Altered Peripheral Blood Gene Expression in Childhood Cancer Survivors With Anthracycline‐Induced Cardiomyopathy – A COG‐ALTE03N1 Report

• Type of Material: Article
• Authors: Purnima Singh, University of Alabama, Birmingham; Disheet A. Shah, Northwestern University; Mariam Jouni, Northwestern University; Romina B. Cejas, Northwestern University; David K. Crossman, University of Alabama, Birmingham; Tarek Magdy, Northwestern University; Shaowei Qiu, University of Alabama, Birmingham; Xuexia Wang, Florida International University; Liting Zhou, University of Alabama, Birmingham; Noha Sharafeldin, University of Alabama, Birmingham; Lindsey Hageman, University of Alabama, Birmingham; Donald E. McKenna, Northwestern University; Saro H. Armenian, City of Hope; Frank M. Balis, Children's Hospital of Philadelphia; Douglas S. Hawkins, Seattle Children's Hospital; Frank G. Keller, Emory University; Melissa M. Hudson, St. Jude Children's Research Hospital; Joseph P. Neglia, University of Minnesota, Minneapolis; A. Kim Ritchey, University of Pittsburgh; Jill P. Ginsberg, Children's Hospital of Philadelphia; Wendy Landier, University of Alabama, Birmingham; Ravi Bhatia, University of Alabama, Birmingham; Paul W. Burridge, Northwestern University; Smita Bhatia, University of Alabama, Birmingham
• Language: English
• Date: 2023-09-26
• Publisher: Wiley
• Publication Version: Final Published Version
• Copyright Statement: © 2023 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley.
• License: Creative Commons Attribution-NonCommercial 4.0 International
• Final Published Version (URL): https://doi.org/10.1161/JAHA.123.029954
• Title of Journal or Parent Work: Journal of the American Heart Association
• Volume: 12
• Issue: 19
• Start Page: e029954
• Grant/Funding Information: National Cancer Institute (NCI) (R35CA220502; Principal Investigator [PI]: S. Bhatia), Leukemia and Lymphoma Society (6563‐19; PI: S. Bhatia), The V Foundation for Cancer Research (DT2019‐010; PI: S. Bhatia), NCI (R01CA220002 and R01CA261898; PI: P.W. Burridge), The Children's Oncology Group study reported here is supported by the National Clinical Trials Network Operations Center Grant (U10CA180886; PI: D.S. Hawkins); the National Clinical Trials Network Statistics & Data Center Grant (U10CA180899; PI: Alonzo); the Children's Oncology Group Chair's Grant (U10CA098543; PI: Adamson); The Children's Oncology Group Statistics & Data Center Grant (U10CA098413; PI: Anderson); the NCI Community Oncology Research Program Grant (UG1CA189955; PI: Pollock); and the Community Clinical Oncology Program Grant (U10CA095861; PI: Pollock), and the St Baldrick's Foundation through an unrestricted grant.
• Keywords: gene expression; childhood cancer survivors; transcriptome; peripheral blood; anthracyclines; cardiomyopathy
• Research Categories: Health Sciences, Oncology; Health Sciences, General; Biology, Genetics

Abstract

Background Anthracycline‐induced cardiomyopathy is a leading cause of premature death in childhood cancer survivors, presenting a need to understand the underlying pathogenesis. We sought to examine differential blood‐based mRNA expression profiles in anthracycline‐exposed childhood cancer survivors with and without cardiomyopathy. Methods and Results We designed a matched case‐control study (Children's Oncology Group‐ALTE03N1) with mRNA sequencing on total RNA from peripheral blood in 40 anthracycline‐exposed survivors with cardiomyopathy (cases) and 64 matched survivors without (controls). DESeq2 identified differentially expressed genes. Ingenuity Pathway Analyses (IPA) and Gene Set Enrichment Analyses determined the potential roles of altered genes in biological pathways. Functional validation was performed by gene knockout in human‐induced pluripotent stem cell‐derived cardiomyocytes using CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR‐associated protein 9) technology. Median age at primary cancer diagnosis for cases and controls was 8.2 and 9.7 years, respectively. Thirty‐six differentially expressed genes with fold change ≥±2 were identified; 35 were upregulated. IPA identified “hepatic fibrosis” and “iron homeostasis” pathways to be significantly modulated by differentially expressed genes, including toxicology functions of myocardial infarction, cardiac damage, and cardiac dilation. Leading edge analysis from Gene Set Enrichment Analyses identified lactate dehydrogenase A (LDHA) and cluster of differentiation 36 (CD36) genes to be significantly upregulated in cases. Interleukin 1 receptor type 1, 2 (IL1R1, IL1R2), and matrix metalloproteinase 8, 9 (MMP8, MMP9) appeared in multiple canonical pathways. LDHA‐knockout human‐induced pluripotent stem cell‐derived cardiomyocytes showed increased sensitivity to doxorubicin. Conclusions We identified differential mRNA expression profiles in peripheral blood of anthracycline‐exposed childhood cancer survivors with and without cardiomyopathy. Upregulation of LDHA and CD36 genes suggests metabolic perturbations in a failing heart. Dysregulation of proinflammatory cytokine receptors IL1R1 and IL1R2 and matrix metalloproteinases, MMP8 and MMP9 indicates structural remodeling that accompanies the clinical manifestation of symptomatic cardiotoxicity.