MRI contrast for molecular and cellular imaging of the brain

人们对开发分子成像方法的兴趣不断增加，这种方法使传统的放射成像技术能够获得有关正常和病变组织中发生的分子和细胞过程的广泛信息。一系列信息被认为是重要的，如监测细胞迁移的能力，能够成像基因表达的报告者的发展，成像受体的强大策略的发展，以及可用于检测特定酶的存在或监测离子状态变化的环境敏感剂的发展。这项工作的长期目标是制定策略，使MRI对比对广泛的分子和细胞过程敏感。这项工作建立在30多年的工作基础上，我们已经展示了第一个检测基因表达的MRI策略，第一个监测钙内流替代物的MRI方法，第一个执行神经元轨迹追踪的MRI方法，以及第一个监测体内单细胞迁移的MRI方法。这些都代表了任何放射成像技术的初步报告，使这些过程能够被测量。这些技术被广泛应用于多种疾病的临床前模型成像。在过去一年中，我们在所有具体目标方面都取得了进展。

There continues to be increasing interest in developing molecular imaging approaches that enable traditional radiological imaging techniques to obtain a wide range of information about molecular and cellular processes that occur in normal and diseased tissue. A range of information is considered important such as the ability to monitor cell migration, the development of reporters that enable imaging of gene expression, the development of robust strategies to image receptors, and the development of environmentally sensitive agents that can be used to detect the presence of specific enzymes or monitor changes in ion status. The long term goals of this work are to develop strategies that enable MRI contrast that is sensitive to a wide range of molecular and cellular processes. This work builds on over 30 years of work where we have demonstrated the first MRI strategy for detecting gene expression, the first MRI approach for monitoring a surrogate of calcium influx, the first MRI approach for performing neuronal track tracing, and the first MRI approach for monitoring the migration of single cells in vivo. These all represented initial reports by any radiological imaging technique which enabled these processes to be measured. These techniques are finding widespread application to imaging pre-clinical models of a broad range of diseases. Over the past year we have made progress in all of the specific aims. Aim 1: Develop iron oxide based contrast for labeling and imaging the migration of endogenous neural stem cells. Over the past few years we have demonstrated the unique advantages of micron sized iron oxide particles for MRI of specific cells. Single cells can be detected and indeed, single particles within single cells can be detected. The main paradigm for MRI of cell migration is to label cells ex vivo and monitor migration after transplantation into an animal. The ability to detect a single particle enables inefficient labeling strategies. In particular, over the past few years we have demonstrated that injection of particles into the ventricles of the rat brain enables particles to be taken up by neural precursors in the subventricular zone and MRI can monitor the migration of cells to the olfactory bulb. A paper got published that measured the changes in migration of new neurons during unilateral nasal occlusion and recovery There was exquisite coupling between bulb anatomy and cell migration both during nasal block and recovery. New neurons were required for bulb re-growth after naris occlusion. We are now studying whether introduction of specific odors after naris oclusion alters the pattern of migration of the cells into the bulb. In a new project inspired by studying these endogenous new neurons we have shown that cortical and mid-brain precursor cells can be grown in the adult CSF to form fully integrated and normally appearing brain tissue. Over the past year we hav shown that this new tissue can integrate into the motor pathway if plasced in CSF near tehstriatum and can integrate into the olfactory pathway when placed in CSF neat the rostral migratory stream to the bulb. A second major project images the entire brain to study immune brain interactions. We have succeeded in detecting T cell migration into the brain at single cell level during virus infection in mouse models. Intreting in many cases T cell accumulation is associated with small bleeding opening the issue of whether T cell infiltration into brain parenchyma is associated with BBB breakdown even when very few cells are involved. Aim 2: Apply microfabrication techniques to manufacture unique metal structures that may be valuable for MRI contrast. Iron oxide particles commonly used for MRI are very potent contrast agents enabling detection of single micron sized particles. However, due to bulk phase manufacture of particles they are not very uniform and they do not contain very high content of metal. A solution to this problem is to use modern microfabrication techniques to manufacture metal based, micron sized contrast agents. Over the past few years we have shown that double doughnut, cylinders, and ellipsoid structures offer unique advantages for distinguishing particles and that these structures can be turned into sensors for pH. We have demonstrated that simple microfabricated gold coated iron discs for cell tracing can be used both for imaging new neurons as well as immune cell infiltration. Finally, our collaborator at NIST, Gary Zabow (former fellow)has developed novel ways to manufacture this class of MRI contrast that are being tested here at NIH. Over the past year, in collabortation with M. Barbic, we have demonstrated that magnetocaloric materials have unique properties that have potential to be very useful for MRI studies of cell tracking. Aim 3: Develop novel delivery mechanisms to extend the applicability of manganese enhanced MRI. Over the past ten years we have demonstrated the remarkable utility of the manganese ion for MRI contrast. Manganese ion enters cells on ligand or voltage gated calcium channels and so can be used as an MRI agent to monitor calcium influx. Once inside of neurons, manganese will move in an anterograde direction and cross functional synapses enabling neuronal networks to be imaged with MRI. Finally, manganese given systemically gives cytoarchitectural information about the rodent brain. These successes have us interested in broadening the ways in which manganese ion can be delivered to cells. A major limitation of manganese enhanced MRI are the concentrations required. This limits translation for use in human imaging. Over the past couple of years we have tackled this problem in two ways, we have published an initial study that demonstrates that manganese positron emitting isotopes will enable PET to obtain similar information that can be obtained with manganese enhanced MRI, including neural tracing and functional activation of tissue. Discussion have begun to decide if to translate this to human studies. We completed our first study in collaboration with Daniel Reich to test if an FDA approved agent that releases Mn might be useful for disease detection. The first manuscript on normal volunteers was published. Studies on MS patients and patients with epilepsy are underway. Please note all human studies are done under the protocols of our collaborators and that is where the relevant human use data is reported for this work. Finally, in collaboration with Dorian McGavern we continue pursuing novel approaches to add Mn2+ to the brain without disruption of the skull using the intricate vessel/marrow system of the skull. Aim 4: Develop strategies that enable cellular processes to alter the relaxivity of MRI contrast agents. We continue to explore ways in which the microfabricated particles produced under Aim 2 can be modulated. Over the past few years we have completed a study that demonstrates that the microfabricated particles can be made into a sensors. Over the past year we have sought ways to make the manufacture of this very interesting class of sensors more robust. In addition, we have developed a novel polymer of Gd chelates that has high relaxivity but relatively low molecular weight. This has been attached to a class of classical neural tracers (CTB and dextrans) to enable new MRI neural tracers. A novel, miniature wireless MRI detector that shows much promise for use where it may be possible to implant an MRI detector has been developed. Two engineering solutions have been demonstrated. Over the past year we have collaborated with a group at NCI to demonstrate the usefulness of this approach for in vivo EPR experiment. Work on an interventional Pituritary imaging coil with Dr. Chittiboina is proceeding and if succesful, we will make this coil wireless to help in surgical procedures.