Corrigendum: Mechanisms by which statins protect endothelial cells from radiation-induced injury in the carotid artery

Karima Ait-Aissa ^* Linette N Leng Nathan Lindsey Xutong Guo Olha M Koval Isabella M Grumbach ^*

• The University of Iowa, Iowa City, United States

Corrigendum: Mechanisms by which statins protect endothelial cells from radiation-induced injury in the carotid artery Karima Ait-Aissa 1,2*, Linette N. Leng 1, Nathanial R. Lindsey 1, Xutong Guo 1, Olha M. Koval 1, Isabella M. Grumbach 1,3,4* 1 Abboud Cardiovascular Research Center, Department of Internal Medicine, Carver College of Medicine, University of Iowa, Iowa City, IA, USA 2 Department of Biomedical Sciences, Dental College of Medicine, Lincoln Memorial University3 Free Radical and Radiation Biology Program, Department of Radiation Oncology, Carver College of Medicine, University of Iowa, Iowa City, IA, USA 4 Iowa City VA Healthcare System, Iowa City, IA, USA Correspondence *: Isabella Grumbach, MD, PhD University of Iowa Carver College of Medicine Division of Cardiovascular Medicine 4336 PBDB 169 Newton Road Iowa City, IA, 52242 Phone: 319-384-4610 Fax: 319-353-5552 e-mail: isabella-grumbach@uiowa.edu Karima Ait-Aissa, PhD Lincoln Memorial University Dental College of Medicine LMU Tower #331 1705 St Mary’s Street Knoxville, TN 37917 e-mail: karima.aitaissa@lmunet.edu Keywords: radiation therapy, carotid stenosis, endothelium, statin, mitochondria, prevention A Corrigendum on Mechanisms by which statins protect endothelial cells from radiation-induced injury in the carotid artery By Karima Ait-Aissa, Linette N Leng, Nathanial R Lindsey, Xutong Guo, Denise Juhr, Olha M Koval, Isabella M Grumbach (2023) Front Cardiovasc Med. 10:1133315. doi: 10.3389/fcvm.2023.1133315   Error in Figure/Table In the published article, there was an error in Figure 1 as published. The labeling of the subfigures 1B-H is incorrect. The corrected Figure 1 and its caption appear below. Figure 1: Pravastatin and atorvastatin preserve endothelial function in vivo following head-and-neck IR. (A–C) Effects of statins on endothelium-dependent relaxation of the carotid artery in response to acetylcholine (ACh). C57BL/6J mice were treated with (A) vehicle, (B) pravastatin (Prava), or (C) atorvastatin (Ator) after head-and-neck irradiation (12Gy) or sham treatment, and relaxation was tested at 24 and 240 hr after irradiation. (D–F) Effects of statins on endothelium-independent relaxation of the carotid artery in response to sodium nitroprusside (SNP), in mice treated as in A, B, and C, respectively. (G–I) Effects of statins on endothelium-dependent relaxation of mesenteric resistance arteries (MRAs), in mice treated as in A, B, and C, respectively. n=5 mice per group. p values were determined using repeated measures 2-way ANOVA followed by Tukey's post-Hoc test. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated. Error in Figure/Table In the published article, there was an error in Figure 3 as published. Images for Vehicle-treated non-irradiated (V-nIR) mitoSOX staining were inadvertently mismatched with those of mitotracker staining and merged images. The corrected Figure 3 and its caption appear below. Figure 3: Pravastatin protects against IR-induced mitochondrial damage or hyperpolarization in vitro. All panels compare cells subjected to irradiation (4Gy) after pretreatment with pravastatin (Prava, 10 M) starting at 18 hr before irradiation. (A, B) Representative images and signal integrated density of mitoSOX fluorescence normalized to mitoTracker fluorescence in HCAECs at (A) 24 and (B) 240 hr after IR. (C, D) mtDNA lesions in DNA extracted from HUVECs at (C) 24 hr and (D) 240 hr after IR. (E, F) Representative images and integrated density of mitochondrial membrane potential in HCAECs, as determined by TMRM fluorescence, at (E) 24 hr and (F) 240 hr after irradiation. Analysis per cell, n=4 independent experiments. p values were determined by Kruskal-Wallis test. