Theranostics 2020; 10(24):11324-11338. doi:10.7150/thno.47893 This issue Cite

Research Paper

Analysis of mesenchymal stem cell proteomes in situ in the ischemic heart

Dunzheng Han1,2*, Junjie Yang1*, Eric Zhang1, Yanwen Liu1, Chan Boriboun1, Aijun Qiao1, Yang Yu1, Jiacheng Sun1, Shiyue Xu1, Liu Yang1, Wenying Yan3, Bihui Luo2, Dongfeng Lu2, Chunxiang Zhang1, Chunfa Jie4, James Mobley5, Jianyi Zhang1, Gangjian Qin1✉

1. Department of Biomedical Engineering, University of Alabama at Birmingham, School of Medicine and School of Engineering, Birmingham, AL 35294, USA.
2. Department of Cardiology, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510120, China.
3. School of Biology and Basic Medical Science, Soochow University, Suzhou, Jiangsu 215123, China.
4. Department of Biochemistry and Nutrition, Des Moines University Medicine and Health Sciences, Des Moines, IA 50312, USA.
5. Department of Anesthesiology and Perioperative Medicine, University of Alabama at Birmingham, School of Medicine, Birmingham, AL 35294, USA.
*These authors contributed equally to this work.

Citation:
Han D, Yang J, Zhang E, Liu Y, Boriboun C, Qiao A, Yu Y, Sun J, Xu S, Yang L, Yan W, Luo B, Lu D, Zhang C, Jie C, Mobley J, Zhang J, Qin G. Analysis of mesenchymal stem cell proteomes in situ in the ischemic heart. Theranostics 2020; 10(24):11324-11338. doi:10.7150/thno.47893. https://www.thno.org/v10p11324.htm
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Abstract

Graphic abstract

Rationale: Cell therapy for myocardial infarction is promising but largely unsuccessful in part due to a lack of mechanistic understanding. Techniques enabling identification of stem cell-specific proteomes in situ in the injured heart may shed light on how the administered cells respond to the injured microenvironment and exert reparative effects.

Objective: To identify the proteomes of the transplanted mesenchymal stem cells (MSCs) in the infarcted myocardium, we sought to target a mutant methionyl-tRNA synthetase (MetRSL274G) in MSCs, which charges azidonorleucine (ANL), a methionine analogue and non-canonical amino acid, to tRNA and subsequently to nascent proteins, permitting isolation of ANL-labeled MSC proteomes from ischemic hearts by ANL-alkyne based click reaction.

Methods and Results: Murine MSCs were transduced with lentivirus MetRSL274G and supplemented with ANL; the ANL-tagged nascent proteins were visualized by bio-orthogonal non-canonical amino-acid tagging, spanning all molecular weights and by fluorescent non-canonical amino-acid tagging, displaying strong fluorescent signal. Then, the MetRSL274G-transduced MSCs were administered to the infarcted or Sham heart in mice receiving ANL treatment. The MSC proteomes were isolated from the left ventricular protein lysates by click reaction at days 1, 3, and 7 after cell administration, identified by LC/MS. Among all identified proteins (in Sham and MI hearts, three time-points each), 648 were shared by all 6 groups, accounting for 82±5% of total proteins in each group, and enriched under mitochondrion, extracellular exosomes, oxidation-reduction process and poly(A) RNA binding. Notably, 26, 110 and 65 proteins were significantly up-regulated and 11, 28 and 19 proteins were down-regulated in the infarcted vs. Sham heart at the three time-points, respectively; these proteins are pronounced in the GO terms of extracellular matrix organization, response to stress and regulation of apoptotic process and in the KEGG pathways of complements and coagulation cascades, apoptosis, and regulators of actin cytoskeleton.

Conclusions: MetRSL274G expression allows successful identification of MSC-specific nascent proteins in the infarcted hearts, which reflect the functional states, adaptive response, and reparative effects of MSCs that may be leveraged to improve cardiac repair.

Keywords: Methionyl-tRNA synthetase, mesenchymal stem cells, myocardial infarction, proteomics, mass spectrometry


Citation styles

APA
Han, D., Yang, J., Zhang, E., Liu, Y., Boriboun, C., Qiao, A., Yu, Y., Sun, J., Xu, S., Yang, L., Yan, W., Luo, B., Lu, D., Zhang, C., Jie, C., Mobley, J., Zhang, J., Qin, G. (2020). Analysis of mesenchymal stem cell proteomes in situ in the ischemic heart. Theranostics, 10(24), 11324-11338. https://doi.org/10.7150/thno.47893.

ACS
Han, D.; Yang, J.; Zhang, E.; Liu, Y.; Boriboun, C.; Qiao, A.; Yu, Y.; Sun, J.; Xu, S.; Yang, L.; Yan, W.; Luo, B.; Lu, D.; Zhang, C.; Jie, C.; Mobley, J.; Zhang, J.; Qin, G. Analysis of mesenchymal stem cell proteomes in situ in the ischemic heart. Theranostics 2020, 10 (24), 11324-11338. DOI: 10.7150/thno.47893.

NLM
Han D, Yang J, Zhang E, Liu Y, Boriboun C, Qiao A, Yu Y, Sun J, Xu S, Yang L, Yan W, Luo B, Lu D, Zhang C, Jie C, Mobley J, Zhang J, Qin G. Analysis of mesenchymal stem cell proteomes in situ in the ischemic heart. Theranostics 2020; 10(24):11324-11338. doi:10.7150/thno.47893. https://www.thno.org/v10p11324.htm

CSE
Han D, Yang J, Zhang E, Liu Y, Boriboun C, Qiao A, Yu Y, Sun J, Xu S, Yang L, Yan W, Luo B, Lu D, Zhang C, Jie C, Mobley J, Zhang J, Qin G. 2020. Analysis of mesenchymal stem cell proteomes in situ in the ischemic heart. Theranostics. 10(24):11324-11338.

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