Phenotype-based Drug Screening: An Strategy to Classify and Identify the Chemical Compounds Modulating Zebrafish M-cell Regeneration

Overview

Journal Front Mol Biosci

Specialty Biology

Date 2022 Nov 10

PMID 36353729

Authors

Ankita Kumari

Xin-An Zeng

Abdul Rahaman

Muhammad Adil Farooq

Yanyan Huang

Mahafooj Alee

Runyu Yao

Murtaza Ali

Ibrahim Khalifa

Omnia Badr

Affiliations

Soon will be listed here.

Abstract

Several disease-modulatory FDA-approved drugs are being used in patients with neurodegenerative diseases. However, information on their toxicity-related profiles is very limited. Therefore, measurement of drug toxicity is essential to increase the knowledge of their side effects. This study aimed to identify compounds that can modulate M-cell regeneration by causing neuro-protection and -toxicity. Here, we developed a simple and efficient assay using Tg (hsp: Gal4FF62A; UAS: nfsB-mCherry) transgenic zebrafish larvae. Interestingly, the phenotype-based drug screening approach, we rapidly investigated 1,260 compounds from the United States drug collection and validated these in large numbers, including 14 compounds, that were obstructing this regeneration process. Next, 4 FDA-approved drugs out of 14 compounds were selected as the lead hits for analysis to clarify their binding patterns with PTEN and SOCS3 signaling due to their significant potential in the inhibition of axon regeneration. Molecular docking studies indicated good binding affinity of all 4 drugs with the respective signaling molecules. This may point to PTEN and SOCS3 as the signaling molecules responsible for reducing axon regeneration. Moreover, the acute effect of compounds in reducing M-cell regeneration delineated their toxic effect. In conclusion, our along with screening strategy will promote the rapid translation of new therapeutics to improve knowledge of the toxicity profile of approved/non-approved drugs efficiently.

References

Webb B, Sali A . Protein Structure Modeling with MODELLER. Methods Mol Biol. 2020; 2199:239-255. DOI: 10.1007/978-1-0716-0892-0_14. View

Zhang J, Yang D, Huang H, Sun Y, Hu Y . Coordination of Necessary and Permissive Signals by PTEN Inhibition for CNS Axon Regeneration. Front Neurosci. 2018; 12:558. PMC: 6104488. DOI: 10.3389/fnins.2018.00558. View

Swinney D, Anthony J . How were new medicines discovered?. Nat Rev Drug Discov. 2011; 10(7):507-19. DOI: 10.1038/nrd3480. View

Friese A, Ursu A, Hochheimer A, Scholer H, Waldmann H, Bruder J . The Convergence of Stem Cell Technologies and Phenotypic Drug Discovery. Cell Chem Biol. 2019; 26(8):1050-1066. DOI: 10.1016/j.chembiol.2019.05.007. View

Rennekamp A, Peterson R . 15 years of zebrafish chemical screening. Curr Opin Chem Biol. 2014; 24:58-70. PMC: 4339096. DOI: 10.1016/j.cbpa.2014.10.025. View

Moffat J, Vincent F, Lee J, Eder J, Prunotto M . Opportunities and challenges in phenotypic drug discovery: an industry perspective. Nat Rev Drug Discov. 2017; 16(8):531-543. DOI: 10.1038/nrd.2017.111. View

Galluzzi L, Joza N, Tasdemir E, Maiuri M, Hengartner M, Abrams J . No death without life: vital functions of apoptotic effectors. Cell Death Differ. 2008; 15(7):1113-23. PMC: 2917777. DOI: 10.1038/cdd.2008.28. View

Lee J, Bogyo M . Target deconvolution techniques in modern phenotypic profiling. Curr Opin Chem Biol. 2013; 17(1):118-26. PMC: 3594516. DOI: 10.1016/j.cbpa.2012.12.022. View

Chung A, Kim P, Kim S, Kim E, Kim D, Jeong I . Generation of demyelination models by targeted ablation of oligodendrocytes in the zebrafish CNS. Mol Cells. 2013; 36(1):82-7. PMC: 3887923. DOI: 10.1007/s10059-013-0087-9. View

10.

Kepp O, Galluzzi L, Lipinski M, Yuan J, Kroemer G . Cell death assays for drug discovery. Nat Rev Drug Discov. 2011; 10(3):221-37. DOI: 10.1038/nrd3373. View

11.

Wagner B . The resurgence of phenotypic screening in drug discovery and development. Expert Opin Drug Discov. 2015; 11(2):121-5. DOI: 10.1517/17460441.2016.1122589. View

12.

Rieger S, Sagasti A . Hydrogen peroxide promotes injury-induced peripheral sensory axon regeneration in the zebrafish skin. PLoS Biol. 2011; 9(5):e1000621. PMC: 3101194. DOI: 10.1371/journal.pbio.1000621. View

13.

Yamanaka I, Miki M, Asakawa K, Kawakami K, Oda Y, Hirata H . Glycinergic transmission and postsynaptic activation of CaMKII are required for glycine receptor clustering in vivo. Genes Cells. 2013; 18(3):211-24. DOI: 10.1111/gtc.12032. View

14.

Pankevich D, Altevogt B, Dunlop J, Gage F, Hyman S . Improving and accelerating drug development for nervous system disorders. Neuron. 2014; 84(3):546-53. PMC: 4254615. DOI: 10.1016/j.neuron.2014.10.007. View

15.

Frantz S . Drug discovery: playing dirty. Nature. 2005; 437(7061):942-3. DOI: 10.1038/437942a. View

16.

Childers W, Elokely K, Abou-Gharbia M . The Resurrection of Phenotypic Drug Discovery. ACS Med Chem Lett. 2020; 11(10):1820-1828. PMC: 7549108. DOI: 10.1021/acsmedchemlett.0c00006. View

17.

Wilson C . Aspects of larval rearing. ILAR J. 2013; 53(2):169-78. DOI: 10.1093/ilar.53.2.169. View

18.

Bremer J, Marsden K, Miller A, Granato M . The ubiquitin ligase PHR promotes directional regrowth of spinal zebrafish axons. Commun Biol. 2019; 2:195. PMC: 6531543. DOI: 10.1038/s42003-019-0434-2. View

19.

Cornet C, Di Donato V, Terriente J . Combining Zebrafish and CRISPR/Cas9: Toward a More Efficient Drug Discovery Pipeline. Front Pharmacol. 2018; 9:703. PMC: 6037853. DOI: 10.3389/fphar.2018.00703. View

20.

Bhatt D, Otto S, Depoister B, Fetcho J . Cyclic AMP-induced repair of zebrafish spinal circuits. Science. 2004; 305(5681):254-8. DOI: 10.1126/science.1098439. View