Fatima El Adili, MD
Mentor: Andrea Bujor, MD, PhD
Trainees Research Project and Progress
SSc is an autoimmune connective tissue disease in which endothelial dysfunction, inflammation and fibroblast activation lead to skin and internal organ fibrosis. This disease carries the highest mortality rate amongst autoimmune diseases, and is frequently complicated by heart involvement. The goal of Dr. Adili’s research project is to determine the role of monocytes and macrophages in SSc cardiomyopathy (CMP). We have recently discovered that the transcription factor Fli1 is expressed at low levels in SSc monocytes. We have deleted Fli1 via siRNA in human Mo/Mø, and via Cre mediated recombination in LysMCre/Fli1fl/fl mouse cells. Our preliminary data shows that deletion of Fli1 in Mo/Mø via si-RNA, or via Cre-mediated recombination using LysMCre mice (LysMCre/Fli1fl/fl), results in upregulation of several pro-inflammatory and chemotactic genes.
To expand on these studies we will leverage an existing repository of human heart tissue from SSc patients and controls to validate the findings from LysMCre/Fli1fl/fl mice. The LysMCre/Fli1fl/fl mice have enhanced inflammatory infiltrates, heart fibrosis and diastolic dysfunction; SSc-CMP will have Mø with low Fli1 that will display a similar phenotype to the LysMCre/Fli1fl/fl heart Mø. This project is innovative because it tests the novel concept that low Fli1 in SSc-Mo/Mø contributes to organ fibrosis, via a comprehensive approach using human samples and conditional mutagenesis and lineage tracing in mice. The significance of this work lies in the potential to identify Fli1 as a new therapeutic target for SSc CMP, which has no current treatment options and high mortality.
Poster presentation at Evans Days Boston University October 18 2019, Boston, MA.
Andreea Bujor, Fatima El adili; Arshi Parvez Olivia Heutlinger; Giuseppina Farina, Flora Sam. Periostin is increased in scleroderma cardiomyopathy. Arthritis Research and Therapy, 2019 November ARRT-D-19-00749 – Under review
Trainee designed and conducted experiments, analyzed data, and reviewed the final manuscript.
Jordan Chambers, PhD
(July 2020 – Present)
Mentor: Wilson Colucci, MD
Trainees Research Project and Progress
The prevalence of heart failure continues to increase and is expected to rise to 8 million people in the U.S by 2030. Heart failure patients face poor prognoses, including reduced quality of life, hospitalization, and premature death. Multiple clinical trials have shown that antidiabetic therapy Sodium-Glucose Linked Transporter 2 (SGLT2) inhibitors are cardioprotective in patients with diabetes, and recently have shown efficacy in improving outcomes in heart failure patients regardless of diabetic status. However, the mechanism(s) of protection remain elusive. The goal of my research project is to understand the mechanism by which SGLT2 inhibitors are cardioprotective in non-diabetics. Our laboratory previously showed that SGLT2 inhibitors prevent cardiac and mitochondrial dysfunction in mice with diet-induced obesity and metabolic syndrome. However, since these mice become diabetic it is unclear if the benefit is due to a cardiac-specific drug target or a consequence of diabetes prevention. To understand how SGLT2 inhibitors protect the heart, we will treat non-diabetic mice that develop dilated cardiomyopathy due to cardiac myocyte-specific over-expression of Gq with the SGLT2 inhibitor ertugliflozin. We will determine the effects of ertugliflozin on cardiac structure, function, and mitochondrial energetics in the Gq hearts, and perform proteomic and gene profile analyses to gain insight into ertugliflozin-mediated cardioprotection. In addition, we will harmonize the omics data generated from this project with samples from our previous study in diabetic mice, enabling assessment of the metabolic effects of SGLT2 inhibition in diabetic vs. non-diabetic hearts. We aim to identify a shared mechanism of action that is independent of glycemia. As the benefits of SGLT2 inhibition in patients with underlying cardiovascular disease and heart failure with or without diabetes were unexpected and the target of action is unknown, these studies have the potential to uncover mechanisms underpinning disease progression and discover new therapeutic avenues in heart failure.
Beatriz Ferran Perez, PhD
Redox therapy approaches to improve ischemic limb vascularization
(July-2018 to Present)
Mentor: Reiko Matsui, MD
Many diseases are associated with increased levels of reactive oxygen and nitrogen species, however it is demonstrated that certain levels of oxidants are necessary to promote ischemic angiogenesis. Oxidants transduce angiogenic signaling by post-translational protein thiol modifications. The reaction of protein thiols with cellular glutathione (GSH) renders stable protein-GSH adducts (S-glutathionylation), which are able to alter the protein function. Protein-GSH adducts are reversed by glutaredoxin-1 (Glrx), a cytosolic enzyme that can inhibit angiogenesis by modulating several proteins including HIF-1a. We found that Glrx inhibition promotes angiogenesis via HIF-1α stabilization.
