Molecular anatomy of ascending aorta in atherosclerosis by MS Imaging: Specific lipid and protein patterns reflect pathology
JOURNAL OF PROTEOMICS
Authors: Martin-Lorenzo, Marta; Balluff, Benjamin; Maroto, Aroa S.; Carreira, Ricardo J.; van Zeijl, Rene J. M.; Gonzalez-Calero, Laura; de la Cuesta, Fernando; Barderas, Maria G.; Lopez-Almodovar, Luis F.; Padial, Luis R.; McDonnell, Liam A.; Vivanco, Fernando; Alvarez-Llamas, Gloria
The molecular anatomy of healthy and atherosclerotic tissue is pursued here to identify ongoing molecular changes in atherosclerosis development. Subclinical atherosclerosis cannot be predicted and novel therapeutic targets are needed. Mass spectrometry imaging (MSI) is a novel unexplored ex vivo imaging approach in CVD able to provide in-tissue molecular maps. A rabbit model of early atherosclerosis was developed and high-spatial-resolution MALDI-MSI was applied to comparatively analyze histologically-based arterial regions of interest from control and early atherosclerotic aortas. Specific protocols were applied to identify lipids and proteins significantly altered in response to atherosclerosis. Observed protein alterations were confirmed by immunohistochemistry in rabbit tissue, and additionally in human aortas. Molecular features specifically defining different arterial regions were identified. Localized in the intima, increased expression of SFA and lysolipids and intimal spatial organization showing accumulation of PI, PG and SM point to endothelial dysfunction and triggered inflammatory response. TG, PA, SM and PE-Cer were identified specifically located in calcified regions. Thymosin beta 4 (TMSB4X) protein was upregulated in intima versus media layer and also in response to atherosclerosis. This overexpression and localization was confirmed in human aortas. In conclusion, molecular histology by MS Imaging identifies spatial organization of arterial tissue in response to atherosclerosis. (C) 2015 Elsevier B.V. All rights reserved.
Thymosin beta 4-Enhancing Therapeutic Efficacy of Human Adipose-Derived Stem Cells in Mouse Ischemic Hindlimb Model
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Authors: Kim, Jong-Ho; Lim, I-Rang; Park, Chi-Yeon; Joo, Hyung Joon; Noh, Ji-Min; Choi, Seung-Cheol; Hong, Soon Jun; Lim, Do-Sun
Thymosin beta 4 (T beta 4) is a G-actin sequestering protein that contributes to diverse cellular activities, such as migration and angiogenesis. In this study, the beneficial effects of combined cell therapy with T beta 4 and human adipose-derived stem cells (hASCs) in a mouse ischemic hindlimb model were investigated. We observed that exogenous treatment with T beta 4 enhanced endogenous TMSB4X mRNA expression and promoted morphological changes (increased cell length) in hASCs. Interestingly, T beta 4 induced the active state of hASCs by up-regulating intracellular signaling pathways including the PI3K/AKT/mTOR and MAPK/ERK pathways. Treatment with T beta 4 significantly increased cell migration and sprouting from microbeads. Moreover, additional treatment with T beta 4 promoted the endothelial differentiation potential of hASCs by up-regulating various angiogenic genes. To evaluate the in vivo effects of the T beta 4-hASCs combination on vessel recruitment, dorsal window chambers were transplanted, and the co-treated mice were found to have a significantly increased number of microvessel branches. Transplantation of hASCs in combination with T beta 4 was found to improve blood flow and attenuate limb or foot loss post-ischemia compared to transplantation with hASCs alone. Taken together, the therapeutic application of hASCs combined with T beta 4 could be effective in enhancing endothelial differentiation and vascularization for treating hindlimb ischemia.