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Researchers from the group of reNEW Associate Investigator Valeria Orlova developed a 3D Vessel-on-Chip model to study CADASIL*, a rare inherited disease affecting the brain’s small vessels. The model robustly recapitulates key disease features in a dish, provides new insight into the underlying mechanism and offers a promising platform for testing potential therapies.
CADASIL is caused by mutations in NOTCH3, and no approved treatments are currently available. Patients face an increased risk of stroke in mid-adulthood and vascular dementia, while research into the disease has long been hampered by the lack of physiologically relevant human model systems.
Dr. Marc Vila Cuenca, a senior researcher in Orlova’s group and first author of the study explains: “Our vessels are complex, multicellular, three-dimensional structures. Many laboratory blood vessel models rely on simple 2D cell cultures, which do not capture this complexity. Given these models don’t faithfully recapitulate the human vasculature and the limited availability of human tissue samples, there is a need for alternative, more predictive laboratory model systems that can be used to study vascular diseases such as CADASIL.”
A 3D Vessel-on-Chip model of CADASIL
In this study, published in Stem Cell Reports, the reNEW team therefore built on a Vessel-on-Chip (VoC) model that was previously developed in the group to study CADASIL. These models consist of two important vascular cell types – vascular smooth muscle cells (VSMCs) and endothelial cells (ECs) – that self-assemble into perfusable 3D structures. Because CADASIL-associated NOTCH3 mutations primarily affect VSMCs, the researchers used the model to compare healthy and diseased VSMCs in a physiologically relevant 3D environment. The team found that CADASIL VSMCs showed altered protein expression and changes in cell shape, accompanied by functional abnormalities.
Dr. Vila Cuenca additionally notes: “Strikingly, these differences were absent in simple 2D cultures. This highlights the importance of complex model systems such as our VoC to study CADASIL and other vascular diseases.”
Stem cells
The researchers initially used VSMCs derived from both patients and healthy donors. However, such tissue samples are rare, and finding well-matched controls for comparison is challenging. Human induced pluripotent stem cells (hiPSCs), by contrast, offer a virtually unlimited and highly versatile source of cellular material. The team therefore generated VSMCs from CADASIL hiPSCs and compared their behavior with that of VSMCs derived from primary tissue.
Encouragingly, the stem cell–derived cells behaved very similarly, showing many of the same CADASIL-related features. This means they can be reliably used in future studies and opens up new possibilities, such as working with perfectly matched (isogenic) controls and applying targeted genetic modifications.
From insights to impact
The validated platform cannot only be used to learn more about the early mechanisms that drive CADASIL, but also as a tool to develop new therapies. As a proof-of-concept, Dr. Vila Cuenca and colleagues successfully reversed CADASIL-related changes in their VoC models through treatment with a drug that targets NOTCH3 protein breakdown.
Dr. Orlova concludes: “Our platform brings exciting and much-needed opportunities to learn more about the early mechanisms that drive CADASIL and develop new drugs. Our hope is this will ultimately contribute to new therapies for patients in need.”
*CADASIL = Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy.
