5-Aminolevulinic acid/sodium ferrous citrate enhanced the antitumor effects of programmed cell death-ligand 1 blockade by regulation of exhausted T cell metabolism in a melanoma model

Cancer Sci. 2021 Jul;112(7):2652-2663. doi: 10.1111/cas.14930. Epub 2021 May 22.

Abstract

Mitochondria are key cytoplasmic organelles. Their activation is critical for the generation of T cell proliferation and cytotoxicity. Exhausted tumor-infiltrating T cells show a decreased mitochondrial function and mass. 5-Aminolevulinic acid (5-ALA), a natural amino acid that is only produced in the mitochondria, has been shown to influence metabolic functions. We hypothesized that 5-ALA with sodium ferrous citrate (SFC) might provide metabolic support for tumor-infiltrating T cells. In a mouse melanoma model, we found that 5-ALA/SFC with a programmed cell death-ligand 1 (PD-L1) blocking Ab synergized tumor regression. After treatment with 5-ALA/SFC and anti-PD-L1 Ab, tumor infiltrating lymphocytes (TILs) were not only competent for the production of cytolytic particles and cytokines (granzyme B, interleukin-2, and γ-interferon) but also showed enhanced Ki-67 activity (a proliferation marker). The number of activated T cells (PD-1+ Tim-3- ) was also significantly increased. Furthermore, we found that 5-ALA/SFC activated the mitochondrial functions, including the oxygen consumption rate, ATP level, and complex V expression. The mRNA levels of Nrf-2, HO-1, Sirt-1, and PGC-1α and the protein levels of Sirt-1 were upregulated by treatment with 5-ALA/SFC. Taken together, our findings revealed that 5-ALA/SFC could be a key metabolic regulator in exhausted T cell metabolism and suggested that 5-ALA/SFC might synergize with anti-PD-1/PD-L1 therapy to boost the intratumoral efficacy of tumor-specific T cells. Our study not only revealed a new aspect of immune metabolism, but also paved the way to develop a strategy for combined anti-PD-1/PD-L1 cancer immunotherapy.

Keywords: PD-1/PD-L1 blockade; cancer immunotherapy; exhausted T cell; mitochondrial function; tumor infiltrating lymphocyte.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Aminolevulinic Acid / pharmacology*
  • Animals
  • B7-H1 Antigen / antagonists & inhibitors*
  • Cell Line, Tumor
  • Citric Acid / pharmacology*
  • Combined Modality Therapy
  • Female
  • Ferrous Compounds / pharmacology*
  • Heme Oxygenase-1 / metabolism
  • Hepatitis A Virus Cellular Receptor 2 / metabolism
  • Ki-67 Antigen / metabolism
  • Lymphocyte Activation / drug effects*
  • Lymphocyte Count
  • Lymphocytes, Tumor-Infiltrating / cytology
  • Lymphocytes, Tumor-Infiltrating / drug effects*
  • Lymphocytes, Tumor-Infiltrating / metabolism
  • Melanoma, Experimental / immunology
  • Melanoma, Experimental / metabolism
  • Melanoma, Experimental / pathology
  • Melanoma, Experimental / therapy
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mitochondria / drug effects*
  • Mitochondria / metabolism
  • Nuclear Respiratory Factor 1 / metabolism
  • Oxygen Consumption / drug effects
  • Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha / metabolism
  • Sirtuin 1 / metabolism

Substances

  • B7-H1 Antigen
  • Ferrous Compounds
  • HAVCR2 protein, human
  • Hepatitis A Virus Cellular Receptor 2
  • Ki-67 Antigen
  • Membrane Proteins
  • Nrf1 protein, mouse
  • Nuclear Respiratory Factor 1
  • Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha
  • Ppargc1a protein, mouse
  • Citric Acid
  • ferrous citrate
  • Aminolevulinic Acid
  • Adenosine Triphosphate
  • Heme Oxygenase-1
  • Hmox1 protein, mouse
  • Sirt1 protein, mouse
  • Sirtuin 1