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.
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.
Jesse Moreira, PhD, MS
(May 2021 – Present)
Mentors: Darrel Kotton, MD and Jessica Fetterman, PhD
Heart failure is increasing rapidly, with the U.S. prevalence projected to reach 8 million by 2030. Five-year survival rates for patients with heart failure are 50%, which is worse than most cancers. Heart failure encompasses a number of disease subtypes with significant heterogeneity in both phenotype and therapeutic responses but the underlying contributors to the clinical heterogeneity in heart failure is not well understood.
Of interest to us, mitochondrial abnormalities have long been noted in heart failure, but whether mitochondrial genetic variation contributes to HF subtypes is unknown. The goal of Dr. Moreira’s research is to identify the underlying mechanisms whereby genetic mutations within the oxidative phosphorylation (OXPHOS) subunits lead to perturbations in metabolism and cardiomyocyte dysfunction.
Under the multi-disciplinary mentorship of Drs. Jessica Fetterman, Darrell Kotton, and Deepa Gopal, Dr. Moreira is developing and optimizing a protocol for the differentiation of cardiomyocytes from induced pluripotent stem cells (iPSC) obtained from Framingham Heart Study patients.
Dr. Moreira will generate iPSC-derived cardiomyocytes with point mutations previously identified as causal variants of mitochondrial cardiomyopathies using CRISPR-Cas9 techniques. He will evaluate cardiomyocyte phenotype and mitochondrial function in the edited and unedited cardiomyocytes in order to gain mechanistic insight into the role of OXPHOS genetic variation in cardiac metabolism and function.
Our studies may reveal novel, targetable mechanistic pathways resulting from aberrant nuclear/mitochondrial interactions in heart failure that can be exploited as treatment options in patients. Moreover, in conjunction with his PhD training in whole animal, integrative cardiovascular pathophysiology, Dr. Moreira’s project will enable him to probe the relations between mitochondrial biology and the progression of heart failure, enhancing his strength as a cardiovascular researcher.
Moreira, JD., Gopal, DM., Kotton, DN., Fetterman, JL. (2021) “Gaining Insight into Mitochondrial Genetic Variation and Downstream Pathophysiology: What Can i(PSCs) Do?” Genes (Basel) 12(11), 1668; https://doi.org/10.3390/genes12111668
Madelyn Ray, PhD
(July 2021 – Present)
Mentors: Ryan Logan, PhD
Opioid use disorder (OUD) diagnoses and deaths from opioid overdoses have skyrocketed in the United States over the recent decade. Recent studies have found links between chronic opioid use and cardiovascular diseases, especially myocardial infarction. In addition, chronic and recurrent opioid use has severe consequences on neurovasculature integrity in the brain. Dr. Logan’s laboratory has recently discovered an enrichment of transcripts related to endothelial cells in the brains of people with OUD in key brain regions involved in addiction. Transcripts were also enriched for processes related to neurovascular integrity, such as blood-brain barrier gap junctions and active transport mechanisms. These findings from the human brain provide further support for changes in neurovasculature associated with OUD.
The goal of Dr. Ray’s research is to answer the following research questions:
1) Does chronic opioid administration in mice lead to alterations in neurovascular integrity? Dr. Ray will perform acute or chronic administration of opioids in mice, after which brains will be harvested to examine markers for neurovascular integrity and blood-brain barrier functions using transcriptomics, RNA-seq.
2) Does neurovascular uncoupling alter dopamine neurotransmission in reward-related brain regions? Dr. Ray will treat mice with epoxygenase inhibitor N-(methylsulfonyl)-2-(2-propynyloxy)-benzenehexanamide (MSPPOH), the NO synthase inhibitor l-NG-Nitroarginine methyl ester (L-NAME), and the COX inhibitor indomethacin to induce a reduction in neurovascular coupling. Mice will then undergo fiber optic implantation to record dopamine transients using fiber photometry between the midbrain and striatum, areas of the brain important for opioid reward and dependence.
3) Does neurovascular uncoupling modulate opioid reward, tolerance, and withdrawal behaviors in mice? Dr. Ray will treat mice with epoxygenase inhibitor N-(methylsulfonyl)-2-(2-propynyloxy)-benzenehexanamide (MSPPOH), the NO synthase inhibitor l-NG-Nitroarginine methyl ester (L-NAME), and the COX inhibitor indomethacin to induce a reduction in neurovascular coupling. Mice will then undergo behavioral testing in opioid self-administration paradigms, including oral operant self-administration and intravenous self-administration to examine opioid seeking, craving, and relapse behaviors.
Leili Behrooz, MD
(July 2021 – Present)
Mentors: Naomi Hamburg, MD
Type 2 diabetes mellitus is a key public health problem worldwide with current trends predicting that 500 million people will have diabetes in 2035. The escalating prevalence of type 2 diabetes worldwide presents a critical cardiovascular challenge. Patients with type 2 diabetes experience accelerated vascular aging, premature atherosclerotic cardiovascular disease, and increased rates of cardiovascular events. There is considerable interest in the clinical cardiovascular benefits of new agents to treat type 2 diabetes including sodium-glucose cotransporter-2 (SGLT2) inhibitors such as dapagliflozin. However, the mechanisms of cardiovascular activity remains incompletely understood. Metabolic disturbances underlie the systemic vascular dysfunction in type 2 diabetes. Elevated glucose and lipids alter signal transduction thereby changing gene expression resulting disturbing endothelial homeostasis. Alterations in endothelial function including inflammation and reduced nitric oxide production promote atherogenesis in diabetes.
Several studies suggest activity of SGLT2 inhibitors on vascular function in animal models and humans. The robust clinical trial evidence with these classes of diabetes medications has not yet been accompanied by complete mechanistic evidence in the vasculature.
Combining vascular functional assessment with targeted endothelial cell gene expression and nitric oxide characterization will expand critical evidence about the mechanisms of action. Prior clinical studies evaluating the effects of SGLT-2 inhibitors have been limited to evaluation of vascular health without measures of endothelial cell phenotype that will provide additional mechanistic information. Dapagliflozin, a SGLT2 inhibitor, has effects on endothelial cell inflammation in cultured cells and improved vasodilation in animal models; however the effect on endothelial cell health in patients is not well-defined. Emerging experimental evidence in preclinical models also links SGLT2 inhibitors to reduction in mitochondrial oxidative stress.
We aim to investigate treatment with a SGLT2 inhibitor for diabetes will alter endothelial cell phenotype and ncRNA or biomarkers. We plan to test this hypothesis by conducting an interventional crossover study with dapagliflozin. We anticipate that evaluating endothelial cell phenotype will provide key additional information regarding the ability of novel treatments for type 2 diabetes to restore vascular health. Our study will provide novel evidence about endothelial cell effects of dapagliflozin that link to both animal models of diabetes and clinical trial data.