Animated Visualization of FOXO3a and SIRT1 Signaling Pathways: Developing Independent Learning Media for Biology Undergraduate Students

Indi Mariana

Authors

Indi Mariana PT. Briliant Knowledge Nexus https://orcid.org/0000-0002-4866-555X

Keywords:

Animated Visualization, FOXO3a, SIRT1, Independent Learning, Molecular Biology Education, Sarcopenic Obesity

Abstract

The complexity of molecular signaling pathways, such as those involving FOXO3a and SIRT1 in sarcopenic obesity, often poses significant conceptual challenges for biology students, necessitating more dynamic instructional tools. This study aims to develop and validate an animated visualization media designed for independent learning to enhance students' understanding of intracellular mechanisms. Employing the Research and Development (R&D) method with the ADDIE model (Analysis, Design, Development, Implementation, Evaluation), the research involved material and media experts for validation, followed by a small-group trial with biology students. The results indicate that the animated media is highly feasible, with expert validation scores exceeding 85%, and students reporting a significant improvement in visualizing the interaction between SIRT1-mediated energy metabolism and FOXO3a-regulated muscle atrophy. Furthermore, the implementation of this media fostered greater learning autonomy compared to conventional static diagrams. In conclusion, the developed animated visualization effectively bridges the gap between complex proteomic data and pedagogical clarity, making it a robust tool for modern biology education. This media provides a practical framework for integrating high-impact biological research into undergraduate curricula to support transformative learning experiences.

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References

Almohaimede, A. A., Bendahmash, M. W., Dhafr, F. M., Awwad, A. F., & Al-Madi, E. M. (2020). Knowledge, Attitude, and Practice (KAP) of Radiographic Protection by Dental Undergraduate and Endodontic Postgraduate Students, General Practitioners, and Endodontists. International Journal of Dentistry, 2020. https://doi.org/10.1155/2020/2728949

Aydin, A., Gürsoy, A., & Karal, H. (2023). Mobile care app development process: using the ADDIE model to manage symptoms after breast cancer surgery (step 1). Discover Oncology, 14(1). https://doi.org/10.1007/s12672-023-00676-5

Bahri, A., Idris, I. S., Muis, H., Arifuddin, M., & Fikri, M. J. N. (2020). Blended Learning Integrated with Innovative Learning Strategy to Improve Self-Regulated Learning. International Journal of Instruction, 14(1), 779 – 794. https://doi.org/10.29333/IJI.2021.14147A

Bawaneh, A. K. (2021). The Satisfaction Level Of Undergraduate Science Students Towards Using E-Learning And Virtual Classes In Exceptional Condition Covid-19 Crisis. Turkish Online Journal of Distance Education, 22(1), 52–65. https://doi.org/10.17718/TOJDE.849882

Beltran, R. S., Marnocha, E., Race, A., Croll, D. A., Dayton, G. H., & Zavaleta, E. S. (2020). Field courses narrow demographic achievement gaps in ecology and evolutionary biology. Ecology and Evolution, 10(12), 5184–5196. https://doi.org/10.1002/ece3.6300

Bindschadler, M., Buddhe, S., Ferguson, M. R., Jones, T., Friedman, S. D., & Otto, R. K. (2022). HEARTBEAT4D: An Open-source Toolbox for Turning 4D Cardiac CT into VR/AR. Journal of Digital Imaging, 35(6), 1759–1767. https://doi.org/10.1007/s10278-022-00659-y

Bogucka, D., Wajda, A., Stypińska, B., Radkowski, M. J., Targowski, T., Modzelewska, E., Kmiołek, T., Ejma-Multański, A., Filipowicz, G., Kaliberda, Y., Dudek, E., & Paradowska-Gorycka, A. (2025). Epigenetic factors and inflammaging: FOXO3A as a potential biomarker of sarcopenia and upregulation of DNMT3A and SIRT3 in older adults. Frontiers in Immunology, 16. https://doi.org/10.3389/fimmu.2025.1467308

Bubou, G. M., & Job, G. C. (2022). Individual innovativeness, self-efficacy and e-learning readiness of students of Yenagoa study centre, National Open University of Nigeria. Journal of Research in Innovative Teaching and Learning, 15(1), 2–22. https://doi.org/10.1108/JRIT-12-2019-0079

