# Investigations of proteome turnover kinetics under cellular differentiation

> **NIH NIH R35** · UNIVERSITY OF COLORADO DENVER · 2023 · $11,338

## Abstract

PROJECT SUMMARY
This administrative supplement application for undergraduate summer research is being submitted to PA-20-272
in accordance to NOT-GM-22-009. The scope of the parent project remains unchanged. Summary of Parent
Project R35-GM146815: Proteins are in a constant flux of continuous synthesis and degradation. Both
components of the protein turnover cycle contribute to protein abundance. Although this fact has been known
for 80 years, currently most biological inquiries are limited to static snapshots of overall transcript and protein
levels, whereas knowledge into the dynamic changes of protein turnover remains severely lagging. This ESI
MIRA proposal seeks to advance the current understanding of how cellular proteomes remodel during cell state
transitions, by incorporating protein turnover kinetics information at key stages of human induced pluripotent
stem cell (iPSC) differentiation into distinct cellular lineages. Working with collaborators, we previously developed
deuterium stable isotope labeling, mass spectrometry, and kinetic modeling methods to quantitate protein
turnover in animal models and in human. In doing so, we found many novel cell states and disease markers may
be discovered from integrating orthogonal protein abundance and kinetics information. In the next five years, we
propose to: (1) apply these methods to acquire a high temporal density map of human iPSC trilineage
differentiation into cardiomyocyte, hepatocyte, and neuroprogenitor cells; (2) interrogate the regulatory principles
that govern turnover flux across different differentiation stages; and (3) assess the functional consequences of
protein degradation on the success and cell maturity of terminal cell production. Finally, a limitation of current
techniques is that the kinetic models used in protein turnover studies largely assume a non-changing protein
pool size at steady state, which does not apply to differentiating cells or progressing diseases. We propose to
expand current models of analyzing heavy water stable isotope label experiments toward dynamical systems
with variable protein pool sizes. If successful, the proposed research would greatly expand the current knowledge
of molecular events that take place during human iPSC differentiation, as well as generate publicly available
data sets and software tools to advance protein turnover studies in diverse areas.

## Key facts

- **NIH application ID:** 10808331
- **Project number:** 3R35GM146815-01S1
- **Recipient organization:** UNIVERSITY OF COLORADO DENVER
- **Principal Investigator:** Edward Lau
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $11,338
- **Award type:** 3
- **Project period:** 2022-09-17 → 2027-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10808331

## Citation

> US National Institutes of Health, RePORTER application 10808331, Investigations of proteome turnover kinetics under cellular differentiation (3R35GM146815-01S1). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10808331. Licensed CC0.

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