Associate Professor

Warren Lee

Department of Medicine


St. Michael’s Hospital: Unity Health Toronto
Keenan Research Centre for Biomedical Science, 30 Bond St., Li Ka Shing Knowledge Institute Room 613, Toronto, Ontario Canada M5B 1W8
Research Interests
Cardiovascular, Infectious Diseases & Immunopathology
Appointment Status
Accepting MSc students, Accepting PhD students

Dr Lee received his M.D. from the University of Toronto and was awarded the Cody Gold Medal.

He completed residencies in Internal Medicine, Respirology, and Critical Care Medicine in Toronto.

He then undertook research training in the Programme in Cell Biology at the Hospital for Sick Children in the laboratory of Dr. Sergio Grinstein, completing a PhD in 2006.

This was followed by postdoctoral training in Microbiology and Immunology at Weill Medical College of Cornell University (New York, NY) in the laboratory of Dr. Carl F. Nathan.

In October 2016, Dr. Lee was named the Canada Research Chair in Mechanisms of Endothelial Permeability.

Research Synopsis

My lab has an interest in how microvascular endothelial permeability is controlled. A number of projects examine how the host-pathogen interaction affects endothelial permeability. In addition, we are investigating how the movement of critical proteins and hormones across the endothelium is regulated.

Every blood vessel in the body is lined with a specialized layer of polarized cells known as endothelium.

An essential function of the endothelial monolayer is the regulation of barrier integrity, which prevents the leakage of plasma and proteins out of the circulation while still permitting the flux of nutrients and immune cells to target tissues.

In principle, permeability of the endothelial monolayer can reflect contributions from leaking between endothelial cells (paracellular leak) and through individual endothelial cells (transcellular leak, or transcytosis).

It is widely accepted that paracellular leak predominates during inflammatory states such as sepsis and acute lung injury. Accordingly, by far the majority of research on endothelial permeability has focused on this route of endothelial permeability: the methods of study are relatively straight-forward and there is obvious relevance to human disease.

In contrast, the contribution of transcytosis to overall endothelial permeability is relatively obscure, particularly in the setting of inflammation. This is largely due to technical difficulties in distinguishing transcellular permeability from intercellular gaps, particularly in a dynamic and quantifiable way.

In addition, endothelial cells grown in culture appear to lose the ability to perform transcytosis as they are passaged.

Much of the initial work on transcytosis used electron microscopy of animal tissues, an expensive and often a mostly descriptive endeavour.

Transcytosis (at least in the apical to basal direction) is best described for the plasma protein albumin and is mediated by caveolae, small vesicles that bud off from the apical endothelial surface and release their cargo at the basal membrane. This process requires the protein caveolin-1 and the large GTPase dynamin; the latter is thought to mediate the scission of internalized caveolae from the apical plasmalemma.

My lab is interested in both routes of permeability and how they are related.

We study paracellular leak during inflammation, using acute lung injury induced by the human influenza A virus as a model system.

We investigate how the virus induces lung endothelial permeability to cause pulmonary edema, a characteristic clinical feature of severe influenza infections in humans.

We have reported effects of the virus on lung endothelial viability and on tight junction integrity; interestingly, at least some of the effect of the virus on endothelial barrier integrity is independent of viral replication and involves degradation of the tight junction constituent claudin-5.

Remarkably, restoration of endothelial barrier integrity using a Tie2 agonist peptide does not impair viral clearance and is sufficient to improve survival from otherwise fatal influenza in a pre-clinical model. This suggests that optimization of the host response to influenza (e.g. strengthening vascular integrity) represents a viable and unappreciated therapeutic approach and this is now a major focus of the lab.

We are also broadly interested in enhancing delivery of drugs and genes to the injured lung.

Another area of study in the lab is the contribution of endothelial transcytosis to the overall permeability of the endothelium to macromolecules.

Interestingly, endothelial transcytosis underlies the first stage of atherosclerosis.