Chandrasekaran, A. R. (2020). Transitioning undergraduate research from wet lab to the virtual in the wake of a pandemic. Biochemistry and Molecular Biology Education, 48(5), 436–438. https://doi.org/10.1002/bmb.21386

Chen, H., Hu, X., Yang, R., Wu, G., Tan, Q., Goltzman, D., & Miao, D. (2020). SIRT1/FOXO3a axis plays an important role in the prevention of mandibular bone loss induced by 1,25(OH)2D deficiency. International Journal of Biological Sciences, 16(14), 2712–2726. https://doi.org/10.7150/ijbs.48169

Ding, H., Liu, D., He, J., Zhou, D., Wang, C., Yang, C., & Xia, Z. (2025). The Role of the Sirt1/Foxo3a Pathway in Mitigating Myocardial Ischemia–Reperfusion Injury by Dexmedetomidine. Chemical Biology and Drug Design, 105(4). https://doi.org/10.1111/cbdd.70100

Du, H., Huangfu, W., Liu, Z., Jia, G., Zhao, F., & Cheng, W. (2024). 5-Demethylnobiletin Ameliorates Isoproterenol-Induced Cardiac Fibrosis and Apoptosis by Repressing the Sirt1/FOXO3a/NF-κB and Wnt/β-Catenin Pathways. Biological and Pharmaceutical Bulletin, 47(10), 1774–1785. https://doi.org/10.1248/bpb.b24-00122

Duan, H., Song, W., Deng, J., Han, Y., Lu, F., Wang, Z., He, Y., & Wu, C. (2025). Muscone ameliorates acute hypobaric hypoxia-induced myocardial injury in rats via regulating the SIRT1/FOXO3a pathway. Phytomedicine, 148. https://doi.org/10.1016/j.phymed.2025.157442

Durden, J. M., Hosking, B., Bett, B. J., Cline, D., & Ruhl, H. A. (2021). Automated classification of fauna in seabed photographs: The impact of training and validation dataset size, with considerations for the class imbalance. Progress in Oceanography, 196. https://doi.org/10.1016/j.pocean.2021.102612

Elmorsy, E. A., Youssef, M. E., Abdel-Hamed, M. R., Amer, M. M., Elghandour, S. R., Alkhamiss, A. S., Mohamed, N. B., Khodeir, M. M., Elsisi, H. A., Alsaeed, T. S., Kamal, M. M., Ellethy, A. T., Elesawy, B. H., & Saber, S. (2024). Activation of AMPK/SIRT1/FOXO3a signaling by BMS-477118 (saxagliptin) mitigates chronic colitis in rats: uncovering new anti-inflammatory and antifibrotic roles. Frontiers in Pharmacology, 15. https://doi.org/10.3389/fphar.2024.1456058

Fu, D., Hu, Z., Xu, X., Dai, X., & Liu, Z. (2022). Key signal transduction pathways and crosstalk in cancer: Biological and therapeutic opportunities. Translational Oncology, 26. https://doi.org/10.1016/j.tranon.2022.101510

Hanafi, Y., Murtadho, N. M., Ikhsan, A., & Diyana, T. N. (2020). Reinforcing public university student’s worship education by developing and implementing mobile-learning management system in the ADDIE instructional design model. International Journal of Interactive Mobile Technologies, 14(2), 215 – 241. https://doi.org/10.3991/ijim.v14i02.11380

Haristiani, N., & Rifai, M. M. (2021). Chatbot-based application development and implementation as an autonomous language learning medium. Indonesian Journal of Science and Technology, 6(3), 561 – 576. https://doi.org/10.17509/ijost.v6i3.39150

Hu, M., Shu, X., Yu, G., Wu, X., Välimäki, M., & Feng, H. (2021). A risk prediction model based on machine learning for cognitive impairment among chinese community-dwelling elderly people with normal cognition: Development and validation study. Journal of Medical Internet Research, 23(2). https://doi.org/10.2196/20298

Jang, E. H., Lee, J. H., & Kim, S. A. (2021). Acute valproate exposure induces mitochondrial biogenesis and autophagy with foxo3a modulation in sh‐sy5y cells. Cells, 10(10). https://doi.org/10.3390/cells10102522

