Functional responses in a patient-specific iPSC-derived vascular model: a novel approach to study migraine

Abstract text (incl. figure legends and references)

Objective Blood vessels from migraine patients are useful for studying migraine pathophysiology and drug development, but are difficult to obtain. Here, we develop a 3D vessel-on-chip model incorporating induced pluripotent stem cell (iPSC)-derived vascular smooth muscle cells (VSMCs) and endothelial cells, allowing to study patient-specific blood vessels. The vascular responses of the cultured blood vessel model are compared to native human blood vessels to validate the model.

Methods In iPSC-derived VSMCs, grown in 2D or in a 3D conformation in a vessel-on-chip model, cAMP responses are measured using the cADDis live cell cAMP assay after stimulation with CGRP and in the presence of phosphodiesterase 3 (PDE3) inhibitors milrinone or cilostazol or the CGRP receptor antagonists rimegepant or olcegepant. Responses to CGRP in the presence of these gepants in iPSC-derived VSMCs were compared to responses in human coronary arteries from heart valve donors (8 F and 7 M, age 48±3 years), as measured in a Mulvany myograph system. Intracellular calcium responses in VSMCs were measured using the calcium dye Cal-520 after stimulation with 10 nM endothelin-1 (ET-1).

Results PDE3 inhibitors milrinone and cilostazol significantly augment the cAMP response to CGRP in iPSC-derived VSMCs (p=0.034 and p=0.002, resp.), while rimegepant inhibits the cAMP response to CGRP, similarly as observed in human isolated arteries. In the 3D cultured blood vessels, olcegepant potently blocked the response to CGRP, while ET-1 potently increased the intracellular calcium concentrations. Comparable results were obtained in human isolated arteries (Labruijere et al. 2013).

Conclusion Functional measurements can be performed in human IPSC-derived VSMCs, in both 2D and 3D conformation, with comparable results in cultured blood vessels and isolated human arteries. These patient-specific vessel-on-chip models could be used in the future to study migraine pathophysiology and improve drug development.