Label-Free Protein Profiling of Adipose-Derived Human Stem Cells under Hyperosmotic Treatment

Oswald, Elizabeth S.; Brown, Lewis M.; Bulinski, J. Chloë; Hung, Clark T.

doi:10.1021/pr200030v

Cited by 25 publications

(16 citation statements)

References 44 publications

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“…Analysis of peptides was performed at the Quantitative Proteomics Center at Columbia University using an ultrahigh-pressure liquid chromatograph and a quadrupole-time-of-flight mass spectrometer using methods described previously (Oswald et al, 2011). This system uses a shotgun proteomics method called MS E in which mass spectra are recorded at alternate low (precursor) and high (product) fragmentation voltages (Silva et al, 2006), resulting in a more complete representation of the intensity of a peptide in a chromatographic peak.…”

Section: Methodsmentioning

confidence: 99%

dMyc expression in the fat body affects DILP2 release and increases the expression of the fat desaturase Desat1 resulting in organismal growth

Parisi

Riccardo

Zola

et al. 2013

Developmental Biology

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Drosophila dMyc (dMyc) is known for its role in cell-autonomous regulation of growth. Here we address its role in the fat body (FB), a metabolic tissue that functions as a sensor of circulating nutrients to control the release of Drosophila Insulin-like peptides (Dilps) from the brain influencing growth and development. Our results show that expression of dMyc in the FB affects development and animal size. Expression of dMyc, but not of CycD/cdk4 or Rheb, in the FB diminishes the ability to retain Drosophila Insulin-like peptide-2 (DILP2) in the brain during starvation, suggesting that expression of dMyc mimics the signal that remotely controls the release of Dilps into the hemolymph. dMyc also affects glucose metabolism and increases the transcription of Glucose-transporter-1 mRNA, and of Hexokinase and Pyruvate-Kinase mRNAs, key regulators of glycolysis. These animals are able to counteract the increased levels of circulating trehalose induced by a high sugar diet leading to the conclusion that dMyc activity in the FB promotes glucose disposal. dMyc expression induces cell autonomous accumulation of triglycerides, which correlates with increased levels of Fatty Acid Synthase and Acetyl CoA Carboxylase mRNAs, enzymes responsible for lipid synthesis. We also found the expression of Stearoyl-CoA desaturase, Desat1 mRNA significantly higher in FB overexpressing dMyc. Desat1 is an enzyme that is necessary for monosaturation and production of fatty acids, and its reduction affects dMyc’s ability to induce fat storage and resistance to animal survival. In conclusion, here we present novel evidences for dMyc function in the Drosophila FB in controlling systemic growth. We discovered that dMyc expression triggers cell autonomous mechanisms that control glucose and lipid metabolism to favor the storage of nutrients (lipids and sugars). In addition, the regulation of Desat1 controls the synthesis of triglycerides in FB and this may affect the humoral signal that controls DILP2 release in the brain.

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Section: Methodsmentioning

confidence: 99%

dMyc expression in the fat body affects DILP2 release and increases the expression of the fat desaturase Desat1 resulting in organismal growth

Parisi

Riccardo

Zola

et al. 2013

Developmental Biology

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“…In this study, 1,016, 790, and 1,104 proteins in bMSCs, ASCs, and fibroblasts were identified, respectively, which suggest that the extraction and labelfree proteomics analysis in our study are more sensitive. Label-free analysis of bone marrow-sourced stem cells has also been utilized previously to study the function of tissues (Han, Hong, & Park, 2019) and in the characterization of cellular responses to chondrogenic treatment regimens (Oswald, Brown, Bulinski, & Hung, 2011). In particular, our PCA results illustrate a significant separation of 2D…”

Section: Proteomics Analysis Reveals the Role Of Isr In Msc Homeostmentioning

confidence: 60%

Aggregation‐induced integrated stress response rejuvenates culture‐expanded human mesenchymal stem cells

