# Top-Down Proteomics of Myofilaments in Heart Failure

> **NIH NIH R01** · UNIVERSITY OF WISCONSIN-MADISON · 2020 · $372,960

## Abstract

Project Summary
Hypertrophic cardiomyopathy (HCM) is a highly prevalent hereditary cardiac disease with over 1400 mutations
identified in 11 genes encoding the protein constituents of sarcomere, the basic contractile units of muscle.
However, the molecular mechanisms by which mutations in sarcomeric proteins lead to cardiac hypertrophy and
failure remain poorly understood. Sarcomeres are composed of myofilaments flanked by dense protein
structures called Z-discs, playing critical roles in controlling contraction and mediating signaling pathways.
Moreover, recent evidence has suggested that sarcomeric protein post-translational modifications (PTMs) play
essential roles in the modulation of contractile function. Our preliminary data convincingly show that the PTMs
of sarcomeric proteins, including multiple key myofilament and Z-disc proteins, are altered in the myocardium of
HCM patients as compared to donor hearts. Changes in the expression of myofilament protein isoforms were
also detected in HCM patient myocardium. Intriguingly, we have observed consistent alterations in sarcomeric
protein PTMs and isoform expression regardless of the mutation types. Thus, we hypothesize that different HCM-
causing mutations activate convergent hypertrophic signaling pathways, yielding comparable alterations in PTMs
and protein isoforms of the sarcomeric proteins that cause abnormal contractile function and pathological
hypertrophy. To test our hypothesis, we will employ a highly interdisciplinary approach featuring top-down
proteomics, induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs), cardiac tissue engineering,
human patient heart tissue samples, and functional studies to delineate the underlying mechanisms of HCM.
Our recently established top-down proteomics platform represents a comprehensive tool to detect and
characterize HCM-related changes in the sarcomeric proteome with high reproducibility and throughput.
Recently, we have made a major methodologic breakthrough that allows us to perform top-down proteomic
analysis of minimal amounts of biopsy tissue, as well as iPSC-CMs cultured in either a 2D monolayer or 3D
engineered cardiac tissue (ECT). In addition, biochemical analyses will be performed to assess changes in
intracellular signaling pathways. Furthermore, we will perform functional measurements to relate sarcomeric
protein PTM alterations and maladaptive signaling to contractile dysfunction. We aim to determine: 1) the
changes in sarcomeric protein PTMs, key hypertrophic signaling pathways, and mechanical properties in
myocardial tissue from HCM patients; 2) the early molecular alterations and functional consequences of different
sarcomeric protein mutations in HCM patient-specific iPSC-CMs; 3) the specific molecular alterations and
functional consequences in iPSC-CM- ECT from multiple HCM patients carrying the same mutation. This
application, enabled by cutting-edge technology, is highly significant with a strong scientific premise ...

## Key facts

- **NIH application ID:** 9838772
- **Project number:** 5R01HL096971-08
- **Recipient organization:** UNIVERSITY OF WISCONSIN-MADISON
- **Principal Investigator:** Ying Ge
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $372,960
- **Award type:** 5
- **Project period:** 2011-08-05 → 2021-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9838772

## Citation

> US National Institutes of Health, RePORTER application 9838772, Top-Down Proteomics of Myofilaments in Heart Failure (5R01HL096971-08). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/9838772. Licensed CC0.

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