Accumulation of LDL-derived cholesterol under the arterial endothelium triggers an inflammatory reaction that culminates in luminal narrowing and eventually an unstable arterial plaque. However, how the LDL gets under the endothelium is poorly understood.

Autopsy studies on young individuals dying of non-cardiac causes reveal a healthy, continuous endothelial layer overlying cholesterol deposits and the average LDL particle is too large to pass through intact cell-cell junctions; thus, LDL is likely to cross the endothelium by transcytosis.

The canonical model of LDL receptor-initiated endocytosis does not explain LDL accumulation in the arterial intima.

We devised an assay to quantify LDL transcytosis by individual cells in a confluent monolayer using total internal reflection fluorescence microscopy, complimented by traditional transwell assays and an ex vivo perfusion assay. The TIRF assay in particular is well suited to mechanistic studies. Using this approach, we have reported an unexpected role for the scavenger receptor SR-BI in LDL transcytosis.

The work in the lab is centred on live cell imaging that is complemented by traditional biochemical and molecular biology approaches. The lab is located in the Keenan Research Centre for Biomedical Science, St. Michael's Hospital (a tertiary care teaching hospital). Dr. Lee and the hospital are affiliated both with the University of Toronto and with Ryerson University.

Selected Publications

Sugiyama MG, Mintsopoulos V, Raheel H, Goldenberg NM, Batt JE, Kuebler WM, Brochard L, Leong-Poi H, Karshafian R, Lee WL. Lung ultrasound and microbubbles enhance aminoglycoside efficacy and delivery to the lung in E. coli-induced pneumonia and ARDS. American Journal of Respiratory and Critical Care Medicine. 2018 (in press)

Fung K, Wang C, Nyegaard S, Heit B, Fairn B, Lee WL. (2017). Scavenger receptor BI mediates the uptake and transcytosis of HDL in brain microvascular endothelial cells independent of caveolin, clathrin and PDZK1. Frontiers in Physiology. 8:841.

Kraehling JR, Chidlow JH, Rajagopal C, Sugiyama MG, Fowler JW, Lee MY, Zhang X, Ramírez CM, Park EJ, Tao B, Chen K, Kuruvilla L, Larriveé B, Folta-Stogniew E, Ola R, Rotllan N, Zhou W, Nagle MW, Herz J, Williams KJ, Eichmann A, Lee WL, Fernández-Hernando C, Sessa WC. (2016). Genome-wide RNAi screen reveals ALK1 mediates LDL uptake and transcytosis in endothelial cells. Nature Communications. 21;7:13516.

Armstrong S, Sugiyama M, Fung YY, Gao Y, Levy A, Wang C, Azizi P, Roufaiel M, Neculai D, Bolz SS, Seidah N, Cybulsky M, Heit B, Lee WL (2015). A novel assay uncovers an unexpected role for SR-BI in LDL transcytosis. Cardiovascular Research. 108(2): 268-77.

Sugiyama MG, Gamage A, Zyla R, Armstrong SM, Advani S, Advani A, Wang C, Lee WL. Influenza Virus Infection Induces Platelet-Endothelial Adhesion Which Contributes to Lung Injury. J Virology. 2015 Dec 4;90(4):1812-23.

Wang C, Armstrong SM, Sugiyama MG, Tabuchi A, Krauszman A, Kuebler WM, Mullen B, Advani S, Advani A, Lee WL. Influenza-Induced Priming and Leak of Human Lung Microvascular Endothelium upon Exposure to Staphylococcus aureus. Am J Respir Cell Mol Biol. 2015 Oct;53(4):459-70.

Sugiyama MG, Armstrong SM, Wang C, Hwang D, Leong-Poi H, Advani A, Advani S, Zhang H, Szaszi K, Tabuchi A, Kuebler WM, Van Slyke P, Dumont DJ, Lee WL. The Tie2-agonist Vasculotide rescues mice from influenza virus infection. Sci Rep. 2015 Jun 5;5:11030.

Honours and Awards

Canada Research Chair in Mechanisms of Endothelial Permeability (2016)