TY - JOUR
T1 - High content Image Analysis to study phenotypic heterogeneity in endothelial cell monolayers
AU - Chesnais, Francois
AU - Hue, Jonas
AU - Roy, Errin
AU - Branco, Marco
AU - Stokes, Ruby
AU - Pellon, Aize
AU - Le Caillec, Juliette
AU - Elbahtety, Eyad
AU - Battilocchi, Matteo
AU - Danovi, Davide
AU - Veschini, Lorenzo
N1 - Funding Information:
We wish to thank Dr V. La Ferla for creating the FIJI importer macro including GUI. A substantial proportion of these methods have built from previous work funded by the Wellcome Trust and the UK Medical Research Council (MRC) through the Human Induced Pluripotent Stem Cell Initiative (WT098503). D.D. also gratefully acknowledges funding from the Department of Health via the National Institute for Health Research comprehensive Biomedical Research Centre award to Guy’s & St. Thomas’ National Health Service Foundation Trust in partnership with King’s College London and King’s College Hospital NHS Foundation Trust.
Funding Information:
This work is supported by an internal King’s College London Dental Institute seed fund awarded to L.V. with D.D. as collaborator. F.C. is supported by a studentship from King’s College London, Faculty of Dentistry Oral and Craniofacial Sciences (FoDOCS).
Funding Information:
We wish to thank Dr V. La Ferla for creating the FIJI importer macro including GUI. A substantial proportion of these methods have built from previous work funded by the Wellcome Trust and the UK Medical Research Council (MRC) through the Human Induced Pluripotent Stem Cell Initiative (WT098503). D.D. also gratefully acknowledges funding from the Department of Health via the National Institute for Health Research comprehensive Biomedical Research Centre award to Guy?s & St. Thomas? National Health Service Foundation Trust in partnership with King?s College London and King?s College Hospital NHS Foundation Trust. This work is supported by an internal King?s College London Dental Institute seed fund awarded to L.V. with D.D. as collaborator. F.C. is supported by a studentship from King?s College London, Faculty of Dentistry Oral and Craniofacial Sciences (FoDOCS).
Publisher Copyright:
© 2022. Published by The Company of Biologists Ltd
PY - 2022/1/15
Y1 - 2022/1/15
N2 - Endothelial cells (ECs) are heterogeneous across and within tissues, reflecting distinct, specialised functions. EC heterogeneity has been proposed to underpin EC plasticity independently from vessel microenvironments. However, heterogeneity driven by contact-dependent or short-range cell-cell crosstalk cannot be evaluated with single cell transcriptomic approaches, as spatial and contextual information is lost. Nonetheless, quantification of EC heterogeneity and understanding of its molecular drivers is key to developing novel therapeutics for cancer, cardiovascular diseases and for revascularisation in regenerative medicine. Here, we developed an EC profiling tool (ECPT) to examine individual cells within intact monolayers. We used ECPT to characterise different phenotypes in arterial, venous and microvascular EC populations. In line with other studies, we measured heterogeneity in terms of cell cycle, proliferation, and junction organisation. ECPT uncovered a previously under-appreciated single-cell heterogeneity in NOTCH activation. We correlated cell proliferation with different NOTCH activation states at the single-cell and population levels. The positional and relational information extracted with our novel approach is key to elucidating the molecular mechanisms underpinning EC heterogeneity.
AB - Endothelial cells (ECs) are heterogeneous across and within tissues, reflecting distinct, specialised functions. EC heterogeneity has been proposed to underpin EC plasticity independently from vessel microenvironments. However, heterogeneity driven by contact-dependent or short-range cell-cell crosstalk cannot be evaluated with single cell transcriptomic approaches, as spatial and contextual information is lost. Nonetheless, quantification of EC heterogeneity and understanding of its molecular drivers is key to developing novel therapeutics for cancer, cardiovascular diseases and for revascularisation in regenerative medicine. Here, we developed an EC profiling tool (ECPT) to examine individual cells within intact monolayers. We used ECPT to characterise different phenotypes in arterial, venous and microvascular EC populations. In line with other studies, we measured heterogeneity in terms of cell cycle, proliferation, and junction organisation. ECPT uncovered a previously under-appreciated single-cell heterogeneity in NOTCH activation. We correlated cell proliferation with different NOTCH activation states at the single-cell and population levels. The positional and relational information extracted with our novel approach is key to elucidating the molecular mechanisms underpinning EC heterogeneity.
KW - Cell Biology
UR - http://www.scopus.com/inward/record.url?scp=85123878523&partnerID=8YFLogxK
U2 - 10.1242/jcs.259104
DO - 10.1242/jcs.259104
M3 - Article
SN - 0021-9533
VL - 135
JO - Journal of Cell Science
JF - Journal of Cell Science
IS - 2
M1 - 259104
ER -
