A Membranome-Centered Approach Defines Novel Biomarkers for Cellular Subtypes in the Intervertebral Disc

Guus G. H. van den Akker; Lars M. T. Eijssen; Stephen M. Richardson; Lodewijk W. van Rhijn; Judith A. Hoyland; Tim J. M. Welting; Jan Willem Voncken

doi:10.1177/1947603518764260

A Membranome-Centered Approach Defines Novel Biomarkers for Cellular Subtypes in the Intervertebral Disc

Guus G. H. van den Akker, Lars M. T. Eijssen, Stephen M. Richardson, Lodewijk W. van Rhijn, Judith A. Hoyland, Tim J. M. Welting, Jan Willem Voncken^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

8 Citations (Web of Science)

Abstract

Objective

Lack of specific marker-sets prohibits definition and functional distinction of cellular subtypes in the intervertebral disc (IVD), such as those from the annulus fibrosus (AF) and the nucleus pulposus (NP).

Design

We recently generated immortalized cell lines from human NP and AF tissues; these comprise a set of functionally distinct clonal subtypes. Whole transcriptome analyses were performed of 12 phenotypically distinct clonal cell lines (4x NP-Responder, 4x NP-nonResponder, 2x AF-Sheet forming, and 2x AF-nonSheet forming). Data sets were filtered for membrane-associated marker genes and compared to literature.

Results

Comparison of our immortal cell lines to published primary NP, AF, and articular chondrocytes (AC) transcriptome datasets revealed preservation of AF and NP phenotypes. NP-specific membrane-associated genes were defined by comparison to AF cells in both the primary dataset (46 genes) and immortal cell-lines (161 genes). Definition of AF-specific membrane-associated genes yielded 125 primary AF cell and 92 immortal cell-line markers. Overlap between primary and immortal NP cells yielded high-confidence NP-specific marker genes for NP-R (CLDN11, TMEFF2, CA12, ANXA2, CD44) and NP-nR (EFNA1, NETO2, SLC2A1). Overlap between AF and immortal AF subtypes yielded specific markers for AF-S (COLEC12, LPAR1) and AF-nS (CHIC1).

Conclusions

The current study provides a reference platform for preclinical evaluation of novel membrane-associated cell type-specific markers in the IVD. Future research will focus on their biological relevance for IVD function in development, homeostasis, and degenerate conditions.

Original language	English
Pages (from-to)	203-220
Number of pages	18
Journal	Cartilage
Volume	11
Issue number	2
DOIs	https://doi.org/10.1177/1947603518764260
Publication status	Published - Apr 2020

Keywords

intervertebral disc
cell lines
nucleus pulposus
annulus fibrosus
biomarkers
membranome
NUCLEUS PULPOSUS CELLS
LOW-BACK-PAIN
EXTRACELLULAR-MATRIX
GENE-EXPRESSION
STEM-CELLS
IN-VITRO
PROGENITOR CELLS
ANNULUS FIBROSUS
CARTILAGE CELLS
FUNCTIONAL-ROLE

Access to Document

10.1177/1947603518764260

Cite this

@article{0d46a5fe2b784213b89ba8348d6eceeb,

title = "A Membranome-Centered Approach Defines Novel Biomarkers for Cellular Subtypes in the Intervertebral Disc",

abstract = "ObjectiveLack of specific marker-sets prohibits definition and functional distinction of cellular subtypes in the intervertebral disc (IVD), such as those from the annulus fibrosus (AF) and the nucleus pulposus (NP).DesignWe recently generated immortalized cell lines from human NP and AF tissues; these comprise a set of functionally distinct clonal subtypes. Whole transcriptome analyses were performed of 12 phenotypically distinct clonal cell lines (4x NP-Responder, 4x NP-nonResponder, 2x AF-Sheet forming, and 2x AF-nonSheet forming). Data sets were filtered for membrane-associated marker genes and compared to literature.ResultsComparison of our immortal cell lines to published primary NP, AF, and articular chondrocytes (AC) transcriptome datasets revealed preservation of AF and NP phenotypes. NP-specific membrane-associated genes were defined by comparison to AF cells in both the primary dataset (46 genes) and immortal cell-lines (161 genes). Definition of AF-specific membrane-associated genes yielded 125 primary AF cell and 92 immortal cell-line markers. Overlap between primary and immortal NP cells yielded high-confidence NP-specific marker genes for NP-R (CLDN11, TMEFF2, CA12, ANXA2, CD44) and NP-nR (EFNA1, NETO2, SLC2A1). Overlap between AF and immortal AF subtypes yielded specific markers for AF-S (COLEC12, LPAR1) and AF-nS (CHIC1).ConclusionsThe current study provides a reference platform for preclinical evaluation of novel membrane-associated cell type-specific markers in the IVD. Future research will focus on their biological relevance for IVD function in development, homeostasis, and degenerate conditions.",

keywords = "intervertebral disc, cell lines, nucleus pulposus, annulus fibrosus, biomarkers, membranome, NUCLEUS PULPOSUS CELLS, LOW-BACK-PAIN, EXTRACELLULAR-MATRIX, GENE-EXPRESSION, STEM-CELLS, IN-VITRO, PROGENITOR CELLS, ANNULUS FIBROSUS, CARTILAGE CELLS, FUNCTIONAL-ROLE",

