TY - JOUR
T1 - Pro-Chondrogenic Effect of miR-221 and Slug Depletion in Human MSCs
AU - Lolli, Andrea
AU - Lambertini, Elisabetta
AU - Penolazzi, Letizia
AU - Angelozzi, Marco
AU - Morganti, Claudia
AU - Franceschetti, Tiziana
AU - Pelucchi, Stefano
AU - Gambari, Roberto
AU - Piva, Roberta
N1 - Publisher Copyright:
© 2014, Springer Science+Business Media New York.
PY - 2014/12
Y1 - 2014/12
N2 - In this study we have inhibited the expression of two negative regulators of chondrogenesis, Slug transcription factor (TF) and the small non-coding single stranded RNA microRNA-221 (miR-221), in human mesenchymal stem cells (MSCs). Our aim was test a new approach to guide the cells toward a chondrocyte – like phenotype, without the employment of differentiating agents, in the prospect of their clinical applications for cell-based cartilage tissue engineering. We have characterized these manipulated cells by gene expression analysis at the RNA and protein levels. We demonstrated that decreased miR-221 or Slug induced an increase of chondrogenic markers, including collagen type II (Col2A1), and the positive chondrogenic TFs Sox9 and TRPS1. Slug and TRPS1 are not direct targets of miR-221 since their expression was not affected by miR-221 content. Further, we showed by gene expression and Chromatin Immunoprecipitation analyses that i. miR-221 is positively regulated by Slug in hMSCs, where Slug and miR-221 high levels hamper cell differentiation, and ii. TRPS1 contributes to maintaining low levels of miR-221, both in hMSCs committed toward chondrogenesis by Slug depletion and in chondrocytes, where the low levels of miR-221 and Slug allow a chondrogenic phenotype.Taken together, our data may be relevant both to understand yet unknown miRNA – TF regulatory loops in cartilage biology and to establish new strategies based on a siRNA approach for cartilage tissue engineering.
AB - In this study we have inhibited the expression of two negative regulators of chondrogenesis, Slug transcription factor (TF) and the small non-coding single stranded RNA microRNA-221 (miR-221), in human mesenchymal stem cells (MSCs). Our aim was test a new approach to guide the cells toward a chondrocyte – like phenotype, without the employment of differentiating agents, in the prospect of their clinical applications for cell-based cartilage tissue engineering. We have characterized these manipulated cells by gene expression analysis at the RNA and protein levels. We demonstrated that decreased miR-221 or Slug induced an increase of chondrogenic markers, including collagen type II (Col2A1), and the positive chondrogenic TFs Sox9 and TRPS1. Slug and TRPS1 are not direct targets of miR-221 since their expression was not affected by miR-221 content. Further, we showed by gene expression and Chromatin Immunoprecipitation analyses that i. miR-221 is positively regulated by Slug in hMSCs, where Slug and miR-221 high levels hamper cell differentiation, and ii. TRPS1 contributes to maintaining low levels of miR-221, both in hMSCs committed toward chondrogenesis by Slug depletion and in chondrocytes, where the low levels of miR-221 and Slug allow a chondrogenic phenotype.Taken together, our data may be relevant both to understand yet unknown miRNA – TF regulatory loops in cartilage biology and to establish new strategies based on a siRNA approach for cartilage tissue engineering.
KW - Chondrogenic potential
KW - Gene silencing
KW - Human mesenchymal stem cells
KW - Slug
KW - miR-221
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U2 - 10.1007/s12015-014-9532-1
DO - 10.1007/s12015-014-9532-1
M3 - Review article
C2 - 24923882
AN - SCOPUS:84930824325
SN - 2629-3269
VL - 10
SP - 841
EP - 855
JO - Stem Cell Reviews and Reports
JF - Stem Cell Reviews and Reports
IS - 6
ER -