Project Description Narrative:
This project aims to confront the significant health challenge posed by cardiovascular disease (CVD), with a particular focus on dilated cardiomyopathy (DCM), a leading cause of death in Wisconsin. The urgency of targeted research efforts is underscored by the state's high obesity rates, which boosts CVD risks. The study will address a critical gap in understanding DCM’s pathophysiology, especially in cases caused by mutations in a gene called Lamin A/C (LMNA). This form of DCM is known as LMNA-deficiency DCM, and is characterized by a particularly severe disease phenotype. Given the limited effectiveness of existing therapies, this highlights the necessity for an in-depth exploration of underlying molecular mechanisms.
There are no accurate human models to study DCM because of the challenges in maintaining primary heart cells in tissue culture over long periods. Moreover, other models are missing the complex communication between two key cell types in the heart—endothelial cells (EC) and cardiomyocytes (CM)—in a process termed EC-CM communication. This barrier necessitates innovative approaches. Induced pluripotent stem cell (iPSC) are generic patient cells that can be induced to specific cell types, including heart, and offers a promising solution by enabling the generation of patient-specific cardiomyocytes and endothelial cells for DCM research.
However, traditional cell culture techniques are limited to monolayers of cells (i.e., flat cells cultured on plastic). Instead, studying EC-CM communication requires three-dimensional cell cultures termed iPSC-derived cardiac organoids (iPSC-COs). These organoids more faithfully represent the three-dimensional structural changes and aberrant cellular communication observed in the hearts of patients carrying mutation in the LMNA gene (i.e., a more accurate model for studying LMNA-deficiency DCM).
Furthermore, by integrating state-of-the art techniques to manipulate the iPSC DNA (called CRISPR gene editing) coupled with high-throughput screening techniques, the researchers aim to identify novel regulators of EC-CM communication, potentially uncovering new therapeutic targets for LMNA-deficiency DCM.
The overall goal of the project is to establish a brand-new research pipeline at MCW to study DCM. To achieve this, the researchers will take a stepwise approach to accomplish the following aims:
- Establish a state-of-the-art iPSC-CO model and investigate LMNA disease (EC-CM communication) using the iPSC-CO model;
- Validate potential communication pathways through DNA editing and EC-CM coculture; and
- Perform a highly novel screening platform (termed CRISPRi/a) using iPSC-CO to find new disease genes in DCM.
With the successful development of this research pipeline, the researchers will stimulate the study DCM and LMNA mutations at MCW through collaborations with clinicians who will recruit new patients and leverage the MCW iPSC Core. They will also be able to broaden the application of our research pipeline to other forms of CVD beyond LMNA-deficiency DCM.