Wanling Huang , Gaohong Fu , Yangeng Wang , Cheng Chen , Yilan Luo , Qiaoqiao Yan , Yang Liu , Chengqiong Mao
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We developed an aerosolized core-shell liposomal nanoplatform (CSNs) complexed with CaP for efficient drug loading, targeting lung macrophages. Various CSNs were synthesized to encapsulate an mRNA based CRISPR/Cas9 system (mCas9/gHK2), and their gene editing efficiency and HK2 knockout were examined at both gene and protein levels in vitro and in vivo. The CSN-mCas9/gHK2 treatment demonstrated a significant reduction in glycolysis and inflammation in macrophages. In an LPS-induced ALI mouse model, inhaled CSN-mCas9/gHK2 mitigated the proinflammatory tumor microenvironment and reprogrammed glucose metabolism in the lung, suggesting a promising strategy for ALI prevention and treatment. This study highlights the potential of combining CRISPR/Cas9 gene editing with inhalation delivery systems for effective, localized pulmonary disease treatment, underscoring the importance of targeted gene modulation and metabolic reprogramming in managing ALI.</p></div><div><h3>Statement of Significance</h3><p>This study investigates an inhalable CRISPR/Cas9 gene editing system targeting pulmonary macrophages, with the aim of modulating glucose metabolism to alleviate Acute Lung Injury (ALI). The research highlights the role of immune cell metabolism in inflammation, as evidenced by changes in macrophage glucose metabolism and a notable reduction in pulmonary edema and inflammation. Additionally, observed alterations in macrophage polarization and cytokine levels in bronchoalveolar lavage fluid suggest potential therapeutic implications. These findings not only offer insights into possible ALI treatments but also contribute to the understanding of immune cell metabolism in inflammatory diseases, which could be relevant for various inflammatory and metabolic disorders.</p></div>","PeriodicalId":237,"journal":{"name":"Acta Biomaterialia","volume":null,"pages":null},"PeriodicalIF":9.4000,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Immunometabolic reprogramming of macrophages with inhalable CRISPR/Cas9 nanotherapeutics for acute lung injury intervention\",\"authors\":\"Wanling Huang , Gaohong Fu , Yangeng Wang , Cheng Chen , Yilan Luo , Qiaoqiao Yan , Yang Liu , Chengqiong Mao\",\"doi\":\"10.1016/j.actbio.2024.03.031\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Acute lung injury (ALI) represents a critical respiratory condition typified by rapid-onset lung inflammation, contributing to elevated morbidity and mortality rates. Central to ALI pathogenesis lies macrophage dysfunction, characterized by an overabundance of pro-inflammatory cytokines and a shift in metabolic activity towards glycolysis. This study emphasizes the crucial function of glucose metabolism in immune cell function under inflammatory conditions and identifies hexokinase 2 (HK2) as a key regulator of macrophage metabolism and inflammation. Given the limitations of HK2 inhibitors, we propose the CRISPR/Cas9 system for precise HK2 downregulation. We developed an aerosolized core-shell liposomal nanoplatform (CSNs) complexed with CaP for efficient drug loading, targeting lung macrophages. Various CSNs were synthesized to encapsulate an mRNA based CRISPR/Cas9 system (mCas9/gHK2), and their gene editing efficiency and HK2 knockout were examined at both gene and protein levels in vitro and in vivo. The CSN-mCas9/gHK2 treatment demonstrated a significant reduction in glycolysis and inflammation in macrophages. In an LPS-induced ALI mouse model, inhaled CSN-mCas9/gHK2 mitigated the proinflammatory tumor microenvironment and reprogrammed glucose metabolism in the lung, suggesting a promising strategy for ALI prevention and treatment. This study highlights the potential of combining CRISPR/Cas9 gene editing with inhalation delivery systems for effective, localized pulmonary disease treatment, underscoring the importance of targeted gene modulation and metabolic reprogramming in managing ALI.</p></div><div><h3>Statement of Significance</h3><p>This study investigates an inhalable CRISPR/Cas9 gene editing system targeting pulmonary macrophages, with the aim of modulating glucose metabolism to alleviate Acute Lung Injury (ALI). The research highlights the role of immune cell metabolism in inflammation, as evidenced by changes in macrophage glucose metabolism and a notable reduction in pulmonary edema and inflammation. Additionally, observed alterations in macrophage polarization and cytokine levels in bronchoalveolar lavage fluid suggest potential therapeutic implications. 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引用次数: 0
