# High-content screening for TPI Deficiency therapeutics

> **NIH NIH R01** · UNIVERSITY OF PITTSBURGH AT PITTSBURGH · 2021 · $578,477

## Abstract

Abstract/Project summary:
TPI Df is a devastating untreatable childhood metabolic disease resulting in anemia,
paralysis, irreversible brain damage and premature death. Numerous subtle amino acid
substitutions in Triosephosphate Isomerase (TPI) are pathogenic and result in rapidly
progressing multisystem disease. Importantly, all known pathogenic TPI Df mutations result
in a protein that retains function and pathogenesis is known to result from increased
turnover of the functioning protein by Protein Quality Control pathways (PQC). We have
developed a human cellular TPI Df assay based on a cellular model of the “common” E104D
mutation and implemented it for high-content, high-throughput imaging. We have used this
model in a pilot screen and validated its utility to identify novel compounds that modulate
mutant TPI protein levels in human cells. We have developed the assay to full HTS
standards, and propose to screen the 225,000 member NIH MLSMR compound library to
identify hit-to-lead compounds to develop into TPI Df small molecule therapies. We will
validate hits in secondary assays for TPI stability and activity in TPI Df patient cells, prioritize
them in a panel of in vitro toxicology and metabolism assays, examine structure activity
relationships (SARs) of the lead compounds and substantially validate them in vivo using
established Drosophila and mouse models that reflect the entire range of TPI Df disease
severities. Overall, this project will discover initial therapies for development and test
efficacy of our lead compounds in TPI Df models, including a newly validated mouse model.

## Key facts

- **NIH application ID:** 10312211
- **Project number:** 1R01HD104346-01A1
- **Recipient organization:** UNIVERSITY OF PITTSBURGH AT PITTSBURGH
- **Principal Investigator:** Michael John Palladino
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $578,477
- **Award type:** 1
- **Project period:** 2021-09-01 → 2026-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10312211

## Citation

> US National Institutes of Health, RePORTER application 10312211, High-content screening for TPI Deficiency therapeutics (1R01HD104346-01A1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10312211. Licensed CC0.

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