South Texas Medical Scientist Training Program, University of Texas Health Science Center San Antonio, San Antonio, Texas.
Integrated Biomedical Sciences Graduate Program, University of Texas Health Science Center San Antonio, San Antonio, Texas.
Am J Physiol Heart Circ Physiol. 2022 Jul 1;323(1):H130-H145. doi: 10.1152/ajpheart.00514.2021. Epub 2022 Jun 3.
Childhood cancer survivors (CCSs) face lifelong side effects related to their treatment with chemotherapy. Anthracycline agents, such as doxorubicin (DOX), are important in the treatment of childhood cancers but are associated with cardiotoxicity. Cardiac toxicities represent a significant source of chronic disability that cancer survivors face; despite this, the chronic cardiotoxicity phenotype and how it relates to acute toxicity remains poorly defined. To address this critical knowledge gap, we studied the acute effect of DOX on murine cardiac nonmyocytes in vivo. Determination of the acute cellular effects of DOX on nonmyocytes, a cell pool with finite replicative capacity, provides a basis for understanding the pathogenesis of the chronic heart disease that CCSs face. To investigate the acute cellular effects of DOX, we present single-cell RNA sequencing (scRNAseq) data from homeostatic cardiac nonmyocytes and compare it with preexisting datasets, as well as a novel CyTOF datasets. SCANPY, a python-based single-cell analysis, was used to assess the heterogeneity of cells detected in scRNAseq and CyTOF. To further assist in CyTOF data annotation, joint analyses of scRNAseq and CyTOF data using an artificial neural network known as sparse autoencoder for clustering, imputation, and embedding (SAUCIE) are performed. Lastly, the panel is tested on a mouse model of acute DOX exposure at two time points (24 and 72 h) after the last dose of doxorubicin and examined with joint clustering. In sum, we report the first ever CyTOF study of cardiac nonmyocytes and characterize the effect of acute DOX exposure with scRNAseq and CyTOF. We describe the first mass cytometry studies of murine cardiac nonmyocytes. The mass cytometry panel is compared with single-cell RNA sequencing data. Homeostatic cardiac nonmyocytes are characterized by mass cytometry to identify and quantify four major cell populations: endothelial cells, fibroblasts, leukocytes, and pericytes. The single-cell acute nonmyocyte response to doxorubicin is studied at 24 and 72 h after doxorubicin exposure given daily for 5 days at a dose of 4 mg/kg/day.
儿童癌症幸存者（CCS）面临与化疗相关的终生副作用。蒽环类药物，如多柔比星（DOX），在儿童癌症治疗中很重要，但与心脏毒性有关。心脏毒性是癌症幸存者面临的慢性残疾的一个重要来源；尽管如此，慢性心脏毒性表型及其与急性毒性的关系仍未得到明确界定。为了解决这一关键的知识空白，我们研究了 DOX 对体内小鼠心脏非心肌细胞的急性作用。确定 DOX 对非心肌细胞的急性细胞作用，为了解 CCS 面临的慢性心脏病的发病机制提供了基础。为了研究 DOX 的急性细胞作用，我们展示了来自稳态心脏非心肌细胞的单细胞 RNA 测序（scRNAseq）数据，并将其与现有数据集以及新的 CyTOF 数据集进行了比较。基于 python 的单细胞分析 SCANPY 用于评估 scRNAseq 和 CyTOF 中检测到的细胞的异质性。为了进一步协助 CyTOF 数据注释，使用稀疏自动编码器聚类、插补和嵌入（SAUCIE）的人工神经网络对 scRNAseq 和 CyTOF 数据进行联合分析。最后，该面板在接受最后一剂多柔比星后 24 和 72 小时两个时间点的急性 DOX 暴露的小鼠模型上进行测试，并与联合聚类一起进行检查。总之，我们报告了心脏非心肌细胞的首次 CyTOF 研究，并使用 scRNAseq 和 CyTOF 描述了急性 DOX 暴露的影响。我们描述了对小鼠心脏非心肌细胞的首次质谱细胞术研究。质谱细胞术面板与单细胞 RNA 测序数据进行了比较。通过质谱细胞术对稳态心脏非心肌细胞进行了特征描述，以鉴定和量化四个主要细胞群体：内皮细胞、成纤维细胞、白细胞和周细胞。研究了多柔比星暴露 5 天，每天 4 毫克/千克/天后，在 24 和 72 小时后单次非心肌细胞对多柔比星的急性反应。
