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 2

Characterization of the SSP in WT mice after activation in vivo.

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Characterization of the SSP in WT mice after activation in vivo. (A) Ana...
(A) Analysis of PHGDH, PSAT1, and PSPH protein levels in resting B cells isolated from mouse spleen (SPL), peripheral blood (PB), and lymph nodes (LN). NIH3T3 murine cells were used as control for high expression of SSP-related enzymes. (B) Representative IHC staining for PNA as GC marker, PHGDH, and PSAT1 on consecutive spleen sections derived from mouse spleens 8 days after sheep RBC immunization (×5 magnification). (C) Expression of PHGDH and PSAT1 in GC B cells and non-GC B cells harvested from mouse spleen 8 days after immunization with sheep RBC. (D) Representative immunoblots of PHGDH, PSAT1, and PSPH proteins levels in murine resting and activated B cells. (E) Mouse B cells were isolated from spleen and left unstimulated (–) or stimulated (+) with anti-IgM/G antibody, CD40L, and IL-4 for 24 hours before protein extraction and quantification of protein levels normalized to HSC70. Individual samples (dots) and means (bars) values are plotted (n = 4). (F) Relative mRNA expression of SSP enzyme genes in resting and activated mouse B cells as determined by qPCR. Isolated mouse B cells were left unstimulated or stimulated with anti-IgM/G antibody, CD40L, and IL-4 for 24 and 48 hours before mRNA extraction. Specific transcript levels were determined relative to β-actin mRNA levels (n = 4). (G) Mass isotopologue distribution of U-[13C6]-glucose–derived serine and glycine from murine resting and activated murine B cells. Cells were left unstimulated or stimulated with anti-IgM/G antibody, CD40L, and IL-4 for 48 hours. Cells were then cultured for 2 hours in serine/glycine-deplete media containing U-[13C]-glucose. 13C isotopologue distribution in serine and glycine was determined by LC-MS. Data are shown as the mean ± SEM. *P < 0.05, **P < 0.01, and ****P < 0.0001, by Mann-Whitney U test (C, E) or by 1-way ANOVA (F).