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Single cell RNA sequencing of intra- and extra- articular ligaments in mice

ABSTRACT: Knee ligaments, the anterior cruciate ligament (ACL) and medial collateral ligament (MCL), have unique functions but distinct regeneration capacities. However, the landscape of cellular heterogeneity in ligaments remains unclear. Here we profile the transcriptional signatures of ACL and MCL cells using single-cell RNA-sequencing. These ligaments consist of three fibroblast types that express Col22a1, Col12a1, or Col14a1 but have distinct localizations in the tissue. In addition, we find substantial heterogeneity in Col12a1 and Col14a1 positive cell clusters when comparing the ACL and MCL. Furthermore, we identified aging related changes in both cell composition and gene expression in ligaments.

ORGANISM(S): Mus musculus

PROVIDER: GSE194427 | GEO | 2022/11/14

REPOSITORIES: GEO

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Project description:The anterior cruciate ligament (ACL) is an essential stabilizer of the tibiofemoral articulation. ACL tears often lead to functional instability and are associated with an increased risk for osteoarthritis. The healing potential of the injured ACL is poorly understood and is considered to be limited. Transcriptome-wide expression profiles of 24 human ACL remnants recovered at the time of surgical reconstruction were analyzed utilizing the Agilent human 8x60K microarray platform. Gene ontology was performed on differentially expressed transcripts based on time-from-injury (acute, <3 months; intermediate, 3-12 months; chronic, >12 months). A subset of transcripts was validated via microfluidic digital polymerase-chain-reaction. Expression of periostin, a highly differentially expressed transcript, was tested by immunohistochemistry. Numerous transcripts covering important functional classifications were differentially expressed by time-from-injury. In acute tears, processes representing angiogenesis were repressed while those representing stem-cell differentiation were elevated. In intermediate tears, processes representing stem-cell proliferation concomitant with cellular component organization/cellular localization were elevated. In chronic tears, processes denoting myosin filament organization were elevated while those representing cellular component organization/cell localization and extracellular matrix organization were repressed. Expression levels of periostin were down-regulated in chronic tears compared to acute (42-fold) and intermediate (29-fold) tears. Immunohistochemistry confirmed a decline in periostin expression in tissues from chronic tears. These findings suggest an initial attempt of the injured ACL to repair, which declines with time-from-injury. These findings have implications for efforts to repair the ACL and may be relevant for reconstruction of the ACL. The functional role of periostin in ACL injuries, and the potential implication for surgical treatment, warrants further investigation. Total RNA obtained frominjured anterior cruciate ligament (ACL) tissues from pateints undergoing ACL surgery.

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Single cell RNA sequencing of intra- and extra- articular ligaments in mice

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