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Pharmacological and genetic perturbation establish SIRT5 as a promising target in breast cancer

Abstract

SIRT5 is a member of the sirtuin family of NAD+-dependent protein lysine deacylases implicated in a variety of physiological processes. SIRT5 removes negatively charged malonyl, succinyl, and glutaryl groups from lysine residues and thereby regulates multiple enzymes involved in cellular metabolism and other biological processes. SIRT5 is overexpressed in human breast cancers and other malignancies, but little is known about the therapeutic potential of SIRT5 inhibition for treating cancer. Here we report that genetic SIRT5 disruption in breast cancer cell lines and mouse models caused increased succinylation of IDH2 and other metabolic enzymes, increased oxidative stress, and impaired transformation and tumorigenesis. We, therefore, developed potent, selective, and cell-permeable small-molecule SIRT5 inhibitors. SIRT5 inhibition suppressed the transformed properties of cultured breast cancer cells and significantly reduced mammary tumor growth in vivo, in both genetically engineered and xenotransplant mouse models. Considering that Sirt5 knockout mice are generally normal, with only mild phenotypes observed, these data establish SIRT5 as a promising target for treating breast cancer. The new SIRT5 inhibitors provide useful probes for future investigations of SIRT5 and an avenue for targeting SIRT5 as a therapeutic strategy.

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Fig. 1: SIRT5 is overexpressed in human breast cancer.
Fig. 2: Sirt5 knock-down inhibits the anchorage-independent growth of human cancer cells.
Fig. 3: Sirt5 deletion impairs tumor growth and reduces metastasis burden in vivo.
Fig. 4: Sirt5 deletion significantly increases succinylation levels of proteins and increases ROS levels.
Fig. 5: Development of thiourea SIRT5 inhibitors JH-I5-2 and DK1-04.
Fig. 6: DK1-04e inhibits the cellular growth of breast cancer cells by targeting SIRT5.
Fig. 7: SIRT5-selective inhibitor prodrug DK1-04e impairs mammary tumor growth in MMTV-PyMT transgenic mice and human breast cancer mouse model.

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Acknowledgements

The authors thank the NCI Physical Sciences-Oncology Network Bioresource Core Facility (PBCF) for MDA-MB-231 cells, and Drs. Ruchika Bhawal and Sheng Zhang from the Cornell Proteomics and Metabolomics Facility for technical assistance. This work was supported in part by NIH R01 grants CA163255 and CA223534. YLNA was supported by NIH/NIGMS grant 5T32GM008500, and IRF was supported by NIH/NIGMS grant T32GM007273, a Cornell Deans Excellence Fellowship, and an HHMI Gilliam Fellowship. JA was supported by grants from the Ecole Polytechnique Federale de Lausanne (EPFL), and the Swiss National Science Foundation (SNSF 31003A_179435). This work utilized the Cornell University NMR facility, which is supported in part by the NSF through MRI award CHE-1531632.

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YLNA, IRF, JYH, RC, HL, and RSW conceptualization; YLNA, IRF, and JYH, formal analysis; YLNA, IRF, JYH, YLC, DAK, QZ, MY, JH, SS, BL, BH, BR, JJB, RD, JM, FW, VM, and TS investigation; YLNA, IRF, JYH, YLC, DAK, QZ, MY, JH, SS, BL, BH, and JJB methodology; YLNA, IRF, and JYH writing-original draft; YLNA, IRF, JYH, HL, and RSW writing-review and editing; JA, RC, HL, and RSW resources; RC, HL, and RSW funding acquisition; HL and RSW supervision; HL and RSW project administration.

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Correspondence to Hening Lin or Robert S. Weiss.

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Cornell University has patents on the SIRT5 inhibitors described in the manuscript. The authors have no additional competing interests to declare.

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Abril, Y.L.N., Fernandez, I.R., Hong, J.Y. et al. Pharmacological and genetic perturbation establish SIRT5 as a promising target in breast cancer. Oncogene 40, 1644–1658 (2021). https://doi.org/10.1038/s41388-020-01637-w

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