Projects

Developing new genetic tools for high-throughput single-cell functional genetics

Based on our recent expertise (Garcia-Gonzalez et al., 2025 Nature Methods), we aim to develop novel RNA/DNA- and protein-based single-cell and spatial barcoding tools. This will involve advanced genetic engineering technologies, using CRISPR-Cas9-assisted genome targeting of very large and expressible DNA elements. Once these novel tools are established, we aim to simultaneously target gene function and track, over long periods, how single-cell lineages with differing gene activity change their proliferation, migration, survival, or differentiation state in a developing living organism or disease conditions. This powerful approach will provide all-in-one spatial and temporal resolution of any mutant or wild-type single-cell-derived lineage.

In the future, one of our lab’s main projects will be to use these new multispectral and DNA/RNA barcoding technologies to characterize the biology and spatial heterogeneity of single cardiovascular progenitors in adult organ homeostasis and disease. We will map how single vascular (and adjacent non-vascular) cells communicate, clonally expand, differentiate, migrate, or die over time within the tissue, and identify which genes are most important and can significantly change their biology when targeted. This will provide exciting insights into the spatial heterogeneity of vascular cells, their niches, and how their collective and social behavior changes after specific genetic or pharmacological interventions.

Another aim of these novel technologies will be to model and quantify at very high spatiotemporal resolution how particular combinations of genetic mutations influence the initiation and development of cancer from single mutant cells.

Identifying novel ways of inducing effective angiogenesis and the development of arteries

We found that genetic or pharmacological induction of angiogenesis leads to cell-cycle arrest in a significant proportion of angiogenic endothelial cells (Pontes-Quero et al., 2019 Nature Communications). This occurs because endothelial cells exhibit a bell-shaped response to mitogenic stimuli. At high levels of VEGF or mitogenic ERK signaling, endothelial cells migrate and sprout but do not effectively proliferate. We intend to identify the mechanisms causing this intrinsic cell-cycle arrest to target it therapeutically when inducing angiogenesis. We envision this as the best way to effectively promote angiogenesis in ischemic tissues.

Arteries are formed by the specification and mobilization of capillary endothelial cells during angiogenesis, a process called arterialization. We have observed that this requires Notch signaling activity, the master regulator of arterialization, and the timely suppression of the cell cycle (Luo et al., 2021 Nature). Arterialization involves integration of numerous genetic and biophysical signals. Our department aims to quantitatively dissect the genetic and biophysical components that regulate arterialization. This knowledge will be crucial for effectively inducing this process. Another goal is to induce arterialization in quiescent or injured organs and assess its potential benefits for overall cardiovascular function.

Modelling and correcting vascular malformations

Modelling and correcting vascular malformations is a critical area of research and clinical practice focused on understanding and treating abnormal blood vessel formations. Vascular malformations—including arteriovenous, venous, and lymphatic malformations—frequently result from somatic genetic mutations that cause errors in vascular development, leading to significant morbidity. We aim to accurately model some of these conditions using advanced genetic technologies, imaging techniques, and in vivo models to replicate complex vascular networks and their pathological changes. These models will help decipher the underlying genetic and molecular mechanisms that drive the growth and pathogenesis of vascular malformations. Current strategies often combine invasive procedures such as embolization, sclerotherapy, and laser therapy with surgical interventions. As we better understand how these mutations alter cell signaling and biology, emerging therapies focus on targeted molecular treatments aimed at normalizing the aberrant signaling pathways responsible for vascular anomalies. Continued research into the pathophysiological mechanisms and pharmacological treatment responses of vascular malformations will enhance therapeutic precision.