Has-miR-30c-1-3p inhibits macrophage autophagy and promotes Mycobacterium tuberculosis survival by targeting ATG4B and ATG9B

Abstract Autophagy is a widespread physiological process in the body, which also protects the host by degrading invading pathogens and harmful substances during pathological conditions. Nevertheless, Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis, has evolved strategies to sub...

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Main Authors: Xianglin Peng, Feifei Pu, Fangzheng Zhou, Xiyong Dai, Feng Xu, Junwen Wang, Jing Feng, Ping Xia
Format: Article
Language:English
Published: Nature Portfolio 2025-03-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-94452-w
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author Xianglin Peng
Feifei Pu
Fangzheng Zhou
Xiyong Dai
Feng Xu
Junwen Wang
Jing Feng
Ping Xia
author_facet Xianglin Peng
Feifei Pu
Fangzheng Zhou
Xiyong Dai
Feng Xu
Junwen Wang
Jing Feng
Ping Xia
author_sort Xianglin Peng
collection DOAJ
description Abstract Autophagy is a widespread physiological process in the body, which also protects the host by degrading invading pathogens and harmful substances during pathological conditions. Nevertheless, Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis, has evolved strategies to subvert autophagy by modulating microRNA (miRNA) expression, enabling its escape from host defenses. In this study, we established an in vitro model using the human macrophage cell line infected with the highly virulent MTB strain H37Rv. Through RNA sequencing and bioinformatic analysis post H37Rv infection, we screened 14 differentially expressed miRNAs. We predicted and demonstrated that miR-30c-1-3p inhibits autophagy and promotes MTB survival by targeting ATG4B and ATG9B during the infection process. The results showed that miR-30c-1-3p expression was gradually increased before 12 h of H37Rv infection, followed by a decrease. Overexpression of miR-30c-1-3p suppressed autophagic activity. We also identified the targeting of miR-30c-1-3p to ATG4B and ATG9B for the first time, and overexpression of both ATG4B and ATG9B, alone or together, on the basis with upregulation of miR-30c-1-3p reversed the inhibition of autophagy. Autophagy levels were analyzed at different levels by western blot, immunofluorescence, and transmission electron microscopy, all of which showed that upregulation of miR-30c-1-3p inhibited autophagy during H37Rv infection. Additionally, the intervention of miR-30c-1-3p mimics resulted in an increased bacterial load in macrophages, suggesting that MTB achieves immune evasion by upregulating miR-30c-1-3p during infection. In conclusion, our study provides a valuable target for the development of host-directed anti-tuberculosis therapy as well as a new diagnostic marker.
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issn 2045-2322
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spelling doaj-art-03082d79fd2544fa824926b7ea3b79432025-08-20T02:10:17ZengNature PortfolioScientific Reports2045-23222025-03-0115111310.1038/s41598-025-94452-wHas-miR-30c-1-3p inhibits macrophage autophagy and promotes Mycobacterium tuberculosis survival by targeting ATG4B and ATG9BXianglin Peng0Feifei Pu1Fangzheng Zhou2Xiyong Dai3Feng Xu4Junwen Wang5Jing Feng6Ping Xia7Department of Orthopedics, Traditional Chinese and Western Medicine Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Orthopedics, Traditional Chinese and Western Medicine Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Orthopedics, Traditional Chinese and Western Medicine Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and TechnologyWuhan Pulmonary Hospital, Wuhan Institute for Tuberculosis ControlWuhan Pulmonary Hospital, Wuhan Institute for Tuberculosis ControlDepartment of Orthopedics, Wuhan Fourth Hospital, Puai HospitalDepartment of Orthopedics, Traditional Chinese and Western Medicine Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Orthopedics, Wuhan Fourth Hospital, Puai HospitalAbstract Autophagy is a widespread physiological process in the body, which also protects the host by degrading invading pathogens and harmful substances during pathological conditions. Nevertheless, Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis, has evolved strategies to subvert autophagy by modulating microRNA (miRNA) expression, enabling its escape from host defenses. In this study, we established an in vitro model using the human macrophage cell line infected with the highly virulent MTB strain H37Rv. Through RNA sequencing and bioinformatic analysis post H37Rv infection, we screened 14 differentially expressed miRNAs. We predicted and demonstrated that miR-30c-1-3p inhibits autophagy and promotes MTB survival by targeting ATG4B and ATG9B during the infection process. The results showed that miR-30c-1-3p expression was gradually increased before 12 h of H37Rv infection, followed by a decrease. Overexpression of miR-30c-1-3p suppressed autophagic activity. We also identified the targeting of miR-30c-1-3p to ATG4B and ATG9B for the first time, and overexpression of both ATG4B and ATG9B, alone or together, on the basis with upregulation of miR-30c-1-3p reversed the inhibition of autophagy. Autophagy levels were analyzed at different levels by western blot, immunofluorescence, and transmission electron microscopy, all of which showed that upregulation of miR-30c-1-3p inhibited autophagy during H37Rv infection. Additionally, the intervention of miR-30c-1-3p mimics resulted in an increased bacterial load in macrophages, suggesting that MTB achieves immune evasion by upregulating miR-30c-1-3p during infection. In conclusion, our study provides a valuable target for the development of host-directed anti-tuberculosis therapy as well as a new diagnostic marker.https://doi.org/10.1038/s41598-025-94452-wATG4BATG9BAutophagymiR-30c-1-3pMycobacterium tuberculosisImmune evasion
spellingShingle Xianglin Peng
Feifei Pu
Fangzheng Zhou
Xiyong Dai
Feng Xu
Junwen Wang
Jing Feng
Ping Xia
Has-miR-30c-1-3p inhibits macrophage autophagy and promotes Mycobacterium tuberculosis survival by targeting ATG4B and ATG9B
Scientific Reports
ATG4B
ATG9B
Autophagy
miR-30c-1-3p
Mycobacterium tuberculosis
Immune evasion
title Has-miR-30c-1-3p inhibits macrophage autophagy and promotes Mycobacterium tuberculosis survival by targeting ATG4B and ATG9B
title_full Has-miR-30c-1-3p inhibits macrophage autophagy and promotes Mycobacterium tuberculosis survival by targeting ATG4B and ATG9B
title_fullStr Has-miR-30c-1-3p inhibits macrophage autophagy and promotes Mycobacterium tuberculosis survival by targeting ATG4B and ATG9B
title_full_unstemmed Has-miR-30c-1-3p inhibits macrophage autophagy and promotes Mycobacterium tuberculosis survival by targeting ATG4B and ATG9B
title_short Has-miR-30c-1-3p inhibits macrophage autophagy and promotes Mycobacterium tuberculosis survival by targeting ATG4B and ATG9B
title_sort has mir 30c 1 3p inhibits macrophage autophagy and promotes mycobacterium tuberculosis survival by targeting atg4b and atg9b
topic ATG4B
ATG9B
Autophagy
miR-30c-1-3p
Mycobacterium tuberculosis
Immune evasion
url https://doi.org/10.1038/s41598-025-94452-w
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