NAD+ supplement potentiates tumor-killing function by rescuing defective TUB-mediated NAMPT transcription in tumor-infiltrated T cells

Cell Rep. 2021 Aug 10;36(6):109516. doi: 10.1016/j.celrep.2021.109516.


Although tumor-infiltrating lymphocytes (TILs) maintain their ability to proliferate, persist, and eradicate tumors, they are frequently dysfunctional in situ. By performing both whole-genome CRISPR and metabolic inhibitor screens, we identify that nicotinamide phosphoribosyltransferase (NAMPT) is required for T cell activation. NAMPT is low in TILs, and its expression is controlled by the transcriptional factor Tubby (TUB), whose activity depends on the T cell receptor-phospholipase C gamma (TCR-PLCγ) signaling axis. The intracellular level of NAD+, whose synthesis is dependent on the NAMPT-mediated salvage pathway, is also decreased in TILs. Liquid chromatography-mass spectrometry (LC-MS) and isotopic labeling studies confirm that NAD+ depletion led to suppressed glycolysis, disrupted mitochondrial function, and dampened ATP synthesis. Excitingly, both adoptive CAR-T and anti-PD1 immune checkpoint blockade mouse models demonstrate that NAD+ supplementation enhanced the tumor-killing efficacy of T cells. Collectively, this study reveals that an impaired TCR-TUB-NAMPT-NAD+ axis leads to T cell dysfunction in the tumor microenvironment, and an over-the-counter nutrient supplement of NAD+ could boost T-cell-based immunotherapy.

Keywords: CAR-T; NAD(+) supplement; NAMPT; PD-1; T cell activation; TUB; cancer immunotherapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism*
  • Adoptive Transfer
  • Animals
  • Cell Death / drug effects
  • Cell Line, Tumor
  • Energy Metabolism / drug effects
  • Humans
  • Lymphocyte Activation / drug effects
  • Lymphocytes, Tumor-Infiltrating / drug effects
  • Lymphocytes, Tumor-Infiltrating / immunology*
  • Mice
  • Mice, Inbred NOD
  • NAD / pharmacology*
  • Neoplasms / genetics
  • Neoplasms / immunology*
  • Neoplasms / pathology*
  • Nicotinamide Phosphoribosyltransferase / genetics*
  • Nicotinamide Phosphoribosyltransferase / metabolism
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / immunology*
  • Transcription, Genetic* / drug effects


  • Adaptor Proteins, Signal Transducing
  • TUB protein, human
  • NAD
  • Nicotinamide Phosphoribosyltransferase