The metabolism of cells regulates their sensitivity to NK cells depending on p53 status

Sana Belkahla,Abrar Ul Haq Khan,Paul Massa,Alexis Fayd’Herbe De Maudave,Joaquin Marco Brualla,Paolo Falvo,Catherine Alexia,Francesco Bertolini,Stefania Orecchioni,Javier Hernandez,Nerea Allende-Vega,Alberto Anel,Lois Coenon,Wissem Mnif,Martin Villalba,Giulia Mitola,Michael Constantinides

doi:10.1038/s41598-022-07281-6

Abstract

Leukemic cells proliferate faster than non-transformed counterparts. This requires them to change their metabolism to adapt to their high growth. This change can stress cells and facilitate recognition by immune cells such as cytotoxic lymphocytes, which express the activating receptor Natural Killer G2-D (NKG2D). The tumor suppressor gene p53 regulates cell metabolism, but its role in the expression of metabolism-induced ligands, and subsequent recognition by cytotoxic lymphocytes, is unknown. We show here that dichloroacetate (DCA), which induces oxidative phosphorylation (OXPHOS) in tumor cells, induces the expression of such ligands, e.g. MICA/B, ULBP1 and ICAM-I, by a wtp53-dependent mechanism. Mutant or null p53 have the opposite effect. Conversely, DCA sensitizes only wtp53-expressing cells to cytotoxic lymphocytes, i.e. cytotoxic T lymphocytes and NK cells. In xenograft in vivo models, DCA slows down the growth of tumors with low proliferation. Treatment with DCA, monoclonal antibodies and NK cells also decreased tumors with high proliferation. Treatment of patients with DCA, or a biosimilar drug, could be a clinical option to increase the effectiveness of CAR T cell or allogeneic NK cell therapies.

Highlights

Leukemic cells proliferate faster than non-transformed counterparts
The immune system recognizes these signals by the expression of activating receptors in cytotoxic lymphocytes (CL), e.g. the Natural Killer G2-D (NKG2D) activating NK receptor recognizes MHC Class I Polypeptide-Related Sequence A/B (MICA/B) and UL16 Binding Proteins (ULBPs) leading to NK cell a ctivation[3]
We have recently shown that the antidiabetic drug metformin favors recognition of tumor cells by cytotoxic lymphocytes by increasing expression of NKG2DL and mainly of intercellular adhesion molecule-1 (ICAM-1)[7]

Summary

Results

DCA regulates expression of stress ligands in leukemic cells. We used DCA concentrations of 1 and 5 mM, which are on the range of those found in plasma of DCA-treated patients that approach 0.5 mM9,30. Inhibiting the binding of lymphocyte function-associated antigen-1 (LFA-1) to ICAM-I using an anti-β2 integrin monoclonal antibody (mAb) blocked eNK cytotoxicity and DCA sensitization in the case of eCTL (Supplemental Fig. 3) This is in agreement with previous results showing that ICAM-I/LFA-1 interaction is essential for target cell recognition by CL36,37 and correlates with p53-dpendent ICAM-1 induction by DCA. Blocking degranulation with EGTA largely decreased eNK cytotoxicity (Supplemental Fig. 4B) These results suggest that DCA made tumor cells more sensitive to CLs by multiple mechanisms, including stress ligand, ICAM-1 and DR5 upregulation. These cells behave as cell lines lacking wt p53 expression or expressing a mut p53 (Fig. 1), rather reducing stress ligand expression This correlated with a lack of increase in MICA/B and ULBP1 proteins at the plasma membrane after DCA treatment (Fig. 4B). The antiCD20 mAb totally abrogated the growth of these cells (data not shown), making irrelevant the cotreatment with DCA and/or eNK

Discussion

B Survival after BCL-P2 cell engraftement

Materials and methods