Dual-Target Therapeutic Prospect Against Huntington’s Disease: In Silico Dissection of HTT and ETV6 Interactions and Ligand-Based Inhibition Strategy
Keywords:
Huntington’s disease, HTT gene, ETV6, transcriptional dysregulation, CAG repeat expansion, Risdiplam, in silico drug design, dual inhibition, protein modeling, virtual screeningAbstract
Huntington's disease (HD) is a hereditary neurodegenerative disorder due to a trinucleotide CAG repeat expansion in the huntingtin (HTT) gene, resulting in the expression of a mutant version of the huntingtin protein (mHTT). The mutation triggers a cascade of harmful cellular processes, such as disrupted transcriptional regulation, mitochondrial dysfunction, imbalance of proteostasis, and progressive neuronal loss, especially in the medium spiny neurons of the striatum. Although HTT has long been the center of HD research, more recent transcriptomic and systems biology research has identified ETV6, a transcriptional repressor of the ETS family, as a potential major contributor to the pathophysiology of the disease. The coincidence of expression patterns between HTT and ETV6, in both murine models and human-derived data, implies a functional interaction that could be responsible for the transcriptional dysregulation characteristic of HD. In the present work, an integrated computational strategy involving gene expression profiling, phylogenetic analysis, protein modeling, virtual ligand screening, and pharmacokinetic analysis was used to investigate the feasibility of simultaneous targeting of both HTT and ETV6. Protein structures were built with Phyre2 and Modeller, and then systematically validated by Ramachandran plot evaluation. A validated library of 38 neuroactive ligands was docked against both protein targets employing AutoDock Vina, integrated into PyRx. Its strongest candidate, Risdiplam—originally designed against spinal muscular atrophy—displayed strong HTT and ETV6 binding affinities. Additional ADME and toxicity experiments validated its suitability for pharmacokinetics as well as its availability in the CNS. Results validate a two-way therapeutic method on the dual approach of suggesting correction of the identical transcriptional imbalances within HD through the simultaneous co-modulation of HTT and ETV6.
Downloads
Metrics
References
Ahamad, S., & Bhat, S. A. (2022). The emerging landscape of small-molecule therapeutics for the treatment of Huntington’s disease. Journal of Medicinal Chemistry, 65(24), 15993-16032.
Ayodele, P. F., Bamigbade, A., Bamigbade, O. O., Adeniyi, I. A., Tachin, E. S., Seweje, A. J., & Farohunbi, S. T. (2023). Illustrated procedure to perform molecular docking using PyRx and biovia discovery studio visualizer: a case study of 10kt with atropine. Progress in Drug Discovery & Biomedical Science, 6(1).
Blumenstock, S., & Dudanova, I. (2020). Cortical and striatal circuits in Huntington’s disease. Frontiers in neuroscience, 14, 82.
Boulila, M., ElSayed, A. I., Rafudeen, M. S., & Omar, A. A. (2020). Investigating molecular evolutionary forces and phylogenetic relationships among melatonin precursor-encoding genes of different plant species. Molecular Biology Reports, 47(3), 1625-1636.
Christodoulou, C. C., & Papanicolaou, E. Z. (2024). Omics and Network-Based Approaches in Understanding HD Pathogenesis.
Christodoulou, C. C., & Papanicolaou, E. Z. (2024). Omics and Network-Based Approaches in Understanding HD Pathogenesis.
Ciurea, A. V., Mohan, A. G., Covache-Busuioc, R. A., Costin, H. P., Glavan, L. A., Corlatescu, A. D., & Saceleanu, V. M. (2023). Unraveling molecular and genetic insights into neurodegenerative diseases: Advances in understanding Alzheimer’s, Parkinson’s, and Huntington’s diseases and amyotrophic lateral sclerosis. International journal of molecular sciences, 24(13), 10809.
De Braekeleer, E., Douet-Guilbert, N., Morel, F., Le Bris, M. J., Basinko, A., & De Braekeleer, M. (2012). ETV6 fusion genes in hematological malignancies: a review. Leukemia research, 36(8), 945-961.
De Braekeleer, E., Douet-Guilbert, N., Morel, F., Le Bris, M. J., Basinko, A., & De Braekeleer, M. (2012). ETV6 fusion genes in hematological malignancies: a review. Leukemia research, 36(8), 945-961.
De, S., Okon, M., Graves, B. J., & McIntosh, L. P. (2016). Autoinhibition of ETV6 DNA binding is established by the stability of its inhibitory helix. Journal of molecular biology, 428(8), 1515-1530.
Gheidari, D., Mehrdad, M., & Hoseini, F. (2024). Virtual screening, molecular docking, MD simulation studies, DFT calculations, ADMET, and drug likeness of Diaza-adamantane as potential MAPKERK inhibitors. Frontiers in Pharmacology, 15, 1360226..
Gutiérrez Ángel, S. (2019). Mutant Huntingtin toxicity modifiers revealed by a spatiotemporal proteomic profiling (Doctoral dissertation, lmu).
Hopkins, A. L. (2008). Network pharmacology: the next paradigm in drug discovery. Nature chemical biology, 4(11), 682-690.
Islam, M. R., Jony, M. H., Thufa, G. K., Akash, S., Dhar, P. S., Rahman, M. M., ... & Venkidasamy, B. (2024). A clinical study and future prospects for bioactive compounds and semi-synthetic molecules in the therapies for Huntington's disease. Molecular Neurobiology, 61(3), 1237-1270.
