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2022-RA-701-ESGO Elucidating mechanisms underlying methotrexate resistance via quantitative proteomics analysis of GTN patient samples and choriocarcinoma cell lines: a crucial role for serine metabolism
  1. Panagiota Ntavelou1,
  2. Marina Georgiou1,
  3. Geoffrey Maher1,2,
  4. Rajat Roy1,
  5. Sijia Yu1,
  6. Paul Ajuh3,
  7. Baljeet Kaur2,
  8. Neil Horowitz4,
  9. Ross S Berkowitz4,
  10. Kevin Elias4,
  11. Michael J Seckl1,2 and
  12. Olivier E Pardo1
  1. 1Division of Cancer, Department of Surgery and Cancer, Imperial College, London, UK
  2. 2Trophoblastic Tumour Screening and Treatment Centre, Charing Cross Hospital, Imperial College Healthcare NHS Trust, London, UK
  3. 3Gemini Biosciences, Liverpool, UK
  4. 4Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, Brigham and Women’s Hospital, Dana Farber Cancer Institute, Harvard Medical School, Boston, MA


Introduction/Background Choriocarcinoma is an aggressive type of Gestational Trophoblastic Neoplasia (GTN). Patients with low-risk GTN following a molar pregnancy frequently commence therapy with single-agent methotrexate (MTX). Unfortunately, many develop resistance (MTX-R) and require either another single agent or more toxic combination agent chemotherapy to achieve remission. Understanding the molecular mechanisms of MTX-R may identify interventions to prevent or reverse this.

Methodology We employed proteomics profiling to identify changes that accompany MTX-R in post molar GTN patient samples and in choriocarcinoma cell lines that were either MTX sensitive (MTX-S) or resistant (MTX-R).

Results Quantitative mass spectrometry (MS) analysis revealed that the de novo serine synthesis pathway was one of the most downregulated pathways both in the MTX-R patients and in the resistant choriocarcinoma cell line. Decreased glucose-derived serine synthesis is supported by our findings that choriocarcinoma MTX-R cells have a less active glycolytic pathway. Concomitant de novo serine synthesis inhibition and serine deprivation from the growth medium promoted MTX-R in cell lines. Moreover, examination of the expression levels of 1-Carbon metabolism enzymes suggested a different utilization of serine, the major 1-Carbon donor, between sensitive and resistant cells. In particular, MTX-R cells may be channeling serine into pathways crucial for cell survival, such as the synthesis of glutathione (GSH), as indicated by the increased levels of this metabolite in resistant cells.

Conclusion Upon MTX-R, choriocarcinoma cells favor redirection of serine to GSH production and this may help with combating chemotherapy-induced reactive oxygen species (ROS) accumulation and hence participate in the resistant phenotype. In contrast, MTX-sensitive cells utilize serine for nucleotide synthesis and the maintenance of proliferation. Hence, targeting upstream pathways or molecules that block the synthesis of serine in combination with or without MTX treatment could improve therapeutic response in patients with MTX resistance.

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