2021
CADASIL is an hereditary small vessel disease caused by mutations in the NOTCH3 gene. These mutations lead to progressive changes in small brain arteries and reduced blood flow to the brain. Patients with CADASIL suffer from strokes and vascular dementia from mid-adulthood. It has recently been shown by our research group, that some NOTCH3 mutations lead to a much earlier onset of CADASIL than other mutations, but why this is the case is not yet understood. CADASIL vessel models representing both severe and mild mutations will enable us to study the molecular mechanisms underlying these differences and will teach us about CADASIL disease pathomechanisms in general. Our university medical center is a CADASIL expert center and for this project we will collaborate with the internationally leading vessel model group in our research center. Together, we will develop 3D CADASIL vessels-on-chip, built up of CADASIL vascular cells. These cells are obtained by harvesting pluripotent stem cells from blood samples of CADASIL patients with different mutations. The stem cells are then differentiated into vascular cells and incorporated into the chips. We will examine structural and functional abnormalities of the vessel wall and the differences between vessels with severe and mild mutations. We aim to share these CADASIL vessel-on-chip with the international CADASIL research community to promote CADASIL research.
***Seed Funding
Collaboration with Penn Medicine Orphan Disease Center
Million Dollar Bike Ride funded
PI: Saskia Lesnik-Oberstein
Institution: Leiden University Medical Center
Grant Amount: $60,228
Funding Period: February 1, 2022 – January 31, 2023
The aim of this project is to understand the reason why CADASIL is produced, and possible factors associated with the severity of the disease. To do this, we will use a new and innovative strategy with omic technology (single-nuclei RNA-seq analysis) to obtain transcripts and pathways associated with the disease and its severity. Based on this information, we purpose to find therapeutic targets overexpressing/inhibiting the molecules found to be significant in the single-nuclei RNA-seq study and those found significant in other omic studies of CADASIL already published, to evaluate later the benefits in our human cellular model (pattern of aggregation of Notch3).
***Seed Funding
Collaboration with Penn Medicine Orphan Disease Center
Million Dollar Bike Ride funded
PI: Israel Fernández Cadenas
Institution: Fundació Privada Institut de Recerca de l’Hospital de la Santa Creu i Sant Pau
Grant Amount: $60,228
Funding Period: February 1, 2022 – January 31, 2023
2020
2020 grant funded by Penn Medicine Million Dollar Bike Ride Team CADASIL
CADASIL, short for Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarct and Leukoencephalopathy, is the most common genetic cause of a pathology known as small vessel disease (SVD) of the brain. During SVDs such as CADASIL, the structure and function of small blood vessels (arteries, arterioles, venules, and capillaries) within the brain become compromised. An early result of this vascular dysfunction is a decrease in blood flow to the brain (cerebral blood flow or CBF), which eventually leads to dementia and/or strokes. This study will elucidate the molecular mechanisms of compromised CBF increases in response to neural activity (“functional hyperemia”) using a clinically relevant CADASIL mouse model. We will specifically focus on examining the impact of CADASIL on capillaries, the smallest and most abundant vessels in the brain, which we have previously shown to be the molecular cornerstone in functional hyperemia responses in healthy animals. Considering that it is known that CADASIL causes an abnormal accumulation of specific proteins around the outside of small vessels in the brain, we propose to examine how two of these proteins, epidermal growth factor receptor (EGFR) and heparin-binding EGF-like growth factor (HB-EGF), contribute to capillary dysfunction in CADASIL. This project, by providing a greater understanding of the cellular pathways contributing to CADASIL pathologies, will create a firm footing for future therapeutic development.
