Chromophobe RCC

Elizabeth (Lisa) Petri Henske, MDDirector, Center for LAM Research and Clinical Care,

Brigham and Women’s Hospital, Boston MAProfessor of Medicine, Harvard Medical School

Medical Oncologist, Dana Farber Cancer InstituteAssociate Member, The Broad Institute of MIT and Harvard

Ehenske@BWH.Harvard.edu

Brigham and Women’s Hospital/Dana-Farber Cancer Institute

Harvard Medical School

Growing chRCC Team!

BWH: Carmen Priolo

David KwiatkowskiDamir Khabibulllin

Long Zhang

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 TSCFilled with abnormal mitochondria Losses of chromosomes 1, 2, 6, 10, 13 and 17Few 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

-log10(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 circlesrepresent features above the threshold. The further the redcircle is away from the (0,0), the more significant the feature is.

Normal kidneyTumor36 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): membranetranspeptidase

• transfers gamma-glutamylmoieties from extracellularGSH, GSSG, or GSHconjugates to an amino acidacceptor (gamma-glutamylamino acid)

• GGT-deficient mice: lowerGSH levels, increased DNAdamage (Rojas et al, 1999),and dysfunctional mitochondria(Will et al, 2000)

GCLC= Glutamate-cysteine ligase catalytic subunit GSS= glutathione synthetaseSLC7A11= cystine-glutamate antiporter

GCLC

GSS

Enhanced expression of glutathione biosynthetic enzymes in ChRCC

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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 sulfoximine48-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

Oxidative stresscaused by an imbalance between production and accumulation of oxygen reactive species (ROS) and the ability to detoxify these reactive products

GlutathioneAn anti-oxidant, helps detoxify reactive oxygen species

MitochondriaCellular powerhouses or “batteries” that make ATP and can generate reactive oxygen species, especially when they are damaged

Major Roadblock: chRCC Models

• Additional cell lines• Additional animal models

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• 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 60 .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

Take Home Hypothesis: Defective glutathione salvage pathway (or gamma-glutamyl cycle)

Chromophobe RCC may have altered sensitivity to oxidative stress

Chromophobe RCC

Elizabeth (Lisa) Petri Henske, MDDirector, Center for LAM Research and Clinical Care,

Brigham and Women’s Hospital, Boston MAProfessor of Medicine, Harvard Medical School

Medical Oncologist, Dana Farber Cancer InstituteAssociate Member, The Broad Institute of MIT and Harvard

Ehenske@BWH.Harvard.edu