 3A4 and 3A5 measured using RT-Q-PCR and western blotting. Luciferase assays will also be employed to measure activity of liganded vitamin d receptor in the presence of the drugs used in this Objectives of the research:

a) Assess Oct-1002 efficacy in vitro and in vivo in androgen sensitive prostate cancer. b) Assess Oct-1002 efficacy in vitro and in vivo in castrate resistant prostate cancer. c) Use vitamin D analogues to manipulate cytochrome levels in prostate cancer cells d) Establish therapeutic gain of manipulating cytochrome P450 levels in Oct-1002 treated Skills required of applicant:
• Good Laboratory skills • Good oral and written presentation skills • Good critical thinking and analytical skills • Good IT skills • Good work ethic and ability to work independently A novel AKT inhibitor for prostate cancer. • Development and characterisation of prostate cancer spheroids.

Source: http://www.science.ulst.ac.uk/sites/gradschool/IMG/pdf/Pre_clinical_eval_of_oct_1002_to_control_prostrate_cancer_malignant_progression.pdf

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