Andreane Cartier

Research Fellow, Vascular Biology Program, Boston Children's Hospital
Research Fellow, Harvard Medical School

I did my PhD in the lab of Dr Jean-Luc Parent in Sherbrooke, Quebec. I characterized a newly identified interacting protein (WDR36) of the beta isoform of the thromboxane A2 receptor (TPβ), a GPCR that regulates platelets aggregation and vascular smooth muscle cells constriction.

My training with Dr Timothy Hla gives me the opportunity to apply to mouse models my knowledge of GPCR signaling, molecular biology and transcriptional gene regulation.

Project 1. Role of endothelial cell S1P1 and S1P2 receptors in tumor angiogenesis and tumor growth.

Project 2. Whole genome siRNA screen to identify new targets of the endocytic pathway of S1P1 induced by Fingolimod, a synthetic functional antagonist of S1P1 and FDA approved molecule, currently used as a treatment for multiple sclerosis.

Project 3. Role of S1P1 and S1P2 in vascular dysfunction caused by a chronic inflammatory environment in vitro and in vivo, and how the imbalance of these two receptors leads to endothelial dysfunction and vascular injury, key processes in cardiovascular diseases development.

Publications Highlights

Binda C, Génier S, Cartier A, Larrivée JF, Stankova J, Young JC and Parent JL. A G protein-coupled receptor and the intracellular synthase of its agonist functionally cooperate. J Cell Biol. 2014 Feb 3;204(3):377-93.

Lachance V, Cartier A, Génier S, Munger S, Germain P, Labrecque P and Parent JL. Regulation of b2-adrenergic receptor maturation and anterograde trafficking by an interaction with Rab geranylgeranyltransferase: modulation of Rab geranylgeranylation by the receptor. J Biol Chem. 2011 Nov 25;286(47):40802-13.

Cartier A., Parent A., Labrecque P, Laroche G and Parent JL. WDR36 acts as a scaffold protein tethering a G protein-coupled receptor, Gaq and a phospholipase Cb in a signaling complex. J Cell Sci. 2011 Oct 1;124(Pt 19):3292-304.

Email: Andreane.Cartier@childrens.harvard.edu

Phone: (617) 919-2404

Reed Crocker

Research Technician, Vascular Biology Program, Boston Children's Hospital

Reed received his Bachelor’s in Biology from Kenyon College, Ohio in the spring of 2020. He completed his honors thesis on the role of long non-coding RNAs in the regulation of the crucial developmental gene klf9 in Xenopus tropicalis. Under the guidance of Dr. Wade Powell, Reed investigated these lncRNAs for their potential in mediating crosstalk between AHR and thyroid hormone signaling.

Reed joined the lab of Dr. Timothy Hla in the summer of 2020. His training has focused on using NGS and bioinformatic approaches to better understand the transcriptome of the lung endothelium. He plans to utilize single-cell RNA sequencing to identify cell-type specific targets of S1P1 signaling and to characterize age-associated differences in acute lung injury.

Email: Reed.Crocker@childrens.harvard.edu

Avishek Ghosh

Research Fellow, Vascular Biology Program, Boston Children's Hospital
Research Fellow, Harvard Medical School

Avishek received his Bachelor degree in Science in Microbiology from St. Xavier’s College, Kolkata under Calcutta University in 2012. After that, he moved to Bangalore to join the National Centre for Biological Sciences, TIFR to do his Integrated MSc-PhD in Biology under the guidance of Dr. Raghu Padinjat, from where he graduated in December, 2020. During his PhD in India, Avishek worked out the biochemical function of a metazoan lipid kinase called PIP4K in the context of regulating cell size, using new LC-MS/MS methods and Drosophila as a model system.

He joined the Hla lab in April, 2021 for post-doctoral training to expand his training on lipid signalling in metazoans. His initial interest in the Hla lab is to understand the fundamental basis of biased signalling outputs of the Sphingosine 1-phosphate receptor 1 (S1P1) using in vitro cell culture models.

