Investigation of novel interaction between microRNA let-7i-5p and.pptx

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1. Investigation of novel interaction between miR-let-7i-5p and HOXA9 gene in acute myeloid leukemia cells Name: Lim Wei Yue ID: 17207473/1 Matric Number: MIC190019 Supervisor: Dr. Shamsul Bin Mohd Zain 1

2. Outline 2

3. Acute myeloid leukemia Cancer of the myeloid line of blood cells, characterized by the rapid growth of abnormal cells that build up in the bone marrow and blood ( Acute Myeloid Leukemia (AML) in Children and Teens - Together , n.d.) Figure 1. Acute myeloid leukemia under microscope 3

4. Blood development ( National Cancer Institute, 2022 ) Figure 2. Blood development 4

5. 6 th most common cancer for male and 8 th for female ! Figure 3. Age-standardized incidence rate for ten common cancers by sex, Malaysia, 2012-2016 (Malaysian National Cancer Registry Report 2012-2016, 2019) 5

6. AML is the highest prevalence among all types of leukemia ! Figure 4. Leukemia types: Ages-specific incidence rate, males, Malaysia 2012-2016 Figure 5. Leukemia types: Ages-specific incidence rate, females, Malaysia 2012-2016 (Malaysian National Cancer Registry Report 2012-2016, 2019) 6

7. MiRNA as cancer regulator (O’Brien et al., 2018) Figure 6. Mature miRNA interact with 3’ untranslated region of target mRNA 7

8. miR-let-7i-5p ( Song et al ., 2018 ) 8

9. ( Esa et al ., 2021 ) 9

10. HOXA9 (Li et al., 2013b) ( Aryal et al., 2021) Figure 7. HOXA9 expression in AML 10

11. Problem statement Lack of a less invasive molecular diagnostic biomarker that can refine the risk of patients with acute myeloid leukemia 11

12. Research Hypothesis miR-let-7i-5p downregulates HOXA9 gene expression, thus suppress the cell proliferation of Acute Myeloid Leukemia 12

13. Research Objectives 13

14. Methodology MV-4-11 THP-1 14

15. To determine the interaction between miR-let-7i-5p and HOXA9 gene expression. Cell culture MV-4-11 (B-myelomonocytic leukemia cell line) THP-1 (Acute monocytic leukemia cell line) (ATCC.org, 2022) 15

16. Transfection Let-7i-5p mimic Negative control Cell plating Transfection of miRNA 20 minutes 72 hours Harvest Lipofectamine 3000 16

17. Data collection qPCR cDNA synthesis RNA Extraction To measure the post-transfection level of miR-let-7i-5p and HOXA9 17

18. To determine effect of miR-let-7i-5p on cell proliferation. MTT assay Spectrophotometer Cell plating + transfection Incubation MTT test 18

19. To investigate the involvement of miR-let-7i-5p and HOXA9 in AML progression pathway using bioinformatics approach. 19

20. 20

21. Expected outcome 21

22. 22

23. References Acute Myeloid Leukemia (AML) in Children and Teens - Together . (n.d.). Retrieved October 23, 2022, from https://together.stjude.org/en-us/about-pediatric-cancer/types/leukemia/acute-myeloid-leukemia-aml.html Chen, K. T. J., Gilabert -Oriol, R., Bally, M. B., & Leung, A. W. Y. (2019). Recent Treatment Advances and the Role of Nanotechnology, Combination Products, and Immunotherapy in Changing the Therapeutic Landscape of Acute Myeloid Leukemia . Pharmaceutical Research , 36 (9). https://doi.org/10.1007/S11095-019-2654-Z Chen, Y., Chen, S., Lu, J., Yuan, D., He, L., Qin, P., Tan, H., & Xu, L. (2021). MicroRNA-363-3p promote the development of acute myeloid leukemia with RUNX1 mutation by targeting SPRYD4 and FNDC3B. Medicine , 100 (18), e25807. https://doi.org/10.1097/MD.0000000000025807 Collins, C. T., & Hess, J. L. (2016). Role of HOXA9 in leukemia : dysregulation, cofactors and essential targets. Oncogene , 35 (9), 1090. https://doi.org/10.1038/ONC.2015.174 Esa , E., Hashim, A. K., Mohamed, E. H. M., Zakaria, Z., Abu Hassan, A. N., Mat Yusoff , Y., Kamaluddin , N. R., Abdul Rahman, A. Z., Chang, K. M., Mohamed, R., Subbiah, I., Jamian , E., Ho, C. S. L., Lim, S. M., Lau, P. C., Pung , Y. F., & Zain, S. M. (2021). Construction of a microRNA-mRNA Regulatory Network in de Novo Cytogenetically Normal Acute Myeloid Leukemia Patients. Genetic Testing and Molecular Biomarkers , 25 (3), 199–210. https://doi.org/10.1089/gtmb.2020.0182 23

