Mir221- and Mir222-enriched adsc-exosomes mitigate PM exposure-exacerbated cardiac ischemia-reperfusion injury through the modulation of the BNIP3-MAP1LC3B-BBC3/PUMA pathway

Tzu-Lin Lee; Wen-Chi Shen; Ya-Chun Chen; Tsai-Chun Lai; Shu-Rung Lin; Shu-Wha Lin; I-Shing Yu; Yen-Hsiu Yeh; Tsai-Kun Li; I-Ta Lee; Chiang-Wen Lee; Yuh-Lien Chen

doi:10.1080/15548627.2024.2395799

Mir221- and Mir222-enriched adsc-exosomes mitigate PM exposure-exacerbated cardiac ischemia-reperfusion injury through the modulation of the BNIP3-MAP1LC3B-BBC3/PUMA pathway

Autophagy. 2025 Feb;21(2):374-393. doi: 10.1080/15548627.2024.2395799. Epub 2024 Sep 8.

Authors

Tzu-Lin Lee¹, Wen-Chi Shen¹, Ya-Chun Chen¹, Tsai-Chun Lai^{1

2

3}, Shu-Rung Lin^{4

5}, Shu-Wha Lin⁶, I-Shing Yu⁷, Yen-Hsiu Yeh⁸, Tsai-Kun Li^{8

9

10}, I-Ta Lee¹¹, Chiang-Wen Lee^{12

13

14}, Yuh-Lien Chen¹

Affiliations

¹ Department of Anatomy and Cell Biology, College of Medicine, National Taiwan University, Taipei, Taiwan.
² Department of Life Sciences, College of Life Sciences, National Chung Hsing University, Taichung, Taiwan.
³ The iEGG and Animal Biotechnology Center, National Chung Hsing University, Taichung, Taiwan.
⁴ Department of Bioscience Technology, College of Science, Chung Yuan Christian University, Taoyuan, Taiwan.
⁵ Center for Nanotechnology, Chung Yuan Christian University, Taoyuan, Taiwan.
⁶ Department of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan University, Taipei, Taiwan.
⁷ Laboratory Animal Center, College of Medicine, National Taiwan University, Taipei, Taiwan.
⁸ Department and Graduate Institute of Microbiology, College of Medicine, National Taiwan University, Taipei, Taiwan.
⁹ Center for Biotechnology, National Taiwan University, Taipei, Taiwan.
¹⁰ Centers for Genomic and Precision Medicine, National Taiwan University, Taipei, Taiwan.
¹¹ School of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei, Taiwan.
¹² Department of Orthopaedic Surgery, Chang Gung Memorial Hospital, Puzi, Chiayi, Taiwan.
¹³ Department of Nursing, Division of Basic Medical Sciences, and Chronic Diseases and Health Promotion Research Center Chang Gung University of Science and Technology, Puzi, Chiayi, Taiwan.
¹⁴ Research Center for Industry of Human Ecology and Research Center for Chinese Herbal Medicine, Chang Gung University of Science and Technology, Taoyuan, Taiwan.

