PMID- 24449574
OWN - NLM
STAT- MEDLINE
DCOM- 20140318
LR  - 20211103
IS  - 2326-5205 (Electronic)
IS  - 2326-5191 (Linking)
VI  - 66
IP  - 1
DP  - 2014 Jan
TI  - Dual role of autophagy in stress-induced cell death in rheumatoid arthritis 
      synovial fibroblasts.
PG  - 40-8
LID - 10.1002/art.38190 [doi]
AB  - OBJECTIVE: To investigate the role of autophagy in the regulation of cell death 
      in rheumatoid arthritis synovial fibroblasts (RASFs). METHODS: RASFs and 
      osteoarthritis synovial fibroblasts (OASFs) were treated with thapsigargin (TG), 
      an inducer of endoplasmic reticulum (ER) stress, and MG132, a proteasome 
      inhibitor. Then, 3-methyladenine was used as an autophagy inhibitor and 
      bafilomycin A1 as a lysosome inhibitor. Polyubiquitinated proteins, p62, and 
      autophagy induction were evaluated by immunoblotting, immunofluorescence 
      microscopy, and immunohistochemistry, respectively. OASFs were transfected with 
      small interfering RNA targeting autophagy-linked FYVE protein (ALFY). Cell death 
      was evaluated by flow cytometry and a caspase 3 activity assay. RESULTS: In 
      RASFs, the induction of autophagy by TG and MG132 was increased compared to that 
      in OASFs. Whereas autophagy promoted a caspase 3-independent induction of cell 
      death under ER stress, autophagy had a protective role in apoptosis induced by 
      proteasome inhibition. Treatment of RASFs with 3-methyladenine blocked TG-induced 
      cell death. ER stress induced a strong accumulation of p62-positive 
      polyubiquitinated protein aggregates, accompanied by the formation of large 
      vacuoles in RASFs but not OASFs. Furthermore, TG-induced p62 protein expression 
      was increased, whereas TG-induced ALFY expression was reduced, in RASFs compared 
      to OASFs. ALFY knockdown promoted the accumulation of p62, the formation of 
      polyubiquitinated protein aggregates, and cell death. CONCLUSION: Our data 
      provide the first evidence of a dual role of autophagy in the regulation of death 
      pathways in RASFs. A reduced expression of ALFY and the formation of p62-positive 
      polyubiquitinated protein aggregates promote cell death in RASFs under severe ER 
      stress.
CI  - Copyright (c) 2014 by the American College of Rheumatology.
FAU - Kato, Masaru
AU  - Kato M
AD  - Center of Experimental Rheumatology, University Hospital Zurich, Zurich, 
      Switzerland.
FAU - Ospelt, Caroline
AU  - Ospelt C
FAU - Gay, Renate E
AU  - Gay RE
FAU - Gay, Steffen
AU  - Gay S
FAU - Klein, Kerstin
AU  - Klein K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Arthritis Rheumatol
JT  - Arthritis & rheumatology (Hoboken, N.J.)
JID - 101623795
RN  - 0 (Adaptor Proteins, Signal Transducing)
RN  - 0 (Autophagy-Related Proteins)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Macrolides)
RN  - 0 (Membrane Proteins)
RN  - 0 (SQSTM1 protein, human)
RN  - 0 (Sequestosome-1 Protein)
RN  - 0 (Transcription Factors)
RN  - 0 (WDFY3 protein, human)
RN  - 120904-94-1 (Polyubiquitin)
RN  - 5142-23-4 (3-methyladenine)
RN  - 88899-55-2 (bafilomycin A1)
RN  - EC 3.4.22.- (CASP3 protein, human)
RN  - EC 3.4.22.- (Caspase 3)
RN  - JAC85A2161 (Adenine)
SB  - IM
CIN - Nat Rev Rheumatol. 2013 Nov;9(11):637. PMID: 24080860
MH  - Adaptor Proteins, Signal Transducing/drug effects/metabolism
MH  - Adenine/analogs & derivatives/pharmacology
MH  - Arthritis, Rheumatoid/*physiopathology
MH  - Autophagy/drug effects/*physiology
MH  - Autophagy-Related Proteins
MH  - Caspase 3/drug effects/metabolism
MH  - Cell Death/physiology
MH  - Cells, Cultured
MH  - Endoplasmic Reticulum Stress/*physiology
MH  - Enzyme Inhibitors/pharmacology
MH  - Fibroblasts/*physiology
MH  - Immunoblotting
MH  - Immunohistochemistry
MH  - Lysosomes/drug effects/physiology
MH  - Macrolides/pharmacology
MH  - Membrane Proteins/drug effects/metabolism
MH  - Microscopy, Fluorescence
MH  - Osteoarthritis/physiopathology
MH  - Polyubiquitin/drug effects/metabolism
MH  - Sequestosome-1 Protein
MH  - Synovial Membrane/*cytology
MH  - Transcription Factors/drug effects/metabolism
EDAT- 2014/01/23 06:00
MHDA- 2014/03/19 06:00
CRDT- 2014/01/23 06:00
PHST- 2013/03/14 00:00 [received]
PHST- 2013/09/03 00:00 [accepted]
PHST- 2014/01/23 06:00 [entrez]
PHST- 2014/01/23 06:00 [pubmed]
PHST- 2014/03/19 06:00 [medline]
AID - 10.1002/art.38190 [doi]
PST - ppublish
SO  - Arthritis Rheumatol. 2014 Jan;66(1):40-8. doi: 10.1002/art.38190.