Targeted Therapy Combined with Immune Modulation Using Gold Nanoparticles for Treating Metastatic Colorectal Cancer
[Technical Report, Annual Report]
Regents of the University of Minnesota
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During this reporting period, we have established a reliable mouse model of CRC LSL-rtTA3shAPCLgr5-CrER. We bred these mice to create a colony of transgenic mice. When experimental mice are 8 weeks of age, they are treated with 4-hydroxytamofixen 4OHT and put on continuous doxycycline via drinking water to initiate tumor formation. We are able to readily detect tumor formation after 4-6 weeks via colonoscopy. We have also detected PD-L1 expression in colon polyps from these mice. We tested the ability of this peptide to be loaded into a MHCI molecule in splenocytes harvested from C57BL6 mice. The control SIINFEKL without a DNA oligo attached is detectable by this antibody, but unfortunately GFP-SIINFEKL added at the same concentration cannot be detected. We are trouble shooting this issue and seeking alternative approaches. We also designed siRNAs to each PVT1 and PD-L1 and successful knockdown of targets was confirmed by RT-qPCR, flow cytometry, andor Western blot. Of the siRNAs for each target, the two most potent siRNAs were selected and ordered with a propylthiol modification at the 3 end and attached to AuNPs. These AuNPs with siRNA attached were delivered to CT26 cells and knockdown of targets was confirmed in the same manner as described above. We are currently working on delivering siRNA-coated AuNPs to our mouse model of CRC.
- Medicine and Medical Research
- Genetic Engineering and Molecular Biology