Session: 16-01-01: Government Agency Student Poster Competition

Paper Number: 149897

149897 - Studying Flow Regulated Pathogenesis of Hypoplastic Left Heart Syndrome in a Three-Dimensional Bioprinted Model of Developing Human Heart 

Introduction: Hypoplastic left heart syndrome (HLHS) is a severe spectrum of congenital heart defect (CHD) marked by underdevelopment of the left heart. Despite the prevalent “no flow, no grow” theory suggesting hemodynamic perturbations as a potential cause of HLHS, molecular mechanisms underlying the onset of HLHS and its progression remain obscure, due to suboptimal experimental models. Recently, we established a human induced pluripotent stem cell (hiPSC)-based 3D bioprinted model of embryonic heart tube that shows recapitulation of robust cardiac function and cellular activities related to early cardiogenesis. In this research, the role of flow hemodynamics in HLHS pathogenesis is investigated via the incorporation of HLHS-derived hiPSCs and flow modulation in the 3D model of human heart tube.

 

Significance and Innovation: 3D bioprinting allows for precise spatial control of biomaterials and cells and incorporation of vasculature. Customized perfusion bioreactor systems facilitate accurate tuning of flow hemodynamics in 3D cultures. Further, hiPSCs provide a patient-specific supply of HLHS-derived cardiac cells. Single cell RNA sequencing (scRNA-seq) offers in-depth analyses of the complex cell heterogeneities and gene expression profiles. By integrating these technologies, our 3D culture platform enables faithful recapitulation of HLHS pathogenesis in terms of dynamic cell-cell and cell-microenvironment crosstalk in vitro. This research can provide new insights into cellular mechanisms underlying human heart development and HLHS. Moreover, it demonstrates the versatility and applicability of this 3D in vitro platform for personalized high-throughput modeling of diseases, interventional procedures, and drug screening.

 

Methods: To examine the role of hemodynamic alterations in HLHS pathogenesis, we constructed 3D bioprinted heart tubes using HLHS hiPSC-derived cardiomyocytes (CMs) and endothelial cells (ECs). Physiological vs. reduced flow was perfused through the cellular heart tube constructs for 5 days. Flow patterns and hemodynamics were analyzed using computational fluid dynamics (CFD) modeling and validated by laser particle imaging velocimetry (PIV). Tissue structure and cell-specific markers were examined by immunohistochemistry. scRNA-seq was performed on the cells harvested from heart tubes under different flow conditions, as well as 2D monocultures (control). Region-specific transcriptome was analyzed in correlation with local wall shear stress (WSS) distributions.

 

Preliminary Results: HLHS iPSC-derived heart tubes under both physiological vs. reduced flow conditions showed long-term viability and cardiac function. Endothelium retention, cardiac tissue compaction, and remodeling were presented over time. Significantly different velocity and WSS profiles were detected within the printed heart tube. Distinct transcriptomes of CMs and ECs related to cardiogenesis at the linear heart tube stage and HLHS-related genes were identified in the presence of reduced flow.

 

Conclusions: This study highlights the association of flow disturbance with HLHS pathogenesis during heart development, teasing out the role of microenvironment as a contributor and potential target for prenatal diagnostics and therapeutics.

Presenting Author: Linqi Jin Emory University & Georgia Institute of Technology

Presenting Author Biography: Linqi received her BSc in Bioengineering from Southwest Jiaotong University (2015-2019) and MSc in Biomedical Engineering from Georgia Institute of Technology (2019-2021). As an AHA Predoctoral Fellow, she is now doing her Ph.D. research at The Wallace H. Coulter Department of Biomedical Engineering at Georgia Tech and Emory University. Linqi's Ph.D. project focuses on 3D in vitro human heart development and disease modeling stands at the intersection of innovation and practical application. Linqi is dedicated to leveraging her educational background and skillset in 3D bioprinting, cardiac tissue engineering, and single-cell sequencing, to contribute to transformative advances in the understanding and treatment of congenital heart defects.

Authors:

Linqi Jin Emory University & Georgia Institute of Technology
Sunder Neelakantan Texas A&M University
Shweta Karnik Georgia Institute of Technology
Arnab Dey Georgia Institute of Technology
Reza Avazmohammadi Texas A&M University
Lakshmi Dasi Georgia Institute of Technology
Holly Bauser-Heaton Emory University
Vahid Serpooshan Emory University & Georgia Institute of Technology