My work is focused on redox-regulation of ischemic vascularization related with aging and type 2 diabetes. Diabetic patients have a worse blood flow recovery in response to ischemia comparing with non-diabetic subjects. Since Glrx expression regulates the ischemia-induced angiogenic responses, my research interest is to find out the molecular mechanisms by which modulating Glrx activity/expression in the skeletal muscle may improve vascular recovery after hindlimb ischemia, and to propose an effective therapy. To achieve this goal, my main objectives are A) generate skeletal muscle-specific Glrx KO mice by using the Cre-Lox system and by adeno-associated virus (AAV) injection; B) contribute to establish an AAV production and purification method for in vivo studies; and C) as a therapeutic approach, apply AAV-mediated Glrx inhibition to poor vascularization models like diabetic mice and middle-age female mice.
A) The colony of muscle-specific Glrx KO mice is already growing and I expect to have the first animal cohort in three months.
B) I contributed to optimize an affordable protocol for producing and purifying AAV in our laboratory, resulted in a manuscript submitted to Scientific Reports, now in the revision process. Tests of the viral suspensions indicated that AAV particles can be used to inhibit Glrx expression in vivo and in vitro. I am working now in the design of the viral DNA to decrease the inflammation caused by the AAV intramuscular injection.
C) I performed pilot experiments inducing hindlimb ischemia in middle-age mice and confirmed that females show worse blood flow recovery than males. Also, I am working on the method to detect Glrx target proteins in skeletal muscle samples by immunoprecipitation with anti-GSH antibodies.
- Poster presentation in the Evans Research Day. October 11, 2018 (Boston University School of Medicine, Boston)
- Poster presentation in the 25th Annual Conference of the Society for Redox Biology and Medicine (SfRBM). November 14-17, 2018 (Chicago)
- Work-in-Progress presentation in the WCVI Seminar Series. January 29th, 2019.
Pre-meeting workshop “Oxidative Stress and Signaling: Methods, Mechanisms, and Therapeutics”. November 14th, 2018 (Chicago), at the 25th SfRBM Annual Conference.
- Weinberg EO, Ferran B, Tsukahara Y, Hatch MMS, Han J, Murdoch CE, Matsui R. “IL-33 induction and signaling are controlled by glutaredoxin-1 in mouse macrophages.” PLoS One. 2019 Jan 25;14(1):e0210827. (PMID: 30682073)
- Toyokazu Kimura, Beatriz Ferran, Yuko Tsukahara, Qifan Shang, Suveer Desai, Ivan Luptak, David Richard Pimentel, Takeshi Adachi, Yasuo Ido, Reiko Matsui, Markus Bachschmid. “Production of adeno-associated virus vectors for in vitro and in vivo applications”. Manuscript in revision, submitted to Scientific Reports.
Jena Goodman, PhD
Vascular Biology and Hypertension
(September 2020 – present)
Mentor: Francesca Seta, PhD
Trainees Research Project and Progress
Recent studies have identified arterial stiffness (AS) as an independent risk factor for cardiovascular disease. AS is characterized by a decrease in arterial/aortic compliance, associated with an increase in the velocity at which blood pressure pulse propagates through the circulatory system, also known as the pulse wave velocity (PWV). A recent genome-wide association study discovered several single nucleotide polymorphisms (SNPs) located within a gene desert on chromosome 14 that are significantly associated with PWV. The PWV-associated, “aortic stiffness” gene desert contains an enhancer for Bcl11b, a transcription factor primarily known for its role in T-cell differentiation and neuronal development. However, the Seta laboratory discovered, for the first time, the presence of Bcl11b in aorta and aortic vascular smooth muscle (VSM) cells. Furthermore, the Seta laboratory discovered Bcl11b in aortic VSM regulates the expression of contractile genes including smooth muscle myosin (MYH11) and smooth muscle actin (α-SMA), further implicating a functional role for Bcl11b in modulating vascular tone. Taken together, the Seta group hypothesizes SNP variants in the BCL11B enhancer locus interferes with Bcl11b expression, disrupting VSM function potentially playing a causative role in AS pathogenesis. Moreover, the Seta group found that when treated with the hypertensive agent angiotensin II (angII), mice lacking Bcl11b in VSM cells (BSMKO) develop aortic aneurysms compared to angII-treated WT, further supporting an important role of Bcl11b in the vasculature.
