Noninvasive site-specific measurement of gene expression in deep tissues with secreted reporters

使用分泌报告基因对深层组织中的基因表达进行无创位点特异性测量

• 批准号: 10432370
• 负责人: Jerzy Olgierd Szablowski
• 金额: $ 22.02万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-13 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10432370
• 关键词: Address; Aging; Animals; Area; Autopsy; Bacterial Proteins; Biochemical; Biological Markers; Biological Models; Blood; Blood Circulation; Blood Tests; Brain; Brain Diseases; Brain Injuries; Brain imaging; Brain region; Cells; Cerebrovascular system; Characteristics; Chemicals; Color; Cytochrome P450; Detection; Development; Diffusion; Disease; Engineering; Equipment; Euthanasia; Eye; FOS gene; Ferritin; Focused Ultrasound; Future; Gene Delivery; Gene Expression; Gene Expression Profiling; Genes; Goals; Histology; Human; Image; Imaging Device; Kinetics; Label; Laboratory Research; Light; Luciferases; Magnetic Resonance Imaging; Measurement; Measures; Methods; Molecular; Monitor; Mus; Neurons; Optical Methods; Organ; Organ Size; Patients; Penetration; Physiological; Process; Production; Public Health; Reporter; Reporting; Research; Serum; Signal Transduction; Site; Specificity; Spinal Cord; Structure; Technology; Testing; Thick; Time; Tissues; Toxic effect; Translations; Vascular Permeabilities; Work; base; blood-brain barrier permeabilization; brain cell; brain size; cell type; cranium; design; gene therapy; immunogenic; improved; in vivo; interstitial; millimeter; multiplexed imaging; new technology; non-invasive monitor; promoter; protein biomarkers; sensor; side effect; therapeutic gene; therapeutic protein; transcytosis; transgene expression; ultrasound

Project Summary Monitoring gene delivery in deep tissues is critical for in vivo studies and translation of gene therapies, but available methods for achieving such measurements have limited capabilities. Ideally, levels of gene expression could be noninvasively measured anywhere in the body of both small and large animals. Ideally, multiple genes could be measured at once, without the need for expensive equipment that would put such a method out of financial reach for academic research groups. However, no such method exists as of now. In this proposal, we will develop technologies for monitoring gene expression that, when fully developed, will be: 1) noninvasive, 2) site-specific, 3) could view multiple types of molecules at once (high-multiplexity of imaging), 4) are not impacted by the depth of penetration, skull thickness, or organ size; 5) will use inexpensive equipment allowing democratized access. To achieve this, we will attempt a drastically different approach – instead of imaging reporters within the deep tissue, we will design reporters that can be easily transported from a known region of the brain into the blood, where they can be easily measured. We will develop two independent methods for achieving this goal. First, will express gene expression reporters that exit the cells into brain interstitial space. Then we will temporarily enhance vascular permeability in selected ~millimeter-sized brain regions to allow for free diffusion of these reporters from the brain into the blood. In the second method, we will develop a new class of gene expression reporters that can cross through an intact brain vasculature to continuously report on gene expression within the transduced brain cells. Spatial precision could be provided for this method using noninvasive gene delivery that genetically labels cells with site-specificity. With each of these methods, we will be able to measure gene expression in a specific brain region with a simple blood draw. Our technologies will be particularly beneficial in the studies of large animal species, where monitoring gene expression in the brain, in most cases, is not feasible without euthanasia. While initially focused on the brain as a test-case with a high need for such methods, the same concepts can be applied to other tissues.

项目摘要 监测深组织中的基因递送对于体内研究和基因疗法的翻译至关重要，但是 实现此类测量的可用方法具有有限的功能。理想情况下，基因表达水平 可以无创测量大小动物体内的任何地方。理想情况下，多个基因 可以立即测量 学术研究小组的财务影响力。但是，到目前为止，还没有这种方法。在这个建议中，我们 将开发用于监测基因表达的技术，当充分发达时将是：1）无创，2） 特定于位点特异性，3）可以一次查看多种类型的分子（成像的高词素性），4）不受影响 通过穿透深度，头骨厚度或组织尺寸； 5）将使用廉价的设备允许 民主化的访问。为了实现这一目标，我们将尝试一种截然不同的方法 - 而不是成像 深层组织中的记者，我们将设计可轻松从已知区域传输的记者 大脑进入血液，可以轻松测量它们。我们将开发两种独立方法 实现这一目标。首先，将表达将细胞退出大脑间质空间的基因表达报告。 然后，我们将暂时增强选定的〜illimeter尺寸大脑区域的血管通透性，以允许 这些记者从大脑中自由扩散到血液中。在第二种方法中，我们将开发一个新类 可以通过完整的脑脉管系统跨越基因的基因表达报告者的连续报告 在转移的脑细胞中的表达。可以使用此方法提供空间精度 非侵入性基因递送，通常用位点特异性标记细胞。使用这些方法，我们将 能够用简单的血液抽血来测量特定大脑区域中的基因表达。我们的技术会 在大型动物物种的研究中特别有益，其中监测大脑中的基因表达， 在大多数情况下，没有安乐死是不可行的。虽然最初是将大脑作为一个高的测试案例 需要这种方法，可以将相同的概念应用于其他组织。