BREAKTHROUGH IN AVM RESEARCH!
Kelleigh’s Cause would like to thank everybody who has participated in our fundraising efforts and those who are taking part in our current events. Your amazing support has allowed us to become a significant voice for raising AVM awareness and for advancing AVM medical research.
Our initial grants to the Medical College of Wisconsin has funded the Kelleigh Gustafson Post-Doctoral AVM Research Fellow for the past two years. The grants have supported translational research [the use of basic research techniques to solve clinical problems] in the field of vascular anomalies, in particular arteriovenous malformations. The funds raised have paid for research supplies, for the tests that analyze proteins produced by AVMs [mass spectroscopy], developed animal models to test new drugs for AVM, as well as, develop and tested quantitative imaging tests for patients with AVM.
We are excited to announce significant breakthroughs.
Kelleigh’s Cause Research Funding; how have your donations been used?
Conducting any kind of medical or biological research is complex and expensive. It requires the use of a well-funded and equipped research laboratory, complex lab equipment, reagents, chemicals, and animals. The human component includes institutional managers, administrators, organized committees and processes for human and animal research, as well as researchers [PhD’s, physicians, technologists, etc.] with appropriate training and interest. Most successful research groups consist of a large group of individuals with a common interest [e.g. AVM] but different types of expertise [e.g. cell biologist, physician specialists, statistician] who all contribute in different ways. Translational research, in which findings generated by a basic science lab can be used to change the medical treatment of patients, is probably the most difficult. This is the type of research that Kelleigh’s Cause has endeavored to support at the Medical College of Wisconsin, in conjunction with Children’s Hospital of Wisconsin. The main leaders in this research have been Patricia Burrows MD, Professor of Radiology, an Interventional radiologist with a long-standing interest in treating children and adults with AVMs, and Ramani Ramchandran, PhD, Professor of Pediatrics and Director of the developmental vascular biology program at the Medical College of Wisconsin. These leaders shared the goals of better understanding vascular malformations and improving the lives of patients with these conditions.
What type of research has been funded by Kelleigh’s Cause?
1. Proteomics of vascular malformations.
Funding from Kelleigh’s Cause was used to pay for lab supplies to process blood and urine samples taken from patients with vascular malformations who are undergoing stage treatment. The project was designed in 2013, before current knowledge about the genetic causes of AVM. The goal was to identify proteins produced uniquely by the vascular malformations that could be used to help identify novel treatments or used to monitor the results of treatment. For example, if a specific protein produced by an AVM could be detected in the blood or urine, a test for that protein could be developed and used to follow the effects of drug treatments. We also hoped that a drug treatment could be developed to target that protein. The project began with writing a protocol and getting it approved by the institutional review board of Children’s Hospital of Wisconsin. Human research, even involving minimal risk such as collecting blood and urine patients undergoing necessary procedures, is heavily regulated for the protection of the patients. Every document, including specific consent forms, had to be written, reviewed, revised, and approved. Every year, the project was reviewed and new revisions needed. Even allocating the donated funds to the research project required input from many specialists. Once the research began, each sample collection required signed consent from the parents, and assent from the child, and then the consent forms had to be carefully stored. The samples were collected by the clinical nurses and anesthesiologists after the children were anesthetized for the procedure, to minimize any discomfort. Blood and urine samples were collected in specific tubes, placed on ice and transported to the research lab in another part of the campus, usually by one of the postdoctoral fellows [PhDs]. They were then processed in the lab, divided into aliquots, documented in a database and stored in a -70°C freezer. By the time the samples were ready for analysis, we no longer had a proteomics expert on our MCW team, so they were shipped to a research proteomics lab in Boston, Massachusetts. Mass spectrometry [a sophisticated test to sort and quantify thousands of tiny proteins] was carried out and a huge volume of computerized data was obtained. This data then had to be translated and interpreted by researchers at MCW, and, in the end, we did not find consistently elevated levels of specific proteins in the samples taken from AVMs. A project like this requires a huge effort. Kelleigh’s Cause paid for reagents and the mass spectrometry cost. The thousands of hours of work involved in managing the protocol, obtaining and processing the samples and analyzing the results were donated by Drs Burrows and Ramchandran, the postdoctoral fellows and other researchers involved, as well as MCW and CHW. Unfortunately, the data did not demonstrate any key proteins that could be used clinically in the management of patients with AVM. The remaining samples are still in storage, available for additional testing.
