Preservation and Vascularization of Cardiac Extracellular Matrix after Myocardial Infarction

心肌梗死后心脏细胞外基质的保存和血管化

基本信息

批准号：
10094074
负责人：
Jianjun Guan
金额：
$ 45.38万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-01-01 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10094074
关键词：
3-Dimensional Affect Antibodies Area Attenuated Blood Vessels Blood capillaries Cardiac Cicatrix Clinical Trials Collagen Deposition Dose-Limiting Drug Delivery Systems Endothelial Cells Extracellular Matrix Extracellular Matrix Degradation FGF2 gene Fibroblasts Gelatinase A Gelatinase B Goals Growth Factor Heart Hydrogels Inflammation Injections Matrix Metalloproteinase Inhibitor Matrix Metalloproteinases Mediating Modeling Monitor Morphogenesis Myocardial Infarction Myofibroblast Outcome Pathway interactions Peptides Pharmaceutical Preparations Principal Investigator Rattus Research Role Signal Pathway Structure System Testing Therapeutic Thick Time Tissues Toxic effect Transforming Growth Factor beta Vascularization Work angiogenesis base cell motility coronary fibrosis density heart function heart preservation implantation improved in vitro Model in vivo inhibitor/antagonist innovation novel therapeutics preservation prevent programs protein expression side effect spatiotemporal

项目摘要

Following myocardial infarction (MI), the degradation of cardiac extracellular matrix (ECM) mainly by upregulated matrix metalloproteinase-2/9 (MMP-2/9), and the progression of cardiac fibrosis after myofibroblast formation, progressively deteriorate cardiac function. As such, impeding MMP-2/9 bioactivity, and inhibiting myofibroblast formation will improve cardiac function. However, the ideal therapeutic strategies to simultaneously achieve both goals remain to be established. Currently, systemic delivery of broad spectrum MMP inhibitors did not show consistent outcomes in clinical trials. MMP-2/9 expression is spatiotemporal in infarcted hearts over the course of post-MI. Yet current systemic delivery approach cannot spatiotemporally deliver MMP inhibitors to the infarcted area. To attenuate cardiac fibrosis, systemic delivery of TGFβ inhibitors or anti-TGFβ antibodies represents a major approach. However, it only decreases the content of active TGFβ. It cannot inhibit TGFβ signaling pathway to prevent myofibroblast formation. Furthermore, the small organic MMP and TGFβ inhibitors have toxicity concerns. The objective of this project is to create drug delivery systems that can be specifically delivered into infarcted hearts to concurrently preserve cardiac ECM, and prevent cardiac fibrosis. Localized delivery will eliminate dose-limiting side effects. The systems will spatiotemporally release MMP-2/9 specific and non-toxic inhibitor, peptide CTTHWGFTLC (CTT), to specifically modulate local MMP-2/9 bioactivity. The systems will also gradually release a multifunctional growth factor bFGF that have anti-fibrotic and proangiogenesis functions. The preserved ECM will thus be vascularized. Vascularization is critical for cardiac ECM as otherwise its structure and composition change over time. In our preliminary work, we have created a fast gelation and degradable hydrogel-based release system capable of efficiently retaining drugs in beating hearts. The system can release CTT for 4 weeks. After being injected into infarcted hearts, the released CTT preserved collagen, increased tissue thickness, and improved cardiac function. Better than many other small organic MMP inhibitors, CTT did not induce cardiac fibrosis. Besides, CTT promoted endothelial cell migration in the presence of TGFβ that is upregulated after MI. These results demonstrate that CTT is potentially a better MMP inhibitor for cardiac therapy than those small organic inhibitors. We have further created a release system that continuously releases both CTT and bFGF. bFGF is known for its angiogenic effect. We found that bFGF is capable of inhibiting TGFβ-induced cardiac fibroblast differentiation into myofibroblast through TGFβ/Erk1/2 pathway. After 4 weeks of implantation, the CTT/bFGF release systems not only increased tissue thickness and preserved collagen composition, but also promoted the formation of a high density of capillaries and remarkably reduced cardiac fibrosis, leading to the increase of cardiac function. Based on our preliminary studies, we hypothesize that localized and spatiotemporal delivery of CTT and bFGF into infarcted hearts, will concurrently attenuate cardiac ECM degradation, vascularize the preserved ECM, and prevent cardiac fibrosis, leading to a significant increase in cardiac function. AIM 1 will test the hypothesis that optimal CTT release profiles will efficiently attenuate MMP-2 bioactivity to prevent MMP-2 mediated ECM degradation. AIM 2 will test the hypothesis that optimal bFGF release profiles will simultaneously promote endothelial cell morphogenesis and prevent cardiac fibroblasts from differentiating into myofibroblasts. AIM 3 will test the hypothesis that delivery of CTT and bFGF release systems after MI will concurrently preserve and vascularize cardiac ECM, and prevent cardiac fibrosis. This project is innovative because it creates translational drug delivery systems to establish: 1) role and efficacy of an efficient MMP-2/MMP-9 inhibitor CTT in cardiac therapy; 2) mechanism and efficacy of bFGF in inhibiting cardiac fibrosis while promoting angiogenesis; and 3) how sustained release of CTT and bFGF simultaneously achieves these three goals. The system is relatively simple and multifunctional. Therefore, it is translational.