Abstract

Epidemiology has shown a strong relationship between fine particulate matter (PM) exposure and cardiovascular disease. However, it remains unknown whether PM aggravates myocardial ischemia-reperfusion (I/R) injury, and the related mechanisms are unclear. Our previous study has shown that adipose stem cell-derived exosomes (ADSC-Exos) contain high levels of Mir221 and Mir222. The present study investigated the effects of PM exposure on I/R-induced cardiac injury through mitophagy and apoptosis, as well as the potential role of Mir221 and Mir222 in ADSC-Exos. Wild-type, mir221- and mir222-knockout (KO), and Mir221- and Mir222-overexpressing transgenic (TG) mice were intratracheally injected with PM (10 mg/kg). After 24 h, mice underwent left coronary artery ligation for 30 min, followed by 3 h of reperfusion (I/R). H9c2 cardiomyocytes were cultured under 1% O₂ for 6 h, then reoxygenated for 12 h (hypoxia-reoxygenation [H/R]). PM aggravated I/R (or H/R) cardiac injury by increasing ROS levels and causing mitochondrial dysfunction, which increased the expression of mitochondrial fission-related proteins (DNM1L/Drp1 and MFF) and mitophagy-related proteins (BNIP3 and MAP1LC3B/LC3B) in vivo and in vitro. Treatment with ADSC-Exos or Mir221- and Mir222-mimics significantly reduced PM+I/R-induced cardiac injury. Importantly, ADSC-Exos contain Mir221 and Mir222, which directly targets BNIP3, MAP1LC3B/LC3B, and BBC3/PUMA, decreasing their expression and ultimately reducing cardiomyocyte mitophagy and apoptosis. The present data showed that ADSC-Exos treatment regulated mitophagy and apoptosis through the Mir221 and Mir222-BNIP3-MAP1LC3B-BBC3/PUMA pathway and significantly reduced the cardiac damage caused by PM+I/R. The present study revealed the novel therapeutic potential of ADSC-Exos in alleviating PM-induced exacerbation of myocardial I/R injury.Abbreviation: ADSC-Exos: adipose-derived stem cell exosomes; AL: autolysosome; ATP: adenosine triphosphate; BBC3/PUMA: BCL2 binding component 3; BNIP3: BCL2/adenovirus E1B interacting protein 3; CASP3: caspase 3; CASP9: caspase 9; CDKN1B/p27: cyclin dependent kinase inhibitor 1B; CVD: cardiovascular disease; DCFH-DA: 2',7'-dichlorodihydrofluorescein diacetate; DHE: dihydroethidium; DNM1L/Drp1: dynamin 1-like; EF: ejection fraction; FS: fractional shortening; H/R: hypoxia-reoxygenation; I/R: ischemia-reperfusion; LDH: lactate dehydrogenase; MAP1LC3B/LC3B: microtubule-associated protein 1 light chain 3 beta; MFF: mitochondrial fission factor; miRNA: microRNA; NAC: N-acetylcysteine; OCR: oxygen consumption rate; PIK3C3/Vps34: phosphatidylinositol 3-kinase catalytic subunit type 3; PM: particulate matter; PRKAA1/AMPK: protein kinase AMP-activated catalytic subunit alpha 1; ROS: reactive oxygen species; SQSTM1/p62: sequestosome 1; TEM: transmission electron microscopy; TRP53/p53: transformation related protein 53; TUNEL: terminal deoxynucleotidyl transferase dUTP nick end labeling.

Keywords: ADSC-exosomes; Mir221 and Mir222; cardiomyocyte; ischemia/reperfusion injury; mitophagy; particulate matter.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis / drug effects
Apoptosis Regulatory Proteins* / metabolism
Autophagy
Cell Line
Exosomes* / metabolism
Male
Membrane Proteins* / metabolism
Mice
Mice, Inbred C57BL
Mice, Transgenic
MicroRNAs* / genetics
MicroRNAs* / metabolism
Microtubule-Associated Proteins / metabolism
Mitochondrial Proteins* / metabolism
Mitophagy / drug effects
Myocardial Reperfusion Injury* / genetics
Myocardial Reperfusion Injury* / metabolism
Myocardial Reperfusion Injury* / pathology
Myocytes, Cardiac / drug effects
Myocytes, Cardiac / metabolism
Myocytes, Cardiac / pathology
Particulate Matter* / adverse effects
Particulate Matter* / toxicity
Proto-Oncogene Proteins / metabolism
Rats
Signal Transduction / drug effects

Substances

MicroRNAs
Microtubule-Associated Proteins
Membrane Proteins
Particulate Matter
Mitochondrial Proteins
BNip3 protein, mouse
Map1lc3b protein, mouse
Apoptosis Regulatory Proteins
MIRN221 microRNA, mouse
Proto-Oncogene Proteins

Grants and funding

This work was supported by research grants from the National Science and Technology Council [MOST 108-2320-B-002-065-MY3; NSTC 111-2320-B-002-022-MY3 to Y.L.C.], Chang Gung University of Science Foundation [grant ZRRPF6N0011 to C.W.L.], and Chang Gung Medical Research Program Foundation [grants CORPF6P0041, CORPF6P0042, CORPF6P0043 to C.W.L.].