Part 1: profiling extra cellular matrix core proteome of human fetal nucleus pulposus in search for regenerative targets

Abstract

Intervertebral disc degeneration is accompanied by a loss of Extra-cellular matrix (ECM) due to an imbalance in anabolic and catabolic pathways. Identifying ECM proteins with anabolic and/or regenerative potential could be the key to developing regenerative therapies. Since human fetal discs grow and develop rapidly, studying these discs may provide valuable insights on proteins with regenerative potential. This study compares core matrisome of 9 fetal and 7 healthy adult (age 22–79) nucleus pulposus (NP), using a proteomic and bioinformatic approach. Of the 33 upregulated proteins in fetus NP’s, 20 of which were involved in ECM assembly pathways: fibromodulin, biglycan, heparan sulfate proteoglycan 2, chondroitin sulfate proteoglycan 4, procollagen C-endopeptidase enhancer and Collagen—type 1a1, 1a2, 6a1, 6a3, 11a1, 11a2, 12a1, 14a1 and 15a1. Moreover, 10 of the upregulated proteins were involved in growth pathways ‘PI3L-Akt signaling’ and ‘regulation of insulin like growth factor transport and uptake.’ Thrombospondin 1,3 and 4, tenascin C, matrilin-3, and collagen- type 1a1, 1a2, 6a1, 6a3 and 9a1. Additionally, matrillin-2 and ‘Collagen triple helix repeat containing 1’ were identified as possible regenerative proteins due to their involvement in ‘Regeneration’ and ‘tissue development’ respectively. In conclusion, the consistency of human fetal NP’s differs greatly from that of healthy adults. In view of these outcomes, the core matrisome of human fetal discs contains an abundant number of proteins that could potentially show regenerative properties, and their potential should be explored in future machinal experiments.

Profiling extra cellular matrix associated proteome of human fetal nucleus pulposus in search for regenerative targets

“…The general proteomic features of the discs were described earlier by Rajasekaran et al 9 . The matrisome consisted for 14% of matrisome associated proteins in fetal NP’s; similar results were seen in adult discs (10%) 11 .…”

“…Written informed consent was obtained, and all methods were carried out according to the Declaration of Helsinki. A detailed description of the sample collection procedure was described earlier by Rajasekaran et al 2020 9 .…”

“…In our previous study, we explored the core matrisome (ECM proteome) of human fetal nucleus pulposus (NP) for proteins of regenerative potential and found a great number of such proteins to be upregulated or uniquely expressed in the fetal NP’s 9 . Nevertheless, only 86% of the matrisome is core matrisome (Collagens, proteoglycans, and glycoproteins) leaving 14% of the fetal matrisome undiscovered.…”

Notochordal Cell-Based Treatment Strategies and Their Potential in Intervertebral Disc Regeneration

“…In the study of Bach and de Vries et al (2016) , only proteins that were identified in both human, canine, and porcine NCCM were included in this analysis. Rajasekaran et al (2020) , Rajasekaran et al (2020b) identified proteins in human fetal matrisome, Caldeira et al (2017) in bovine fetal matrisome, and Kudelko et al (2021) in murine matrisome. The study by Tam et al (2020) was not included in this comparative proteomic overview focusing on NC-rich tissues, since it used NPC-containing adolescent human NP tissue.…”

Influence of endplate avulsion and Modic changes on the inflammation profile of herniated discs: a proteomic and bioinformatic approach

“…The peptides with rank one and having a q-value < 0.01 were considered for protein identification. A more detailed description of bioinformatic analysis was published earlier [13].…”