Department of Cell Biology and Molecular Medicine, New Jersey Medical School, Rutgers Biomedical and Health Sciences, 185 South Orange Avenue/Medical Science Building G-624, Newark, NJ, 07103-2501, United States of America.
Sci Rep. 2017 Jun 8;7(1):3079. doi: 10.1038/s41598-017-03160-7.
Inhibitor of DNA binding (Id) proteins play important roles in regulating cardiac development via paracrine signaling. Id1/Id3 knockout mice die at mid-gestation with multiple cardiac defects. Single Id knockout studies have not reported cardiomyopathies. To bypass embryonic lethality we used Tie2CRE-mediated recombination to conditionally delete Id1 against global Id3 ablation (Id cDKOs), which develops adult-onset dilated cardiomyopathy. We confirm upregulation of thrombospondin-1 (TSP1) in Id cDKO hearts. Colocalization studies reveal increased TSP1 expression in the vicinity of endothelial cells and near regions of endocardial fibrosis/disruption. Downstream fibrotic molecules were upregulated. Endocardial capillary density was reduced with evidence of vascular distention. Treatment of Id cDKO cardiac explants with LSKL, a peptide antagonist of TSP1 activation of TGFβ, reversed the increased expression of fibrotic molecules. We conducted bone marrow transplant experiments in which we transferred bone marrow cells from Id cDKO mice into lethally irradiated WT mice. The majority of WT recipients of Id cDKO bone marrow cells phenocopied Id cDKO cardiac fibrosis 4 months post-transplantation. Injection of LSKL into adult Id cDKO mice led to downregulation of fibrotic molecules. The results prompt caution when bone marrow transfers from individuals potentially carrying mutations in the Id axis are applied in clinical settings.
DNA 结合抑制因子(Id)蛋白通过旁分泌信号在调节心脏发育中发挥重要作用。Id1/Id3 基因敲除小鼠在妊娠中期死亡,伴有多种心脏缺陷。单一 Id 基因敲除研究并未报道心肌病。为了避免胚胎致死性,我们使用 Tie2CRE 介导的重组来条件性删除 Id1,以消除全局的 Id3 消融(Id cDKO),这会导致成年期扩张型心肌病。我们证实 Id cDKO 心脏中血栓素-1(TSP1)的上调。共定位研究显示 TSP1 在内皮细胞附近和心内膜纤维化/破坏附近区域的表达增加。下游纤维化分子上调。心内膜毛细血管密度降低,伴有血管扩张的证据。用 LSKL(TSP1 激活 TGFβ的肽拮抗剂)处理 Id cDKO 心脏外植体,逆转了纤维化分子的表达增加。我们进行了骨髓移植实验,将 Id cDKO 小鼠的骨髓细胞转移到致死性辐射的 WT 小鼠中。大多数 WT 受体接受 Id cDKO 骨髓细胞移植 4 个月后,表现出 Id cDKO 心脏纤维化的表型。LSKL 注射到成年 Id cDKO 小鼠中导致纤维化分子的下调。这些结果提示,在临床环境中应用可能携带 Id 轴突变的个体的骨髓移植时需要谨慎。