Publications Highlights:

1. Mathre, S., Reddy, K.B., Ramya, V., Krishnan, H., Ghosh, A., and Raghu, P. (2019). Functional analysis of the biochemical activity of mammalian phosphatidylinositol 5 phosphate 4-kinase enzymes. Biosci Rep. 2019 Feb 19;39(2):BSR20182210. PMID: 30718367.

2. Ghosh A, Sharma S, Shinde D, Ramya V, Raghu P. A novel mass assay to measure phosphatidylinositol-5-phosphate from cells and tissues. Biosci Rep. 2019 Oct 30;39(10):BSR20192502. PMID: 31652444.

3. Ghosh, A., Raghu, P. Label-Free Quantification of Phosphoinositides in Drosophila by Mass Spectrometry. Methods Mol Biol. (2021); 2251:19-37. PMID: 33481229.

Email: avishek.ghosh@childrens.harvard.edu

Anjali Gupta

Research Fellow, Vascular Biology Program, Boston Children's Hospital
Research Fellow, Harvard Medical School

I received my Bachelor's and Master's degrees in Biological Sciences from the Indian Institute of Science Education and Research, Mohali (IISER Mohali). Then I pursued my Ph.D. degree in the laboratory of Prof. Thorsten Wohland at the National University of Singapore. My doctorate studies focussed on understanding the role of membrane lipid composition in regulating plasma membrane dynamics and organization. I utilized advanced quantitative microscopy methods to measure lipid diffusion dynamics in live cell membranes and artificial model membranes. A part of my thesis research demonstrated the role of long acyl chain ceramides in mediating plasma membrane cholesterol and actin cytoskeleton dynamics. In Dr. Timothy Hla's lab, I intend to extend my understanding of sphingolipid biology in in vivo and in vitro systems. I joined the Hla lab in October 2020 to investigate the role of S1P receptor 1 in the recently discovered glymphatic transport pathway.

Publication Highlights:

1. Gupta A, Phang IY, Wohland T. To Hop or not to Hop: Exceptions in the FCS Diffusion Law. Biophys J. 2020 05 19; 118(10):2434-2447. PMID: 32333863.

2. Gupta A, Korte T, Herrmann A, Wohland T. Plasma membrane asymmetry of lipid organization: fluorescence lifetime microscopy and correlation spectroscopy analysis. J Lipid Res. 2020 02; 61(2):252-266. PMID: 31857388.

3. Gupta A, Muralidharan S, Torta F, Wenk MR, Wohland T. Long acyl chain ceramides govern cholesterol and cytoskeleton dependence of membrane outer leaflet dynamics. Biochim Biophys Acta Biomembr. 2020 03 01; 1862(3):183153. PMID: 31857071.

4. Gupta A, Marzinek JK, Jefferies D, Bond PJ, Harryson P, Wohland T. The disordered plant dehydrin Lti30 protects the membrane during water-related stress by cross-linking lipids. J Biol Chem. 2019 04 19; 294(16):6468-6482. PMID: 30819802.

5. Huang S*, Lim SY*, Gupta A*, Bag N, Wohland T. Plasma membrane organization and dynamics is probe and cell line dependent. Biochim Biophys Acta Biomembr. 2017 Sep; 1859(9 Pt A):1483-1492. PMID: 27998689.

E-mail: anjali.gupta@childrens.harvard.edu

Bongnam Jung

Research Associate, Vascular Biology Program, Boston Children's Hospital
Research Associate, Harvard Medical School

Bongnam did her PhD in Cellular and Developmental Biology at Cornell University, New York. During her study, she worked on the role of Sphingosine 1-phosphate (S1P) receptor 1 in retinal vascular development using in vitro and in vivo systems under the guidance of Dr. Timothy Hla. She went to Sweden to do her Post-doctoral training with Dr. Christer Betsholtz to learn more about blood vessels. She was interested in characterizing pericytes, the cells surrounding the endothelium, over the course of brain development in mouse. She also investigated the involvement of fibroblast growth factor binding protein 1 (FGFBP1) in pericyte deficiency-mediated blood brain barrier breakdown. She joined back the Hla lab in Feb, 2019 and wants to explore the function of S1P receptors in the central nervous system and their underlying mechanisms.

