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

基本信息

批准号：
10704050
负责人：
Jerzy Olgierd Szablowski
金额：
$ 18.29万
依托单位：
RICE UNIVERSITY
依托单位国家：
美国
项目类别：
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/10704050
关键词：
Address 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 Democracy 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 Immobilization 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 blood-brain barrier permeabilization brain cell brain size cell type cranium design gene therapy immunogenic improved in vivo interstitial millimeter new technology non-invasive monitor promoter protein biomarkers sensor side effect therapeutic gene therapeutic protein transcytosis transgene expression ultrasound

项目摘要

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.

项目概要监测深层组织中的基因传递对于体内研究和基因疗法的转化至关重要，但是理想情况下，实现这一目标的现有方法的能力有限。理想情况下，可以无创地测量小型和大型动物体内的多个基因。可以立即测量，而不需要昂贵的设备，这会使这种方法失效然而，在本提案中，目前还没有这样的方法。将开发监测基因表达的技术，该技术完全开发后将：1）非侵入性，2）特定位点，3) 可以同时查看多种类型的分子（成像的高度多重性），4) 不受影响根据穿透深度、颅骨厚度或器官大小；5) 将使用允许的廉价设备；为了实现这一目标，我们将尝试一种截然不同的方法——而不是想象。在深层组织内生产，我们将设计可以轻松从已知区域运输的产品大脑进入血液，我们将开发两种独立的方法来测量它们。首先，将表达产生离开细胞进入大脑间隙的基因。然后，我们将暂时增强选定的毫米级大脑区域的血管通透性，以允许在第二种方法中，我们将开发一个新的类别。产生的基因表达可以穿过完整的脑脉管系统来持续报告基因使用该方法可以提供转导脑细胞内的表达空间精度。通过这些方法中的每一种，我们都可以通过非侵入性基因传递对细胞进行位点特异性标记。我们的技术将能够通过简单的抽血来测量特定大脑区域的基因表达。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，如果不实施安乐死是不可行的，而最初将大脑作为具有高风险的测试案例。需要这样的方法，相同的概念可以应用于其他组织。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Engineered serum markers for non-invasive monitoring of gene expression in the brain.

用于无创监测大脑基因表达的工程血清标记物。

DOI：
发表时间：
2024-01-10
期刊：
影响因子：
46.9
作者：
Lee, Sangsin;Nouraein, Shirin;Kwon, James J;Huang, Zhimin;Wojick, Jessica A;Xia, Boao;Corder, Gregory;Szablowski, Jerzy O
通讯作者：
Szablowski, Jerzy O