Jie, Z., Dalal, S., Bakar, M. S. B. A., & Yahaya, W. A. W. (2025). Cultural Spectacle and Media Interference: Scientific Aesthetic and Humanistic Meditation in Interactive Animation. Studies in Systems, Decision and Control, 291, 385–395. https://doi.org/10.1007/978-3-031-87372-0_35

Junk, P., & Kiel, C. (2021). Engineering of Biological Pathways: Complex Formation and Signal Transduction. Methods in Molecular Biology, 2315, 59–70. https://doi.org/10.1007/978-1-0716-1468-6_4

Kalleny, N. (2020). Advantages of Kahoot! Game-based formative assessments along with methods of its use and application during the COVID-19 Pandemic in various live learning sessions. Journal of Microscopy and Ultrastructure, 8(4), 175–185. https://doi.org/10.4103/JMAU.JMAU_61_20

Kasem, E. A., Hamza, G., El-Shafai, N. M., Ghanem, N. F., Mahmoud, S., Sayed, S. M., Alshehri, M. A., Al-Shuraym, L. A., Ghamry, H. I., Mahfouz, M. E., & Shukry, M. (2025). Thymoquinone-Loaded Chitosan Nanoparticles Combat Testicular Aging and Oxidative Stress Through SIRT1/FOXO3a Activation: An In Vivo and In Vitro Study. Pharmaceutics, 17(2). https://doi.org/10.3390/pharmaceutics17020210

Kavadella, A., Da Silva, M. A. D., Kaklamanos, E. G., Stamatopoulos, V., & Giannakopoulos, K. (2024). Evaluation of ChatGPT’s Real-Life Implementation in Undergraduate Dental Education: Mixed Methods Study. JMIR Medical Education, 10(1). https://doi.org/10.2196/51344

Kavanaugh, L. G., Dey, D., Shafer, W. M., & Conn, G. L. (2024). Structural and functional diversity of Resistance-Nodulation-Division (RND) efflux pump transporters with implications for antimicrobial resistance. Microbiology and Molecular Biology Reviews, 88(3). https://doi.org/10.1128/mmbr.00089-23

Kim, S. M., Suh, H. J., Lee, W. C., Choi, Y., & Chang, Y. B. (2025). Amino acid (Leu, Gln, Cys)–EGCG mixture mitigates dexamethasone-induced muscle atrophy via PI3K-Akt-mTOR and SIRT1-PGC1α pathway regulation. Food Bioscience, 73. https://doi.org/10.1016/j.fbio.2025.107665

Lee, G. G., Mun, S., Shin, M. K., & Zhai, X. (2025). Collaborative Learning with Artificial Intelligence Speakers: Pre-service Elementary Science Teachers’ Responses to the Prototype. Science and Education, 34(2), 847–875. https://doi.org/10.1007/s11191-024-00526-y

Li, H., Chou, P., Du, F., Sun, L., Liu, J., & Wang, W. (2021). Depleting microRNA-183-3p improves renal tubulointerstitial fibrosis after acute kidney injury via SIRT1/PUMA/FOXO3a deacetylation. Life Sciences, 269. https://doi.org/10.1016/j.lfs.2021.119017

Lin, C., Zeng, J., Zhang, S., Xu, X., Chen, L., Yang, Z., Wu, W., Hu, C., & Zhao, Y. T. (2024). Remedial effects of tilapia skin peptides against dexamethasone-induced muscle atrophy in mice by modulation of AKT/FOXO3a and Sirt1/PGC-1α signaling pathways. Journal of Functional Foods, 113. https://doi.org/10.1016/j.jff.2023.105954

Liu, C., Wong, P. Y., Wang, Q., Wong, H. Y., Huang, T., Cui, C., Zhang, N., Cheung, W. H., & Wong, R. M. Y. (2024). Short-chain fatty acids enhance muscle mass and function through the activation of mTOR signalling pathways in sarcopenic mice. Journal of Cachexia, Sarcopenia and Muscle, 15(6), 2387–2401. https://doi.org/10.1002/jcsm.13573

