FDA approved L-type channel blocker Nifedipine reduces cell death in hypoxic A549 cells through modulation of mitochondrial calcium and superoxide generation

Manohar, Kuruba and Gupta, Rishikesh Kumar and Gupta, Parth and Saha, Debasmita and Gare, Suman and Sarkar, Rahuldeb and Misra, Ashish and Giri, Lopamudra (2021) FDA approved L-type channel blocker Nifedipine reduces cell death in hypoxic A549 cells through modulation of mitochondrial calcium and superoxide generation. Free Radical Biology and Medicine, 177. pp. 189-200. ISSN 0891-5849

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Abstract

As hypoxia is a major driver for the pathophysiology of COVID-19, it is crucial to characterize the hypoxic response at the cellular and molecular levels. In order to augment drug repurposing with the identification of appropriate molecular targets, investigations on therapeutics preventing hypoxic cell damage is required. In this work, we propose a hypoxia model based on alveolar lung epithelial cells line using chemical inducer, CoCl2 that can be used for testing calcium channel blockers (CCBs). Since recent studies suggested that CCBs may reduce the infectivity of SARS-Cov-2, we specifically select FDA approved calcium channel blocker, nifedipine for the study. First, we examined hypoxia-induced cell morphology and found a significant increase in cytosolic calcium levels, mitochondrial calcium overload as well as ROS production in hypoxic A549 cells. Secondly, we demonstrate the protective behaviour of nifedipine for cells that are already subjected to hypoxia through measurement of cell viability as well as 4D imaging of cellular morphology and nuclear condensation. Thirdly, we show that the protective effect of nifedipine is achieved through the reduction of cytosolic calcium, mitochondrial calcium, and ROS generation. Overall, we outline a framework for quantitative analysis of mitochondrial calcium and ROS using 3D imaging in laser scanning confocal microscopy and the open-source image analysis platform ImageJ. The proposed pipeline was used to visualize mitochondrial calcium and ROS level in individual cells that provide an understanding of molecular targets. Our findings suggest that the therapeutic value of nifedipine may potentially be evaluated in the context of COVID-19 therapeutic trials. © 2021

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IITH Creators:

IITH Creators	ORCiD
Misra, Ashish	https://orcid.org/0000-0002-3042-8302
Giri, Lopamudra	http://orcid.org/0000-0002-2352-7919

Item Type:

Article

Additional Information:

The authors acknowledge the research facilities provided by the Indian Institute of Technology Hyderabad, India, and Department of Science and Technology for the fellowship of Kuruba Manohar, and Ministry of Education for the fellowship support for Parth Gupta. We also thank Vaibhav Dhyani, Shibu Chameettachal and Jagadeesh Mahadevan for their suggestions during RT-PCR and IF experiments. This study was supported by the Department of Biotechnology, Ministry of Science & Technology, India (Grant number: BT/PR21261/MED/31/348/2016 and BT/PR22239/NNT/28/1269/2017 ).

Uncontrolled Keywords:

Calcium channel blocker; Hypoxia; Mitochondrial calcium; Nifedipine; Reactive oxygen species (ROS); SARS-Cov-2

Subjects:

Others > Biotechnology
Chemical Engineering

Divisions:

Department of Biotechnology
Department of Chemical Engineering

Depositing User:

. LibTrainee 2021

Date Deposited:

23 Aug 2022 08:45

Last Modified:

23 Aug 2022 08:45