# Administrative support to R01 HL148104: Understanding Cardiac C-Looping Using Microscale In Vitro Models

> **NIH NIH R01** · RENSSELAER POLYTECHNIC INSTITUTE · 2022 · $67,767

## Abstract

During normal cardiac development of vertebrates, the embryonic heart starts as a straight tube along the
midline of the embryo. It subsequently transforms into a c-shaped heart loop reliably toward the right side of
the body. This cardiac c-looping is the earliest evident event of left-right asymmetry breaking (also called
chirality or handedness) of a human organ. The inversed lateralization (i.e., toward the left side) of cardiac
looping often leads to severe clinical outcomes, including dextrocardia, septum defects, double outlet right
ventricle, and even death of fetuses and infants. In the past, we have been able to recapitulate multicellular
chiral morphogenesis in vitro and shown that cardiac cells from chick hearts right before looping have an
intrinsic rightward bias. Also, protein kinase C activators, newly identified regulators of cardiac cell chirality, can
reverse chick cardiac c-looping. In the funded R01 grant (HL148104), we proposed to investigate how intrinsic
cell chirality determines asymmetric cardiac looping biochemically and biomechanically. We now request a
research supplement to address a Reviewers’ concern (relevance to human diseases) by using human cardiac
cells and investigating the role of the planar cell polarity (PCP; also called noncanonical wnt) signaling, a
pathway known for cardiac laterality defects. We hypothesize that PCP regulates the scale of multicellular
chiral morphogenesis and cardiac c-looping. To test the hypothesis, we propose to evaluate the expression of
PCP proteins and assess the role of PCP proteins in forming 2D in vitro multicellular chiral patterns and ex ovo
looping hearts in chick embryonic development. Specifically, we will first assess the polarity and chirality of
PCP markers in patterned human cardiac cells. We will pattern cardiomyocytes in 2D, examine the location
and morphology of the PCP markers, and determine how PCP signals biomechanically regulate chiral
morphogenesis. Then, we will determine the role of PCP signaling in chick cardiac c looping. With the chick ex
ovo culture, we will evaluate the localization of PCP proteins during the looping stages, interfere with PCP
signaling to examine the effects of PCP, and use cell vertex-based numerical simulation to assess the
biomechanical role of PCP signaling in the asymmetric heart looping. Overall, the proposed research will
delineate the role of PCP signaling in chiral multicellular morphogenesis and in early cardiac asymmetric
looping.

## Key facts

- **NIH application ID:** 10630645
- **Project number:** 3R01HL148104-02S1
- **Recipient organization:** RENSSELAER POLYTECHNIC INSTITUTE
- **Principal Investigator:** Leo Q. Wan
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $67,767
- **Award type:** 3
- **Project period:** 2022-09-01 → 2025-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10630645

## Citation

> US National Institutes of Health, RePORTER application 10630645, Administrative support to R01 HL148104: Understanding Cardiac C-Looping Using Microscale In Vitro Models (3R01HL148104-02S1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10630645. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*