Ma, B., Zhu, Z., Zhang, J., Ren, C., & Zhang, Q. (2020). Aucubin alleviates diabetic nephropathy by inhibiting NF-κB activation and inducing SIRT1/SIRT3-FOXO3a signaling pathway in high-fat diet/streptozotocin-induced diabetic mice. Journal of Functional Foods, 64. https://doi.org/10.1016/j.jff.2019.103702

Martin, A., Rechs, A., Landerholm, T., & McDonald, K. (2021). Course-Based Undergraduate Research Experiences Spanning Two Semesters of Biology Impact Student Self-Efficacy but not Future Goals. Journal of College Science Teaching, 50(4), 33–47. https://doi.org/10.1080/0047231X.2021.12290516

McDonald, A. R., Roberts, R., Koeppe, J. R., & Hall, B. L. (2022). Undergraduate structural biology education: A shift from users to developers of computation and simulation tools. Current Opinion in Structural Biology, 72, 39–45. https://doi.org/10.1016/j.sbi.2021.07.012

Meng, F., Zhang, Z., Chen, C., Liu, Y., Yuan, D., Hei, Z., & Luo, G. (2021). PI3K/AKT activation attenuates acute kidney injury following liver transplantation by inducing FoxO3a nuclear export and deacetylation. Life Sciences, 272. https://doi.org/10.1016/j.lfs.2021.119119

Panagopoulos, D. J. (2022). Electromagnetic Fields of Wireless Communications: Biological and Health Effects. Electromagnetic Fields of Wireless Communications: Biological and Health Effects, 1–532. https://doi.org/10.1201/9781003201052

Pandaram, A., Paul, J., Wankhar, W., Thakur, A., Verma, S., Vasudevan, K., Wankhar, D., Kammala, A. K., Sharma, P., Jaganathan, R., Iyaswamy, A., & Rajan, R. (2024). Aspartame Causes Developmental Defects and Teratogenicity in Zebra Fish Embryo: Role of Impaired SIRT1/FOXO3a Axis in Neuron Cells. Biomedicines, 12(4). https://doi.org/10.3390/biomedicines12040855

Peng, F., Huang, X., Shi, W., Xiao, Y., Jin, Q., Li, L., Xu, D., & Wu, L. (2021). 5,7,3′,4′-tetramethoxyflavone ameliorates cholesterol dysregulation by mediating SIRT1/FOXO3a/ABCA1 signaling in osteoarthritis chondrocytes. Future Medicinal Chemistry, 13(24), 2153–2166. https://doi.org/10.4155/fmc-2021-0247

Qin, H., Zhang, H., Zhang, X., Zhang, S., Zhu, S., & Wang, H. (2021). Resveratrol attenuates radiation enteritis through the SIRT1/FOXO3a and PI3K/AKT signaling pathways. Biochemical and Biophysical Research Communications, 554, 199–205. https://doi.org/10.1016/j.bbrc.2021.03.122

Qiu, C. W., Chen, B., Zhu, H. F., Liang, Y. L., & Mao, L. S. (2024). Gastrodin alleviates cisplatin nephrotoxicity by inhibiting ferroptosis via the SIRT1/FOXO3A/GPX4 signaling pathway. Journal of Ethnopharmacology, 319. https://doi.org/10.1016/j.jep.2023.117282

Rahmayanti, H., Oktaviani, V., & Syani, Y. (2020). Development of sorting waste game android based for early childhood in environmental education. Journal of Physics: Conference Series, 1434(1). https://doi.org/10.1088/1742-6596/1434/1/012029

Retraction: Involvement of FOXO Transcription Factors, TRAIL-FasL/Fas, and Sirtuin Proteins Family in Canine Coronavirus Type II-Induced Apoptosis (PLoS One. (2011) 6:11 (e27313) DOI: 10.1371/journal.pone.0027313). (2025). PLoS ONE, 20(5 May). https://doi.org/10.1371/journal.pone.0324091

Ristanto, R. H., Rusdi, Mahardika, R. D., Darmawan, E., & Ismirawati, N. (2020). Digital Flipbook Imunopedia (DFI) A Development in Immune System e-Learning Media. International Journal of Interactive Mobile Technologies, 14(19), 140 – 162. https://doi.org/10.3991/ijim.v14i19.16795

