Identification of therapeutic targets and novel therapies for prostate cancer
Aim: The aim of this project is to evaluate the potential of interference with stress signaling pathways in prostate cancer (PCa) cells as a novel form of therapy. In that context, we aim to evaluate if MTHFD2 and HSF1 can function as therapeutic targets for prostate cancer, as well as test the efficacy of a small molecule stress signaling inhibitor as an agent that can block disease progression and drug resistance. In addition, we will also aim to elucidate novel regulators of endoplasmic reticulum stress (ER) pathways in PCa.
Approach: To test the validity of our hypothesis, that MTHFD2 and HSF1 may have key functions in PCa progression, we will establish xenograft tumors in nude mice and inhibit specific gene expression using nanoliposomal siRNA injection. To test the hypothesis that MKC8866 inhibits PCa progression and drug resistance, we will establish xenografts of human cancer cells in nude mice and treat the mice with MKC8866 either alone, or in combination with drugs used in the clinic in advanced PCa models. We will also use CRISPR-Cas9 edited PCa cells and knockout screen to identify synthetic lethality pathways to improve the utility of MKC8866 using the xenograft models. Tumor growth will be monitored with calipers and tissue samples will be collected for subsequent analysis.
Use of animals and application of 3R: This project will use 576 nude mice in total. The number and use of the animals are planned and designed strictly following the guidelines of 3R principles to minimize the use of animals. The experimental procedures may cause moderate and severe distress to the animals. To enhance the animal welfare and minimize the severity, enrichments in cages, detailed clinical/severity score sheet and necessary anesthesia will be applied.
Approach: To test the validity of our hypothesis, that MTHFD2 and HSF1 may have key functions in PCa progression, we will establish xenograft tumors in nude mice and inhibit specific gene expression using nanoliposomal siRNA injection. To test the hypothesis that MKC8866 inhibits PCa progression and drug resistance, we will establish xenografts of human cancer cells in nude mice and treat the mice with MKC8866 either alone, or in combination with drugs used in the clinic in advanced PCa models. We will also use CRISPR-Cas9 edited PCa cells and knockout screen to identify synthetic lethality pathways to improve the utility of MKC8866 using the xenograft models. Tumor growth will be monitored with calipers and tissue samples will be collected for subsequent analysis.
Use of animals and application of 3R: This project will use 576 nude mice in total. The number and use of the animals are planned and designed strictly following the guidelines of 3R principles to minimize the use of animals. The experimental procedures may cause moderate and severe distress to the animals. To enhance the animal welfare and minimize the severity, enrichments in cages, detailed clinical/severity score sheet and necessary anesthesia will be applied.