Targeted migration of stem cells to improve cellular therapy after TB

干细胞的靶向迁移可改善结核病后的细胞治疗

• 批准号: 8399471
• 负责人: PAUL J YAROWSKY
• 金额: --
• 依托单位: BALTIMORE VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8399471
• 关键词: Address; Adult; Affect; Alloys; Animal Model; Animals; Antigens; Area; Blood - brain barrier anatomy; Brain; Brain region; Caliber; Cell Survival; Cells; Cerebral cortex; Contralateral; Coupled; Cyclosporine; Distress; Engraftment; Ensure; Extravasation; Fluorescence; Genes; Head; Healthcare Systems; Heat-Shock Proteins 70; Hemorrhage; Hippocampus (Brain); Hour; Human; Immune; Implant; In Vitro; Incidence; Injection of therapeutic agent; Injury; Intra-Arterial Injections; Intracarotid; Intraventricular; Ipsilateral; Iron; Knowledge; Label; Laboratories; Lesion; Life; Long-Term Care; Magnetic Resonance Imaging; Magnetism; Metalloproteases; Methods; Microglia; Microscopic; Military Personnel; Modeling; Movement; Needles; Neocortex; Neodymium; Nerve Degeneration; Neuraxis; Nickel; Outcome Measure; Patients; Penetration; Population; Positioning Attribute; Preparation; Procedures; Proteins; Rattus; Reagent; Research; Rhodamine; Rhodamine B; Risk; Safety; Severities; Side; Signal Transduction; Site; Skin; Staining method; Stains; Stem cell transplant; Stem cells; Stroke; Structure; Techniques; Technology; Testing; Therapeutic; Therapeutic Intervention; Thick; Time; Tissues; Transfection; Transplantation; Traumatic Brain Injury; Up-Regulation; Veterans; Xenograft procedure; base; care burden; cell motility; cranium; design; functional outcomes; human stem cells; improved; injured; insight; intravenous injection; iron oxide; lateral ventricle; magnetic field; migration; morris water maze; nanoparticle; nerve stem cell; neurogenesis; neuron loss; neurorestoration; neurotrophic factor; novel therapeutics; overexpression; preclinical study; prevent; primary outcome; public health relevance; research study; stem cell therapy; therapeutic target; therapy design; Address; Adult; Affect; Alloys; Animal Model; Animals; Antigens; Area; Blood - brain barrier anatomy; Brain; Brain region; Caliber; Cell Survival; Cells; Cerebral cortex; Contralateral; Coupled; Cyclosporine; Distress; Engraftment; Ensure; Extravasation; Fluorescence; Genes; Head; Healthcare Systems; Heat-Shock Proteins 70; Hemorrhage; Hippocampus (Brain); Hour; Human; Immune; Implant; In Vitro; Incidence; Injection of therapeutic agent; Injury; Intra-Arterial Injections; Intracarotid; Intraventricular; Ipsilateral; Iron; Knowledge; Label; Laboratories; Lesion; Life; Long-Term Care; Magnetic Resonance Imaging; Magnetism; Metalloproteases; Methods; Microglia; Microscopic; Military Personnel; Modeling; Movement; Needles; Neocortex; Neodymium; Nerve Degeneration; Neuraxis; Nickel; Outcome Measure; Patients; Penetration; Population; Positioning Attribute; Preparation; Procedures; Proteins; Rattus; Reagent; Research; Rhodamine; Rhodamine B; Risk; Safety; Severities; Side; Signal Transduction; Site; Skin; Staining method; Stains; Stem cell transplant; Stem cells; Stroke; Structure; Techniques; Technology; Testing; Therapeutic; Therapeutic Intervention; Thick; Time; Tissues; Transfection; Transplantation; Traumatic Brain Injury; Up-Regulation; Veterans; Xenograft procedure; base; care burden; cell motility; cranium; design; functional outcomes; human stem cells; improved; injured; insight; intravenous injection; iron oxide; lateral ventricle; magnetic field; migration; morris water maze; nanoparticle; nerve stem cell; neurogenesis; neuron loss; neurorestoration; neurotrophic factor; novel therapeutics; overexpression; preclinical study; prevent; primary outcome; public health relevance; research study; stem cell therapy; therapeutic target; therapy design

