Analysis of Keratinocytic Exosomes from Diabetic and Nondiabetic Mice by Charge Detection Mass Spectrometry.
Analysis of Keratinocytic Exosomes from Diabetic and Nondiabetic Mice by Charge Detection Mass Spectrometry.
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DOI：
10.1021/acs.analchem.2c00453
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发表时间：
2022-06-28
影响因子：
7.4
通讯作者：
Clemmer, David E.
中科院分区：
文献类型：
--
作者：
Brown, Brooke A.;Guda, Poornachander R.;Zeng, Xuyao;Anthony, Adam;Couse, Andrew;Barnes, Lauren F.;Sharon, Edie M.;Trinidad, Jonathan C.;Sen, Chandan K.;Jarrold, Martin F.;Ghatak, Subhadip;Clemmer, David E.
未解决的炎症损害糖尿病伤口愈合。最近，我们报道了小鼠创面角化细胞源性外显体（Exoκ）的RNA包装不足导致持续炎症[Zhou， X.et al.ACS, 2020, 14(10), 12732-12748]。在此，我们使用电荷检测质谱（CDMS）分析了从5日龄糖尿病小鼠伤口边缘组织和杂合非糖尿病小鼠对照组中分离的完整Exoκ。在CDMS中，可以同时测量单个外泌体粒子的电荷(z)和质量电荷比（m/z），从而可以直接分析外泌体在1 - 200mda范围内的质量。这些测量结果显示Exoκ的质量范围从~10到bbb100 MDa。这些外泌体的m和z值似乎属于科（亚种群）；统计建模分析部分解析了~ 10-20个Exoκ亚群。互补蛋白质组学、免疫荧光和电子显微镜研究支持CDMS结果，即糖尿病小鼠和非糖尿病小鼠的Exoκ存在很大差异。高z （bbbb650）和高m （bbbb44mda）亚群在非糖尿病动物中更为丰富。我们认为这些高m和高z粒子可能是由于货物包装的不同而产生的。这一观点的准确性在其他最近的CDMS结果，包括基因组包装疫苗，以及外泌体成像实验的讨论。基于m和z的物理性质的完整外泌体颗粒的表征提供了一种研究伤口愈合的新方法，并表明CDMS可能对其他病理有用。
Unresolved inflammation compromises diabetic wound healing. Recently, we reported that inadequate RNA packaging in murine wound-edge keratinocyte-originated exosomes (Exoκ) leads to persistent inflammation [Zhou, X.et al.ACS Nano 2020, 14(10), 12732–12748]. Herein, we use charge detection mass spectrometry (CDMS) to analyze intact Exoκ isolated from a 5 day old wound-edge tissue of diabetic mice and a heterozygous nondiabetic littermate control group. In CDMS, the charge (z) and mass-to-charge ratio (m/z) of individual exosome particles are measured simultaneously, enabling the direct analysis of masses in the 1–200 MDa range anticipated for exosomes. These measurements reveal a broad mass range for Exoκ from ~10 to >100 MDa. The m and z values for these exosomes appear to fall into families (subpopulations); a statistical modeling analysis partially resolves ~10–20 Exoκ subpopulations. Complementary proteomics, immunofluorescence, and electron microscopy studies support the CDMS results that Exoκ from diabetic and nondiabetic mice vary substantially. Subpopulations having high z (>650) and high m (>44 MDa) are more abundant in nondiabetic animals. We propose that these high m and z particles may arise from differences in cargo packaging. The veracity of this idea is discussed in light of other recent CDMS results involving genome packaging in vaccines, as well as exosome imaging experiments. Characterization of intact exosome particles based on the physical properties of m and z provides a new means of investigating wound healing and suggests that CDMS may be useful for other pathologies.
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影响因子：
17.1
作者：
Eming SA;Martin P;Tomic-Canic M
通讯作者：
Tomic-Canic M
DOI：
10.1007/s13361-018-2038-3
发表时间：
2018-11-01
影响因子：
3.2
作者：
Draper, Benjamin E.;Anthony, Staci N.;Jarrold, Martin F.
通讯作者：
Jarrold, Martin F.
影响因子：
20.3
作者：
Bai, Ming;Grieshaber-Bouyer, Ricardo;Nigrovic, Peter A.
通讯作者：
Nigrovic, Peter A.
DOI：
10.1007/s13361-016-1362-8
发表时间：
2016-06-01
影响因子：
3.2
作者：
Keifer, David Z.;Motwani, Tina;Jarrold, Martin F.
通讯作者：
Jarrold, Martin F.
影响因子：
4.1
作者：
Chopra, Neha;Arya, Braham Dutt;Choudhury, Sangeeta
通讯作者：
Choudhury, Sangeeta