Multiscale drug screening for cardiac fibrosis identifies MD2 as a therapeutic target

Affiliations

• Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA
• Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA
• Department of Medicine, Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA
• Department of Internal Medicine, Division of Cardiovascular Medicine, University of California, Davis, Davis, CA 95616, USA
• David Geffen School of Medicine at University of California, Los Angeles, CA 90095, USA
• Greenstone Biosciences, Palo Alto, CA 94305, USA
• Department of Experimental Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA
• Department of Basic Medical Sciences and Translational Cardiovascular Research Center, University of Arizona College of Medicine, Phoenix, AZ 85004, USA

PMID:  #  DOI: 10.1016/j.cell.2024.09.034

Abstract

Cardiac fibrosis impairs cardiac function, but no effective clinical therapies exist. To address this unmet need, we employed a high-throughput screening for antifibrotic compounds using human induced pluripotent stem cell (iPSC)-derived cardiac fibroblasts (CFs). Counter-screening of the initial candidates using iPSC-derived cardiomyocytes and iPSC-derived endothelial cells excluded hits with cardiotoxicity. This screening process identified artesunate as the lead compound. Following profibrotic stimuli, artesunate inhibited proliferation, migration, and contraction in human primary CFs, reduced collagen deposition, and improved contractile function in 3D-engineered heart tissues. Artesunate also attenuated cardiac fibrosis and improved cardiac function in heart failure mouse models. Mechanistically, artesunate targeted myeloid differentiation factor 2 (MD2) and inhibited MD2/Toll-like receptor 4 (TLR4) signaling pathway, alleviating fibrotic gene expression in CFs. Our study leverages multiscale drug screening that integrates a human iPSC platform, tissue engineering, animal models, in silico simulations, and multiomics to identify MD2 as a therapeutic target for cardiac fibrosis.

心脏纤维化会损害心脏功能，但目前尚无有效的临床疗法。为了解决这一未满足的需求，我们采用高通量筛选技术，通过人诱导多能干细胞（iPSC）衍生的心脏成纤维细胞（CFs）筛选抗纤维化化合物。在使用iPSC衍生的心肌细胞和内皮细胞进行的反向筛选中，排除了具有心脏毒性的候选物。该筛选过程确定了青蒿琥酯（artesunate）作为领先化合物。在促纤维化刺激后，青蒿琥酯（artesunate）抑制了人原代CFs的增殖、迁移和收缩，减少了胶原沉积，并改善了三维工程心脏组织的收缩功能。青蒿琥酯（artesunate）还在心力衰竭小鼠模型中减轻了心脏纤维化并改善了心脏功能。从机制上看，青蒿琥酯（artesunate）靶向骨髓分化因子2（MD2），抑制MD2/TLR4信号通路，减轻CFs中的纤维化基因表达。我们的研究利用多尺度药物筛选，整合了人iPSC平台、组织工程、动物模型、计算机模拟和多组学，以确定MD2作为心脏纤维化的治疗靶点。