Metabolic reprogramming in tumors represents a potential therapeutic target. Herein we used shRNA depletion and a novel lactate dehydrogenase (LDHA) inhibitor, GNE-140, to probe the role of LDHA in tumor growth in vitro and in vivo. In MIA PaCa-2 human pancreatic cells, LDHA inhibition rapidly affected global metabolism, although cell death only occurred after 2 d of continuous LDHA inhibition. Pancreatic cell lines that utilize oxidative phosphorylation (OXPHOS) rather than glycolysis were inherently resistant to GNE-140, but could be resensitized to GNE-140 with the OXPHOS inhibitor phenformin. Acquired resistance to GNE-140 was driven by activation of the AMPK–mTOR–S6K signaling pathway, which led to increased OXPHOS, and inhibitors targeting this pathway could prevent resistance. Thus, combining an LDHA inhibitor with compounds targeting the mitochondrial or AMPK–S6K signaling axis may not only broaden the clinical utility of LDHA inhibitors beyond glycolytically dependent tumors but also reduce the emergence of resistance to LDHA inhibition.

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We would like to thank the Discovery Chemistry Small Molecule analytical group for their support; the in vivo cell culture and dosing groups for their support of in vivo studies; R. Bourgon for bioinformatic assistance; the Genentech gCell group (R. Neve, M. Yu and M. Liang-Chu) for cell line banking and maintenance; and members of Genentech gCSI group (E. Lin, B. Lam, Y. Yu and J. Tan) for assistance on cell-based drug screens; S. Sriraman for assistance with the 18FDG-uptake assays; and A. Bruce for assistance with generating figures.

Author information

Author notes

    • Peter K Jackson

    Present address: Department of Microbiology and Immunology, Stanford University, Stanford, California, USA.

    • Aaron Boudreau
    •  & Hans E Purkey

    These authors contributed equally to this work.


  1. Discovery Oncology, Genentech, South San Francisco, California, USA.

    • Aaron Boudreau
    • , David Peterson
    • , Raju Pusapati
    •  & Marie Evangelista
  2. Discovery Chemistry, Genentech, South San Francisco, California, USA.

    • Hans E Purkey
    • , Kirk Robarge
    • , Sharada Labadie
    • , Aihe Zhou
    •  & Binqing Wei
  3. Translational Oncology, Genentech, South San Francisco, California, USA.

    • Anna Hitz
    • , Mandy Kwong
    • , Rebecca Hong
    • , Min Gao
    • , Christopher Del Nagro
    • , Laura Corson
    • , Marcia Belvin
    • , Peter K Jackson
    • , Georgia Hatzivassiliou
    • , Deepak Sampath
    •  & Thomas O'Brien
  4. Drug Metabolism and Pharmacokinetics, Genentech, South San Francisco, California, USA.

    • Shuguang Ma
    • , Laurent Salphati
    •  & Jodie Pang
  5. Chemistry, WuXi AppTec Co., Ltd., Shanghai, China.

    • Tommy Lai
  6. Structural Biology, WuXi AppTec Co., Ltd., Shanghai, China.

    • Yingjie Li
    •  & Zhongguo Chen
  7. Biochemical and Cellular Pharmacology, Genentech, South San Francisco, California, USA.

    • Ivana Yen
    • , Steve Sideris
    •  & Shiva Malek
  8. Department of Pathology, Genentech, South San Francisco, California, USA.

    • Mark McCleland
    •  & Ron Firestein
  9. Biomedical Imaging, Genentech, South San Francisco, California, USA.

    • Alex Vanderbilt
    •  & Simon Williams
  10. Bioinformatics, Genentech, South San Francisco, California, USA.

    • Anneleen Daemen
  11. Structural Biology, Genentech, South San Francisco, California, USA.

    • Charles Eigenbrot


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A.B. contributed in the generation of, and characterized, resistant cell lines and to manuscript writing; H.E.P. and K.R. designed and directed compound generation and characterization of lead molecules and assisted in generating co-crystal structure; A.H. contributed through the generation and characterization shLDHA cell lines and by performing metabolic profiling experiments; D.P. characterized the induction of ROS, performed cell line assays and characterized lactate levels in cell lines; S.L. designed and assisted in compound generation and characterization of lead molecules; M.K. and M.G. performed all MS-based assays; R.H. designed and carried out in vivo studies; C.D.N. performed cell cycle analysis of cells; R.P. assisted in understanding the mechanism of resistance; M.M. and R.F. assisted with knockdown construct design and characterization; S. Ma, L.S. and J.P. designed and carried out pharmacokinetic studies in mice; A.Z. and T.L. designed and assisted in compound generation and characterization of lead molecules; Y.L. and Z.C. obtained a co-crystal structure; B.W. designed molecules and assisted in obtaining a co-crystal structure; I.Y., S.S. and S. Malek designed and performed biochemical and selectivity assays; L.C. designed and generated shLDHA cells; A.V. and S.W. performed cellular glucose uptake assays; A.D. assisted in analysis of metabolite profiling in cells; M.B. performed characterization of lead molecule in cells; C.E. directed protein production and generation of a co-crystal structure; P.K.J. directed the design and characterization of shLDHA cells; G.H. contributed intellectually to the design of all metabolic assays and helped draft the manuscript; D.S. directed in vivo studies and helped draft the manuscript; M.E. directed the generation and characterization of resistant clones, as well as cell line screening and in vivo studies; T.O'B. directed the characterization of the role of LDHA in cells and helped design and direct metabolic and cell cycle and in vivo studies; M.E. and T.O'B. drafted and revised the manuscript for intellectual content and final approval.

Competing interests

All authors were full-time employees of either Genentech or WuXi at the time these studies were conducted.

Corresponding authors

Correspondence to Marie Evangelista or Thomas O'Brien.

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    Supplementary Text and Figures

    Supplementary Results, Supplementary Tables 1–3 and Supplementary Figures 1–9.

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    Supplementary Dataset 1

    Complete list of the fold change of all metabolites detected in MIA PaCa-2 cells following treatment with 2 μM GNE-140.

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