Quantitative Proteomics Evaluation of Human Multipotent Stromal Cell for β Cell Regeneration

Kuljanin, Miljan; Elgamal, Ruth M.; Bell, Gillian I.; Xenocostas, Dimetri; Xenocostas, Anargyros; Hess, David A.; Lajoie, Gilles

doi:10.1016/j.celrep.2018.10.107

Cited by 7 publications

(4 citation statements)

References 47 publications

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“…In addition to the secretion of potent chemotactic or angiogenic stimuli, the secretome of the BM-MSCs can stimulate β-cell regeneration in NOD/SCID mice with STZ-induced hyperglycemia. 42,57 To this end, we sought to use this robust in vivo model to determine if the secretome of Panc-MSCs provides tissue regenerative benefits comparable to BM-MSCs (Figure 7A). Hyperglycemia was significantly reduced (Figure 7B that prospectively identifies heterogeneous MSCs from various tissues.…”

Section: Intrapancreatic Injection Of Panc-msc CM Reduces Hyperglycemia In Stz-treated Micementioning

confidence: 99%

Characterization of a Vimentinhigh/Nestinhigh proteome and tissue regenerative secretome generated by human pancreas-derived mesenchymal stromal cells

Cooper

Sherman

Bell³

et al. 2020

Stem Cells

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Multipotent/mesenchymal stromal cells (MSCs) exist within a variety of postnatal tissues; however, global proteomic analyses comparing tissue-specific MSC are limited. Using human bone marrow (BM)-derived MSCs as a gold standard, we used label-free mass spectrometry and functional assays to characterize the proteome, secretome, and corresponding function of human pancreas-derived MSCs (Panc-MSCs) with a classical phenotype (CD90+/CD73+/CD105+/CD45−/CD31−). Both MSC subtypes expressed mesenchymal markers vimentin, α-SMA, and STRO-1; however, expression of nestin was increased in Panc-MSCs. Accordingly, these Vimentin high /Nestin high cells were isolated from fresh human pancreatic islet and non-islet tissues. Next, we identified expression of >60 CD markers shared between Panc-MSCs and BM-MSCs, including validated expression of CD14. An additional 19 CD markers were differentially expressed, including reduced pericyte-marker CD146 expression on Panc-MSCs. Panc-MSCs also showed reduced expression of proteins involved in lipid and retinoid metabolism. Accordingly, Panc-MSCs showed restricted responses to adipogenic stimuli in vitro, although both MSC types demonstrated trilineage differentiation. In contrast, Panc-MSCs demonstrated accelerated growth kinetics and competency to proneurogenic stimuli in vitro. The secretome of Panc-MSCs was highly enriched for proteins associated with vascular development, wound healing and chemotaxis. Similar to BM-MSCs, Panc-MSCs conditioned media augmented endothelial cell survival, proliferation, and tubule formation in vitro. Importantly, the secretome of both MSC types was capable of stimulating chemotactic infiltration of murine endothelial cells in vivo and reduced hyperglycemia in STZ-treated mice following intrapancreatic injection. Overall, this study provides foundational knowledge to develop Panc-MSCs as a unique MSC subtype with functional properties beneficial in regenerative medicine for diabetes and vascular disease. K E Y W O R D S adipogenesis, multipotent stromal cells, nestin, pancreas, proteomics, regenerative medicine, secretome

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Section: Intrapancreatic Injection Of Panc-msc CM Reduces Hyperglycemia In Stz-treated Micementioning

confidence: 99%

Characterization of a Vimentinhigh/Nestinhigh proteome and tissue regenerative secretome generated by human pancreas-derived mesenchymal stromal cells

Cooper

Sherman

Bell³

et al. 2020

Stem Cells

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“…A study from Kuljanin et al expanded donor-derived human multipotent stromal cells and collected the conditioned media ex vivo . Almost all 16 secreted proteins identified that could be used to predict the regenerative potency of the cells are glycoproteins . For in vivo studies, proteins may be collected from several types of bodily fluids, such as blood, plasma, urine, saliva, nipple aspirate fluid, and cerebrospinal fluids. − Reduction of the protein complexity by a trap column or immunoaffinity depletion to remove abundant proteins before further analysis may be performed to increase the coverage of secreted glycoproteins .…”

