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 3

Genetic loss and pharmacological inhibition of PHGDH impairs GC responses.

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Genetic loss and pharmacological inhibition of PHGDH impairs GC response...
(A) Mice were injected with sheep RBCs and spleens examined by IHC and flow cytometry 8 days after immunization. (B) Representative flow cytometric analysis of splenic B cells from Phgdh+/+;Cd19-Cre (WT) or Phgdhfl/fl;Cd19-Cre (F/F) before and after immunization to identify GC B cells (CD19+B220+CD38loCD95hi), as well as DZ (CD86loCXCR4hi) and LZ (CD86hiCXCR4lo) B cells, within GC splenic population. (C) Flow cytometric analysis of absolute numbers of B cell subsets within CD19+B220+CD38loCD95hi splenic population from Phgdhfl/fl;Cd19-Cre (n = 6) and Phgdh+/+;Cd19-Cre (n = 4) after immunization. (D) Average GC area (left) and proportion (%) of GC area per spleen area (right) from Phgdhfl/fl;Cd19-Cre (F/F) and Phgdh+/+;Cd19-Cre (WT) mice after immunization. (E) WT mice were immunized with sheep RBCs 1 day before PH-755 treatment (300 mg/kg PH-755 orally twice daily for 7 days). Spleens were analyzed 8 days after immunization. (F) Representative flow cytometric analysis of splenic B cells from mice immunized with sheep RBCs and treated with vehicle/PH-755 to identify GC, DZ, and LZ B cells within GC splenic population. (G) Flow cytometric analysis of absolute number of GC, DZ, and LZ B cells within B220+ splenocytes collected from mice 8 days after sheep RBCs immunization; mice were treated with vehicle (n = 5) or PH-755 (n = 5). (H) Average GCs area (left) and proportion (%) of GC area per spleen area (right) from vehicle- and PH-755–treated mice 8 days after sheep RBC immunization. (I) Summary of NP2-specific plasma cells (PCs; left) and NP2-specific IgG1 PCs (right; total number per popliteal lymph nodes). WT mice were treated with either vehicle (n = 5) or PH-755 (n = 5) for 7 days. Animals were injected with NP-CGG 1 day before PH-755 treatment. Popliteal lymph nodes were collected 8 days after NP-CGG immunization. (J) Serum antibody titers for NP2-specific IgG1 8 days after NP-CGG immunization. Data are shown as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, by Mann-Whitney U test (C, G, I, and J) or unpaired t test (D and H).