Publications:
PIP2 corrects cerebral blood flow deficits in small vessel disease by rescuing capillary Kir2.1 activity
***Seed Funding
Collaboration with Penn Medicine Orphan Disease Center
Million Dollar Bike Ride funded
PI: Massayo Koide, PhD
Institution: University of Vermont Larner College of Medicine
Award Amount: $82,795
Funding Period: February 1, 2021 – January 31, 2022
2019
With the support of cureCADASIL funding in 2019, my laboratory has now successfully trained a postdoctoral fellow, Dr. Dorothee Schoemaker, to conduct blood biomarkers analysis. The larger aim of this work is to identify an accessible, sensitive and reliable strategy to track the progression of CADASIL over time. Dr. Schoemaker is currently optimizing an enzyme-linked immunosorbent assay (ELISA) to quantify the concentration of the Notch 3 ectodomain (N3ECD) in the blood. Previous work in animal models from our research group has revealed that blood levels of N3ECD are affected in CADASIL and are related to disease severity, suggesting that this could represent a clinically useful biomarker.
Over the next period, we will continue optimizing assays for quantification of blood levels of N3ECD, as to obtain a protocol that is highly sensitive to variations in N3ECD levels, and reliable. We will also work on optimizing assays for other potentially relevant blood biomarkers for CADASIL. Finally, we aim to examine associations between levels of these biomarkers and clinical symptoms of the disease, including cerebrovascular burden on magnetic resonance imaging (MRI) and cognitive changes. These analyses would allow determining whether these biomarkers are accurately mirroring symptoms of CADASIL and could be used to estimate disease progression.
**Research Alliance
Joseph Arboleda-Velasquez MD, PhD
Schepens Eye Research Institute of Mass. Eye and Ear
Harvard Medical School
2019 grant funded by Penn Medicine Million Dollar Bike Ride Team CADASIL
$81,000
The emerging view from our recent work on CADASIL is that fundamental alterations in extracellular matrix proteins drive early functional deficits in the reactivity of the brain vasculature. In the smooth muscle cells that constitute the arterial wall, voltage-dependent potassium (Kv) channels act as a brake on contractility. We have found that a significant increase in the number of Kv channels underlies cerebrovascular dysfunction in CADASIL syndrome.
Importantly, CADASIL is caused by mutations in the NOTCH3 receptor, which is expressed not only in vascular smooth muscle cells, but also in pericytes. Pericytes are mural cells that wrap around the endothelial cells forming the capillaries. They are an under-studied cell type while increasing evidence suggests that contractile pericytes regulate capillary blood flow. Virtually nothing is known about the impact of CADASIL on pericytes and whether extracellular matrix dysregulation impairs pericyte contractility, and therefore cerebral blood flow regulation. The overall goal of this project is to provide a better understanding of the impact of the CADASIL-causing mutation on cerebral blood flow regulation by pericytes at the capillary level.
***Seed Funding
Collaboration with Penn Medicine Orphan Disease Center
Million Dollar Bike Ride funded
PI: Fabrice Dabertrand, PhD
The Dabertrand Lab
University of Colorado at Denver
In the Fall 2018 funded research focused on the generation of in vivo models for testing the preclinical efficacy of our therapeutic antibody on different animal models. Our previous published work reported on the treatment of models carrying the C455R mutation in Notch 3. We are currently conducting studies in models carrying the R1031C mutation. Validation of our findings in models with different CADASIL mutations in Notch3 will demonstrate how general the results are and also the extent to which our therapeutic antibody may impact disease in the larger population of individuals with CADASIL.
**Research Alliance
Joseph Arboleda-Velasquez MD, PhD
Schepens Eye Research Institute of Mass. Eye and Ear
Harvard Medical School
Patient Data Sharing of Genetic and Health information Informs Genetics Discovery and Fuels Research
Patient Data Sharing of Genetic and Health information Informs Genetics Discovery and Fuels Research
Broad sharing of genomic and phenotypic data is needed to inform variant classification, gene-disease validity assessments, and genomic medicine. The Clinical Genome Resource (ClinGen) has engaged 2340 patients in data sharing through its patient registry, GenomeConnect, since 2014. The registry is open to anyone who has had genetic testing, and, to date, has shared 969 sequence variants with ClinVar. These efforts have resulted in 491 (51%) previously unsubmitted sequence variants being added to ClinVar, additional case-level evidence being made available, 25 variant classification updates being identified and shared, and numerous requests for additional information from clinicians and researchers. Given GenomeConnect’s success in patient data sharing, we sought to broaden efforts by working with external registries that may be collecting data but not sharing publicly. We initiated a pilot to engage patient registries in data sharing and assess the utility of data shared from these groups. Here, we report on a pilot of 100 individuals from five external registries in the Patient-Data Sharing Program.
