Phenotype-based drug screening: An in vivo strategy to classify and identify the chemical compounds modulating zebrafish M-cell regeneration

Ankita Kumari; Xin-An Zeng; Abdul Rahaman; Muhammad Adil Farooq; Yanyan Huang; Mahafooj Alee; Runyu Yao; Murtaza Ali; Ibrahim Khalifa; Omnia Badr

doi:10.3389/fmolb.2022.984461

Phenotype-based drug screening: An in vivo strategy to classify and identify the chemical compounds modulating zebrafish M-cell regeneration

Front Mol Biosci. 2022 Oct 24:9:984461. doi: 10.3389/fmolb.2022.984461. eCollection 2022.

Authors

Ankita Kumari^{1

2

3}, Xin-An Zeng^{1

2

3}, Abdul Rahaman^{1

2

3}, Muhammad Adil Farooq⁴, Yanyan Huang², Mahafooj Alee¹, Runyu Yao^{1

3}, Murtaza Ali^{1

2

3}, Ibrahim Khalifa^{4

5}, Omnia Badr^{4

6}

Affiliations

¹ School of Food Science and Engineering, South China University of Technology, Guangzhou, China.
² Guangdong Key Laboratory of Food Intelligent Manufacturing, Foshan University, Foshan, Guangdong, China.
³ Overseas Expertise Introduction Centre for Discipline Innovation of Food Nutrition and Human Health (111 Centre), Guangzhou, China.
⁴ Department of Food Science and Technology, Khwaja Fareed University of Engineering and Information Technology, Rahimyar Khan, Punjab, Pakistan.
⁵ Food Technology Department, Faculty of Agriculture, Benha University, Qalyubia, Egypt.
⁶ Department of Genetics and Genetic Engineering, Faculty of Agriculture, Benha University, Qalyubia, Egypt.

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 in vivo assay using Tg (hsp: Gal4FF62A; UAS: nfsB-mCherry) transgenic zebrafish larvae. Interestingly, via 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 in silico 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 in vivo along with in silico screening strategy will promote the rapid translation of new therapeutics to improve knowledge of the toxicity profile of approved/non-approved drugs efficiently.

Keywords: M-cell; axon; phenotype drug screening (PDS); regeneration; zebrafish.