2. Development of an animal model of AVM [using zebrafish]
Funding from Kelleigh’s Cause was used to hire a postdoctoral fellow [Dr. Shahram Eisa-Beygei] and purchase supplies and animals for the research. Initially, zebrafish embryos with RASA1 mutations were purchased from a commercial source. This mutation is known to be associated with a high frequency of AVM in humans. The zebrafish were bred and treated in ways that enhance the visualization of developing blood vessels. They developed defects in the body and blood vessels consistent with what was expected with RASA1 mutation. However, after many months of waiting for the numbers to increase, genetic screening showed that they did not in fact have the RASA1 mutation! In order to obtain a suitable animal model, a different technique [CRISPR] was used, which is currently under development by a medical student Scott Self who is pursuing this project as part of his honors in research aspect of the medical summer research program. We hope that by inducing mutation in rasa1 with CRISPR method (a genomic engineering tool), we will recapitulate the defects in vasculature that we have already observed with rasa1 morpholino(a method to knockdown rasa1 gene)-injected embryos. Dr. Eisa-Beygei and collaborators from around the world performed extremely sophisticated tests to learn how the abnormal blood vessels developed and led to AVM formation.
3. Drug testing using zebrafish AVM model
Funding from Kelleigh’s Cause was used to purchase drugs, animals and lab supplies in order to test the altered zebrafish to different dosages of five different medications that had potential to reverse the defect. Scott Self, the MCW medical student carrying out this research during his free time, developed a grading system to analyze the fish that were injected with rasa1 morpholino, a technique frequently used in zebrafish to knockdown genes. Rasa1 knockdown fish showed numerous vascular defects, which were evaluated under a special microscope to identify defects. FDA-approved compounds were tested for toxicity first, and non-toxic doses were used to evaluate which drug and dosage was the most effective in “rescuing” the vascular defect. Scott’s salary [stipend] was paid by MCW, and lab space and equipment was provided by Dr. Ramchandran’s lab at MCW. As part of his Honors in Research aspect of the medical school summer research project, Scott Self is committed to pursuing this project to completion.
4. Characterization of endothelial cilia distribution during cerebral-vascular development in zebrafish (Danio rerio).
Funding from Kelleigh’s Cause was used to hire a postdoctoral fellow, Dr. Shahram Eisa-Beygei and to pay for animals and lab supplies. Lab space and equipment, such as a state-of-the-art confocal microscope, were provided by Dr. Ramchandran, Children’s Hospital of Wisconsin and MCW. The purpose of this work was to obtain insights into the earliest aspects of blood vessel development and determine the role of a tiny endothelial cell appendage, the cilia, in this development. In a very short period of time, and starting essentially from scratch, Dr. Eisa Beygei was able to develop a zebrafish model to study cilia under various conditions. He found that cilia were involved in the very earliest part of blood vessel formation, when a mass of cells develops a lumen or channel. These findings were completely new and were published rapidly, meriting a cover page illustration and complementary editorial in a prominent research journal. Almost simultaneously, another group working from Europe found that some of the genes known to be causative for AVM are localized on the cilia. So the next steps in this research will be to study the effects of these gene mutations on blood vessel development using the same cilia imaging techniques, as well as to study the integrity of cilia in surgical specimens of human AVMs.
SEEKING ADDITIONAL RESEARCH GRANT APPLICANTS
We are currently seeking additional grant applicants that can help us obtain this ultimate goal of finding a better treatment or cure for AVM. For more information about our research or grant application process, please reach out to us by email at firstname.lastname@example.org