Biology subjects

Davaapil, H.

Publications and source records attributed to Davaapil, H..

3 recordsLinked to original sources

bioRxiv2023

Differentiation and quality control of smooth muscle cells from human pluripotent stem cells via the neural crest lineage

The Sinha laboratory has developed protocols for differentiating human pluripotent stem cells (hPSCs) into vascular smooth muscle cells along developmental lineage-specific pathways. In development, paraxial mesoderm (PM), lateral plate mesoderm (LM) and neural crest (NC) linages each give rise to smooth muscle cells significant in a location-specific manner. Induced PSCs derived from patients enduring disease provide a platform from which disease-relevant cell models can be established in the laboratory. Here we describe a robust protocol for differentiating hPSCs into vascular smooth muscle cells via a neural crest lineage and the control steps required to ensure consistently high-quality differentiated cells.

cell biology↗

bioRxiv2022

A phenotypic screen of Marfan syndrome iPSC-derived vascular smooth muscle cells uncovers GSK3β as a new target

Marfan syndrome (MFS) is a rare connective tissue disorder caused by mutations in FBN1. Patients with MFS notably suffer from aortic aneurysm and dissection. Despite considerable effort, animal models have proven to be poorly predictive for therapeutic intervention in human aortic disease. Using a "humanised" model system may be more appropriate in identifying new therapeutic targets. Patient-derived induced pluripotent stem cells can be differentiated into vascular smooth muscle cells (VSMCs) and recapitulate major features of MFS. We have screened 1,022 small molecules in our in vitro model, exploiting the highly-proteolytic nature of MFS-VSMCs, and identified 36 effective compounds. Further analysis identified GSK3{beta} as a recurring target in the compound screen. GSK3{beta} inhibition/knockdown did not ameliorate the proliferation defect in MFS-VSMCs but improved MFS-VSMC apoptosis and proteolysis. To conclude, we have identified GSK3{beta} as a novel target for MFS, forming the foundation for future work in MFS and other aortic diseases.

cell biology↗

bioRxiv2021

Regenerative and non-regenerative transcriptional states of the human epicardium: from foetus to adult and backagain

Epicardial activation appears to be required for cardiac regeneration. Although reverting quiescent adult epicardium to an active neonatal or foetal state will likely represent a key therapeutic approach for human cardiac regeneration, the exact molecular differences between human adult and foetal epicardium are not understood. We used single-cell RNA sequencing to compare epicardial cells from both foetal and adult hearts. We found two foetal epicardial cell types, mesothelial and fibroblast-like, with only the mesothelial population present in adults. We also identified foetal-specific epicardial genes associated with regeneration and angiogenesis, and found that adult epicardium may be primed for immune and inflammatory responses. We predict that restoring the foetal epicardial state in human hearts would increase adult angiogenic potential. Finally, we demonstrated that human embryonic stem-cell derived epicardium is a valid model for the foetal epicardium and for investigating epicardial-mediated cardiac regeneration in humans. Our study defines regenerative programs in human foetal epicardium that are absent in the adult, brings human context to animal studies, and provides a roadmap for directing the epicardium in human heart regeneration.

cell biology↗