Lipid droplets can promote drug accumulation and activation

ABSTRACT Genetic screens in cultured human cells represent a powerful unbiased strategy to identify cellular pathways that determine drug efficacy, providing critical information for

clinical development. We used insertional mutagenesis-based screens in haploid cells to identify genes required for the sensitivity to lasonolide A (LasA), a macrolide derived from a marine

sponge that kills certain types of cancer cells at low nanomolar concentrations. Our screens converged on a single gene, _LDAH_, encoding a member of the metabolite serine hydrolase family

that is localized on the surface of lipid droplets. Mechanistic studies revealed that LasA accumulates in lipid droplets, where it is cleaved into a toxic metabolite by LDAH. We suggest that

selective partitioning of hydrophobic drugs into the oil phase of lipid droplets can influence their activation and eventual toxicity to cells. Access through your institution Buy or

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INTERROGATION OF CANCER VULNERABILITY USING A MULTIPLEXED CELL LINE SCREENING PLATFORM Article Open access 02 July 2021 METABOLIC DRUG SURVEY HIGHLIGHTS CANCER CELL DEPENDENCIES AND

VULNERABILITIES Article Open access 14 December 2021 SYNTHETIC LETHAL STRATEGIES FOR THE DEVELOPMENT OF CANCER THERAPEUTICS Article 03 December 2024 DATA AVAILABILITY The complete lists of

the hits from the genetic screens are given in Supplementary Data 1. RNA-seq data from Hap1 cells is freely available at NCBI GEO, under accession no. GSE75515. The GI50 data for LasA and

the RNA-seq data for cancer cell lines is publicly available (accession numbers given in the appropriate Methods section). Software for analysis of screen results has been described

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Download references ACKNOWLEDGEMENTS We thank D. Herschlag for bringing the LasA project to our attention, C. Pataki and R. Kopito for comments and advice on lipid droplet fractionation

experiments and A. Lebensohn for advice on the project. The work was funded by DP2 GM105448 (R.R.), R35 GM118082 (R.R.), DP2 AI104557 (J.E.C.), American Heart Association Transformational

Research Projects no. 18TPA34230103 (A.P.) and no. 18TPA34230086 (Y.-H.G.), and Dominic Ferraioli Foundation (A.P.). R.R. is a Josephine Q. Berry Faculty Scholar in Cancer Research at

Stanford, J.E.C. is a David and Lucile Packard Foundation fellow and R.D. was supported by fellowships from the Stanford Dean’s Fund and Alex’s Lemonade Stand Foundation. AUTHOR INFORMATION

AUTHORS AND AFFILIATIONS * Department of Biochemistry, Stanford University School of Medicine, Stanford, CA, USA Ramin Dubey & Rajat Rohatgi * Department of Chemistry, Stanford

University, Stanford, CA, USA Craig E. Stivala & Barry M. Trost * Genentech, South San Francisco, CA, USA Craig E. Stivala & Huy Quoc Nguyen * Department of Molecular and Cellular

Physiology, Albany Medical College, Albany, NY, USA Young-Hwa Goo & Antoni Paul * Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA, USA Jan

E. Carette * Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA Rajat Rohatgi Authors * Ramin Dubey View author publications You can also search for this

author inPubMed Google Scholar * Craig E. Stivala View author publications You can also search for this author inPubMed Google Scholar * Huy Quoc Nguyen View author publications You can also

search for this author inPubMed Google Scholar * Young-Hwa Goo View author publications You can also search for this author inPubMed Google Scholar * Antoni Paul View author publications

You can also search for this author inPubMed Google Scholar * Jan E. Carette View author publications You can also search for this author inPubMed Google Scholar * Barry M. Trost View author

publications You can also search for this author inPubMed Google Scholar * Rajat Rohatgi View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS

R.R. and R.D. designed the project. B.M.T. and C.E.S. designed and synthesized LasA, LasF, Ces-73, Ces-24a and Ces-24b. R.D. and J.E.C. executed the haploid genetic screens. R.D. and H.Q.N.

performed the mass spectrometry experiments. R.D., A.P. and Y.-H.G. designed and constructed the LDAH variants. R.D. performed all other experiments and analyses presented in the paper. R.R.

and R.D. wrote the paper and all the authors edited and commented on the paper. CORRESPONDING AUTHOR Correspondence to Rajat Rohatgi. ETHICS DECLARATIONS COMPETING INTERESTS The authors

declare no competing interests. ADDITIONAL INFORMATION PUBLISHER’S NOTE Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION Supplementary Figs. 1–12. REPORTING SUMMARY SUPPLEMENTARY VIDEO 1 Movie of live Hap1 cells expressing LDAH-GFP. SUPPLEMENTARY DATA 1

Compiled data from the haploid screens (see Fig. 1c and Supplementary Figs. 1 and 2a). RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Dubey, R.,

Stivala, C.E., Nguyen, H.Q. _et al._ Lipid droplets can promote drug accumulation and activation. _Nat Chem Biol_ 16, 206–213 (2020). https://doi.org/10.1038/s41589-019-0447-7 Download

citation * Received: 13 February 2019 * Accepted: 02 December 2019 * Published: 13 January 2020 * Issue Date: February 2020 * DOI: https://doi.org/10.1038/s41589-019-0447-7 SHARE THIS

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