chromophobe rcc - kidneycan · 2019. 12. 20. · gss= glutathione synthetase slc7a11=...
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Chromophobe RCC
Elizabeth (Lisa) Petri Henske, MDDirector, Center for LAM Research and Clinical Care,
Brigham and Women’s Hospital, Boston MA
Professor of Medicine, Harvard Medical School
Medical Oncologist, Dana Farber Cancer Institute
Associate Member, The Broad Institute of MIT and Harvard
Ehenske@BWH.Harvard.edu
Carmen Priolo, MD PhDAssistant Professor of Medicine, BWH, Harvard Medical School
Brigham and Women’s Hospital/Dana-Farber Cancer Institute
Harvard Medical School
Growing chRCC Team!
BWH: Carmen Priolo
David KwiatkowskiDamir Khabibulllin
Long ZhangRachel Yu
DFCI:David BraunToni Choueiri
Sabina SignorettiMatthew Freedman
+ Others!
Wei Shi (MGH)John Asara (Beth Israel, Boston)
Ed Reznik (MSKCC)
National Cancer Institute/NIH (RO1)
US Department of Defense Kidney Cancer Program (Hypothesis Award)
The Tuttle Family
Lymphangioleiomyomatosis (LAM)
Angiomyolipomas
ChRCC can occur in Tuberous Sclerosis Complex and Birt-Hogg-Dube Syndrome
Renal Cell Carcinoma
Tuberous Sclerosis Complex
Birt-Hogg-Dube
5% of all sporadic renal cancers
Occur in BHD and TSC
Filled with abnormal mitochondria
Losses of chromosomes 1, 2, 6, 10, 13 and 17
Few driver mutations (TCGA: ~50%: p53, Tsc1, PIK3CA)
4
Chromophobe RCC
• 367 measured metabolites
• 188 differential metabolites
TumorNormal
*Glucose metabolism
*Glutathione metabolism
Metabolomics: Nine chRCC vs. matched normal kidney
7.00
6.00
5.00
4.00
3.00
2.00
1.00
0.00
-1.00
-lo
g10
(p)
log2(FC)-8.00 -7.00 -6.00 -5.00 -4.00 -3.00 -2.00 -1.00 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00
5-oxoproline
Ɣglutamylleucine
Ɣglutamylglutamate
Top differential metabolites in ChRCC compared to normal kidney
Features selected by volcano plot with fold change threshold 2
(FC; x) and t-tests threshold p <0.01 (y). The red circles
represent features above the threshold. The further the red
circle is away from the (0,0), the more significant the feature is.
Normal kidney
Tumor36 metabolites decreased in tumor vs normal
Decreased expression of gamma-glutamyl
transferase 1 (GGT1) in ChRCC
nsp<0.001
GGT1 gene locus= Chr 22q11.23 RNA seq data from TCGA
n=25 n=66 n=18 n=82
The glutathione salvage pathway (or gamma-glutamyl cycle)
• gamma-glutamyl transferase 1
(GGT1): membrane
transpeptidase
• transfers gamma-glutamyl
moieties from extracellular
GSH, GSSG, or GSH
conjugates to an amino acid
acceptor (gamma-glutamyl
amino acid)
• GGT-deficient mice: lower
GSH levels, increased DNA
damage (Rojas et al, 1999),
and dysfunctional mitochondria
(Will et al, 2000)
GCLC= Glutamate-cysteine ligase catalytic subunit
GSS= glutathione synthetase
SLC7A11= cystine-glutamate antiporter
GCLC
GSS
Enhanced expression of glutathione biosynthetic enzymes in ChRCC
10
GGT1 downregulation increases De Novo Glutathione Synthesis
CB-839
Priolo C, Khabibullin D,… Henske EP. PNAS 2018HEK293,72h
GGT1 down-regulation sensitizes HEK203 to inhibition
of glutathione biosynthesis
Damir Khabibullin
BSO=buthionine sulfoximine
48-hr treatment after 72-hr transfection with siGGT1 or siControl
Working model and Future Directions
Hypothesis: Defects in the Ɣ-glutamyl cycle lead to accumulation of damaged
mitochondria and tumorigenesis
Compensatory metabolic reprogramming Impaired response to oxidative stress
• Role in renal tumorigenesis• Increased sensitivity to agents that inhibit glutathione synthesis
p-AMPK(Thr172)
GGT1
p-ULK1(S555)
GGT1 downregulation in HEK293: Erk activation and suppression of LKB1, AMPK and ULK1
13
siCtrl siGGT1
siCtrl siGGT1
siCtrl siGGT1
p-LKB1(Ser428)
actin
p-Erk1/2(Thr202/Tyr204)
ATG7
ATG5
β-actin
Conan GK. Nature Medicine. 2019
HEK293T, 72hAmbion siGGT1 5725
Rachel Yu, unpublished
Major Roadblock: chRCC Models
• Additional cell lines
• Additional animal models
15
• Chromophobe RCC with sarcomatoid differentiation
• Clonal losses of chromosomes 1, 3, 4, 11, 13, 14, and 19 and gains of X, Y, 6, 8, and 20.
– (vs ChRCC losses of chromosomes 1, 2, 6, 10, 13 and 17)
• TP53 missense mutation
Genes Chromosomes and Cancer, 2017
H E K 2 9 3 U O K 2 7 6
0 .0
0 .5
1 .0
1 .5
Re
lati
ve
GG
T1
mR
NA
Low GGT1 in UOK276 relative to HEK293
GGT1Chromo1039 cellsChromo1039 Tissue
GGT1
ChromoA cells
ChRCC-derived primary cell lines: low GGT1 expression
A
B
(Gerharz et al., Am J Path, 1995)
SNP arrays, 250K
Big Questions
Additional mouse models of chRCC
Which nephron segment?
Ksp (Kidney specific cadherin)
Aqp2 (Aquaporin 2, Principal cells of collecting duct)
ATP6V1B1 (alpha-Intercalated cells of collecting duct )
Which genes?
GGT1 and PTEN (20% ChRCC has PTEN mutation)
GGT1 and TSC2
Take Home Hypothesis:
Defective glutathione salvage pathway (or gamma-glutamyl cycle)
Role in Pathogenesis and Therapy of chRCC
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