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated. Error in Figure/Table In the published article, there was an error in Figure 4 as published. In Figure 4A, images for mitoSox and mtTracker staining of atorvastatin-treated cells in the left upper panels were mistakenly switched. We confirmed that the analysis was performed in the correct panels. Figure 4C, D: The labeling for the samples treated with atorvastatin and/or IR is incorrect. The correct description for radiation treatment (IR) should be “- + - +” instead of “- + + +”. Figure 4E, F: Images for TMRM in the left lower panel are mislabeled. The correct labeling of the images should read: Vehicle, Vehicle + IR, Atorva, Atorva + IR (instead of Vehicle, Vehicle + IR, Prava, Prava + IR). The corrected Figure 4 and its caption appear below. Figure 4: Atorvastatin does not protect against IR-induced mitochondrial damage or hyperpolarization in vitro. All panels compare HCAECs subjected to irradiation (4 Gy) after pretreatment with atorvastatin (Ator, 5 M) or vehicle. Parameters assessed are: (A, B) Representative images and signal integrated density of MitoSOX fluorescence normalized to MitoTracker fluorescence at (A) 24 and (B) 240 hr after irradiation, in cells treated with atorvastatin (5M) or vehicle starting 18hr before irradiation. (C, D) Damage to mtDNA as assessed by PCR assay. mtDNA lesions at (C) 24 and (D) 240 hr after irradiation, in HUVECs treated with atorvastatin or vehicle starting 18 hr before IR. (E, F) Representative images and integrated density of mitochondrial membrane potential, as determined by TMRM fluorescence, at (E) 24 and (F) 240 hr after irradiation. Analysis per cell, n=4 independent experiments, p values by Kruskal-Wallis test. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated. Error in Figure/Table In the published article, there was an error in Figure 7 as published. The labeling for the samples treated with atorvastatin and/or IR in Figure 7E - H is incorrect. The correct description for radiation treatment (IR) should be “- + - +” instead of “- + + +”. The corrected Figure 7 and its caption appear below. Figure 7: Pravastatin, but not atorvastatin, prevents irradiation-induced reduction of mtDNA transcription and ETC activity. All panels compare HCAECs subjected to irradiation (4Gy) after pretreatment with atorvastatin (Ator, 5M, overnight), pravastatin (Prava, 10M, overnight) or vehicle. (A-D) Effects of pretreatment with pravastatin (Prava, 10M, overnight) on transcriptional activity. (A, B) Quantitative (q)RT-PCR for cytochrome c oxidase I (MT-COI) at (A) 24 and (B) 240 hr after irradiation. (C-D) qRT-PCR for NADH-ubiquinone oxidoreductase chain 1 (MT-ND1) at (C) 24 and (D) 240 hr after irradiation. (E-H) Effects of pretreatment with atorvastatin (Ator, 5 M, overnight) on transcriptional activity. (E, F) qRT-PCR for MT-COI at (E) 24 and (F) 240 hr after irradiation. (G, H) qRT-PCR for MT-ND1 at (G) 24 and (H) 240 hr after irradiation. (I, J) Activity of ETC complex 1, as assessed by fluorometric assay at (I) 24 and (J) 240 hr after irradiation. p values by Kruskal-Wallis test. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.  Incorrect Supplementary Material In the published article, there was an error in Supplemental Figure 3. The labeling for the samples treated with atorvastatin and/or IR in Supplemental Figure 3E - H is incorrect. The correct description for radiation treatment (IR) should be “- + - +” instead of “- + + +”. The corrected figure and its caption appear below. Supplemental Figure 3: Neither pravastatin nor atorvastatin affects IR-induced transcription of nuclear DNA. (A-D) Effects of pretreatment with pravastatin (Prava, 10 μM, 1 hr) on nucDNA damage in HCAECs after irradiation (IR, 4 Gy). (A, B) Quantitative (q)RT-PCR for NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 1 (B, NDUF1), with cDNA normalized to 100 ng at 24 and 240 hr after IR. (C, D) qRT-PCR for cytochrome c oxidase 11 (D, COX11), with cDNA normalized to 100 ng at 24 and 240 hr after IR. (E-H) Effects of pretreatment with atorvastatin (5 μM, overnight) on nucDNA damage in HCAECs subjected to IR. (E, F) qRT-PCR for NDUF1 (B), with cDNA normalized to 100 ng at 24 and 240 hr after IR. (G, H) qRT-PCR for COX11 (D), with cDNA normalized to 100 ng at 24 and 240 hr after IR. Statistical analysis by Kruskal-Wallis test. Ns indicates not significant. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.