Proteasome inhibition up-regulates p53 and apoptosis-inducing factor in chondrocytes causing severe growth retardation in mice.
Proteasome inhibitors (PI), a novel class of anticancer drugs, are relatively well tolerated and have recently been introduced into the clinic for the treatment of multiple myeloma. The tumor selectivity and low toxicity of PIs are surprising, given the crucial role of the ubiquitin/proteasome system in a multitude of cellular processes. Here, we show that systemic administration of PIs specifically impairs the ubiquitin/proteasome system in growth plate chondrocytes. Importantly, young mice displayed severe growth retardation during treatment as well as 45 days after the cessation of treatment with clinically relevant amounts of MG262 (0.2 micromol/kg body weight/injection) or bortezomib (1.0 mg/kg body weight/injection). Dysfunction of the ubiquitin/proteasome system was accompanied by the induction of apoptosis of stem-like and proliferative chondrocytes in the growth plate. These results were recapitulated in cultured fetal rat metatarsal bones and chondrocytic cell lines (rat, human). Apoptosis was associated with up-regulation of the proapoptotic molecules, p53 and apoptosis-inducing factor (AIF), both in vitro and in vivo. In addition to the observation that AIF is expressed in the growth plate, we also provide evidence that AIF serves as a direct target protein for ubiquitin, thus explaining its prominent up-regulation upon proteasome inhibition. Suppression of p53 or AIF expression with small interfering RNAs partly rescued chondrocytes from proteasome inhibition-induced apoptosis (35% and 41%, respectively). Our observations show that proteasome inhibition may selectively target essential cell populations in the growth plate causing significant growth failure. These findings could have important implications for the use of proteasome inhibitors in the treatment of childhood cancer.
Zaman
Farasat
F
Department of Woman and Child Health, Pediatric Endocrinology Unit, Astrid Lindgren Children's Hospital, Stockholm, Sweden. Farasat.Zaman@ki.se
Menendez-Benito
Victoria
V
Eriksson
Emma
E
Chagin
Andrei S
AS
Takigawa
Masaharu
M
Fadeel
Bengt
B
Dantuma
Nico P
NP
Chrysis
Dionisios
D
Sävendahl
Lars
L
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D051033:Apoptosis Inducing Factor / Q000096:biosynthesis / Q000378:metabolism*; D001897:Boronic Acids / Q000494:pharmacology; D000069286:Bortezomib; D019902:Chondrocytes / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism*; D006130:Growth Disorders / Q000139:chemically induced / Q000201:enzymology* / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D011480:Protease Inhibitors / Q000494:pharmacology*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D061988:Proteasome Inhibitors; D011719:Pyrazines / Q000494:pharmacology; D051381:Rats; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis* / Q000235:genetics; D025801:Ubiquitin / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
D051033:Apoptosis Inducing Factor; D001897:Boronic Acids; C423917:MG 262; C494830:AIFM1 protein, mouse; D011480:Protease Inhibitors; D061988:Proteasome Inhibitors; D011719:Pyrazines; D016159:Tumor Suppressor Protein p53; D025801:Ubiquitin; D000069286:Bortezomib; D046988:Proteasome Endopeptidase Complex