6th Commonwealth Chemistry Posters

Main Objective:

CRISPR/Cas9-mediated silencing of PFK-1 allosteric activation using two independent approaches,

1)Knocking out the regulatory domain of PFK-1

2)Knocking out the PFK-2 by disrupting the PFKFB3 gene

to decrease the rate of glycolysis and proliferation of MCF-7 breast cancer cells, for designing a therapeutic tool kit to treat breast cancer

Warburg effect of cancer cells:

Cancer cells consume high glucose level compared to normal cells through “Aerobic glycolysis” even in the presence of high oxygen level and functioning Mitochondria

Conclusion:

This study aimed to disrupt the allosteric regulation of PFK-1 using two independent approaches- disruption of the PFKFB3 gene by targeting exon 2 and disruption of the regulatory domain of the PFK-1 enzyme by targeting exon 13 using specifically designed sgRNA- as they would allow healthy cells to maintain sufficient levels of ATP via the basal function of PFK-1 while significantly impacting energy-centric metabolism in cancer cells.

Future work:

•Detection of truncated protein

•Sequencing to verify the precise genome modification

•Proliferation assay and downstream analysis to investigate the effect of modified gene

Acknowledgement:

Undergraduate research funds, Department of Chemistry, University of Colombo

References:

1.Gatenby, R. A., & Gillies, R. J. (2004). Why do cancers have high aerobic glycolysis? Nature Reviews Cancer, 4(11), 891–899. https://doi.org/10.1038/nrc1478

2.Jiang, F., & Doudna, J. A. (2017). CRISPR–Cas9 Structures and Mechanisms. Annual Review of Biophysics, 46(1), 505–529. https://doi.org/10.1146/annurev-biophys-062215-010822

3.Lu, L., Chen, Y., & Zhu, Y. (2017). The molecular basis of targeting PFKFB3 as a therapeutic strategy against cancer. Oncotarget, 8(37), 62793–62802. https://doi.org/10.18632/oncotarget.19513

4.Luqmani, Y. A. (2005). Mechanisms of Drug Resistance in Cancer Chemotherapy. Medical Principles and Practice, 14(Suppl. 1), 35–48. https://doi.org/10.1159/000086183

5.Moreno-Sánchez, R., Rodríguez-Enríquez, S., Marín-Hernández, A., & Saavedra, E. (2007). Energy metabolism in tumor cells. The FEBS Journal, 274(6), 1393–1418. https://doi.org/10.1111/j.1742-4658.2007.056

6th Commonwealth Chemistry Posters

CRISPR/Cas9-mediated silencing of PFK-1 allosteric activation as a novel therapeutic approach for cancer

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