Cell-directed gene therapy for pain recovery after surgery and inflammation

用于手术和炎症后疼痛恢复的细胞定向基因疗法

基本信息

批准号：
10546458
负责人：
THOMAS JEFFREY MARTIN
金额：
$ 45.9万
依托单位：
WAKE FOREST UNIVERSITY HEALTH SCIENCES
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：
2022-01-15 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10546458
关键词：
3-Dimensional Acute Pain Address Anti-Inflammatory Agents Arthritis Attention Behavior Behavior assessment Behavioral Bioinformatics Biological Assay Cells Chronic Chronic inflammatory pain Clinical Complementary DNA Complex Critical Pathways Data Development Dimensions Disease Progression Dose Dose Limiting Drug abuse Freund&apos s Adjuvant Gene Expression Genes Genetic Transcription Human Immune response Immune system Immunobiology Immunologics Infiltration Inflammation Inflammation Mediators Inflammatory Injury Interleukin-1 beta Intervention Knee Light Link Macrophage Measures Mechanics Mediator Modeling Molecular Morphine Muscle Nanotechnology Nucleic Acids Operative Surgical Procedures Opioid Outcome Pain Pain management Patients Peripheral Pharmaceutical Preparations Phenotype Plasmids Polyethyleneimine Positioning Attribute Postoperative Pain Process Production Rattus Recovery Recovery of Function Research Resolution Signal Pathway Signal Transduction Skin Skin Tissue Small Interfering RNA Speed Surgical Models Surgical incisions TNF gene Technology Testing Therapeutic Tissues Traction Trauma Weight-Bearing state addiction chronic pain chronic pain management clinically relevant cytokine design gain of function gene induction gene therapy genetic signature genome-wide high risk human data improved inflammatory pain injured innovation insight knock-down loss of function multidisciplinary nanoparticle non-opioid analgesic novel opioid abuse opioid epidemic opioid sparing overexpression pain behavior pain model pain reduction postoperative recovery prevent programs response side effect single-cell RNA sequencing surgical pain therapeutic target tissue repair tool transcriptome sequencing transcriptomics translational potential wound wound healing

3-Dimensional Acute Pain Address Anti-Inflammatory Agents Arthritis Attention Behavior Behavior assessment Behavioral Bioinformatics Biological Assay Cells Chronic Chronic inflammatory pain Clinical Complementary DNA Complex Critical Pathways Data Development Dimensions Disease Progression Dose Dose Limiting Drug abuse Freund&apos s Adjuvant Gene Expression Genes Genetic Transcription Human Immune response Immune system Immunobiology Immunologics Infiltration Inflammation Inflammation Mediators Inflammatory Injury Interleukin-1 beta Intervention Knee Light Link Macrophage Measures Mechanics Mediator Modeling Molecular Morphine Muscle Nanotechnology Nucleic Acids Operative Surgical Procedures Opioid Outcome Pain Pain management Patients Peripheral Pharmaceutical Preparations Phenotype Plasmids Polyethyleneimine Positioning Attribute Postoperative Pain Process Production Rattus Recovery Recovery of Function Research Resolution Signal Pathway Signal Transduction Skin Skin Tissue Small Interfering RNA Speed Surgical Models Surgical incisions TNF gene Technology Testing Therapeutic Tissues Traction Trauma Weight-Bearing state addiction chronic pain chronic pain management clinically relevant cytokine design gain of function gene induction gene therapy genetic signature genome-wide high risk human data improved inflammatory pain injured innovation insight knock-down loss of function multidisciplinary nanoparticle non-opioid analgesic novel opioid abuse opioid epidemic opioid sparing overexpression pain behavior pain model pain reduction postoperative recovery prevent programs response side effect single-cell RNA sequencing surgical pain therapeutic target tissue repair tool transcriptome sequencing transcriptomics translational potential wound wound healing

