Development of Brain MRI Contrast Agents

脑MRI造影剂的开发

基本信息

批准号：
10916002
负责人：
Alan Koretsky
金额：
$ 108.06万
依托单位：
NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10916002
关键词：
2019-nCoV Acceleration Aerosols Arizona Binding Biological Assay Blood - brain barrier anatomy Blood capillaries Brain Brain Diseases COVID-19 detection COVID-19 therapeutics Collaborations Contrast Media Copper Detection Development Diagnosis Diagnostic Dose Exhalation Family Goals Health Sciences Hour Human Image In Vitro Individual Injections Intravenous Investigation Joints Kinetics Laboratories Lead Magnetic Resonance Imaging Maryland Methods Modeling Molecular Mus Mutant Strains Mice National Institute of Neurological Disorders and Stroke Neurologic Neurosciences Pathologic Patients Pharmacodynamics Pharmacologic Substance Positron Positron-Emission Tomography Post-Traumatic Headaches Radiation exposure Radiology Specialty Regenerative Medicine Research Research Personnel Research Project Grants Resolution Signal Transduction Site Stroke Supervision TFRC gene Testing Therapeutic Time Transgenic Mice United States National Institutes of Health Universities University Health Services Visualization Washington Work blood-brain barrier crossing brain magnetic resonance imaging contrast imaging cost design detection method dimer drug discovery imaging agent imaging study improved in vivo intravenous injection iron oxide nanoparticle mathematical model molecular modeling molecular pathology mouse model nanobodies nervous system disorder novel therapeutics receptor receptor binding response therapeutic candidate transcytosis

项目摘要

We have made substantial progress towards Aims 1-5, 7&8. Specifically: a) We have demonstrated that a mutant mouse transferrin receptor binding nanobody with strong pH dependent unbinding crosses the BBB in vivo approximately 500 times more effectively than a control nanobody. b) We have demonstrated that the mutant mouse transferrin receptor binding nanobody can carry other nanobodies as cargo across the mouse BBB. A tandem dimer of nanobodies that bind to the P2X7 receptor has been used as a model cargo. c) We have shown that cargo transported across the BBB can be detected in capillary depleted brain lysates and histochemically at their target sites between 1 and 16 hours after injection. d) We have demonstrated that there is a strong nonlinearity in the dose-response curve for the BBB crossing nanobodies. At doses greater than 30 nmol/kg injected intravenously, there is no greater brain accumulation of cargo. The maximum brain concentration achieved was 4.5 nM. This concentration is an order of magnitude too low for MRI detection of iron oxide nanoparticles. We therefore have decided to switch to PET imaging for further development molecular contrast imaging agents. e) We are developing methods to conjugate nanobodies to the positron emitting Copper-64 for PET scans. f) We have established a new collaboration with Dr. Dale Kiesewetter and his research group to perform PET scans in living mice at multiple time points after injection. g) We have initiated new collaborations with M Janoswki at U Maryland, D Sehlin and S. Syvanen at Uppsala U, and I Huggins at Ionis Pharmaceuticals to perform additional imaging studies using our BBB crossing nanobody constructs. h) We have established new collaborations with Dr F. Nugent at USUHS, Dr. N. Plesnila at U Munich, and Dr. F. Porreca at U Arizona to test our nanobody constructs for efficacy as BBB crossing P2X7 receptor blocking therapeutics in mouse models of TBI, stroke, and post-traumatic headache. i) Ongoing work by our collaborators at Washington University indicates that our SARS-CoV-2 nanobodies have potential for use in real time environmental surveillance of aerosols and for detection of SARS-CoV-2 in exhaled breath.

我们已经在目标1-5、7和8方面取得了重大进展。具体来说： a）我们已经证明，与对照纳米机构相比，具有强pH依赖性的突变小鼠转铁蛋白受体结合纳米植物的体内横穿BBB约500倍。 b）我们已经证明，突变小鼠转移蛋白受体结合纳米病毒可以携带其他纳米体作为跨小鼠BBB的货物。与P2X7受体结合的纳米型的串联二聚体已被用作模型货物。 c）我们已经证明，可以在注射后1至16小时之间在毛细管耗尽的脑裂解物中检测到跨BBB的货物。 d）我们已经证明了BBB穿越纳米体的剂量反应曲线中存在很强的非线性。在静脉注射的剂量大于30 nmol/kg时，货物的大脑积累没有更大的货物。达到的最大脑浓度为4.5 nm。该浓度是一个数量级，无法检测到氧化铁纳米颗粒的MRI检测。因此，我们已决定改用PET成像以进一步开发分子对比成像剂。 e）我们正在开发将纳米偶联的方法与铜扫描发射铜-64的纳米结合。 f）我们已经与Dale Kiesewetter博士及其研究小组建立了新的合作，以在注射后多个时间点在活着的老鼠中进行PET扫描。 g）我们已经在Uppsala U的U Janoswki，D Sehlin和S. Syvanen与M Janoswki启动了新的合作，在Ionis Pharmaceuticals的I Huggins使用我们的BBB交叉交叉纳米体构建体进行了其他成像研究。 h）我们已经与USUHS的F. Nugent博士，U慕尼黑的N. Plesnila博士建立了新的合作TBI，中风和创伤后头痛。 i）我们在华盛顿大学的合作者正在进行的工作表明，我们的SARS-COV-2纳米构造有可能用于实时环境监视气溶胶，并在呼出的呼吸中检测SARS-COV-2。