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

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 基因过度表达是一种新型且安全的疼痛我们建议开发一种大手术大鼠模型中局部外周免疫系统的治疗靶点。细胞定向基因疗法可以纠正由以下原因引起的疼痛的潜在局部免疫学原因炎症过程，特别是亚慢性术后疼痛或炎症状况。对人类原代巨噬细胞进行无偏倚的全基因组转录组分析，我们发现 ED2/CD163 基因诱导调节肿瘤坏死因子 α (TNFa) 和白细胞介素 (IL)-1 β (IL-1b)。使用经过临床测试的纳米颗粒过度表达，旨在靶向巨噬细胞，促进更快 3D 人体器官型皮肤组织的伤口愈合并防止亚慢性术后疼痛行为并减少了接受皮肤肌肉切开和牵开 (SMIR) 手术的大鼠的局部 TNFa 和 IL-1b。巨噬细胞中的 ED2/CD163 是治疗炎性疼痛的安全靶标我们提出了一项多维研究计划，包括：1）亚慢性手术疼痛。模型、SMIR 手术和膝关节炎症模型、完全弗氏佐剂 (CFA) 诱导的膝关节关节炎；2) 使用巨噬细胞定向纳米技术获得和丧失 ED2/CD163 功能；3) 新颖的、临床的相关且复杂的操作性疼痛相关行为；4) 细胞/分子、组织和转录组结果机械目标参与；5) 研究 ED2/CD163 对阿片类药物需求的影响。通过以下具体目标实施我们的计划：1) 确定巨噬细胞特异性 ED2/CD163 基因诱导有效促进炎症疼痛的消退。旨在将核酸特异性递送至巨噬细胞的纳米颗粒（Man-PEI）将用于进行 ED2/CD163 功能的获得（过度表达）或丧失（敲低）我们将评估经典行为（von Frey）。和负重），以及新颖的复杂且临床相关的功能活动和注意力相关行为由我们的团队开发和验证 2) 定义 ED2/CD163 目标参与度，即 ED2/CD163 作为决定巨噬细胞表型变化和细胞重编程动态的信号驱动因素此外，我们还将使用 TNFa 和 IL-1b 作为下游靶点参与度。单细胞 RNAseq (scRNAseq)、聚类和表型轨迹分析，以确定 ED2/CD163 如何影响浸润发炎组织的巨噬细胞中的基因表达程序 3) 建立巨噬细胞。 ED2/CD163 基因诱导可减少手术疼痛和炎症疼痛中的阿片类药物用量。构建 SMIR 或关节炎和 ED2/CD163 过表达大鼠中吗啡的剂量反应以测量我们的项目将建立 ED2/CD163 作为针对术后疼痛的细胞定向基因疗法。这将减少阿片类药物的需求并揭示它如何影响免疫反应和炎症疼痛我们的多学科团队拥有独特的能力来成功完成这些研究。