Analysis of mesenchymal stem cell proteomes in situ in the ischemic heart.
Analysis of mesenchymal stem cell proteomes in situ in the ischemic heart.
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作者：
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
基本原理：心肌梗死的细胞治疗是有希望的，但在很大程度上是不成功的，部分原因是缺乏对机制的理解。能够在受伤心脏中原位识别干细胞特异性蛋白质组的技术可能会揭示所给予的细胞如何对受伤的微环境做出反应并发挥修复作用。目的：为了鉴定梗死心肌中移植的间充质干细胞（MSC）的蛋白质组，我们试图靶向MSC中的突变型甲硫氨酰-tRNA合成酶（MetRSL 274 G），其将叠氮正亮氨酸（ANL）（甲硫氨酸类似物和非经典氨基酸）充电至tRNA，随后充电至新生蛋白质，从而允许通过基于ANL-炔的点击反应从缺血心脏分离ANL标记的MSC蛋白质组。方法和结果：用慢病毒MetRSL 274 G转导鼠MSC并补充ANL;通过跨越所有分子量的生物正交非规范氨基酸标记和通过显示强荧光信号的荧光非规范氨基酸标记，使ANL标记的新生蛋白可视化。然后，将MetRSL 274 G转导的MSC施用于接受ANL治疗的小鼠的梗塞心脏或假心脏。在细胞施用后第1、3和7天通过点击反应从左心室蛋白裂解物中分离MSC蛋白质组，通过LC/MS鉴定。（在Sham和MI心脏中，各三个时间点），648个由所有6组共享，占每组中总蛋白的82±5%，并且在细胞外泌体，氧化还原过程和poly（A）RNA结合。值得注意的是，在三个时间点，在梗死心脏与假心脏中分别有26、110和65个蛋白质显著上调，11、28和19个蛋白质下调;这些蛋白质在细胞外基质组织、应激反应和凋亡过程调节的GO术语中以及在补体和凝血级联、凋亡和肌动蛋白细胞骨架的调节剂。结论：MetRSL 274 G表达允许成功鉴定梗死心脏中的MSC特异性新生蛋白，其反映了MSC的功能状态、适应性反应和修复作用，这些可以用于改善心脏修复。
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.
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影响因子：
5.6
作者：
Lee, Sang Hun;Lee, Yu Jin;Han, Ho Jae
通讯作者：
Han, Ho Jae
影响因子：
37.8
作者：
Natarajan N;Abbas Y;Bryant DM;Gonzalez-Rosa JM;Sharpe M;Uygur A;Cocco-Delgado LH;Ho NN;Gerard NP;Gerard CJ;MacRae CA;Burns CE;Burns CG;Whited JL;Lee RT
通讯作者：
Lee RT
影响因子：
3.4
作者：
Kojima, Kyoko;Bowersock, Gregory J.;Kojima, Chinatsu;Klug, Christopher A.;Grizzle, William E.;Mobley, James A.
通讯作者：
Mobley, James A.
影响因子：
48
作者：
Dieck, Susanne Tom;Kochen, Lisa;Hanus, Cyril;Heumueller, Maximilian;Bartnik, Ina;Nassim-Assir, Belquis;Merk, Katrin;Mosler, Thorsten;Garg, Sakshi;Bunse, Stefanie;Tirrell, David A.;Schuman, Erin M.
通讯作者：
Schuman, Erin M.
影响因子：
14.8
作者：
Ngo, John T.;Champion, Julie A.;Tirrell, David A.
通讯作者：
Tirrell, David A.