# Exploiting metabolic vulnerabilities of CD4 T cell subsets to control inflammatory disease

> **NIH NIH R01** · VANDERBILT UNIVERSITY MEDICAL CENTER · 2024 · $390,666

## Abstract

SUMMARY
Inflammatory diseases are often driven by inappropriate responses of effector CD4 T cells (Teff). IL17, IFNγ, or
dual-producing polyfunctional effector Th1, Th17, or Th17.1 T cells can become imbalanced with suppressive
Treg CD4 T cells in a variety of disease settings, including inflammatory bowel diseases (IBD). A key therapeutic
objective in efforts to shift the immunologic balance towards tolerance, therefore, is to selectively inhibit Teff and
promote Treg. We have shown that Teff and Treg subsets utilize different metabolic programs that represent
fundamental features of T cell biology. Here we explore one carbon (1C) metabolism and related
microenvironmental factors to modulate CD4 T cells in inflammatory diseases. 1C metabolism integrates multiple
nutrient inputs to provide intermediates for de novo methionine and purine synthesis and is commonly targeted
with anti-folate drugs. An in vivo CRISPR screen of primary T cells in IBD with a 1C metabolism enzyme-focused
gRNA library identified the mitochondrial enzyme Methylene-tetrahydrofolate Dehydrogenase 2 (MTHFD2) as
conditionally essential for effector T cell proliferation and inflammation. MTHFD2 was upregulated in T cells a
variety of inflammatory conditions and while MTHFD2-deficiency impaired Teff, MTHFD-deficient Treg had
increased FoxP3 expression in both mouse and human T cells. Consistent with a role as a metabolic checkpoint
on inflammation, MTHFD2-deficiency protected against IBD and other inflammatory diseases. Mechanistically,
MTHFD2 inhibition suppressed mTORC1 activity, possibly through reduced methionine and/or interrupted purine
synthesis. Importantly, local nutrients play key roles in 1C metabolism and T cell fate. To quantify T cell access
to nutrients in vivo, we established Positron Emission Tomography (PET) tracer-based methods to directly image
and measure nutrient uptake in vivo. These studies showed a sharp increase in T cell glucose uptake in
inflammation. In addition, IBD is often associated with folate-deficiency. The effects of dietary folate on T cell
1C metabolism, mTORC1 signaling, and fate, however, are unclear. Because inflammation is associated with
fevers and enzymes are temperature-dependent, we also tested fever conditions on T cell metabolism. We found
fever led to increased cytokine production from Teff and mitochondrial Reactive Oxygen Species (ROS)
specifically in Th1 cells that, surprisingly, led to Tp53-dependent apoptosis. These findings support the
hypothesis that 1C metabolism is limiting and serves as a metabolic checkpoint to integrate local nutrients and
physical conditions through MTHFD2 and mTORC1 signaling to provide new immunometabolic targets to
modulate effector and regulatory T cells. To test this, we will: (1) Test the role and mechanism of MTHFD2 as a
limiting enzyme in methionine and nucleotide synthesis essential for mTORC1 signaling and effector T cells; and
(2) Determine how nutrient and microenvironmental factors such ...

## Key facts

- **NIH application ID:** 10795080
- **Project number:** 5R01DK105550-12
- **Recipient organization:** VANDERBILT UNIVERSITY MEDICAL CENTER
- **Principal Investigator:** Jeffrey C Rathmell
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $390,666
- **Award type:** 5
- **Project period:** 2015-04-01 → 2027-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10795080

## Citation

> US National Institutes of Health, RePORTER application 10795080, Exploiting metabolic vulnerabilities of CD4 T cell subsets to control inflammatory disease (5R01DK105550-12). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10795080. Licensed CC0.

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