PHGDH is required for germinal center formation and is a therapeutic target in MYC-driven lymphoma
Annalisa D’Avola, … , Karen H. Vousden, John C. Riches
Annalisa D’Avola, … , Karen H. Vousden, John C. Riches
Published March 22, 2022
Citation Information: J Clin Invest. 2022;132(9):e153436. https://doi.org/10.1172/JCI153436.
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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 4

PHGDH inhibition impairs B cell proliferation and de novo serine and glycine synthesis.

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PHGDH inhibition impairs B cell proliferation and de novo serine and gly...
(A) Mass isotopologue distribution of U-[13C6]-glucose–derived serine and glycine in B220+ B cells isolated from Phgdh+/+;Cd19-Cre (WT) or Phgdhfl/fl;Cd19-Cre (F/F) or WT C57BL/6J mice. Isolated cells were stimulated with anti-IgM/G antibody, CD40L, and IL-4 for 48 hours and were then cultured for 2 hours in serine/glycine-deplete media with U-[13C]-glucose (and treated with/without PH-755 for WT B cells). (B and C) Representative proliferation profiles and quantification of B220+ B cells from either Phgdh+/+;Cd19-Cre (WT) or Phgdhfl/fl;Cd19-Cre (F/F) mice. Cells were labeled with the Cell Proliferation Dye eFluor 670 and then cultured for 3 days with anti-IgM/G antibody, CD40L, and IL-4 in complete media, serine/glycine-free medium, or serine/glycine-free medium containing glycine and formate (n = 4 per genotype). (D) Cell-cycle analysis of B220+ cells isolated from Phgdh+/+;Cd19-Cre (WT) or Phgdhfl/fl;Cd19-Cre (F/F) mice. Cells were cultured for 48 hours with anti-IgM/G antibody, CD40L, and IL-4 in complete media, serine/glycine-free medium, or serine/glycine-free medium containing glycine and formate; they then underwent BrdU labeling and 7-AAD staining to assess cell cycle. (E and F) Representative proliferation profiles and quantification of B220+ B cells from WT mice. Cells were labeled with the Cell Proliferation Dye eFluor 670 and were then cultured for 3 days with anti-IgM/G antibody, CD40L, and IL-4 in complete media, serine/glycine-free medium, or serine/glycine-free medium containing glycine and formate in combination with DMSO or 10 μM PH-755 (n = 5 per group). (G) Cell-cycle analysis of B220+ B from WT mice. Cells were cultured for 48 hours with anti-IgM/G, CD40L, and IL-4 in complete medium, serine/glycine-free medium, or serine/glycine-free medium containing glycine and formate in combination with DMSO or 10 μM PH-755 before cell-cycle analysis. (H) Activate Caspase-3 on B220+ B cells from either Phgdh+/+;Cd19-Cre (WT) or Phgdhfl/fl;Cd19-Cre (left panel; n = 4 per group) and treated as described in D, and on B cells isolated from WT mice (right panel; n = 3 per group) and treated as described in G. Data are shown as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001, by 1-way ANOVA (C and F).