Peptidome analysis of umbilical cord mesenchymal stem cell (hUC-MSC) conditioned medium from preterm and term infants

Abstract

Background The therapeutic role of mesenchymal stem cells (MSCs) has been widely confirmed in several animal models of premature infant diseases. Micromolecule peptides have shown promise for the treatment of premature infant diseases. However, the potential role of peptides secreted from MSCs has not been studied. The purpose of this study is to help to broaden the knowledge of the hUC-MSC secretome at the peptide level through peptidomic profile analysis. Methods We used tandem mass tag (TMT) labeling technology followed by tandem mass spectrometry to compare the peptidomic profile of preterm and term umbilical cord MSC (hUC-MSC) conditioned medium (CM). Gene Ontology (GO) enrichment analysis and ingenuity pathway analysis (IPA) were conducted to explore the differentially expressed peptides by predicting the functions of their precursor proteins. To evaluate the effect of candidate peptides on human lung epithelial cells stimulated by hydrogen peroxide (H2O2), quantitative real-time PCR (qRT-PCR), western blot analysis, and enzyme-linked immunosorbent assay (ELISA) were, respectively, adopted to detect inflammatory cytokines (TNF-α, IL-1β, and IL-6) expression levels at the mRNA and protein levels. Results A total of 131 peptides derived from 106 precursor proteins were differentially expressed in the preterm hUC-MSC CM compared with the term group, comprising 37 upregulated peptides and 94 downregulated peptides. Bioinformatics analysis showed that these differentially expressed peptides may be associated with developmental disorders, inflammatory response, and organismal injury. We also found that peptides 7118TGAKIKLVGT7127 derived from MUC19 and 508AAAAGPANVH517 derived from SIX5 reduced the expression levels of TNF-α, IL-1β, and IL-6 in H2O2-treated human lung epithelial cells. Conclusions In summary, this study provides further secretomics information on hUC-MSCs and provides a series of peptides that might have antiinflammatory effects on pulmonary epithelial cells and contribute to the prevention and treatment of respiratory diseases in premature infants.

Peptide KLF14-P1 secreted from mesenchymal stem cells protects against bronchopulmonary dysplasia both in vivo and in vitro

“…39,40 KLF14-P1, as a small molecule active polypeptide paracrine secreted by MSCs, is located on the zinc finger (ZNF) domain. 23 ZNF is a relatively small and structurally stable protein motif, which contains multiple finger protrusions, which can be in series contact with target molecules, including DNA, RNA, protein or lipid substrates, and there are few conformational changes when binding to targets. 24 At the same time, FITC-labeled KLF14-P1 were cultured with MLE-12 cells for 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 and 24 h, and FITC-labeled KLF14-P1 could enter MLE-12 cell cytoplasm smoothly.…”

“…And using protparam on-line analysis, KLF14-P1 consists of 8 amino acids with a molecular weight of 942.3Da, conforming to the characteristics of peptide drugs. 23,24,25 Therefore, this study was designed to observe KLF14-P1's therapeutic effects on BPD progression in vivoand in vitro , and to explore the possible mechanisms, so as to provide a new experimental basis for KLF14-P1 in BPD treatment.…”

DNA Methylation Pattern of CALCA and CALCB in Extremely Premature Infants with Monochorionic Triplets after Single‐Embryo Transfer

“…Premature infants are susceptible to many complications due to impaired immunity, inflammation, and vascular regulation [ 13 , 14 ]. The discovery with regard to the relationship between calcitonin/calcitonin gene-related peptide (CT/CGRP) and pregnancy highlighted new signal transmission mediators in various physiological processes including reproduction [ 15 , 16 ].…”

Mass spectrometry for the study of adipocyte cell secretome in cardiovascular diseases

“…The correlation with antibody‐based methods, such as enzyme‐linked immunosorbent assay (ELISA), is high and the quantitative differences can be measured up to a concentration range of four/five orders of magnitude (Frauenstein & Meissner, 2018). Accurate quantitative results can be also obtained with labeling methods such as SILAC (Kashyap et al, 2010), isotope‐coded affinity tag (ICAT) (H. Zhou et al, 2009), isobaric tags for relative and absolute quantification (iTRAQ) (Brandi et al, 2016; Creaney et al, 2017; Ma et al, 2016), and TMT (Wang et al, 2020), which are widely applied in secretomics.…”