# Investigations of proteome turnover kinetics under cellular differentiation

> **NIH NIH R35** · UNIVERSITY OF COLORADO DENVER · 2022 · $388,750

## Abstract

PROJECT SUMMARY
Investigations of Proteome Turnover Kinetics Under Cellular Differentiation: Proteins are in a constant flux
of continuous synthesis and degradation, both of which contribute to regulate protein levels in the cell. 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:** 10499189
- **Project number:** 1R35GM146815-01
- **Recipient organization:** UNIVERSITY OF COLORADO DENVER
- **Principal Investigator:** Edward Lau
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $388,750
- **Award type:** 1
- **Project period:** 2022-09-17 → 2027-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10499189

## Citation

> US National Institutes of Health, RePORTER application 10499189, Investigations of proteome turnover kinetics under cellular differentiation (1R35GM146815-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10499189. Licensed CC0.

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