Targeted Systemic Delivery of SDF-1 DNA for the Treatment of Chronic Heart Disease

项目摘要/摘要 基于基质衍生因子-1(SDF-1)的基因治疗可促进间充质干细胞(MSC)归巢 心肌梗死区治疗急性心肌梗死或慢性心力衰竭 (CHF)在临床前和临床试验中显示出相当大的前景。然而，最成熟的 技术要求使用特定的设备进行经心内膜注射，这只能由 专家。为了方便管理和具有成本效益的治疗，有针对性的全身给药策略 有待开发。在这一应用中，编码SDF-1的质粒DNA将被系统地和特异性地 通过形成SDF-1/缺血心肌靶向多肽(IMTP)将SDF-1传递到梗死部位 很复杂。为了生成SDF-1/IMTP络合物，我们首次利用了配体到金属的电荷转移 允许将靶向多肽直接掺入到载体SDF-1 DNA中而不需要 对于任何基因携带者来说。我们推测，锌离子与巯基之间的LMCT跃迁 靶向多肽的半胱氨酸可以自发地推动多肽与载体SDF-1的整合 它已经被修改为含有锌离子。已知二价金属离子，如锌离子， 通过插入DNA碱基将正常的B-DNA转化为金属结合的DNA(M-DNA 对在pH为7.0-8.5的范围内。在初步研究中，我们用锌离子产生了M-DNA，证实 通过LMCT转变形成M-DNA/靶向多肽复合体，并证明了 M-DNA/靶向多肽复合体可增强基因在靶细胞中的转染率。在这 在CHF的动物模型中，我们打算展示靶向系统的治疗效果。 通过LMCT产生的SDF-1/IMTP复合体的形成来传递SDF-1 DNA 过渡。为了实现最终目标，我们打算演示以下内容：1)将IMTP直接整合到 2)将SDF-1基因靶向导入低氧细胞 动物模型中的原代心肌细胞和梗死灶；3)促进MSCs向 SDF-1在动物模型中的梯度。

PROJECT SUMMARY / ABSTRACT Stromal-derived factor-1 (SDF-1)-based gene therapy that can improve mesenchymal stem cell (MSC) homing to the region of myocardial infarct for the treatment of acute myocardial infarction (MI) or chronic heart failure (CHF) has shown considerable promise in preclinical and clinical trials. However, the most established technology requires trans-endocardial injection using a specific device, which can only be handled by an expert. For convenience administration and cost-effective treatment, a targeted systemic delivery strategy has to be developed. In this application, a plasmid DNA encoding SDF-1 will be systemically and specifically delivered to the infarct site by the formation of the SDF-1/ischemic myocardium targeting peptide (IMTP) complex. To generate the SDF-1/IMTP complex, for the first time, we utilize the ligand-to-metal charge transfer transition allowing for direct incorporation of the targeting peptide to the plasmid SDF-1 DNA without the need for any gene carriers. We hypothesized that the LMCT transition between Zn2+ ions and the sulfhydryl group in cysteine of the targeting peptide could spontaneously drive the integration of the peptide to the plasmid SDF-1 that has already been modified to contain Zn2+ ions. It has been known that divalent metal ions, such as Zn2+, lead to the conversion of normal B-DNA to metal-bound DNA (M-DNA) through intercalation into the DNA base pairs in the pH range of 7.0-8.5. In the preliminary studies, we generated M-DNA using Zn2+ ions, confirmed the formation of the M-DNA/targeting peptide complex through the LMCT transition, and demonstrated that the M-DNA/targeting peptide complex led to the enhancement in the gene transfection in the target cells. In this project, in an animal model of CHF, we intend to demonstrate therapeutic efficacy of the targeted systemic delivery of a plasmid SDF-1 DNA by the formation of the SDF-1/IMTP complex generated through the LMCT transition. To achieve the final goal, we intend to demonstrate the followings: 1) direct integration of IMTP into the SDF-1 plasmid through the LMCT transition; 2) targeted transfection of the SDF-1 gene into hypoxic primary cardiomyocytes and the infarct site in the animal model; and 3) facilitated migration of MSCs towards the SDF-1 gradient in the animal model.