Authors and Affiliations:
Juliann M. Savatt1, Danielle R. Azzariti2, W. Andrew Faucett1, Jon Florin3, David H. Ledbetter1, Vanessa Rangel Miller3, Emily Palen1, Heidi L. Rehm2,5,6 Jud Rhode3, Les Rogers7, Sandra Talbird8, Laura Trutoiu9, Jo Anne Vidal3, Christine Waggoner10, Erin Rooney Riggs1, Christa Lese Martin1
1Geisinger, Danville, Pennsylvania; 2The Broad Institute of MIT and Harvard, Cambridge, Massachusetts; 3No Stomach For Cancer, Middleton, Wisconsin, 4Invitae, San Francisco, California; 5Center for Genomic Medicine, Massachusetts General Hospital, Boston, Massachusetts; 6Harvard Medical School, Boston, Massachusetts, 7CFC International, St. Petersburg, Florida, 8CureCADASIL, Plainsboro, New Jersey, 9Association for Creatine Deficiencies, Carlsbad, California, 10Cure GM1 Foundation, Albany, California
From July 2018 and May 2019, five organizations began engaging participants in data sharing – Association for Creatine Deficiencies, Cardio-Facio-Cutaneous Syndrome International, CureCADASIL, No Stomach For Cancer, and GM1 Patient Network. Registry participants control their participation via an online consent within their registries. Those who opt to share data are asked to upload a copy of their genetic testing report to their account to allow for uniform data mining, may be invited to complete additional surveys to collect additional phenotype data, and are provided with the option to receive updates regarding their results. Fifty-six percent of participants uploaded their report. Genetic counselors review reports to ensure standardized data collection and submitted de-identified data to ClinVar. A total of 81 unique variants were shared, of which 45.6% (n=37/81) were novel to ClinVar demonstrating that condition specific registries are a source of novel variants. Participants also provided additional health and case-level information that could inform future gene and variant curation efforts, such as those performed by ClinGen. Of variants that were previously submitted by the reporting laboratory, 12% (n=3/25) were out of date compared to the current classification. These pilot data show that registries can empower patients to contribute valuable information to the public knowledge base, benefiting patients and the genetics community.
2018
Suning Ping, a post-doctoral fellow in Dr. Li-Ru Zhao’s lab at SUNY Upstate Medical University in Syracuse, received a student travel grant from cureCADASIL to present her work entitled “STEM CELL FACTOR AND GRANULOCYTE COLONY-STIMULATING FACTOR PROMOTE BRAIN REPAIR AND COGNITIVE FUNCTION THROUGH VEGF-MEDIATED ANGIOGENESIS IN A MOUSE MODEL OF CADASIL”. Dr. Li-Ru Zhao’s lab has recently demonstrated the efficacy of combining two hematopoietic growth factors. Stem cell factor (SCF) and granulocyte colony-stimulating factor (G-CSF) improve cognitive function, increase brain blood vessel density and increase neural network rewiring in a transgenic mouse model of CADASIL. Dr. Ping demonstrated that a third growth factor, vascular epithelial growth factor (VEGF), is necessary for SCF and G-CSF to function. By blocking VEGF with Avastin, the improvement in cognitive function, enhanced blood vessel density and enhanced neural network rewiring that was observed with SCF and G-CSF does not occur. Dr. Ping’s results shed light on a key mechanism by which hematopoietic growth factors may restrict CADASIL pathology. Read the Abstract.