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项目摘要

Summary We have recently discovered macrophage ED2/CD163 gene overexpression as a novel and safe pain therapeutic target in the local peripheral immune system in a major surgery rat model. We propose to develop a cell-directed gene therapy that would correct the underlying local immunological cause of pain resulting from inflammatory processes, specifically in sub-chronic postoperative pain or inflammatory conditions. Through unbiased genome-wide transcriptomic analyses in human primary macrophages we identified that ED2/CD163 gene induction modulates tumor necrosis factor alpha (TNFa) and interleukin (IL)-1 beta (IL-1b). CD163 overexpression using a clinically tested nanoparticle designed to target macrophages promoted a more rapid wound healing in 3D human organotypic skin tissues and prevented sub-chronic postoperative pain behaviors and reduced local TNFa and IL-1b in rats with skin-muscle incision and retraction (SMIR) surgery. We hypothesize that ED2/CD163 in macrophages is a safe target for the treatment of inflammatory pain with opioid sparing effects. We propose a multidimensional research plan including, 1) a sub-chronic surgical pain model, the SMIR surgery, and a knee inflammatory mode, the Complete Freund Adjuvant (CFA)-induced knee arthritis; 2) ED2/CD163 gain and loss of function using macrophage-directed nanotechnology; 3) novel, clinically relevant, and complex operant pain-related behaviors; 4) cellular/molecular, tissue, and transcriptomic outcomes for mechanistic target engagement; and 5) studies for ED2/CD163’s effects on opioid requirements. We will implement our plan through these specific aims: 1) Determine that macrophage specific ED2/CD163 gene induction effectively promotes resolution of inflammatory pain. A mannosilated polyethyleneimine nanoparticles (Man-PEI) designed to deliver nucleic acids specifically to macrophages will be used to conduct ED2/CD163 gain (overexpression) or loss (knock down) of function. We will assess classic behaviors (von Frey and weight bearing), and novel complex and clinically relevant functional activity and attention-related behaviors developed and validated by our team. 2) Define ED2/CD163 target engagement, i.e. ED2/CD163 as a signaling driver that dictates the dynamics of macrophage phenotype change and cellular reprogramming in inflammatory pain. TNFa and IL-1b will be measured as downstream target engagement. Also, we will use single-cell RNAseq (scRNAseq), cluster, and phenotype trajectory analysis to define how ED2/CD163 impacts gene expression programs in macrophages infiltrating the inflamed tissue. 3) Establish that macrophage ED2/CD163 gene induction results in opioid-sparing effects in surgical and inflammatory pain. We will construct dose responses of morphin in rats with SMIR or arthritis and ED2/CD163 overexpression to measure opioid-sparing effects. Our project will establish ED2/CD163 as a cell-directed gene therapy for postsurgical pain that will reduce opioid requirements and disinter how it influences immune responses and inflammatory pain recovery. Our multidisciplinary team is uniquely equipped to successfully complete these studies.

概括我们最近发现巨噬细胞ED2/CD163基因过表达是一种新颖而安全的疼痛主要手术大鼠模型中局部外周免疫系统中的治疗靶标。我们建议开发一个细胞定向的基因疗法将纠正因炎症过程，特别是在术后疼痛或炎症情况下进行的炎症过程。通过在人类原代巨噬细胞中，无偏基因组的转录组分析我们确定ED2/CD163 基因诱导调节肿瘤坏死因子α（TNFA）和白介素（IL）-1β（IL-1B）。 CD163 使用临床测试的纳米颗粒旨在靶向巨噬细胞的过表达促进了更快的 3D人体有机皮肤组织中的伤口愈合并预防后慢性后疼痛行为并减少了皮肤肌肉切口和缩回（SMIR）手术的大鼠中局部TNFA和IL-1B。我们假设巨噬细胞中的ED2/CD163是治疗炎症性疼痛与阿片类药物的保留作用。我们提出了一项多维研究计划，包括，1）亚气术疼痛模型，Smir手术和膝盖炎症模式，完整的Freund辅助（CFA）诱导的膝盖关节炎; 2）ED2/CD163使用巨噬细胞指导的纳米技术增益和功能丧失； 3）新颖，临床上相关且复杂的操作疼痛相关行为； 4）细胞/分子，组织和转录组结果用于机械目标参与； 5）研究ED2/CD163对阿片类药物需求的影响。我们将通过这些特定目的实施我们的计划：1）确定巨噬细胞特异性ED2/CD163基因诱导有效地促进了炎症性疼痛的分辨率。甘露的聚乙烯亚胺旨在将核酸专门传递到巨噬细胞的纳米颗粒（MAN-PEI）将用于进行 ED2/CD163增益（过表达）或功能的损失（击倒）。我们将评估经典行为（冯·弗雷和负重轴承），新型复杂且与临床相关的功能活动以及与注意力相关的行为由我们的团队开发和验证。 2）定义ED2/CD163目标参与，即ED2/CD163为A 决定巨噬细胞表型变化和细胞重编程的动态的信号驱动器在炎症性疼痛中。 TNFA和IL-1B将被测量为下游目标参与。另外，我们将使用单细胞RNASEQ（SCRNASEQ），群集和表型轨迹分析，以定义ED2/CD163如何影响巨噬细胞中的基因表达程序渗入发炎的组织。 3）确定巨噬细胞 ED2/CD163基因诱导会导致阿片类药物对手术和炎症性疼痛的影响。我们将在患有SMIR或关节炎和ED2/CD163过表达的大鼠中吗啡的构建剂量反应以测量阿片类药物的效果。我们的项目将建立ED2/CD163作为术后疼痛的细胞指导基因疗法这将减少阿片类药物的要求并删除其如何影响免疫复杂和炎症性疼痛恢复。我们的多学科团队与成功完成这些研究完全相当。