DESCRIPTION (provided by applicant): Stem cell transplantation following traumatic brain injury (TBI) has been suggested as part of a therapeutic intervention to mitigate the severity of the injury. To date, preclinical studies of cel based therapeutics have focused on stereotactic injection of cells into, or adjacent to regions of injury. However, this delivery approach is problematic due to frequently limited and unpredictable migratory capacity of most of these cells. Coupled with variability in the cells ability to localize to the injury area, multiple injections are commonly performed. The cumulative effect of TBI, together with multiple penetrations/injections into the brain to delivery cells, increases the risk of bleeding/direct tissue injury counterproductive to the benefit the cells may offer. To address this issue, we propose to develop a practical and clinically applicable method to enhance the transplantation efficacy of stem cells following TBI. This approach involves loading stem cells, human neuroprogenitor cells (NPCs), with the ferromagnetic compound MION, combined with externally positioned magnets to 'pull' loaded stem cells towards the site of cortical injury. We propose to also enhance the ability of the cells to migrate through the brai by the use of transient metalloproteinase overexpression. One of the key advantages to our proposed stem cell migration strategy is that the stem cells can be delivered by injection either into the lateral ventricle or intra-arterially and directed towards the injured region. Currently tere are no technologies available to improve the migration, retention, and survival of stem cells transplanted into the central nervous system. Our hypothesis is that MION labeling and magnetic targeting and upregulation of Heat shock protein 70 (Hsp70) in stem cells will enhance their migration and survival into the lesion in an animal model of traumatic brain injury (TBI). Specific Aim 1A: To determine whether MION- labeled NPCs transfected with HSP-70 can be localized at the site of lesion in the ipsilateral cortex by injection into the ipsilateral lateral ventricle. Specific Aim 1B: To determine whether transplanted MION-labeled NPCs can be localized in the cerebral cortex at the site of lesion with intra-arterial injection of stem cells n TBI animals. Specific Aim 1C: To determine whether transplanted MION-labeled NPCs can be localized in the cerebral cortex at the site of lesion with intravenous injection of stem cells in BI animals. Specific Aim 2) With these proof-of-principle experiments verifying our hypothesis that an applied magnetic field can target the migration of stem cells within the brain, we will use MIONRB-labeled NPCs in a rat model of TBI and determine whether we can localize SPIO- labeled NPCs at the sites of neuropathological changes using either form of injection. Functional outcome following transplantation will be evaluated with the Morris water maze test. Following the injury there will also be a significant increase in the number of activated microglia in the ara of the cortex adjacent to the site of the lesion. Treatment with NPCs will evaluate if the increase in the number of activated microglia is decreased following transplantation and no additional microgliosis occurs due to the procedure. Using magnets (neodymium) on one side of the cortex, the contralateral cortex will serve as the control side. We have already demonstrated that in non-TBI animals, MION-labeled NPCs are localized in the ipsilateral cortex following injection into the ipsilateral lateral ventricle. We will assess the retention of the NPCs at the lesion site over short intervals (hours) after transplantation and over longer intervals (days). In TBI, the brain blood barrier is compromised, thus as a sub aim, we will assess if the magnetic field will enhance the extravasation of stem cells. Under all these conditions, we will augment the survival of the stem cells migrating into the lesion, by their transfection with pro-survival gene, HSP70. The same studies will be carried out without HSP70 transfection. Using these techniques, we will optimize stem cell transplantation so that it will be a more effective targeted therapeutic method. PUBLIC HEALTH RELEVANCE: The incidence of traumatic brain injury (TBI) among the U.S. military veteran population is increasing and will impose a long-term care burden on the VA health care system. It is ever more important to develop new therapeutics that will facilitate the neurorestorative capacity of the central nervous system in TBI. Recent research has proposed using stem cell therapy. The lack of basic knowledge concerning stem cell migration following a transplant remains a bottleneck for attempting to design therapies. We have designed a method by using directed magnetic fields and labeling stem cells with MIONRB to deliver stem cells to cortical regions damaged in TBI. We will test our methods of targeting stem cells in an animal model of TBI. These results could shed further insight into using stem cell therapy as a neurorestorative therapy.