Jablonka, S., Hennlein, L., & Sendtner, M. (2022). Therapy development for spinal muscular atrophy: perspectives for muscular dystrophies and neurodegenerative disorders. Neurological research and practice, 4(1), 2.
Kamat, P. K., Kalani, A., Rai, S., Swarnkar, S., Tota, S., Nath, C., & Tyagi, N. (2016). Mechanism of oxidative stress and synapse dysfunction in the pathogenesis of Alzheimer’s disease: understanding the therapeutics strategies. Molecular neurobiology, 53, 648-661.
Koriath, C. A. M. (2020). Next‐generation sequencing in the diagnosis of Dementia and Huntington’s disease Phenocopy Syndromes (Doctoral dissertation, UCL (University College London)).
KUSHWAHA, S. P., GUPTA, S. K., & KUMAR, S. (2021). Amended homology model intended for T1R2 subunit of human sweet taste receptor using Phyre 2.0 and ModLoop. International Journal of Pharmaceutical Research (09752366), 13(2).
Li, Y., Li, S., & Wu, H. (2022). Ubiquitination-proteasome system (UPS) and autophagy two main protein degradation machineries in response to cell stress. Cells, 11(5), 851.
Li, Z., Wu, W., Li, Q., Heng, X., Zhang, W., Zhu, Y., ... & Yan, Y. (2024). BCL6B-dependent suppression of ETV2 hampers endothelial cell differentiation. Stem Cell Research & Therapy, 15(1), 226.
Lontay, B., Kiss, A., Virág, L., & Tar, K. (2020). How do post-translational modifications influence the pathomechanistic landscape of Huntington’s disease? A comprehensive review. International journal of molecular sciences, 21(12), 4282.
Malla, B., Guo, X., Senger, G., Chasapopoulou, Z., & Yildirim, F. (2021). A systematic review of transcriptional dysregulation in Huntington’s disease studied by RNA sequencing. Frontiers in genetics, 12, 751033.
McInnis, M. G. (1996). Anticipation: an old idea in new genes. American journal of human genetics, 59(5), 973.
Mi, J., Wu, F., & Yang, N. (2024). Transformations and Challenges in Cancer Treatment. In New Anti-cancer Drug Development and Evaluation (pp. 1-25). Springer, Singapore.
Rouaux, C., Loeffler, J. P., & Boutillier, A. L. (2004). Targeting CREB-binding protein (CBP) loss of function as a therapeutic strategy in neurological disorders. Biochemical pharmacology, 68(6), 1157-1164.
Sajjad, M. U. (2010). Regulation of the redox homeostasis during polyglutamine misfolding in Huntington’s Disease (Doctoral dissertation, University of Southampton).
Salokas, K., Dashi, G., & Varjosalo, M. (2023). Decoding oncofusions: unveiling mechanisms, clinical impact, and prospects for personalized cancer therapies. Cancers, 15(14), 3678.
Sawant, N., Morton, H., Kshirsagar, S., Reddy, A. P., & Reddy, P. H. (2021). Mitochondrial abnormalities and synaptic damage in Huntington’s disease: a focus on defective mitophagy and mitochondria-targeted therapeutics. Molecular neurobiology, 1-28.
Shafie, A., Ashour, A. A., Anwar, S., Anjum, F., & Hassan, M. I. (2024). Exploring molecular mechanisms, therapeutic strategies, and clinical manifestations of Huntington’s disease. Archives of Pharmacal Research, 47(6), 571-595.
SUDARMANTO, B. S. A., YUSWANTO, A., SUSIDARTI, R. A., & NOEGROHATI, S. (2017). Molecular Modeling of Human 3β-Hydroxysteroid Dehydrogenase Type 2: Combined Homology Modeling, Docking and QSAR Approach. JURNAL ILMU KEFARMASIAN INDONESIA, 15(1), 7-16.
Tabrizi, S. J., Flower, M. D., Ross, C. A., & Wild, E. J. (2020). Huntington disease: new insights into molecular pathogenesis and therapeutic opportunities. Nature Reviews Neurology, 16(10), 529-546.
Testa CM, Jankovic J. Huntington disease: a quarter century of progress since the gene discovery. Journal of the neurological sciences. 2019 Jan 15;396:52-68.
Wei, Y., Han, S., Wen, J., Liao, J., Liang, J., Yu, J., ... & Zhang, B. (2023). E26 transformation-specific transcription variant 5 in development and cancer: modification, regulation and function. Journal of biomedical science, 30(1), 17.
Zulfat, M., Hakami, M. A., Hazazi, A., Mahmood, A., Khalid, A., Alqurashi, R. S., ... & Huang, X. (2024). Identification of novel NLRP3 inhibitors as therapeutic options for epilepsy by machine learning-based virtual screening, molecular docking and biomolecular simulation studies. Heliyon, 10(15).
Downloads
Published
How to Cite
Issue
Section
License
This work is licensed under a Creative Commons Attribution 4.0 International License.
You are free to:
- Share — copy and redistribute the material in any medium or format
- Adapt — remix, transform, and build upon the material for any purpose, even commercially.
Terms:
- Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.
- No additional restrictions — You may not apply legal terms or technological measures that legally restrict others from doing anything the license permits.
