# Characterization of TMEM164 as novel multi-pass transmembrane enzyme and its role in ferroptosis

> **NIH NIH F32** · SCRIPPS RESEARCH INSTITUTE, THE · 2024 · $76,756

## Abstract

Project Summary/Abstract
The goal of this proposal is to understand how transmembrane protein 164 (TMEM164) attenuates
polyunsaturated fatty acid (PUFA) metabolism and ferroptotic signaling. Ferroptosis is a novel iron-dependent
form of cell death implicated in a broad range of human diseases, including cancer and type 2 diabetes mellitus
(T2DM). In T2DM, the severity of diabetic state increases risk for renal tubular injury related to iron overload and
lipid peroxidation, two key features of ferroptosis. Characterizing enzymes that regulate the PUFA content of
human cells would not only deepen our understanding of ferroptosis, but also identify therapeutic targets for
treating diseases like T2DM where this form of cell death is dysregulated. Recent genome editing screens have
identified TMEM164, a multi-pass transmembrane protein of uncharacterized function, as a key regulator of
ferroptosis. We hypothesize, based on its Alphafold-predicted structure, that TMEM164 is a novel type of
cysteine-dependent transmembrane enzyme that regulates cellular PUFA content. Here we seek to test this
hypothesis and identify the specific enzymatic functions performed by TMEM164 in PUFA lipid metabolism, as
well as their contribution to conferring ferroptosis sensitivity to human cells. In Aim 1, I will determine the lipid
profiles of cells genetically deleted for TMEM164 and test whether wild type, but not a putative catalytic cysteine
mutant form of TMEM164 can rescue these lipid perturbations. From these data, I will define candidate
physiological substrates for TMEM164, which will be tested in recombinant protein systems. In Aim 2, I will
leverage our lab’s longstanding expertise in covalent inhibitor discovery to identify electrophilic compounds that
block TMEM164 activity through modifying the putative catalytic cysteine of the protein. Successful completion
of this project will broaden our fundamental understanding of the lipid metabolic pathways involved in ferroptosis
and identify new candidate drug targets for suppressing the contribution of ferroptosis to human degenerative
diseases like T2DM.

## Key facts

- **NIH application ID:** 10884159
- **Project number:** 5F32DK135331-02
- **Recipient organization:** SCRIPPS RESEARCH INSTITUTE, THE
- **Principal Investigator:** Timothy Brandon Ware
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $76,756
- **Award type:** 5
- **Project period:** 2023-07-01 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10884159

## Citation

> US National Institutes of Health, RePORTER application 10884159, Characterization of TMEM164 as novel multi-pass transmembrane enzyme and its role in ferroptosis (5F32DK135331-02). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10884159. Licensed CC0.

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