Asian Spine J.  2020 Aug;14(4):409-420. 10.31616/asj.2019.0217.

Proteomic Signature of Nucleus Pulposus in Fetal Intervertebral Disc

• Rajasekaran S
• Soundararajan DCR
• Tangavel C
• K. S. SVA
• Nayagam SM
• Matchado MS
• Muthurajan R
• Shetty AP
• Kanna RM
• Dharmalingam K

• KMID: 2505234
• DOI: http://doi.org/10.31616/asj.2019.0217

Abstract

Study Design: Profiling proteins expressed in the nucleus pulposus of fetal intervertebral disc (IVD). Purpose: To evaluate the molecular complexity of fetal IVDs not exposed to mechanical, traumatic, inflammatory, or infective insults to generate improved knowledge on disc homeostasis. Overview of Literature: Low back pain is the most common musculoskeletal disorder, causing a significant reduction in the quality of life, and degenerative disc disorders mainly contribute to the increasing socioeconomic burden. Despite extensive research, the causative pathomechanisms behind degenerative disc disorders are poorly understood. Precise molecular studies on the intricate biological processes involved in maintaining normal disc homeostasis are needed.
Methods
IVDs of nine fetal specimens obtained from medical abortions were used to dissect out the annulus fibrosus and nucleus pulposus under sterile operating conditions. Dissected tissues were transferred to sterile Cryovials and snap frozen in liquid nitrogen before transporting to the research laboratory for protein extraction and further liquid chromatography tandem mass spectrometry (LC-MS/ MS) analysis. Collected data were further analyzed using Gene Functional Classification Tool in DAVID and STRING databases.
Results
A total of 1,316 proteins were identified through LC-MS/MS analysis of nine fetal IVD tissues. Approximately 247 proteins present in at least four fetal discs were subjected to further bioinformatic analysis. The following 10 clusters of proteins were identified: collagens, ribosomal proteins, small leucine-rich proteins, matrilin and thrombospondin, annexins, protein disulfide isomerase family proteins and peroxiredoxins, tubulins, histones, hemoglobin, and prolyl 4-hydroxylase family proteins.
Conclusions
This study provides fundamental information on the proteome networks involved in the growth and development of healthy fetal discs in humans. Systematic cataloging of proteins involved in various structural and regulatory processes has been performed. Proteins expressed most abundantly (collagen type XIV alpha 1 chain, biglycan, matrilin 1, and thrombospondin 1) in their respective clusters also elucidate the possibility of utilizing these proteins for potential regenerative therapies.

Keyword

Intervertebral disc degeneration; Low back pain; Disc degeneration; Fetal intervertebral disc proteome