摘要
急性肺损伤(ALI)是一种严重的呼吸系统疾病,其典型特征是快速发作的肺部炎症,导致发病率和死亡率升高。急性肺损伤发病机制的核心是巨噬细胞功能障碍,其特点是促炎细胞因子过多和代谢活动转向糖酵解。这项研究强调了在炎症条件下葡萄糖代谢在免疫细胞功能中的关键作用,并确定己糖激酶 2(HK2)是巨噬细胞代谢和炎症的关键调节因子。鉴于 HK2 抑制剂的局限性,我们建议使用 CRISPR/Cas9 系统精确下调 HK2。我们开发了一种与 CaP 复合物的气溶胶核壳脂质体纳米平台(CSNs),用于高效载药,靶向肺巨噬细胞。我们合成了多种CSNs来封装基于mRNA的CRISPR/Cas9系统(mCas9/gHK2),并在体外和体内的基因和蛋白质水平上检测了它们的基因编辑效率和HK2基因敲除。经 CSN-mCas9/gHK2 处理后,巨噬细胞中的糖酵解和炎症明显减少。在 LPS 诱导的 ALI 小鼠模型中,吸入 CSN-mCas9/gHK2 可减轻促炎性肿瘤微环境,并重塑肺部葡萄糖代谢,这为预防和治疗 ALI 提供了一种前景广阔的策略。这项研究凸显了将 CRISPR/Cas9 基因编辑与吸入给药系统结合起来进行有效的局部肺病治疗的潜力,强调了靶向基因调控和代谢重编程在控制 ALI 方面的重要性。这项研究调查了针对肺巨噬细胞的可吸入CRISPR/Cas9基因编辑系统,目的是调节葡萄糖代谢以缓解急性肺损伤(ALI)。研究强调了免疫细胞代谢在炎症中的作用,巨噬细胞葡萄糖代谢的变化以及肺水肿和炎症的明显减轻就是证明。此外,在支气管肺泡灌洗液中观察到的巨噬细胞极化和细胞因子水平的变化也表明了潜在的治疗意义。这些发现不仅为可能的 ALI 治疗提供了见解,而且有助于了解炎症性疾病中的免疫细胞代谢,这可能与各种炎症性和代谢性疾病有关。
Immunometabolic reprogramming of macrophages with inhalable CRISPR/Cas9 nanotherapeutics for acute lung injury intervention
Acute lung injury (ALI) represents a critical respiratory condition typified by rapid-onset lung inflammation, contributing to elevated morbidity and mortality rates. Central to ALI pathogenesis lies macrophage dysfunction, characterized by an overabundance of pro-inflammatory cytokines and a shift in metabolic activity towards glycolysis. This study emphasizes the crucial function of glucose metabolism in immune cell function under inflammatory conditions and identifies hexokinase 2 (HK2) as a key regulator of macrophage metabolism and inflammation. Given the limitations of HK2 inhibitors, we propose the CRISPR/Cas9 system for precise HK2 downregulation. We developed an aerosolized core-shell liposomal nanoplatform (CSNs) complexed with CaP for efficient drug loading, targeting lung macrophages. Various CSNs were synthesized to encapsulate an mRNA based CRISPR/Cas9 system (mCas9/gHK2), and their gene editing efficiency and HK2 knockout were examined at both gene and protein levels in vitro and in vivo. The CSN-mCas9/gHK2 treatment demonstrated a significant reduction in glycolysis and inflammation in macrophages. In an LPS-induced ALI mouse model, inhaled CSN-mCas9/gHK2 mitigated the proinflammatory tumor microenvironment and reprogrammed glucose metabolism in the lung, suggesting a promising strategy for ALI prevention and treatment. This study highlights the potential of combining CRISPR/Cas9 gene editing with inhalation delivery systems for effective, localized pulmonary disease treatment, underscoring the importance of targeted gene modulation and metabolic reprogramming in managing ALI.
Statement of Significance
This study investigates an inhalable CRISPR/Cas9 gene editing system targeting pulmonary macrophages, with the aim of modulating glucose metabolism to alleviate Acute Lung Injury (ALI). The research highlights the role of immune cell metabolism in inflammation, as evidenced by changes in macrophage glucose metabolism and a notable reduction in pulmonary edema and inflammation. Additionally, observed alterations in macrophage polarization and cytokine levels in bronchoalveolar lavage fluid suggest potential therapeutic implications. These findings not only offer insights into possible ALI treatments but also contribute to the understanding of immune cell metabolism in inflammatory diseases, which could be relevant for various inflammatory and metabolic disorders.
期刊介绍:
Acta Biomaterialia is a monthly peer-reviewed scientific journal published by Elsevier. The journal was established in January 2005. The editor-in-chief is W.R. Wagner (University of Pittsburgh). The journal covers research in biomaterials science, including the interrelationship of biomaterial structure and function from macroscale to nanoscale. Topical coverage includes biomedical and biocompatible materials.