Rizal, R., Rusdiana, D., Setiawan, W., & Siahaan, P. (2021). Development of a problem-based learning management system-supported smartphone (PBLMS3) application using the ADDIE model to improve digital literacy. International Journal of Learning, Teaching and Educational Research, 20(11), 115–131. https://doi.org/10.26803/ijlter.20.11.7

Rizal, R., Rusdiana, D., Setiawan, W., & Siahaan, P. (2022). Learning Management System Supported Smartphone (Lms3): Online Learning Application in Physics for School Course To Enhance Digital Literacy of Pre-Service Physics Teachers. Journal of Technology and Science Education, 12(1), 191–203. https://doi.org/10.3926/JOTSE.1049

Şahin, B., Bural, İ. B., Özcan, H., Yildirim, S., Tanriöver, G., & Yoldaş, Ş. B. (2026). Association of miR-34a with metastatic progression through the FoxO3a–SIRT1 regulatory axis in breast cancer. Turkish Journal of Medical Sciences, 56(1), 326–332. https://doi.org/10.55730/1300-0144.6166

Setiyani, Waluya, S. B., Sukestiyarno, Y. L., & Cahyono, A. N. (2022). E-Module Design Using Kvisoft Flipbook Application Based on Mathematics Creative Thinking Ability for Junior High Schools. International Journal of Interactive Mobile Technologies, 16(4), 116 – 136. https://doi.org/10.3991/ijim.v16i04.25329

Shirpoor, A., & Naderi, R. (2022). Maternal Ethanol Exposure-Induced Cardiac Fibrosis is Associated with Changes in TGF-β and SIRT1/FOXO3a Signaling in Male Rat Offspring: A Three-Month Follow-up Study. Cardiovascular Toxicology, 22(9), 858–865. https://doi.org/10.1007/s12012-022-09761-7

Song, T., Pabst, F., Eck, U., & Navab, N. (2025). Enhancing Patient Acceptance of Robotic Ultrasound through Conversational Virtual Agent and Immersive Visualizations. IEEE Transactions on Visualization and Computer Graphics, 31(5), 2901–2911. https://doi.org/10.1109/TVCG.2025.3549181

Spatioti, A. G., Kazanidis, I., & Pange, J. (2022). A Comparative Study of the ADDIE Instructional Design Model in Distance Education. Information (Switzerland), 13(9). https://doi.org/10.3390/info13090402

Sun, D. M., Yao, J. J., Dong, Z. L., Qian, J., Huang, Q. F., Sun, Y. T., & Liu, X. R. (2021). Effects of Sirt1 on proliferation, migration, and apoptosis of endothelial progenitor cells in peripheral blood of SD rats with chronic obstructive pulmonary disease. Asian Pacific Journal of Tropical Biomedicine, 11(10), 429–439. https://doi.org/10.4103/2221-1691.326097

Sun, L., Wang, H., Meng, N., Wang, Z., Li, G., Jiao, X., & Wang, J. (2023). Distribution and Spread of the Mobilized RND Efflux Pump Gene Cluster tmexCD-toprJ in Klebsiella pneumoniae from Different Sources . Microbiology Spectrum, 11(4). https://doi.org/10.1128/spectrum.05364-22

Suprapto, N., Ibisono, H. S., & Mubarok, H. (2021). THE USE OF PHYSICS POCKETBOOK BASED ON AUGMENTED REALITY ON PLANETARY MOTION TO IMPROVE STUDENTS’ LEARNING ACHIEVEMENT. Journal of Technology and Science Education, 11(2), 526 – 540. https://doi.org/10.3926/jotse.1167

Tang, H., Du, H., Kuang, X., Huang, H., Zeng, J., Long, C., Zhu, B., Fu, L., Wang, H., Zhang, Q., Lin, S., Yan, J., & Shen, H. (2022). Arbutin Protects Retinal Pigment Epithelium Against Oxidative Stress by Modulating SIRT1/FOXO3a/PGC-1α/β Pathway. Frontiers in Genetics, 13. https://doi.org/10.3389/fgene.2022.922807

Tang, H., Li, K., Zhang, S., Lan, H., Liang, L., Huang, C., & Li, T. (2021). Inhibitory Effect of Paeonol on Apoptosis, Oxidative Stress, and Inflammatory Response in Human Umbilical Vein Endothelial Cells Induced by High Glucose and Palmitic Acid Induced through Regulating SIRT1/FOXO3a/NF-κB Pathway. Journal of Interferon and Cytokine Research, 41(3), 111–124. https://doi.org/10.1089/jir.2019.0236