描述（由申请人提供）： 脑部损伤（TBI）后的干细胞移植已被建议作为治疗干预的一部分，以减轻损伤的严重程度。迄今为止，基于CEL的治疗剂的临床前研究集中于对细胞的立体定向注射到损伤区域或附近。但是，由于大多数这些细胞的经常有限且不可预测的迁移能力，这种传递方法是有问题的。结合细胞定位到损伤区域的能力的可变性，通常进行多次注射。 TBI的累积作用，以及对大脑递送细胞的多次穿透/注射，增加了出血/直接组织损伤适得其反的风险，以适得其反。为了解决这个问题，我们建议开发一种实用且临床上适用的方法，以增强TBI后干细胞的移植功效。这种方法涉及加载干细胞，人类神经元素细胞（NPC），具有铁磁化合物mion，并与外部定位的磁体结合使用，将载荷干细胞“拉动”皮质损伤部位。我们建议还通过使用瞬态金属蛋白酶过表达来增强细胞通过BRAI迁移的能力。我们提出的干细胞迁移策略的关键优势之一是，可以通过注射到侧心室或动脉内并针对受伤区域来递送干细胞。目前，尚无用于改善移植到中枢神经系统的干细胞的迁移，保留和存活的技术。我们的假设是，在干细胞中热休克蛋白70（HSP70）的mion标记和磁靶向以及上调将增强其迁移和存活到脑损伤动物模型（TBI）中。具体目标1A：确定是否可以通过注射到同侧侧向侧面，将用HSP-70转染的MION标记的NPC定位在同侧皮质的病变部位 心室。具体目标1B：确定是否可以将移植的MION标记的NPC定位在病变部位的大脑皮层中，并在动脉内注射干细胞N TBI动物。具体目标1C：确定是否可以将移植的Mion标记的NPC定位在病变部位的大脑皮层中，并在BI动物中静脉注射干细胞。 Specific Aim 2) With these proof-of-principle experiments verifying our hypothesis that an applied magnetic field can target the migration of stem cells within the brain, we will use MIONRB-labeled NPCs in a rat model of TBI and determine whether we can localize SPIO- labeled NPCs at the sites of neuropathological changes using either form of injection.移植后的功能结果将通过莫里斯水迷宫检验进行评估。损伤后，在病变部位附近的皮质的ARA中，活化的小胶质细胞的数量也将显着增加。 NPC治疗将评估是否增加 在移植后，活化的小胶质细胞的数量减少，并且由于该过程而没有发生其他小胶质细胞增多。使用皮质一侧的磁铁（新近山），对侧皮层将用作控制侧。我们已经证明，在非TBI动物中，注射到同侧外侧心室后，MION标记的NPC位于同侧皮层中。我们将在移植后短时（小时）和更长的间隔（天）评估NPC在病变部位的保留。在 TBI，脑血屏障被损害，因此作为子目标，我们将评估磁场是否会增强干细胞的渗出。在所有这些条件下，我们将通过促寿使基因HSP70的转染来增强干细胞迁移到病变中的存活。在没有HSP70转染的情况下将进行相同的研究。使用这些技术，我们将优化干细胞移植，以便它是更有效的目标 治疗方法。 公共卫生相关性： 美国军事退伍军人人口中创伤性脑损伤（TBI）的发生率正在增加，并将对VA医疗保健系统施加长期护理负担。开发新的治疗剂将促进中枢神经系统在TBI中的神经养生能力。最近的研究提出了使用干细胞疗法。缺乏关于移植后干细胞迁移的基本知识仍然是试图设计疗法的瓶颈。我们通过使用定向磁场并用MionRB将干细胞标记为将干细胞传递到在TBI中受损的皮质区域中设计了一种方法。我们将测试在TBI动物模型中靶向干细胞的方法。这些结果可能会进一步深入了解使用干细胞疗法作为神经训练疗法。