Liver Kinase B1 Fine-Tunes Lineage Commitment of Human Fetal Synovium-Derived Stem Cells.
AMP-Activated Protein Kinase Kinases
Base Sequence
CD146 Antigen
/ metabolism
Cell Differentiation
Cells, Cultured
Fetal Stem Cells
/ physiology
Genetic Vectors
Humans
Kruppel-Like Factor 4
Lentivirus
Protein Serine-Threonine Kinases
/ genetics
Stage-Specific Embryonic Antigens
/ metabolism
Synovial Membrane
/ cytology
CRISPR-Cas9
adipogenesis
chondrogenesis
lentivirus
liver kinase B1
synovium-derived stem cell
Journal
Journal of orthopaedic research : official publication of the Orthopaedic Research Society
ISSN: 1554-527X
Titre abrégé: J Orthop Res
Pays: United States
ID NLM: 8404726
Informations de publication
Date de publication:
02 2020
02 2020
Historique:
received:
12
02
2019
accepted:
25
07
2019
pubmed:
21
8
2019
medline:
4
3
2020
entrez:
21
8
2019
Statut:
ppublish
Résumé
Liver kinase B1 (LKB1), a serine/threonine protein, is a key regulator in stem cell function and energy metabolism. Herein, we describe the role of LKB1 in modulating the differentiation of synovium-derived stem cells (SDSCs) toward chondrogenic, adipogenic, and osteogenic lineages. Human fetal SDSCs were transduced with CRISPR associated protein 9 (Cas9)-single-guide RNA vectors to knockout or lentiviral vectors to overexpress the LKB1 gene. Analyses including ICE (Inference of CRISPR Edits) data from Sanger sequencing and quantitative polymerase chain reaction (qPCR) as well as Western blot demonstrated successful knockout (KO) or overexpression (OE) of LKB1 in human fetal SDSCs without any detectable side effects in morphology, proliferation rate, and cell cycle. LKB1 KO increased CD146 expression; interestingly, LKB1 OE increased SSEA4 level. The qPCR data showed that LKB1 KO upregulated the levels of SOX2 and NANOG while LKB1 OE lowered the expression of POU5F1 and KLF4. Furthermore, LKB1 KO enhanced, and LKB1 OE inhibited, chondrogenic and adipogenic differentiation potential. However, perhaps due to the inherent inability to achieve osteogenesis, LKB1 did not obviously affect osteogenic differentiation. These data demonstrate that LKB1 plays a significant role in determining human SDSCs' adipogenic and chondrogenic differentiation, which might provide an approach for fine-tuning the direction of stem cell differentiation in tissue engineering and regeneration. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 38:258-268, 2020.
Identifiants
pubmed: 31429977
doi: 10.1002/jor.24449
pmc: PMC7294510
mid: NIHMS1046858
doi:
Substances chimiques
CD146 Antigen
0
KLF4 protein, human
0
Kruppel-Like Factor 4
0
MCAM protein, human
0
Stage-Specific Embryonic Antigens
0
stage-specific embryonic antigen-4
0
Protein Serine-Threonine Kinases
EC 2.7.11.1
STK11 protein, human
EC 2.7.11.1
AMP-Activated Protein Kinase Kinases
EC 2.7.11.3
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
258-268Subventions
Organisme : NIAMS NIH HHS
ID : R01 AR067747
Pays : United States
Organisme : Musculoskeletal Transplant Foundation
ID : Established Investigator Award
Pays : International
Organisme : NIGMS NIH HHS
ID : U54 GM104942
Pays : United States
Organisme : NIGMS NIH HHS
ID : P20 GM121322
Pays : United States
Organisme : NIAMS NIH HHS
ID : AR067747
Pays : United States
Informations de copyright
© 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.
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