# A Multi-Omics Approach to Discover Metabolic Critical Quality Attributes for Cardiomyocyte Biomanufacturing

> **NIH NIH R01** · UNIVERSITY OF WISCONSIN-MADISON · 2021 · $375,430

## Abstract

PROJECT SUMMARY
Human induced pluripotent stem cells (iPSCs) provide a promising source of patient-specific cardiac cells. Our
team has pioneered development of protocols to differentiate iPSCs to cardiomyocytes (iPSC-CMs) but these
cells lack mature, adult-like phenotypes. One hallmark of CM maturation is a transition from glycolysis and
glucose oxidation to fatty acid oxidation as the dominant metabolic pathway, among other metabolic changes.
Our premise is that identifying metabolic critical quality attributes (CQAs) of maturity will provide fundamental
insight into phenotypic maturation of iPSC-CMs, new tools to facilitate discovery of effective strategies to mature
iPSC-CMs, and novel technologies to monitor maturation state during iPSC-CM biomanufacturing. To achieve
this premise, we will employ an integrative quantitative metabolomics and proteomics approach to profile
metabolite and metabolic enzyme concentrations, and metabolic pathway utilization, in iPSC-CMs undergoing
maturation by extended time in culture or biochemical/biomechanical stimulation. Comparing metabolic
transitions during in vitro iPSC-CM maturation to metabolic transitions during development in vivo and to
acquisition of maturation phenotypes will allow us to map metabolic transitions to developmental processes. We
will perform multivariate data analyses to predict metabolic CQAs of maturation phenotypes and build novel tools
to monitor these CQAs during iPSC-CM biomanufacturing. Thus, the proposed study will provide fundamental
new insight into metabolic pathway utilization during iPSC-CM maturation and cardiac development, and will
predict metabolic CQAs that will facilitate monitoring progression of maturation in iPSC-CMs during
biomanufacturing. Our specific aims are:
1. Profile metabolic transitions during iPSC-CM differentiation and maturation. At different iPSC-CM
 maturation stages induced by extended culture, micropatterned substrates, carbon source availability, and
 electromechanical stimulation, we will quantify metabolite and protein concentrations via metabolomics and
 proteomics and targeted metabolic assays. We will assess maturation via molecular and functional assays
 to relate changes in metabolites and metabolic pathway utilization to acquisition of maturation phenotypes.
2. Assess metabolic pathway enrichment in developing murine cardiomyocytes. We will use
 metabolomics and proteomics to profile metabolite and protein concentrations in murine CMs at different
 developmental stages and compare to metabolic transitions that occur during iPSC-CM maturation.
3. Identify metabolic CQAs and develop tools for assessment of iPSC-CM maturity during
 biomanufacturing. We will use multivariate analysis to predict metabolic CQAs that identify maturation state
 of iPSC-CMs. We will then develop assays to monitor these CQAs (combinations of metabolite and metabolic
 enzymes) during biomanufacturing via targeted metabolomics/proteomics, and by use of CRISPR-Cas9
...

## Key facts

- **NIH application ID:** 10218267
- **Project number:** 5R01HL148059-03
- **Recipient organization:** UNIVERSITY OF WISCONSIN-MADISON
- **Principal Investigator:** Sean P Palecek
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $375,430
- **Award type:** 5
- **Project period:** 2019-07-01 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10218267

## Citation

> US National Institutes of Health, RePORTER application 10218267, A Multi-Omics Approach to Discover Metabolic Critical Quality Attributes for Cardiomyocyte Biomanufacturing (5R01HL148059-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10218267. Licensed CC0.

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