Reversine enhances generation of progenitor-like cells by dedifferentiation of annulus fibrosus cells.
The aim of this study was to determine if treatment with reversine, a purine analog, promoted generation of skeletal progenitor cells from lineage-committed annulus fibrosus cells. Reversine modulated cell growth, morphology, and the actin cytoskeleton of annulus fibrosus cells. Microarray profiling coupled with Ingenuity Pathway Analysis revealed that reversine treatment resulted in a significant expression change in many genes including those required for cell-cell interaction, cell movement, cell growth, and development. Further analysis revealed that there was involvement of gene networks concerned with cellular assembly and organization, DNA replication and repair, tissue morphology, and cell-to-cell signaling. The gene expression profile was dependent on reversine concentration. In osteogenic media, cells pretreated with 300 nM reversine exhibited an increased induction in alkaline phosphatase activity and enhanced expression of alkaline phosphatase, bone sialoprotein, osteocalcin, and collagen type I mRNA. Maintained in adipogenic media, the reversine-pretreated annulus cells displayed evidence of adipogenic differentiation: accumulation of cytosolic lipid droplets and increased expression of PPAR-gamma2, LPL, and Fabp mRNA. In chondrogenic media, cells pretreated with reversine exhibited marked increase in the induction of aggrecan, collagen types II, IX, and XI, and versican. It is concluded that reversine treatment induced annulus fibrosus cell plasticity and promoted their differentiation along mesenchymal lineages. This agent could be used to generate skeletal progenitor cells to orchestrate the repair of the intervertebral disc.
Saraiya
Mansi
M
Department of Orthopaedic Surgery, Thomas Jefferson University , Philadelphia, PA, USA.
Nasser
Rena
R
Zeng
Yan
Y
Addya
Sankar
S
Ponnappan
Ravi Kumar
RK
Fortina
Paolo
P
Anderson
David Greg
DG
Albert
Todd J
TJ
Shapiro
Irving M
IM
Risbud
Makarand V
MV
D050156:Adipogenesis / Q000187:drug effects / Q000235:genetics; D053687:Adult Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D054337:Cell Dedifferentiation / Q000187:drug effects* / Q000235:genetics; D003094:Collagen / Q000378:metabolism; D020869:Gene Expression Profiling; D053263:Gene Regulatory Networks / Q000187:drug effects; D007403:Intervertebral Disc / Q000166:cytology* / Q000187:drug effects* / Q000293:injuries; D009025:Morpholines / Q000494:pharmacology*; D039902:Multipotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D024510:Muscle Development / Q000187:drug effects / Q000235:genetics; D032448:Myoblasts, Skeletal / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics; D011687:Purines / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D023822:Tissue Engineering / Q000379:methods
D009025:Morpholines; D011687:Purines; D003094:Collagen; D000469:Alkaline Phosphatase; C484369:2-(4-morpholinoanilino)-6-cyclohexylaminopurine