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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

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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 7

Genetic loss and pharmacological inhibition of PHGDH reduces lymphoma progression in vivo.

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Genetic loss and pharmacological inhibition of PHGDH reduces lymphoma pr...
(A) Immunoblot of PHGDH, PSAT1, and PSPH expression in splenic B cells from WT (n = 3) and Eμ-Myc mice (n = 6). HSC70 was used as loading control. (B) Representative IHC staining (×20 magnification) for B220, Ki67, MYC, PHGDH, and PSAT1 abundance in sections of spleens from either WT (n = 3) or Eμ-Myc (n = 3) mice. (C) Isotope tracing analysis in splenic B cells isolated from either C57BL/6J WT mice or Eμ-MYC mice and cultured for 2 hours with 13C6-labeled glucose. Serine and glycine levels were measured by LC-MS. The percent distribution of each isotopologue of their respective metabolite pool is represented as mean ± SEM of triplicate cultures and is representative of 3 independent experiments. (D) Schematic showing lymphoma transplantation model, in which Myc/+;Rosa26-CreERT2/+;Phgdhfl/fl lymphoma cells are injected via the tail vein into 9-week-old male C57BL/6J mice. Three days after lymphoma engraftment, mice were randomized to receive either vehicle or tamoxifen treatment by oral gavage for 4 days. Samples were collected 20 days after injection. (E) Representative pictures of spleens from mice (n = 3 per group) sacrificed 20 days after transplantation (left), and quantification of the spleen weight (right). (F) Schematic showing lymphoma transplantation model, in which Myc/+ lymphoma cells are injected via the tail vein into 9-week-old male C57BL/6J mice. Two days after lymphoma engraftment, mice were randomized to be treated with either vehicle or PH-755 by oral gavage for 14 days. (G) Representative pictures of spleens from mice (n = 3 per group) sacrificed after 7 or 14 days after transplantation (left), and quantification of the spleen weight (right). Data are shown as the mean ± SEM. *P < 0.05 and ***P < 0.001, by 2-tailed Student’s t test (E and G).

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