Wang, C., Chen, W. gui, Chen, H., Zhao, J., He, J., Hu, Y., He, M., Zhang, Y., Lin, C. shou, Chang, J., & Liu, X. (2025). Exercise improves hypobaric hypoxia-induced skeletal muscle dysfunction via Sirt1-Mediated myotube and mitochondrial remodeling. Journal of Advanced Research, 83, 423–437. https://doi.org/10.1016/j.jare.2025.08.022

Wang, F., Jia, X., Zhang, J., Gao, P., Song, J., Chang, W., Hu, K., Zhang, X., An, F., Yan, C., Geng, G., Yuan, W., & Zhao, Y. (2025). Bushen Huoxue Fufang Tenghuang Jiangu Capsule Attenuates Postmenopausal Osteoporosis via Activating Sirt1/FoxO3a in Ovariectomized Rats. Chinese Journal of Modern Applied Pharmacy, 42(24), 4277–4284. https://doi.org/10.13748/j.cnki.issn1007-7693.20241883

Wetzel, G. M., & Sanchez, D. T. (2022). Heterosexual Young Adults’ Experience With and Perceptions of the Orgasm Gap: A Mixed Methods Approach. Psychology of Women Quarterly, 46(2), 131–146. https://doi.org/10.1177/03616843221076410

Xie, L., Li, J., Peng, Q., Liu, X., Lin, F., Dai, X., & Ling, B. (2025). Contribution of RND superfamily multidrug efflux pumps AdeABC, AdeFGH, and AdeIJK to antimicrobial resistance and virulence factors in multidrug-resistant Acinetobacter baumannii AYE. Antimicrobial Agents and Chemotherapy, 69(7). https://doi.org/10.1128/aac.01858-24

Yang, F., Cai, M., Mortenson, C., Fakhari, H., Lokmanoglu, A. D., Hullman, J., Franconeri, S., Diakopoulos, N., Nisbet, E. C., & Kay, M. (2024). Swaying the Public? Impacts of Election Forecast Visualizations on Emotion, Trust, and Intention in the 2022 U.S. Midterms. IEEE Transactions on Visualization and Computer Graphics, 30(1), 23–33. https://doi.org/10.1109/TVCG.2023.3327356

Zha, C., Qi, Y., Xing, F., & Li, J. (2024). Astragaloside IV Inhibits the Pyroptosis in the Acute Kidney Injury through Targeting the SIRT1/FOXO3a Axis. Chemical and Pharmaceutical Bulletin, 72(10), 923–931. https://doi.org/10.1248/cpb.c24-00151

Zhang, F., Gao, F., Wang, K., Liu, X., & Zhang, Z. (2021). MiR-34a inhibitor protects mesenchymal stem cells from hyperglycaemic injury through the activation of the SIRT1/FoxO3a autophagy pathway. Stem Cell Research and Therapy, 12(1). https://doi.org/10.1186/s13287-021-02183-2

Zhang, N., & Liu, Y. (2023). Design and implementation of virtual laboratories for higher education sustainability: a case study of Nankai University. Frontiers in Education, 8. https://doi.org/10.3389/feduc.2023.1322263

Zhu, D., Lu, Y., Jamil, S., Ashfaq, H., Al-Emam, A., & Hassan, H. M. (2026). Ayanin combats against barium sulphate nanoparticles induced hepatotoxicity via modulating SIRT1/FOXO3a and HO-1/ferritin pathways: A biochemical, histopathological and computational approaches. Journal of Trace Elements in Medicine and Biology, 94. https://doi.org/10.1016/j.jtemb.2026.127836

Zhu, M., Miao, S., Zhou, W., Elnesr, S. S., Dong, X., & Zou, X. (2021). MAPK, AKT/FoxO3a and mTOR pathways are involved in cadmium regulating the cell cycle, proliferation and apoptosis of chicken follicular granulosa cells. Ecotoxicology and Environmental Safety, 214. https://doi.org/10.1016/j.ecoenv.2021.112091