Efficient Targeting of Therapeutic Cells in Stroke and EAE

中风和 EAE 治疗细胞的有效靶向

• 批准号: 8654367
• 负责人: Piotr Walczak
• 金额: $ 35.08万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8654367
• 关键词: Adhesions; Adverse effects; Animals; Area; Binding; Biodistribution; Biological Assay; Blood - brain barrier anatomy; Blood flow; Brain; Brain Injuries; Bypass; CCR; CXCR4 gene; Carotid Arteries; Cell Adhesion; Cell Adhesion Molecules; Cell Respiration; Cell Therapy; Cell model; Cells; Cerebrovascular Circulation; Cerebrum; Clinical; Clinical Trials; Devices; Diffusion Magnetic Resonance Imaging; Disadvantaged; Disease; Docking; Endothelial Cells; Endothelium; Engraftment; Ensure; Exhibits; Experimental Autoimmune Encephalomyelitis; Extravasation; Failure; Genetic Engineering; Histology; Homing; Hot Spot; Human; Image; In Vitro; Inflammation; Inflammatory; Injection of therapeutic agent; Integrin alpha4beta1; Intra-Arterial Injections; Intracarotid; Intravenous; Ipsilateral; Ischemia; Label; Lesion; Ligands; Lipopolysaccharides; Magnetic Resonance Imaging; Mediating; Medicine; Metabolism; Microfluidics; Modeling; Molecular; Monitor; Multiple Sclerosis; Neurons; Outcome; Parkinson Disease; Pathology; Patients; Pilot Projects; Rattus; Relative (related person); Research Personnel; Risk; Route; Safety; Science; Source; Spin Labels; Stem cells; Stroke; Surface; Techniques; Testing; Therapeutic; Time; Transgenes; Vascular Cell Adhesion Molecule-1; Weight; base; brain tissue; cellular engineering; cellular targeting; chemokine receptor; clinical application; cytokine; design; effective therapy; improved; in vitro testing; in vivo; induced pluripotent stem cell; injection/infusion; nervous system disorder; novel strategies; overexpression; preclinical study; precursor cell; prevent; progenitor; receptor; research study; response; stem cell biology; targeted delivery; therapeutic target; time use; tissue oxygenation; vascular bed

DESCRIPTION (provided by applicant): The prospect of using stem cells for therapeutic purposes has been one of the most promising fields of science and medicine in recent years. Progress in this area has been substantial, including a better understanding of stem cell biology, the identification of new sources of stem cells, and encouraging therapeutic results in a variety of diseases. Neurological disorders remain one of the greatest challenges in medicine, with little or no effective treatments available. Preclinical studies using stem cells have been encouraging and have led to the initiation of a few clinical trials for Parkinson's disease, multiple sclerosis and stroke. Unfortunately, none of these trials demonstrated a satisfactory therapeutic outcome. There are many suggested reasons for that failure, with one of the primary reasons being an inefficient biodistribution and targeting of stem cells. Intraarterial delivery could potentially bypass this limitation, and a few attempts have been made to use this approach for direct targeting of brain lesions. The major obstacle limiting this approach is the lack of techniques tha enable efficient binding of cells to endothelium, as well as the risk of microembolism as a result of excessive cell binding. Our preliminary results indicate that overexpression of the docking receptor VLA-4 greatly improves the targeting efficiency of human, glial progenitors towards areas of inflammation. Using a microfluidics in vitro adhesion assay, cell binding to activated brain endothelial cells greatly increased as compared to non-VLA-4 controls (71.5¿11.7 vs. 36.4¿3.3 cells/FOV, respectively, p=0.045). In a LPS-induced rat global inflammatory brain model, cells containing the VLA-4 transgene demonstrated much enhanced homing in vivo following intraarterial injection. Real-time, quantitative serial whole brain MR imaging of magnetically labeled cells revealed that, VLA-4+ cells docked exclusively within the vascular bed of the ipsilateral carotid artery indicating a first pass adhesion mechanism. Pixel-by-pixel analysis revealed that injection of VLA-4+ cells in LPS-treated animals resulted in 3,979¿705 hypointense pixels as compared to 868¿317 in VLA-4- LPS-treated controls (p=0.014). With these encouraging results, the overall aim of this proposal is to induce pluripotent stem cells-derived glial precursors to overexpress the adhesion molecules VLA-4 and LFA-1 and the chemokine receptors CXCR-4 and CCR2. Combined with intracarotid delivery, we hypothesize that a highly efficient and specific engraftment within inflammatory brain lesions will occur. The ability of cells to bind to endothelium and extravasate into the brain parenchyma will be initially tested in vitro using a microfluidics model blood brain barrier. Experiments will then be performed in vivo in rat models of stroke and autoimmune encephalomyelitis. To ensure the safety of this approach, we will monitor cell delivery, cerebral blood flow, and oxygenation in real-time using MRI. Upon successful completion of our studies, this new targeting approach could significantly improve the efficacy of cell-based therapy with applications in many areas of medicine.

描述（由适用提供）：近年来，将干细胞用于治疗目的的前景一直是科学和医学最有前途的领域之一。该领域的进展是很大的，包括更好地了解干细胞生物学，对干细胞的新来源的鉴定以及鼓励治疗会导致各种疾病。神经系统疾病仍然是医学中最大的挑战之一，几乎没有有效的治疗方法。使用干细胞的临床前研究令人鼓舞，并导致了一些帕金森氏病，多发性硬化症和中风的临床试验。不幸的是，这些试验都没有表现出令人满意的治疗结果。有许多建议的原因，其失败的原因之一是主要原因之一是干细胞的生物分布效率低下和靶向。内部分娩可能有可能绕过这一限制，并尝试使用这种方法直接靶向脑部病变。限制这种方法的主要障碍是缺乏能够有效的细胞与森林的结合，以及由于过量的细胞结合而导致微栓塞的风险。我们的初步结果表明，对接受体VLA-4的过表达极大地提高了人类胶质祖细胞对炎症区域的靶向效率。与非VLA-4对照组相比，使用体外粘附测定法中的微流体学分别增加了与活化的脑内皮细胞结合的细胞大大增加（71.5¿11.7vs.36.4¿3.3细胞/FOV，P = 0.045）。在LPS诱导的大鼠全球炎症大脑模型中，含有VLA-4变换的细胞在注射后体内显示出大量增强的归巢。磁性标记细胞的实时定量串行整体大脑MR成像表明，VLA-4+细胞仅在同侧颈动脉的血管床内停靠，表明第一次通过的粘合剂机制。像素像素分析表明，在LPS治疗的动物中注射VLA-4+细胞的注射导致3,979€705低渗性像素，而VLA-4-4-LPS处理的对照中的868？317（p = 0.014）。通过这些令人鼓舞的结果，该建议的总体目的是诱导多能干细胞衍生的神经胶质前体，以过表达粘合分子VLA-4和LFA-1以及趋化因子受体CXCR-4和CCR2。结合核内递送，我们假设会发生炎症性脑部病变内的高效和特异性植入。细胞与内皮结合并外外向脑paranchyma结合的能力最初将是 使用微流体模型的血脑屏障进行体外测试。然后将在中风和自身免疫性脑脊髓炎的大鼠模型中进行实验。为了确保这种方法的安全性，我们将使用MRI实时监测细胞输送，脑血流量和氧合。成功完成研究后，这种新的靶向方法可以通过在许多医学领域的应用中显着提高基于细胞的治疗的效率。