Anne Cao Le, Poh-Yi Gan, Daniel Koo Yuk Cheong, Virginie Oudin, Jonathan Dick, Maliha Alikhan, Mawj Mandwie, Ian Alexander, A. Richard Kitching, Grant J Logan, Kim Maree O'Sullivan
{"title":"腺相关病毒基因疗法增强了使用重组脱氧核糖核酸酶 I 治疗抗髓过氧化物酶肾小球肾炎的效果","authors":"Anne Cao Le, Poh-Yi Gan, Daniel Koo Yuk Cheong, Virginie Oudin, Jonathan Dick, Maliha Alikhan, Mawj Mandwie, Ian Alexander, A. Richard Kitching, Grant J Logan, Kim Maree O'Sullivan","doi":"10.1101/2024.09.09.612148","DOIUrl":null,"url":null,"abstract":"Extracellular DNA (ecDNA) released from injured and dying cells powerfully induces injurious inflammation. In this study we show ecDNA renal presence in patients and experimental mice with myeloperoxidase anti-neutrophil cytoplasmic antibody-associated glomerulonephritis (MPO-ANCA GN). Twice daily administration of intravenous DNase I (ivDNase I) in two models of anti-MPO GN was effective at reducing glomerular deposition of ecDNA, histological injury, leukocyte infiltration and NETosis. Comprehensive investigation into DNase I modes of action revealed the enzyme reduced lymph node DC numbers and their activation status, resulting in decreased frequency of MPO-specific CD4 effector T cells (IFN-gamma and IL17A producing), reductions in dermal anti-MPO delayed type hypersensitivity responses and increased frequency of MPO-specific T regulatory cells. Renal expression of inflammatory chemokines were also decreased. To overcome the translational obstacle of the short half-life of DNase I (<5 hours), we tested an adeno-associated viral vector encoding DNase I in one of the models. Along with the endpoint changes described above, a single vector treatment also enhanced therapeutic benefit as seen by reductions in MPO-ANCA and albuminuria. These results indicate ecDNA is a potent driver of anti-MPO GN and that DNase I is a potential therapeutic that can be delivered using gene technology.","PeriodicalId":501182,"journal":{"name":"bioRxiv - Immunology","volume":"207 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Treatment of anti-myeloperoxidase glomerulonephritis using recombinant deoxyribonuclease I is enhanced by adeno-associated virus gene therapy\",\"authors\":\"Anne Cao Le, Poh-Yi Gan, Daniel Koo Yuk Cheong, Virginie Oudin, Jonathan Dick, Maliha Alikhan, Mawj Mandwie, Ian Alexander, A. Richard Kitching, Grant J Logan, Kim Maree O'Sullivan\",\"doi\":\"10.1101/2024.09.09.612148\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Extracellular DNA (ecDNA) released from injured and dying cells powerfully induces injurious inflammation. In this study we show ecDNA renal presence in patients and experimental mice with myeloperoxidase anti-neutrophil cytoplasmic antibody-associated glomerulonephritis (MPO-ANCA GN). Twice daily administration of intravenous DNase I (ivDNase I) in two models of anti-MPO GN was effective at reducing glomerular deposition of ecDNA, histological injury, leukocyte infiltration and NETosis. Comprehensive investigation into DNase I modes of action revealed the enzyme reduced lymph node DC numbers and their activation status, resulting in decreased frequency of MPO-specific CD4 effector T cells (IFN-gamma and IL17A producing), reductions in dermal anti-MPO delayed type hypersensitivity responses and increased frequency of MPO-specific T regulatory cells. Renal expression of inflammatory chemokines were also decreased. To overcome the translational obstacle of the short half-life of DNase I (<5 hours), we tested an adeno-associated viral vector encoding DNase I in one of the models. Along with the endpoint changes described above, a single vector treatment also enhanced therapeutic benefit as seen by reductions in MPO-ANCA and albuminuria. These results indicate ecDNA is a potent driver of anti-MPO GN and that DNase I is a potential therapeutic that can be delivered using gene technology.\",\"PeriodicalId\":501182,\"journal\":{\"name\":\"bioRxiv - Immunology\",\"volume\":\"207 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-09-16\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"bioRxiv - Immunology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1101/2024.09.09.612148\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"bioRxiv - Immunology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1101/2024.09.09.612148","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
受伤和死亡细胞释放的细胞外 DNA(ecDNA)可强力诱发损伤性炎症。在这项研究中,我们发现在髓过氧化物酶抗中性粒细胞胞浆抗体相关性肾小球肾炎(MPO-ANCA GN)患者和实验小鼠肾脏中存在 ecDNA。在两种抗 MPO GN 模型中,每天两次静脉注射 DNase I(ivDNase I)可有效减少肾小球的 ecDNA 沉积、组织学损伤、白细胞浸润和 NETosis。对 DNase I 作用模式的全面研究表明,该酶能减少淋巴结 DC 的数量及其活化状态,从而降低 MPO 特异性 CD4 效应 T 细胞(产生 IFN-gamma 和 IL17A)的频率,减少真皮抗 MPO 迟发型超敏反应,增加 MPO 特异性 T 调节细胞的频率。肾脏中炎症趋化因子的表达也有所减少。为了克服 DNase I 半衰期短(5 小时)这一转化障碍,我们在其中一个模型中测试了编码 DNase I 的腺相关病毒载体。除了上述终点变化外,单一载体治疗还能提高治疗效果,这体现在 MPO-ANCA 和白蛋白尿的减少上。这些结果表明,ecDNA 是抗 MPO GN 的强大驱动力,而且 DNase I 是一种可利用基因技术提供的潜在疗法。
Treatment of anti-myeloperoxidase glomerulonephritis using recombinant deoxyribonuclease I is enhanced by adeno-associated virus gene therapy
Extracellular DNA (ecDNA) released from injured and dying cells powerfully induces injurious inflammation. In this study we show ecDNA renal presence in patients and experimental mice with myeloperoxidase anti-neutrophil cytoplasmic antibody-associated glomerulonephritis (MPO-ANCA GN). Twice daily administration of intravenous DNase I (ivDNase I) in two models of anti-MPO GN was effective at reducing glomerular deposition of ecDNA, histological injury, leukocyte infiltration and NETosis. Comprehensive investigation into DNase I modes of action revealed the enzyme reduced lymph node DC numbers and their activation status, resulting in decreased frequency of MPO-specific CD4 effector T cells (IFN-gamma and IL17A producing), reductions in dermal anti-MPO delayed type hypersensitivity responses and increased frequency of MPO-specific T regulatory cells. Renal expression of inflammatory chemokines were also decreased. To overcome the translational obstacle of the short half-life of DNase I (<5 hours), we tested an adeno-associated viral vector encoding DNase I in one of the models. Along with the endpoint changes described above, a single vector treatment also enhanced therapeutic benefit as seen by reductions in MPO-ANCA and albuminuria. These results indicate ecDNA is a potent driver of anti-MPO GN and that DNase I is a potential therapeutic that can be delivered using gene technology.