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Single‐cell RNA‐sequencing atlas of bovine caudal intervertebral discs: Discovery of heterogeneous cell populations with distinct roles in homeostasis
Author(s) -
Panebianco Christopher J.,
Dave Arpit,
Charytonowicz Daniel,
Sebra Robert,
Iatridis James C.
Publication year - 2021
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.202101149r
Subject(s) - adamts , biology , intervertebral disc , microbiology and biotechnology , extracellular matrix , cell type , gene expression , gene , cell , genetics , computational biology , anatomy , matrix metalloproteinase , thrombospondin , metalloproteinase
Back and neck pain are significant healthcare burdens that are commonly associated with pathologies of the intervertebral disc (IVD). The poor understanding of the cellular heterogeneity within the IVD makes it difficult to develop regenerative IVD therapies. To address this gap, we developed an atlas of bovine ( Bos taurus ) caudal IVDs using single‐cell RNA‐sequencing (scRNA‐seq). Unsupervised clustering resolved 15 unique clusters, which we grouped into the following annotated partitions: nucleus pulposus (NP), outer annulus fibrosus (oAF), inner AF (iAF), notochord, muscle, endothelial, and immune cells. Analyzing the pooled gene expression profiles of the NP, oAF, and iAF partitions allowed us to identify novel markers for NP ( CP, S100B, H2AC18, SNORC, CRELD2, PDIA4, DNAJC3, CHCHD7 , and RCN2 ), oAF ( IGFBP6, CTSK, LGALS1 , and CCN3 ), and iAF ( MGP, COMP, SPP1, GSN, SOD2, DCN, FN1, TIMP3, WDR73 , and GAL ) cells. Network analysis on subpopulations of NP and oAF cells determined that clusters NP1, NP2, NP4, and oAF1 displayed gene expression profiles consistent with cell survival, suggesting these clusters may uniquely support viability under the physiological stresses of the IVD. Clusters NP3, NP5, oAF2, and oAF3 expressed various extracellular matrix (ECM)‐associated genes, suggesting their role in maintaining IVD structure. Lastly, transcriptional entropy and pseudotime analyses found that clusters NP3 and NP1 had the most stem‐like gene expression signatures of the NP partition, implying these clusters may contain IVD progenitor cells. Overall, results highlight cell type diversity within the IVD, and these novel cell phenotypes may enhance our understanding of IVD development, homeostasis, degeneration, and regeneration.