author = "{van den Akker}, {Guus G. H.} and Eijssen, {Lars M. T.} and Richardson, {Stephen M.} and {van Rhijn}, {Lodewijk W.} and Hoyland, {Judith A.} and Welting, {Tim J. M.} and Voncken, {Jan Willem}",

year = "2020",

month = apr,

doi = "10.1177/1947603518764260",

language = "English",

volume = "11",

pages = "203--220",

journal = "Cartilage",

issn = "1947-6035",

publisher = "SAGE Publications Inc.",

number = "2",

}

TY - JOUR

T1 - A Membranome-Centered Approach Defines Novel Biomarkers for Cellular Subtypes in the Intervertebral Disc

AU - van den Akker, Guus G. H.

AU - Eijssen, Lars M. T.

AU - Richardson, Stephen M.

AU - van Rhijn, Lodewijk W.

AU - Hoyland, Judith A.

AU - Welting, Tim J. M.

AU - Voncken, Jan Willem

PY - 2020/4

Y1 - 2020/4

N2 - ObjectiveLack of specific marker-sets prohibits definition and functional distinction of cellular subtypes in the intervertebral disc (IVD), such as those from the annulus fibrosus (AF) and the nucleus pulposus (NP).DesignWe recently generated immortalized cell lines from human NP and AF tissues; these comprise a set of functionally distinct clonal subtypes. Whole transcriptome analyses were performed of 12 phenotypically distinct clonal cell lines (4x NP-Responder, 4x NP-nonResponder, 2x AF-Sheet forming, and 2x AF-nonSheet forming). Data sets were filtered for membrane-associated marker genes and compared to literature.ResultsComparison of our immortal cell lines to published primary NP, AF, and articular chondrocytes (AC) transcriptome datasets revealed preservation of AF and NP phenotypes. NP-specific membrane-associated genes were defined by comparison to AF cells in both the primary dataset (46 genes) and immortal cell-lines (161 genes). Definition of AF-specific membrane-associated genes yielded 125 primary AF cell and 92 immortal cell-line markers. Overlap between primary and immortal NP cells yielded high-confidence NP-specific marker genes for NP-R (CLDN11, TMEFF2, CA12, ANXA2, CD44) and NP-nR (EFNA1, NETO2, SLC2A1). Overlap between AF and immortal AF subtypes yielded specific markers for AF-S (COLEC12, LPAR1) and AF-nS (CHIC1).ConclusionsThe current study provides a reference platform for preclinical evaluation of novel membrane-associated cell type-specific markers in the IVD. Future research will focus on their biological relevance for IVD function in development, homeostasis, and degenerate conditions.

AB - ObjectiveLack of specific marker-sets prohibits definition and functional distinction of cellular subtypes in the intervertebral disc (IVD), such as those from the annulus fibrosus (AF) and the nucleus pulposus (NP).DesignWe recently generated immortalized cell lines from human NP and AF tissues; these comprise a set of functionally distinct clonal subtypes. Whole transcriptome analyses were performed of 12 phenotypically distinct clonal cell lines (4x NP-Responder, 4x NP-nonResponder, 2x AF-Sheet forming, and 2x AF-nonSheet forming). Data sets were filtered for membrane-associated marker genes and compared to literature.ResultsComparison of our immortal cell lines to published primary NP, AF, and articular chondrocytes (AC) transcriptome datasets revealed preservation of AF and NP phenotypes. NP-specific membrane-associated genes were defined by comparison to AF cells in both the primary dataset (46 genes) and immortal cell-lines (161 genes). Definition of AF-specific membrane-associated genes yielded 125 primary AF cell and 92 immortal cell-line markers. Overlap between primary and immortal NP cells yielded high-confidence NP-specific marker genes for NP-R (CLDN11, TMEFF2, CA12, ANXA2, CD44) and NP-nR (EFNA1, NETO2, SLC2A1). Overlap between AF and immortal AF subtypes yielded specific markers for AF-S (COLEC12, LPAR1) and AF-nS (CHIC1).ConclusionsThe current study provides a reference platform for preclinical evaluation of novel membrane-associated cell type-specific markers in the IVD. Future research will focus on their biological relevance for IVD function in development, homeostasis, and degenerate conditions.

KW - intervertebral disc

KW - cell lines

KW - nucleus pulposus

KW - annulus fibrosus

KW - biomarkers

KW - membranome

KW - NUCLEUS PULPOSUS CELLS

KW - LOW-BACK-PAIN

KW - EXTRACELLULAR-MATRIX

KW - GENE-EXPRESSION

KW - STEM-CELLS

KW - IN-VITRO

KW - PROGENITOR CELLS

KW - ANNULUS FIBROSUS

KW - CARTILAGE CELLS

KW - FUNCTIONAL-ROLE

U2 - 10.1177/1947603518764260

DO - 10.1177/1947603518764260

M3 - Article

C2 - 29629573

VL - 11

SP - 203

EP - 220

JO - Cartilage

JF - Cartilage

SN - 1947-6035

IS - 2

ER -