As a T-32 post-doctorate fellow I will identify the molecular mechanisms by which Bcl11b-dependent signaling pathways relate to AS, aortic aneurysms and vascular function by addressing mainly three aims:
(1) to identify transcriptional regulation associated with Bcl11b in the vasculature; I will perform chromatin immunoprecipitation (ChIP) sequencing on VSM cells and aortas of WT and BSMKO mice; this study will be the first to identify DNA binding targets of Bcl11b. specifically within the vasculature;
(2) to identify the Bcl11b-dependent mechanisms of aortic aneurysms; I will prepare libraries for single-cell RNA sequencing using digested aortic tissue from WT and BSMKO mice 7-days after ang II challenge; I will then validate promising targets from single-cell RNA sequencing by qRT-PCR and Western Blot; this will allow me also to identify cell subpopulations within the aneurysmal aorta with distinctive gene profiles which may contribute to disease pathogenesis;
(3) to develop and characterize an ex vivo VSM cell culture model in order to preserve the phenotype of isolated VSM cells; when cultured with standard methods, VSM cell are known to become rapidly senescent and lose their contractile phenotype limiting their utility in in vitro studies; the new 3D culture approach I will develop could overcome these limitations.
Zhou Y.; Wan X.; Seidel K.; Zhang M.; Li Z.; Goodman J.B.; Seta F.; Hamburg N.; Han J.; (2020) “Aging and Hypercholesterolemia Differentially Affect the Unfolded Protein Response in the Vasculature of ApoE−/− Mice” Cardiovascular Research. In Review.
Valisno J.A.*; May J.M.*; Singh K.; Venegas L.; Budbazar E.; Goodman J.B.; Helm E.Y.; Nicholson C.J.; Avram D.; Cohen R.A.; Mitchell G.F.; Morgan K.G.; Seta F.; (2020) “BCL11B is a newly identified regulator of arterial stiffness and related target organ damage” Circulation Research. In Review.
Morgan R.; Qin F.; Goodman J.B.; Croteau D.; Siwik D.A.; Tong X.; Pimentel D.R.; Cohen R.A.; Colucci W. S. (2020) “Reversible oxidation of sarco/endoplasmic reticulum calcium ATPase C674 is required for Raf/MEK/ERK hypertrophic signaling in cardiac myocytes” Journal of Biological Chemistry. In Review.
Schneider E.R.; Mastrotto M.; Laursen W.J.; Schulz V.P.; Goodman J.B.; Funk O. H.; Gallager P.G.; Gracheva E.O.; Bagriantsev S.N. (2014) “Neuronal Mechanism for acute mechanosensation in tactile-foraging waterfowl” PNAS.
Laursen W.J.; Mastrotto M.; Pesta D.; Funk O.; Goodman J.B.; Merriman D.; Ignolia N.T.; Shulman G.I.; Bagriantsev S.N.; Gracheva E.O. (2014) “Neuronal UCP1 Expression Supports Cranial Endothermy in Mammalian Hibernators” PNAS.
Sana Majid, MD
Determining Cardiovascular Injury due to the Use of Novel Tobacco Products
(July 2018 – Present)
Mentor: Naomi M. Hamburg, MD, MS
Electronic cigarettes (e-cigarettes) are marketed as safer alternatives to combustible tobacco products; however, whether e-cigarettes are a harm reduction tool with reduced cardiovascular (CV) toxicity is unknown. Dr. Majid seeks to identify e-cigarette product characteristics associated with CV injury. Specifically, Dr. Majid will evaluate the effects of Juul, the top selling e-cigarette in the US, on endothelial cell phenotype. She hypothesizes that Juul e-liquids will induce endothelial cell toxicity and that toxicity will vary based on the flavoring additives and nicotine levels. In order to test this hypothesis, she will evaluate the effects of available e-liquids at varying concentrations on cellular viability, nitric oxide production, and oxidative stress in commercially available endothelial cells. She has learned how to perform the assays and will start cell exposures in the coming weeks. Dr. Majid has additional work evaluating the relations of plasma lipids with e-cigarette use in the Cardiovascular Injury due to Tobacco Use (CITU) cohort. She hypothesizes that specific volatile organic compounds may mediate the association of alterations in plasma lipids associated with e-cigarette use. She has written the analytic plan and has begun the analyses. Through her efforts, Dr. Majid will identify e-cigarette product characteristics associated with cardiovascular toxicity. Her work will lend additional support for regulations on product characteristics associated with cardiovascular injury such as setting permissible limits on flavoring additives.
AHA Tobacco Regulation and Addiction Center (A-TRAC) 2.0 Fellowship (2018-2020)
American Heart Association (AHA) Scientific Sessions 2018, Chicago, IL (November 10-12)
- Majid, J. L. Fetterman, R. J. Keith, R. M. Weisbrod, M. Holbrook, M. M. Stathos, R. Breton-Romero, R. Bastin, B. Feng, R. M. Robertson, A. Bhatnagar, N. M. Hamburg. The Impact of Combustible and Electronic Cigarette Use on Vascular Health: An Observational Study. Boston University Evans Department of Medicine Research Days (October 11).
Introduction to Statistical Computing – Boston University, Boston, MA (Fall 2018)