**Young CADASIL Investigators
PI: Li-Ru Zhao MD, PhD
Postdoc: Ping Suning
Lab of Dr. Li-Ru Zhao
SUNY Upstate Medical University in Syracuse
The purpose of this summer fellowship funded by cureCADASIL was to understand the dysregulation of NOTCH signaling in CADASIL through immunohistological characterization of brain vasculature from CADASIL subjects and controls. Given the diversity in CADASIL-causing NOTCH3 mutations, our approach was to use a diverse cohort of NOTCH3 mutations to address the hypothesis that CADASIL is a disease caused by loss of function in NOTCH signaling. Utilizing a combination of markers for distinct vascular cell types, classical and novel NOTCH3 signaling targets we characterized the cellular consequences of CADASIL causing NOTCH3 mutations on brain vasculature. This fellowship aided to further advance our understanding of the consequences of NOTCH3 dysregulation in brain vasculature of CADASIL patients and provide preliminary data to support a grant submission to continue this work.
***Seed Funding
PI: Luisa Iruela Arispe MSc, PhD
Postdoc: Milagros Romay, PhD
Student: Margaret Ramirez
The Arispe Lab
University of California at Los Angeles
Funding to initiate experiments with animals carrying the R1031C mutation. This analysis is critical for us to examine how many patients could potentially benefit from our modality of treatment: only those with a specific mutation or patients with different mutations associated to different mechanisms.
**Research Alliance
Joseph Arboleda-Velasquez MD, PhD
Schepens Eye Research Institute of Mass. Eye and Ear
Harvard Medical School
Dr. Dorothee Schoemaker is a Post-Doctoral fellow in the lab of Dr. Joseph Arboleda- Velasquez. She will work with cureCADASIL to grow and enrich the registry of CADASIL subjects and will work with members of the medical and research community to increase our understanding of the vascular contributions to cognitive decline.
**Research Alliance
Joseph Arboleda-Velasquez MD, PhD
Schepens Eye Research Institute of Mass. Eye and Ear
Harvard Medical School
2017
Funds provided by cureCADASIL have been instrumental in addressing additional experiments suggested by reviewers from the journal considering publication of our in vivo testing of a potential treatment for CADASIL. Specifically, we were able to conduct additional analyses of brain integrity to evaluate vascular leakage and how it can be prevented. We were also able to fund additional experiments to enable another animal trial. Funds were used to generate more animal model mice, determine whether or not they carry the CADASIL mutation and support their maintenance. The latter will include animals with established brain pathology to determine the potential effect of the drug. This will be informative of potential efficacy in patients with moderate to advanced CADASIL.
**Research Alliance
Joseph Arboleda-Velasquez MD, PhD
Schepens Eye Research Institute of Mass. Eye and Ear
Harvard Medical School
2016
Supplemental funding assisted in identifying a roster of blood biomarkers that have the potential to report whether a therapy for CADASIL would be effective in clinical trials. These biomarkers include proteins that are significantly more or less abundant in the presence of CADASIL and revert to their normal levels after treatment with an experimental drug in laboratory models. We are launching a major effort to further develop and validate these biomarkers in blood from CADASIL patients as a necessary step towards the goal of launching a clinical trial that will test the safety and efficacy of experimental treatments for CADASIL. As part of the plan we will 1) develop assays specific for the human proteins, 2) analyze samples already collected from research sites from around the world, and 3) establish the foundation for collection of additional samples in the US. Validation of these biomarkers will enable the design of clinical studies to evaluate modalities of treatment that Dr. Arboleda-Velasquez found to be promising in laboratory models of CADASIL.
**Research Alliance
Joseph Arboleda-Velasquez MD, PhD
Schepens Eye Research Institute of Mass. Eye and Ear
Harvard Medical School
The aim of this summer internship funded by cureCADASIL was to image and quantify these defects extensively and begin describing the mechanism involved. Determine if notch functions cell autonomously in mechanosensory neurons. Begin investigating the mechanism of action of Notch in mechanosensory neuron connectivity and address the potential link to CADASIL. This research is described in a publication presented in a poster at the 2017 Society for Neuroscience. Quin Brown Poster Summer Research 2k16 (3)
**Young CADASIL Investigators
PI: Rachid El Bejjani, PhD
Student: Quin Brown
Davidson College
Blood biomarkers in a mouse model of CADASIL, 2016 Brain Research publication