The [2Fe‐2S] protein CISD2 plays a key role in preventing iron accumulation in cardiomyocytes

Abstract

Considered a key aging gene, CISD2, encoding CDGSH iron-sulfur domaincontaining protein 2, plays a central role in regulating calcium homeostasis, preventing mitochondrial dysfunction, and the activation of autophagy and apoptosis in different cells. Here, we show that cardiomyocytes from CISD2null mice accumulate high levels of iron and contain high levels of transferrin receptor and ferritin. Using proteomics and transmission electron microscopy, we further show that the lack of CISD2 induces several features of the aging process in young mice, but other features are not induced. Taken together, our findings suggest that CISD2 protects cardiomyocytes from overaccumulation of iron, which is common in aging hearts and can contribute to the pathogenesis of heart failure.

Propagation of a rapid cell-to-cell H2O2 signal over long distances in a monolayer of cardiomyocyte cells

“…A day before the experiment the media was replaced with FluoroBrite DMEM (ThermoFisher) to reduce background fluorescence [ 24 , 31 ]. For roGFP2 and roGFP2-Orp1 expressing cells, H9c2 cells were transfected with pMF1707 or pMOS016, obtained from Addgene ( https://www.addgene.org/ ), using the Sigma's gene juice protocol, and selected using a Cytek Aurora cell sorter (Tecan) followed by growth in the presence of the appropriate antibiotics [ [32] , [33] , [34] ]. Cells were plated at an initial density of 500,000 per well for the 6-well plate, 25,000 per well for the 96-well plate, and 2,500,000 per dish for the 100 mm dishes and imaged using the IVIS (Revvity) or Lionheart FX (Agilent) apparatuses [ 24 ].…”

Propagation of a rapid cell-to-cell H2O2signal over long distances in a monolayer of cardiomyocyte cells

“…Injury, imaging, and data analyses were performed as described in 20 . Protein extraction was performed from H9c2(2-1) cells grown on 100 mm plates treated and imaged with the IVIS platform as described in 20,28,29 . Cells were scrapped from the plates and proteins were extracted, identified (Uniprot ID-UP000002494), and analyzed as described in 20,28,29 .…”

“…Protein extraction was performed from H9c2(2-1) cells grown on 100 mm plates treated and imaged with the IVIS platform as described in 20,28,29 . Cells were scrapped from the plates and proteins were extracted, identified (Uniprot ID-UP000002494), and analyzed as described in 20,28,29 . Proteomics data are available vin ProteomeXchange with identifier PXD047131.…”

ROS are evolutionary conserved cell-to-cell signals

“…Mice used in this study were housed under pathogen-free conditions in animal care facilities and received humane care in compliance with the Guide for the Care and Use of Laboratory Animals (https://www.ncbi.nlm.nih.gov/books/NBK54050/) 71 . All experimental procedures were approved by the Institutional Animal Care and Use Committee of the University of Missouri (https://research.missouri.edu/acqa/acuc).…”

“…C57BL/6J mice were obtained from Jackson Laboratory (https://www.jax.org/) at 8 weeks of age and used between the age of 9 and 10 weeks of age. Mice were placed under anesthesia using 4% Isoflurane, sacrificed, and the hearts were immediately collected 71 . The hearts were rinsed in FluoroBrite DMEM (ThermoFisher, Waltham, MA) to remove excess blood.…”