asparagine starvation inhibits rhabdomyosarcoma growth connective tissue oncology society 2014...

Asparagine starvation inhibits rhabdomyosarcoma

growth

Connective Tissue Oncology Society 2014 Annual Meeting in Berlin

Simone Hettmer, Anna C Schinzel, Daria Tchessalova,Nigel GJ Richards, William C Hahn, Amy J Wagers

Ras pathway activationin fusion-negative rhabdomyosarcoma

Skeletal muscle

Ras pathway activation

Sarcoma induction in skeletal muscle

Transcriptional profiling identified 141 discrete genesthat are upregulated

in Kras; p16p19null mouse sarcomas and in human rhabdomyosarcomascompared to skeletal muscle.

Negative controlshRNA

shRNASarcoma

genes

Positive controlshKras

shASNS

Functional screening ofsarcoma-relevant candidate genes

q<0.001

Customized shRNA proliferation screen (in vitro)

Asparagine Synthetase (ASNS)

The growth-inhibitory effects of ASNS silencing are due to Asparagine starvation

Kras;p16p19null mouse RMS cell line

Kras;p16p19null mouse RMS cell line

Asparagine starvation reduces the synthesis of nascent polypeptide chains in rhabdoyosarcoma cells

Asparagine starvation reduces human rhabdomyosarcoma growth and peptide synthesis

Human RMS cell line RH30 (PAX3:FOXO1a +)

Asparagine starvation reduces mouse rhabdomyosarcoma growth in vivo

shASNS

shLuc

100 cells per injection

Asparagine starvation reduces mouse rhabdomyosarcoma growth in vivo

shASNS

shLuc

Aspara-ginase

Aspara-ginase

100 cells per injection

Asparagine homeostasis is anactionable cellular process

Amino Sulfoximines

Asparaginase and Amino Sulfoximine 5 inhibit rhabdomyosarcoma growth

Amino Sulfoximine 5Asparaginase

Kras;p16p19null mouse RMS cell line

Asparaginase effects onrhabdomyosarcoma growth in vivo

Aspara-ginase

Mouse RMS Rh30 (Hu RMS) Rh41 (Hu RMS)

Aspara-ginase

Control Aspara-ginase

Control Aspara-ginase

Control

p=0.02 p=0.09 p=0.0004

Asparagine homeostasis is an actionable metabolic vulnerability in sarcoma

Sarcoma induction by Ras pathway activation in skeletal muscle identified a cluster of sarcoma-relevant genes.

Functional screening identified Asparagine homeostasis as a metabolic vulnerability in sarcoma.

Asparagine starvation inhibits growth and nascent peptide synthesis in mouse and human sarcomas.

Chemical targeting of Asparagine homeostasis reduces mouse and human rhabdomyosarcoma growth.

Acknowledgements

Amy Wagers

Wagers lab:Daria TchessalovaTata Nageswara Rao

DFCI shRNA screening core:Anna SchinzelWilliam Hahn

David Langenau

Nigel Richards

Peter HoughtonRichard Lock

Joslin Flow Cytometry Core

Funding:PALS/St Baldrick’s FoundationAlex’s Lemonade StandBear NecessitiesSARC