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.
Etterevaluering
The outcome of genome-wide gene modification is unknown in the new cell line, 22Rv1.
It is possible that the cell line has a metastatic potential, and this could increase the severity of the experiments. The applicant has updated the score sheet for registering clinical signs, taking into account the possibility of metastatic growth.
Because of the change, we classify the project overall as severe, cf. the Regulation annex B.
The Norwegian Food Safety Authority must retrospectively assess all severe experiments. You are therefore required to submit necessary information making it possible for us to evaluate 1) whether the objectives of the project were achieved, 2) the harm inflicted on the animals, and 3) any elements that may contribute to further implementation of 3 R in future studies. This information must be submitted via FOTS.
It is possible that the cell line has a metastatic potential, and this could increase the severity of the experiments. The applicant has updated the score sheet for registering clinical signs, taking into account the possibility of metastatic growth.
Because of the change, we classify the project overall as severe, cf. the Regulation annex B.
The Norwegian Food Safety Authority must retrospectively assess all severe experiments. You are therefore required to submit necessary information making it possible for us to evaluate 1) whether the objectives of the project were achieved, 2) the harm inflicted on the animals, and 3) any elements that may contribute to further implementation of 3 R in future studies. This information must be submitted via FOTS.
Begrunnelse for etterevalueringen
The aim of this research project has been to evaluate the potential of interfering with stress signaling pathways in prostate cancer cells as a novel form of therapy.
A total of 230 mice have been reported as utilized in the project, which was approved for the use of up to 572 mice with a moderate severity.
Two applications for changes have been approved in the project period. In connection with the changes, a revision of the severity level was made, and the original score sheet was replaced with a new, better adapted score sheet.
Due to the variety of experiments within this project, the actual severity scores can vary significantly. The responsible researcher states that the procedures for surveillance could be more flexible according to the conditions of individual animals.
There has been a close cooperation with the animal facility staff regarding scoring of clinical signs, mitigating measures and humane endpoints.
Important mitigating measures have been addition of soft bedding material, increased surveillance frequency and timely euthanasia. The humane endpoints were determined through a thorough discussion with local evaluation personnel.
Severity was categorized as severe for 38 mice with a substantial tumor burden and the occurrence of necrosis within the tumor.
163 mice have been reported with moderate severity, and 29 with mild severity.
There is recent development of ex vivo organoid models for assessment of cancer therapeutics and basic studies, which could partially reduce the use of animals.
The responsible researcher states that the group have successfully assessed the major parts of their goals, which led to validation of novel targets for prostate cancer therapy and some novel findings in the aspect of mechanisms. However, due to time constraints, some experiments were not completed as planned.
The research group have validated the targetability of certain targets, providing a solid basis for further investigation, and have launched a follow-up project (FOTS#30531).
A total of 230 mice have been reported as utilized in the project, which was approved for the use of up to 572 mice with a moderate severity.
Two applications for changes have been approved in the project period. In connection with the changes, a revision of the severity level was made, and the original score sheet was replaced with a new, better adapted score sheet.
Due to the variety of experiments within this project, the actual severity scores can vary significantly. The responsible researcher states that the procedures for surveillance could be more flexible according to the conditions of individual animals.
There has been a close cooperation with the animal facility staff regarding scoring of clinical signs, mitigating measures and humane endpoints.
Important mitigating measures have been addition of soft bedding material, increased surveillance frequency and timely euthanasia. The humane endpoints were determined through a thorough discussion with local evaluation personnel.
Severity was categorized as severe for 38 mice with a substantial tumor burden and the occurrence of necrosis within the tumor.
163 mice have been reported with moderate severity, and 29 with mild severity.
There is recent development of ex vivo organoid models for assessment of cancer therapeutics and basic studies, which could partially reduce the use of animals.
The responsible researcher states that the group have successfully assessed the major parts of their goals, which led to validation of novel targets for prostate cancer therapy and some novel findings in the aspect of mechanisms. However, due to time constraints, some experiments were not completed as planned.
The research group have validated the targetability of certain targets, providing a solid basis for further investigation, and have launched a follow-up project (FOTS#30531).