Publication Highlights

1. Jung B, Arnold TD, Raschperger E, Gaengel K, Betsholtz C.. Visualization of vascular mural cells in developing brain using genetically labeled transgenic reporter mice. J Cereb Blood Flow Metab. (2017)

2. Ding BS, Liu CH, Sun Y, Chen Y, Swendeman SL, Jung B, Chavez D, Cao Z, Christoffersen C, Nielsen LB, Schwab SR, Rafii S, Hla. HDL activation of endothelial sphingosine-1-phosphate receptor-1 (S1P1) promotes regeneration and suppresses fibrosis in the liver. JCI Insight. (2016)

3. He L, Vanlandewijck M, Raschperger E, Andaloussi Mäe M, Jung B, Lebouvier T, Ando K, Hofmann J, Keller A, Betsholtz C. Analysis of the brain mural cell transcriptome. Sci Rep. (2016)

Email: Bongnam.Jung@childrens.harvard.edu

Phone: 617-919-2166

Andrew Kuo

Research Fellow, Boston Children's Hospital
Research Fellow, Harvard Medical School

Andrew received his Bachelor Degree in Zoology from National Taiwan University. Further, he completed his PhD training in Cell Biology under the laboratory of Dr. William Sessa, where he became interested in lipid metabolism in the vasculature system, particular triglyceride metabolism in the endothelium. His thesis project characterized Lipid Droplets (LD) for the first time in endothelial cells and demonstrated endothelium LD function as a buffer for lipid homeostasis and transport lipids to subendothelial layer.

Andrew joined the Hla lab in May, 2017 to continue his postdoctoral training studying sphingolipid biology. His current research interests include understanding S1P compartmentalization and availability in the blood brain barrier and investigating the functions of S1P bound to different chaperones in vitro and in vivo.

Publications Highlights

Kuo, A., Lee, M. Y., Sessa, W. C.. Lipid Droplet Biogenesis and Function in the Endothelium. Circ Res. (2017)

Jozsef, L., Tashiro, K., Kuo, A., Park, E., Skoura, A., Albinsson, S., Rivera-Molina, F., Harrison, K. D., Iwakiri Y, Toomre D, Sessa WC. Reticulon 4 is necessary for endoplasmic reticulum tubulation, STIM1-Orai1 coupling, and store-operated calcium entry. J Biol. Chem. (2014)

Harrison, K. D., Park, E., Gao, N., Kuo, A., Rush, J. S., Waechter, C. J., Lehrman, M. A. and Sessa, W. C.. NgBR is essential for cellular dolichol synthesis and protein glycosylation. EMBO J. (2011).

Email: Andrew.Kuo@childrens.harvard.edu

Phone: (617) 919-2403

Michel Levesque

Research Fellow, Vascular Biology Program, Boston Children's Hospital
Research Fellow, Harvard Medical School

The focus of my research as a graduate student was the application of a molecular tool for gene targeting, previously engineered in the laboratory of my PhD mentor, Dr. Jean-Pierre Perreault. More precisely, I have used highly specific ribozymes to block gene expression and the replication of two different RNA viruses (HCV and HIV). I have also developed an efficient design process to select active ribozymes against genes of interest. Through this work, I have gained experience on working with RNA in different contexts and in the characterization of gene expression in mammalian cells using techniques such as RNAse protection assays, Northern blots and luciferase assays. I have also developed an expertise in gene targeting and nucleic acid based technology design.

