Preclinical study of Pancreatic Cancer sonoporation in patient derived samples and study of the role of the immune system.
The purpose of this experiment is: to determine the efficacy of ultrasound- and microbubble- guided drug delivery (sonoporation) in patient derived orthotopic and heterotopic xenograft models of pancreatic ductal adenocarcinoma (PDAC) both in immunedeficient and immuneproficient mouse models.
Anticipated adverse effects on the animals: Our planned procedures can be classified as procedures of moderate severity. For the heterotopic models engraftment will be performed under a minor surgical procedure and development of disease will be limited to the subcutaneous compartment. The primary distress that the animals may undergo throughout the treatment duration is ruffled fur and weight loss. For the orthotopic models a minimal abdominal incision will be performed to ensure fast recovery and analgesia will be applied to minimise pain suffering. Locoregional and distant metastasis are expected to occur with the most severe complication being ascites.
The expected scientific benefits or benefits for society:
In these experiments we aim to confirm the higher effectiveness (in comparison to current standards of care) of ultrasound- and microbubble- guided drug delivery suggested by our previous application (nr. 9059). In addition, the usage of patient derived material and the evaluation of the immune system will allow us to identify putative biomarkers of response to this kind of therapy.
Survival from PDAC is only anecdotal, with a 5-year survival rate of only 3%. Effectiveness of novel therapeutic modalities and identification of biomarkers of response will have a high impact in patients survival.
Number of animals
In total 948 animals will be used. 84 from C57BL/6J strain and 864 from NSG strain.
How will the requirements for 3R be accomplished by the experiment:
Replacement
We are working in setting up a bioreactor system to grow pancreatic cancer organoids so that the next steps of the project can be performed ex vivo. However, this system is in its early stages and it is not expected to be fully functional until the end of 2019. As soon as this system is fully functional it will produce a large reduction in the number of animals used in research for this project.
Reduction
By the usage of prestablished patient derived xenograft models through a collaboration with the Catalan Institute of Oncology and Oslo Hospital. the number of animals required for model generation will be reduced, allowing to focus the majority of the animal use for actual efficacy studies.
Refinement
Follow up of orthotopic PDAC models has been hampered by the deep location of the tumors in the abdominal cavity, difficulting early detection of tumoral volumes and forcing mice to undergo long courses of disease before treatment efficacy could be detected (tumoral volumes higher than 1500 mm3). The application of ultrasound imaging will allow the evaluation of tumoral volumes through the course of treatment cycles allowing to decrease the length of the disease course required to detect treatment efficacy. Tumoral volumes will be limited to 800 mm3.
Experienced technicians and veterinarians will monitor the animals and they will very early discover any sign of illness and take the decision of euthanasia.
Anticipated adverse effects on the animals: Our planned procedures can be classified as procedures of moderate severity. For the heterotopic models engraftment will be performed under a minor surgical procedure and development of disease will be limited to the subcutaneous compartment. The primary distress that the animals may undergo throughout the treatment duration is ruffled fur and weight loss. For the orthotopic models a minimal abdominal incision will be performed to ensure fast recovery and analgesia will be applied to minimise pain suffering. Locoregional and distant metastasis are expected to occur with the most severe complication being ascites.
The expected scientific benefits or benefits for society:
In these experiments we aim to confirm the higher effectiveness (in comparison to current standards of care) of ultrasound- and microbubble- guided drug delivery suggested by our previous application (nr. 9059). In addition, the usage of patient derived material and the evaluation of the immune system will allow us to identify putative biomarkers of response to this kind of therapy.
Survival from PDAC is only anecdotal, with a 5-year survival rate of only 3%. Effectiveness of novel therapeutic modalities and identification of biomarkers of response will have a high impact in patients survival.
Number of animals
In total 948 animals will be used. 84 from C57BL/6J strain and 864 from NSG strain.
How will the requirements for 3R be accomplished by the experiment:
Replacement
We are working in setting up a bioreactor system to grow pancreatic cancer organoids so that the next steps of the project can be performed ex vivo. However, this system is in its early stages and it is not expected to be fully functional until the end of 2019. As soon as this system is fully functional it will produce a large reduction in the number of animals used in research for this project.
Reduction
By the usage of prestablished patient derived xenograft models through a collaboration with the Catalan Institute of Oncology and Oslo Hospital. the number of animals required for model generation will be reduced, allowing to focus the majority of the animal use for actual efficacy studies.
Refinement
Follow up of orthotopic PDAC models has been hampered by the deep location of the tumors in the abdominal cavity, difficulting early detection of tumoral volumes and forcing mice to undergo long courses of disease before treatment efficacy could be detected (tumoral volumes higher than 1500 mm3). The application of ultrasound imaging will allow the evaluation of tumoral volumes through the course of treatment cycles allowing to decrease the length of the disease course required to detect treatment efficacy. Tumoral volumes will be limited to 800 mm3.
Experienced technicians and veterinarians will monitor the animals and they will very early discover any sign of illness and take the decision of euthanasia.