Bijonowski

Yuan

et al. 2020

Biotech & Bioengineering

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Protein homeostasis is critical for cellular function, as loss of homeostasis is attributed to aging and the accumulation of unwanted proteins. Human mesenchymal stem cells (MSCs) have shown promising therapeutic potential due to their impressive abilities to secrete inflammatory modulators, angiogenic, and regenerative cytokines. However, there exists the problem of human MSC expansion with compromised therapeutic quality. Duringin vitro expansion, human MSCs are plated on stiff plastics and undergo culture adaptation, which results in aberrant proliferation, shifts in metabolism, and decreased autophagic activity. It has previously been shown that three‐dimensional (3D) aggregation can reverse some of these alterations by heightening autophagy and recovering the metabolic state back to a naïve phenotype. To further understand the proteostasis in human MSC culture, this study investigated the effects of 3D aggregation on the human MSC proteome to determine the specific pathways altered by aggregation. The 3D aggregates and 2D cultures of human MSCs derived from bone marrow (bMSC) and adipose tissue (ASC) were analyzed along with differentiated human dermal fibroblasts (FB). The proteomics analysis showed the elevated eukaryotic initiation factor 2 pathway and the upregulated activity of the integrated stress response (ISR) in 3D aggregates. Specific protein quantification further determined that bMSC and ASC responded to ISR, while FB did not. 3D aggregation significantly increased the ischemic survival of bMSCs and ASCs. Perturbation of ISR with small molecules salubrinal and GSK2606414 resulted in differential responses of bMSC, ASC, and FB. This study indicates that aggregation‐based preconditioning culture holds the potential for improving the therapeutic efficacy of expanded human MSCs via the establishment of ISR and homeostasis.

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“…Source: Suppl. Table 1 of Oswald et al (2011) . ΩfAⒶ ΩgAⒶ Mannitol-balanced 5.5 (control), 25 or 100 mM d -glucose media.…”

Section: Methodsmentioning

confidence: 99%

Chemical composition and the potential for proteomic transformation in cancer, hypoxia, and hyperosmotic stress

Dick¹

2017

PeerJ

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The changes of protein expression that are monitored in proteomic experiments are a type of biological transformation that also involves changes in chemical composition. Accompanying the myriad molecular-level interactions that underlie any proteomic transformation, there is an overall thermodynamic potential that is sensitive to microenvironmental conditions, including local oxidation and hydration potential. Here, up- and down-expressed proteins identified in 71 comparative proteomics studies were analyzed using the average oxidation state of carbon (ZC) and water demand per residue (), calculated using elemental abundances and stoichiometric reactions to form proteins from basis species. Experimental lowering of oxygen availability (hypoxia) or water activity (hyperosmotic stress) generally results in decreased ZC or of up-expressed compared to down-expressed proteins. This correspondence of chemical composition with experimental conditions provides evidence for attraction of the proteomes to a low-energy state. An opposite compositional change, toward higher average oxidation or hydration state, is found for proteomic transformations in colorectal and pancreatic cancer, and in two experiments for adipose-derived stem cells. Calculations of chemical affinity were used to estimate the thermodynamic potentials for proteomic transformations as a function of fugacity of O2 and activity of H2O, which serve as scales of oxidation and hydration potential. Diagrams summarizing the relative potential for formation of up- and down-expressed proteins have predicted equipotential lines that cluster around particular values of oxygen fugacity and water activity for similar datasets. The changes in chemical composition of proteomes are likely linked with reactions among other cellular molecules. A redox balance calculation indicates that an increase in the lipid to protein ratio in cancer cells by 20% over hypoxic cells would generate a large enough electron sink for oxidation of the cancer proteomes. The datasets and computer code used here are made available in a new R package, canprot.

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Label-Free Protein Profiling of Adipose-Derived Human Stem Cells under Hyperosmotic Treatment

Cited by 25 publications

References 44 publications

dMyc expression in the fat body affects DILP2 release and increases the expression of the fat desaturase Desat1 resulting in organismal growth

dMyc expression in the fat body affects DILP2 release and increases the expression of the fat desaturase Desat1 resulting in organismal growth

Aggregation‐induced integrated stress response rejuvenates culture‐expanded human mesenchymal stem cells

Chemical composition and the potential for proteomic transformation in cancer, hypoxia, and hyperosmotic stress

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