24. How does qPCR work (technology basics)? - BioSistemika . (n.d.). Retrieved October 25, 2022, from https://biosistemika.com/blog/qpcr-technology-basics/ Li, D. P., Li, Z. Y., Sang, W., Cheng, H., Pan, X. Y., & Xu, K. L. (2013a). HOXA9 Gene Expression in Acute Myeloid Leukemia . Cell Biochemistry and Biophysics 2013 67:3 , 67 (3), 935–938. https://doi.org/10.1007/S12013-013-9586-8 Li, D. P., Li, Z. Y., Sang, W., Cheng, H., Pan, X. Y., & Xu, K. L. (2013b). HOXA9 Gene Expression in Acute Myeloid Leukemia . Cell Biochemistry and Biophysics 2013 67:3 , 67 (3), 935–938. https://doi.org/10.1007/S12013-013-9586-8 Li, D. P., Li, Z. Y., Sang, W., Cheng, H., Pan, X. Y., & Xu, K. L. (2013c). HOXA9 gene expression in acute myeloid leukemia . Cell Biochemistry and Biophysics , 67 (3), 935–938. https://doi.org/10.1007/S12013-013-9586-8 Liu, Z., Zhong, L., Dan, W., Chu, X., Liu, C., Luo, X., Zhang, Z., Lu, Y., Wan, P., Wang, X., & Liu, B. (2022). miRNA-222-3p enhances the proliferation and suppresses the apoptosis of acute myeloid leukemia cells by targeting Axin2 and modulating the Wnt /β-catenin pathway. Biochemical and Biophysical Research Communications , 620 , 83–91. https://doi.org/10.1016/J.BBRC.2022.06.054 24

25. Most commonly found mutations in AML. | Download High-Quality Scientific Diagram . (n.d.). Retrieved October 23, 2022, from https://www.researchgate.net/figure/Most-commonly-found-mutations-in-AML_fig1_333989149 MV-4-11 - CRL-9591 | ATCC . (n.d.). Retrieved October 25, 2022, from https://www.atcc.org/products/crl-9591 O’Brien, J., Hayder , H., Zayed, Y., & Peng, C. (2018). Overview of microRNA biogenesis, mechanisms of actions, and circulation. Frontiers in Endocrinology, 9(AUG), 402. https://doi.org/10.3389/FENDO.2018.00402/BIBTEX Purification of RNA from cells and tissues— NucleoSpin RNA Plus . (n.d.). Retrieved October 25, 2022, from https://www.takarabio.com/products/nucleic-acid-purification/rna-purification-kits/total-rna-from-cells-and-tissues/nucleospin-rna-plus Song, J., Wang, L., Ma, Q., Yang, Y., Yang, Z., Wang, B., & He, N. (2018). Let-7i-5p inhibits the proliferation and metastasis of colon cancer cells by targeting kallikrein-related peptidase 6. Oncology Reports, 40(3), 1459–1466. https://doi.org/10.3892/OR.2018.6577/HTML 25

26. I wish to acknowledge Dr. Shamsul Bin Mohd Zain for supervising & Farhan Syafiq for guiding. Acknowledgements 26