After I received my PhD, I moved to MSKCC to study, as research fellow, the roles of alternative polyadenylation of genes related to cancer (CD47 and PTEN). I have used protein (GFP fusion and immunofluorescence) and mRNA (FISH) localisation to assess the role of short and long 3’UTR isoforms. I have also developed western blot and immunoprecipitation conditions to evaluate the isoforms expression and post-translational modifications of a key element of the polyadenylation complex (FIP1L1).

Then I moved to Weill Cornell Medicine to work with Dr. Timothy Hla. My research has been focusing on developing a method to study gene expression profile in mouse aortic endothelial cells. Specifically, I have been trying to use the S1PR1 GFP signaling mice model to collect separately endothelial cells exposed to laminar shear stress and disturbed blood flow to characterize their transcriptome. I have also started a project focusing on the transcriptional regulation of S1PR1.

Publication Highlights

Lévesque MV, Lévesque D, Brière FP, Perreault J-P. (2010) Investigating a New Generation of Ribozymes in Order to Target HCV. PLoS One 5: e9627. PMID: 20224783

Lévesque MV, Perreault J-P. (2012) Target Induced SOFA-HDV Ribozyme. Methods Mol Biol. 848:369-84 PMID: 22315081

Scarborough RJ, Lévesque MV, Boudrias-Dalle E, Chute IC, Daniels SM, Ouellette RJ, Perreault JP, and Gatignol A. (2014) A conserved target site in HIV-1 Gag RNA is accessible to both HDV Ribozyme and short hairpin RNA activities. Mol Ther Nucleic Acids. 3:e178 PMID: 25072692

Email: Michel.Levesque@childrens.harvard.edu

Phone: (617) 919-2167

Daniel Lin

Research Fellow, Vascular Biology Program, Boston Children's Hospital
Research Fellow, Harvard Medical School

Daniel did his Ph.D. training by joining the double Ph.D. program at both University of Tsukuba, Japan and National Taiwan University, Taiwan. He received the first Ph.D. at University of Tsukuba under Dr. Yasunori Kanaho, where he characterized the lymphangiogenesis role of small GTPase Arf6 in embryonic development and cancer progression and how Arf6 regulates intracellular trafficking of b1 integrin (Lin et al., Scientific Report, 2017). In addition, he received the second Ph.D. from National Taiwan University under Dr. Hsinyu Lee, where he found the VEGF-C signaling pathway mediated by lipid mediator lysophosphatidic acid (LPA) in human prostate cancer, and how it regulates tumor lymphangiogenesis and lymphatic metastasis (Lin et al., BBA-Moll Cell Biol Lipids, 2018).

Daniel joined the Hla lab in September 2019 to investigate the role of S1P receptor 2 in lymphatic vessel development at embryonic stages and pathological conditions.

Publication Highlights

Lin YC, Chen CC, Chen WM, Lu KY, Shen TL, Jou YC, Shen CH, Ohbayashi N, Kanaho Y, Huang YL, Lee H. (2018). LPA_1/3 Signaling Mediates Tumor Lymphangiogenesis through Promoting CRT Expression in Prostate Cancer. Biochimica et Biophysica Acta – Molecular and Cell Biology of Lipids.
Huang YL, Lin YC, Lin CC, Chen WM, Chen B, Lee H. (2018). High Glucose Induces VEGF-C Expression via the LPA1/3-Akt-ROS-LEDGF Signaling Axis in Human Prostate Cancer PC-3 Cells. Cellular Physiology and Biochemistry.
Lin YC, Ohbayashi N, Hongu T, Katagiri N, Funakoshi Y, Lee H, Kanaho Y. (2017). Arf6 in Lymphatic Endothelial Cells Regulates Lymphangiogenesis by Controlling Directional Cell Migration. Scientific Reports.

