# Study of mitochondria-depleted pluripotent stem cells in  development and disease

> **NIH NIH F31** · UT SOUTHWESTERN MEDICAL CENTER · 2022 · $37,674

## Abstract

PROJECT SUMMARY/ABSTRACT
Mitochondria are critical for the healthy development of an individual. When dysfunctional, mitochondria give rise
to a devastating group of developmental disorders that include several myopathies and neuropathies and affect
an estimated 1:5,000 people. Many of these disorders are caused by mutations in mitochondrial DNA (mtDNA),
and despite decades of research, many mtDNA diseases lack effective treatments. We have adapted a system
of mitochondria-depletion to generate pluripotent stem cells (PSCs) that lack detectable levels of mitochondria
(henceforth referred to as mito-depleted PSCs). We have found that mito-depleted PSCs survive in culture for
several days and, moreover, by fusing healthy PSCs with mito-depleted PSCs, we were successful in restoring
mitochondria to mito-depleted PSCs. These findings suggest mito-depleted PSCs provide a unique platform to
uncover and study mechanisms by which mitochondria regulate pluripotency and early development. In addition,
they may constitute an exciting biotechnology that can be harnessed in a potentially universal strategy for
replacing mutant mitochondria with healthy mitochondria in mtDNA disease patient cells. This project aims to
utilize the exciting and unstudied platform of mito-depleted PSCs to study mitochondrial roles in early
development, as well as devise a new strategy to treat mtDNA disease. We will accomplish this by first
determining the effects mito-depletion on mouse embryonic stem cells (mESCs). Using both targeted and
unbiased methods, we will determine the overarching role of mitochondria in the pluripotent state and early
embryonic development. Specifically, we will determine changes in the transcriptional, epigenetic, and metabolic
states of mito-depleted mESCs. In addition, we will model the developmental ability of mito-depleted mESCs in
vitro through pluripotent state changes and differentiation. We will also generate mtDNA-corrected hiPSCs
reprogrammed from Mitochondrial Encephalopathy, Lactic acidosis, and Stroke-like episodes (MELAS)
syndrome patients via mito-depletion and subsequent cytoplast fusion. We will determine reversal of metabolic
and developmental phenotypes associated with MELAS through directed differentiation to neural and cardiac
lineages, combined with metabolic Seahorse assays. Deciphering the biological mechanisms by which
mitochondria regulate development and disease are of fundamental importance to advancing human health and
developing innovative therapeutic strategies to treat mtDNA disease like MELAS syndrome.

## Key facts

- **NIH application ID:** 10535808
- **Project number:** 1F31NS125906-01A1
- **Recipient organization:** UT SOUTHWESTERN MEDICAL CENTER
- **Principal Investigator:** Daniel Andrew Schmitz
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $37,674
- **Award type:** 1
- **Project period:** 2022-09-01 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10535808

## Citation

> US National Institutes of Health, RePORTER application 10535808, Study of mitochondria-depleted pluripotent stem cells in  development and disease (1F31NS125906-01A1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10535808. Licensed CC0.

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