High throughput synthetic lethality screen reveals a tumorigenic role of adenylate cyclase in fumarate hydratase-deficient cancer cells

Michael Boettcher,Johannes Fredebohm,Jörg D Hoheisel,Christian Frezza,Viola Ladenburger,Andrew Lawson,Jonas Wolf,Tomer Shlomi

doi:10.1186/1471-2164-15-158

Abstract

BackgroundSynthetic lethality is an appealing technique for selectively targeting cancer cells which have acquired molecular changes that distinguish them from normal cells. High-throughput RNAi-based screens have been successfully used to identify synthetic lethal pathways with well-characterized tumor suppressors and oncogenes. The recent identification of metabolic tumor suppressors suggests that the concept of synthetic lethality can be applied to selectively target cancer metabolism as well.ResultsHere, we perform a high-throughput RNAi screen to identify synthetic lethal genes with fumarate hydratase (FH), a metabolic tumor suppressor whose loss-of-function has been associated with hereditary leiomyomatosis and renal cell carcinoma (HLRCC). Our unbiased screen identified synthetic lethality between FH and several genes in heme metabolism, in accordance with recent findings. Furthermore, we identified an enrichment of synthetic lethality with adenylate cyclases. The effects were validated in an embryonic kidney cell line (HEK293T) and in HLRCC-patient derived cells (UOK262) via both genetic and pharmacological inhibition. The reliance on adenylate cyclases in FH-deficient cells is consistent with increased cyclic-AMP levels, which may act to regulate cellular energy metabolism.ConclusionsThe identified synthetic lethality of FH with adenylate cyclases suggests a new potential target for treating HLRCC patients.

Highlights

Synthetic lethality is an appealing technique for selectively targeting cancer cells which have acquired molecular changes that distinguish them from normal cells
Loss-of-function mutations in fumarate hydratase (FH) have been associated with a number of diseases including hereditary leiomyomatosis and renal cell carcinoma (HLRCC), a cancer syndrome characterized by a malignant form of renal cancer [12]
Utilizing pooled RNAi screen to identify synthetic lethality with FH To identify genes that are synthetic lethal with FH, we applied an approach that combines the pooled short hairpin RNA vectors (shRNA) silencing of 10,000 different genes with short-interfering RNA (siRNA) silencing of FH in a FH-proficient embryonic kidney cell-line HEK293T (Figure 1A)

Summary

Introduction

Synthetic lethality is an appealing technique for selectively targeting cancer cells which have acquired molecular changes that distinguish them from normal cells. In forward genetics, gene knockdown effects are measured in multiple, The recent identification of somatic mutations in cancer that affect metabolic enzymes suggests that the concept of synthetic lethality may be successfully applied to target cancer metabolism [8,9,10,11]. Such genetic mutations include the identification of two metabolic tumor suppressors in TCA cycle, fumarate hydratase (FH) and succinate dehydrogenase (SDH), and oncogene, isocitrate dehydrogenase (IDH), as well as other passenger loss-of-function mutations [8,9,10]. While this represents a promising application of metabolic synthetic lethality, no high-throughput screening for synthetic lethality has been performed so far

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