Email: Daniel.Lin@childrens.harvard.edu

Colin Niaudet

Research Associate, Vascular Biology Program, Boston Children's Hospital

During his PhD, Colin studied the protective role of sphingosine-1-phosphate upon radiation of the endothelium, under the direction of Francois PARIS (INSERM, Nantes, France). He then joined Pr. Christer BETSHOLTZ’s lab, where he helped understanding S1PR1 function in the developing vascular system (Karolinska institute, Stockholm, Sweden). He also characterized the physiological roles of another vascular GPCR, the adhesion receptor Gpr116, especially in the endothelium of the central nervous system (Uppsala university, Sweden). He joined Pr. Timothy HLA’s lab in 2019, and continue to characterize the role of S1P signaling, focusing on vascular maintenance in the retina, while pursuing its study of Gpr116 functions.

Publication highlights:

Niaudet C, Petkova M, Jung B, Lu S, Laviña B, Offermanns S, Brakebusch C, Betsholtz C. Adgrf5 contributes to patterning of the endothelial deep layer in retina. Angiogenesis.(2019)

Gaengel K, Niaudet C, Hagikura K, Laviña B, Muhl L, Hofmann JJ, Ebarasi L, Nyström S, Rymo S, Chen LL, Pang MF, Jin Y, Raschperger E, Roswall P, Schulte D, Benedito R, Larsson J, Hellström M, Fuxe J, Uhlén P, Adams R, Jakobsson L, Majumdar A, Vestweber D, Uv A, Betsholtz C. The sphingosine-1-phosphate receptor S1PR1 restricts sprouting angiogenesis by regulating the interplay betweenVE-cadherin and VEGFR2. Dev Cell. (2012)

Bonnaud S, Niaudet C, Legoux F, Corre I, Delpon G, Saulquin X, Fuks Z, Gaugler MH, Kolesnick R, Paris F. Sphingosine-1-phosphate activates the AKT pathway to protect small intestines from radiation-induced endothelial apoptosis. Cancer Res. (2010

Colin.Niaudet@childrens.harvard.edu

Steven Swendeman

Research Associate, Vascular Biology Program, Boston Children's Hospital
Research Associate, Harvard Medical School

I am currently investigating the role of the HDL-associated lipid-chaperone, Apolipoprotein M, in vascular function and physiology in normal homeostasis and in human diseases. I did my PhD in Immunology at the Tufts University School of Medicine and Post-doctoral fellowships in Human Genetics at the Whitehead Institute at MIT and in Oncology at the Memorial Sloan-Kettering Cancer Center. I did further research in Vascular Biology at Weill-Cornell Medical College and The Hospital for Special Surgery.

Publication Highlights

Ding BS, Liu CH, Sun Y, Chen Y, Swendeman SL, Jung B, Chavez D, Cao Z, Christoffersen C, Nielsen LB, Schwab SR, Rafii S, Hla T HDL activation of endothelial sphingosine-1-phosphate receptor-1 (S1P1) promotes regeneration and suppresses fibrosis in the liver. JCI Insight. 2016 Dec 22;1(21):e87058. doi: 10.1172/jci.insight.87058.

Christensen PM, Liu CH, Swendeman SL, Obinata H, Qvortrup K, Nielsen LB, Hla T, Di Lorenzo A, Christoffersen C. Impaired endothelial barrier function in apolipoprotein M-deficient mice is dependent on sphingosine-1-phosphate receptor 1. FASEB J. 2016 Jun;30(6):2351-9. doi: 10.1096/fj.201500064. Epub 2016 Mar 8.

Galvani S, Sanson M, Blaho VA, Swendeman SL, Obinata H, Conger H, Dahlbäck B, Kono M, Proia RL, Smith JD, Hla T. HDL-bound sphingosine 1-phosphate acts as a biased agonist for the endothelial cell receptor S1P1 to limit vascular inflammation. Sci Signal. 2015 Aug 11;8(389):ra79. doi: 10.1126/scisignal.aaa2581.

Email: Steven.Swendeman@childrens.harvard.edu

Phone: (617) 919-2166

The Hla Lab

Dr. Timothy Hla's laboratory at Boston Children's Hospital

Lab Members