Section: Applications Of Ms-based Glycoproteomics In Biological Systemsmentioning

confidence: 99%

Recent Advances in Glycoproteomic Analysis by Mass Spectrometry

2019

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Glycosylation is one of the most common protein modifications and is essential for cells. This modification is exceptionally complex because glycans are highly diverse and can be covalently bound to several amino acid residues in proteins through various configurations. There are two major types of protein glycosylation, i.e., N-linked glycosylation in which glycans are attached to the side chain of asparagine and O-linked glycosylation referring to glycans being bound to the side chains of serine and threonine. 1,2 Glycosylation plays vital roles in cells, including determination of protein folding, trafficking and stability, and regulation of nearly every extracellular activity such as cell-cell communication and cellmatrix interactions. 3,4 Aberrant protein glycosylation is directly related to multiple diseases, including cancer, neurodegenerative disorders, pulmonary diseases, blood disorders, and genetic diseases. 5,6 Due to the importance and complexity of protein glycosylation in biological systems, there is a longstanding interest to develop innovative methods to study glycoproteins and apply them for biomedical research. Investigation of protein glycosylation has become more popular with the development of modern instrumentation and computational methods. According to a PubMed search using the keyword "glycosylation", 16 publications were listed during 1960-1970 while over 20 000 studies were reported in the past 10 years. With the growing interests in protein glycosylation, this trend is expected to continue in the next decades.Mass spectrometry (MS)-based proteomics provides an excellent opportunity to globally analyze proteins and their modifications. [7][8][9][10][11][12][13][14][15][16][17][18][19] Nonetheless, it is still extremely challenging to comprehensively analyze protein glycosylation. 20 Unlike many other modifications with a fixed structure for the modified group, such as phosphorylation, the diversity of glycans makes it more challenging to employ the commonly used database searching methods such as SEQUEST and Mascot to identify glycopeptides in bottom-up proteomics. Lowabundance glycoproteins in complex biological samples are also hindered for detection by many high-abundance nonglycoproteins. Furthermore, glycans can interfere with the fragmentation of the peptide backbone. 20,21 Innovative and effective methods are critical to overcome these hurdles and to allow for comprehensive analysis of glycoproteins using MS.

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“…Herein, we describe a protocol for parallel proteomic and metabolomic analysis of mouse IVD tissues, building from the combined expertise of our collaborative team ( Figure 1). Our group has lead technical development in proteomics [76][77][78][79][80] and applied these methodologies to develop biomarker panels to improve classifications F I G U R E 1 Schematic overview of protocol for simultaneous assessment of proteomic and metabolomic changes of ovarian carcinomas, 81 evaluate the potential of multipotent stromal cells for pancreas regeneration, 82 and developed optimized protocols to characterize extracellular matrices, [83][84][85] or increase the detection of low-abundant proteins in extracellular matrix-rich samples. 86 These studies led to the development of the current protocol for label-free quantitative proteomics of murine IVD tissue.…”

Section: Introductionmentioning

confidence: 99%

Protocol for parallel proteomic and metabolomic analysis of mouse intervertebral disc tissues

et al. 2020

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The comprehensiveness of data collected by "omics" modalities has demonstrated the ability to drastically transform our understanding of the molecular mechanisms of chronic, complex diseases such as musculoskeletal pathologies, how biomarkers are identified, and how therapeutic targets are developed. Standardization of protocols will enable comparisons between findings reported by multiple research groups and move the application of these technologies forward. Herein, we describe a protocol for parallel proteomic and metabolomic analysis of mouse intervertebral disc (IVD) tissues, building from the combined expertise of our collaborative team. This protocol covers dissection of murine IVD tissues, sample isolation, and data analysis for both proteomics and metabolomics applications. The protocol presented below was optimized to maximize the utility of a mouse model for "omics" applications, accounting for the challenges associated with the small starting quantity of sample due to small tissue size as well as the extracellular matrix-rich nature of the tissue.

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Quantitative Proteomics Evaluation of Human Multipotent Stromal Cell for β Cell Regeneration

Cited by 7 publications

References 47 publications

Characterization of a Vimentinhigh/Nestinhigh proteome and tissue regenerative secretome generated by human pancreas-derived mesenchymal stromal cells

Characterization of a Vimentinhigh/Nestinhigh proteome and tissue regenerative secretome generated by human pancreas-derived mesenchymal stromal cells

Recent Advances in Glycoproteomic Analysis by Mass Spectrometry

Protocol for parallel proteomic and metabolomic analysis of mouse intervertebral disc tissues

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