Fumarate hydratase deficiency in renal cancer induces glycolytic addiction and hypoxia-inducible transcription factor 1alpha stabilization by glucose-dependent generation of reactive oxygen species.
Hereditary leiomyomatosis and renal cell cancer (HLRCC) is an inherited cancer syndrome linked to biallelic inactivation of the gene encoding the tricarboxylic acid cycle enzyme fumarate hydratase (FH). Individuals with HLRCC are at risk to develop cutaneous and uterine leiomyomas and an aggressive form of kidney cancer. Pseudohypoxic drive-the aberrant activation of cellular hypoxia response pathways despite normal oxygen tension-is considered to be a likely mechanism underlying the etiology of this tumor. Pseudohypoxia requires the oxygen-independent stabilization of the alpha subunit of the hypoxia-inducible transcription factor (HIF-1alpha). Under normoxic conditions, proline hydroxylation of HIF-1alpha permits VHL recognition and subsequent targeting for proteasomal degradation. Here, we demonstrate that inactivating mutations of FH in an HLRCC-derived cell line result in glucose-mediated generation of cellular reactive oxygen species (ROS) and ROS-dependent HIF-1alpha stabilization. Additionally, we demonstrate that stable knockdown of FH in immortalized renal epithelial cells results in ROS-dependent HIF-1alpha stabilization. These data reveal that the obligate glycolytic switch present in HLRCC is critical to HIF stabilization via ROS generation.
Sudarshan
Sunil
S
Urologic Oncology Branch, National Cancer Institute/NIH, Bethesda, MD 20892, USA. neckersl@mail.nih.gov
Sourbier
Carole
C
Kong
Hye-Sik
HS
Block
Karen
K
Valera Romero
Vladimir A
VA
Yang
Youfeng
Y
Galindo
Cynthia
C
Mollapour
Mehdi
M
Scroggins
Bradley
B
Goode
Norman
N
Lee
Min-Jung
MJ
Gourlay
Campbell W
CW
Trepel
Jane
J
Linehan
W Marston
WM
Neckers
Len
L
D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D005649:Fumarate Hydratase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology*; D006019:Glycolysis / Q000187:drug effects*; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism*; D007680:Kidney Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D009097:Multienzyme Complexes / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D018384:Oxidative Stress; D051745:Protein Kinase C-delta / Q000378:metabolism; D034622:RNA Interference; D017382:Reactive Oxygen Species / Q000378:metabolism*
D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D009097:Multienzyme Complexes; D017382:Reactive Oxygen Species; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases; D051745:Protein Kinase C-delta; D005649:Fumarate Hydratase; D005947:Glucose