PHGDH is required for germinal center formation and is a therapeutic target in MYC-driven lymphoma
Annalisa D’Avola, Nathalie Legrave, Mylène Tajan, Probir Chakravarty, Ryan L. Shearer, Hamish W. King, Katarina Kluckova, Eric C. Cheung, Andrew J. Clear, Arief S. Gunawan, Lingling Zhang, Louisa K. James, James I. MacRae, John G. Gribben, Dinis P. Calado, Karen H. Vousden, John C. Riches
Annalisa D’Avola, Nathalie Legrave, Mylène Tajan, Probir Chakravarty, Ryan L. Shearer, Hamish W. King, Katarina Kluckova, Eric C. Cheung, Andrew J. Clear, Arief S. Gunawan, Lingling Zhang, Louisa K. James, James I. MacRae, John G. Gribben, Dinis P. Calado, Karen H. Vousden, John C. Riches
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Research Article Immunology Metabolism

PHGDH is required for germinal center formation and is a therapeutic target in MYC-driven lymphoma

Abstract

The synthesis of serine from glucose is a key metabolic pathway supporting cellular proliferation in healthy and malignant cells. Despite this, the role that this aspect of metabolism plays in germinal center biology and pathology is not known. Here, we performed a comprehensive characterization of the role of the serine synthesis pathway in germinal center B cells and lymphomas derived from these cells. We demonstrate that upregulation of a functional serine synthesis pathway is a metabolic hallmark of B cell activation and the germinal center reaction. Inhibition of phosphoglycerate dehydrogenase (PHGDH), the first and rate-limiting enzyme in this pathway, led to defective germinal formation and impaired high-affinity antibody production. In addition, overexpression of enzymes involved in serine synthesis was a characteristic of germinal center B cell–derived lymphomas, with high levels of expression being predictive of reduced overall survival in diffuse large B cell lymphoma. Inhibition of PHGDH induced apoptosis in lymphoma cells, reducing disease progression. These findings establish PHGDH as a critical player in humoral immunity and a clinically relevant target in lymphoma.

Authors

Annalisa D’Avola, Nathalie Legrave, Mylène Tajan, Probir Chakravarty, Ryan L. Shearer, Hamish W. King, Katarina Kluckova, Eric C. Cheung, Andrew J. Clear, Arief S. Gunawan, Lingling Zhang, Louisa K. James, James I. MacRae, John G. Gribben, Dinis P. Calado, Karen H. Vousden, John C. Riches

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Figure 6

PHGDH inhibition impairs proliferation and promotes apoptosis in Burkitt lymphoma cells.

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PHGDH inhibition impairs proliferation and promotes apoptosis in Burkitt...
(A) Western blot analysis of PHGDH, PSAT1, and PSPH protein expression in B cell–derived lymphoma cell lines (mantle cell lymphoma [MCL]). Representative of 3 independent experiments. HSC70 was used as loading control. (B) Cell cycle profile of Ramos (top), Raji (center), and Daudi (bottom) cells. Cells were plated either in complete medium or equivalent medium lacking serine and glycine supplemented or not with 0.5 mM sodium formate and 0.4 mM glycine and treated with DMSO (as a solvent control) or 10 μM PH-755, followed by incubation with 10 μM BrdU and by staining with anti-BrdU and 7-ADD. Data are presented as mean ± SEM and are representative of 3 independent experiments, with value for DMSO-treated cells and cultured in complete medium set to 1.0. (C) Ramos (left), Raji (center), and Daudi (right) cells were cultured in the same conditions specified in B for 48 hours. Cells were then permeabilized, fixed, and stained for active Caspase-3. Positive cells for active Caspase-3 were analyzed by flow cytometry. Graph shows the mean derived from 3 independent experiments, with value for DMSO-treated cells and cultured in complete medium set to 1.0. Data are shown as the mean ± SEM. **P < 0.01, ***P < 0.001, and ****P < 0.0001, by 1-way ANOVA with Tukey’s post hoc test. (D) Mass isotopologue distribution of U-[13C6]-glucose–derived serine and glycine for Ramos (top) and Daudi (bottom) cells cultured for 2 and 24 hours in medium lacking serine and glycine in presence of U-[13C6]-glucose (10 mM) and treated with DMSO or 10 μM PH-755. Serine, glycine, ATP, and GTP levels were measured by LC-MS. The percentage distribution of each isotopologue for their respective metabolite pool is shown. Data are presented as mean ± SEM of 6 repeats and are representative of 3 independent experiments.