心肌梗死（MI）后，心脏细胞外基质（ECM）的降解主要通过基质金属蛋白酶-2/9（MMP-2/9）的上调，以及肌成纤维细胞形成后心脏纤维化的进展，导致心脏功能逐渐恶化。，阻碍 MMP-2/9 的生物活性，并抑制肌成纤维细胞的形成将改善心脏功能。然而，目前仍需建立同时实现这两个目标的理想治疗策略。广谱 MMP 抑制剂在临床试验中并未表现出一致的结果，即梗塞后的心脏中 MMP-2/9 的表达是时空的。 TGFβ抑制剂或抗TGFβ抗体的全身给药是治疗纤维化的主要方法，但它只能降低活性TGFβ的含量，不能抑制TGFβ信号通路。此外，小型有机 MMP 和 TGFβ 抑制剂存在毒性问题，该项目的目标是创建可以特异性递送至梗塞心脏以同时保护心脏 ECM 并防止心脏纤维化的药物递送系统。将消除剂量限制性副作用。该系统将时空释放 MMP-2/9 特异性无毒抑制剂、肽 CTTHWGFTLC (CTT)，以特异性调节局部。 MMP-2/9 生物活性。该系统还将逐渐释放具有抗纤维化和促血管生成功能的多功能生长因子，因此血管化对于心脏 ECM 至关重要，否则其结构和成分会随着时间的推移而发生变化。在我们的前期工作中，我们创建了一种基于快速凝胶化和可降解的水凝胶的释放系统，该系统能够有效地将药物保留在跳动的心脏中，该系统在注射到心脏后可以释放CTT 4周。与许多其他小型有机 MMP 抑制剂相比，CTT 释放的 CTT 不会诱导心肌纤维化，此外，CTT 在 TGFβ 存在下促进内皮细胞迁移，而 TGFβ 则上调。 MI。这些结果表明，CTT 可能是比那些小型有机抑制剂更好的 MMP 抑制剂。我们进一步创建了一种能够持续释放 CTT 和 bFGF 的释放系统。我们发现bFGF能够通过TGFβ/Erk1/2途径抑制TGFβ诱导的心肌成纤维细胞分化。植入4周后，CTT/bFGF释放系统不仅增加了组织厚度并保留了胶原成分。还促进了高密度毛细血管的形成并显着减少了心脏纤维化，从而导致心脏功能的增强。根据我们的初步研究，我们发现了这种局部性和时空性。将 CTT 和 bFGF 递送至梗塞心脏，将同时减弱心脏 ECM 降解，使保留的 ECM 血管化，并防止心脏纤维化，从而导致心脏功能显着增强，AIM 1 将检验 CTT 释放曲线将有效减弱 MMP 的最佳假设。 -2 防止 MMP-2 介导的 ECM 降解的生物活性 AIM 2 将测试最佳 bFGF 释放曲线将同时促进内皮细胞形态发生并预防心脏的假设。 AIM 3 将测试 MI 后 CTT 和 bFGF 释放系统的递送将同时保护心脏 ECM 和血管化并预防心脏纤维化这一假设，因为它创建了转化药物递送系统以建立：1。 ) 高效 MMP-2/MMP-9 抑制剂 CTT 在心脏治疗中的作用和功效 2) bFGF 在抑制心脏纤维化同时促进心脏纤维化的机制和功效；血管生成；3) CTT 和 bFGF 如何同时实现这三个目标。因此，它是转化性的。