Established in 2016, the RVCL Research Center at Washington University is leading an effort to coordinate global research on retinal vasculopathy with cerebral leukoencephalopathy (RVCL), provide comprehensive and multidisciplinary patient care and initiate clinical studies and drug trials.
Our aims are to serve as a resource to help families, educate physicians about the disease and increase overall awareness of RVCL.
Together with collaborators from around the world, studies are underway to better understand and treat this very rare disease.
Approximately 35 unrelated families worldwide have been identified with RVCL. Sixteen families are located in the USA and others have been identified in Australia, Germany, France, the Netherlands, Spain, Mexico, Japan, Turkey, Switzerland and Italy.
This autosomal dominant disorder features frame-shift mutations in the carboxy terminus of TREX1 that produces a truncated and mislocalized protein. We call the latter process “TREX on the loose.”
The wild-type protein translated from the TREX1 gene consists of 314 amino acids (~ 32 kDa) and is encoded by a single exon on chromosome 3p21. TREX1 consists of three domains important for its role as a DNA repair enzyme (i.e., exonuclease domains I, II, and III). TREX1 also has an extended C-terminal “tail” domain of ~ 70 amino acids containing a leucine-rich sequence required for its endoplasmic reticulum (ER) localization.
As described above, RVCL results from a mutation in the tail domain that changes the protein reading frame, introduces a stop codon, and thus causes a premature termination leading to partial loss of the tail. As a result, TREX1 loses it ability to remain in the ER and becomes mislocalized throughout the cell. Of the 35 identified families worldwide with RVCL, 16 kindreds have a particular mutation identified on the protein level as V235G fs*6. This means that in TREX1 (in particular, variant 4), the amino acid valine changes to a glycine at amino acid #235. Although normal TREX1 protein is 314 amino acids long, this mutation causes the protein to terminate shortly after amino acid #235.
For reasons that are as yet unclear, most people who carry these heterozygous TREX1 tail mutations live a fairly normal life until middle age, when vision and brain damage begin. Progressive deterioration proceeds over a 5 to 15 year period. New studies conducted by our laboratory have identified that TREX1 is expressed by a subset of human brain microglia cells that also are often close to blood vessels. However, TREX1 clusters densely in microglia cells in proximity to brain lesions of RVCL patients. Further studies are needed to determine how clustered TREX1 may impact the symptoms of RVCL.
In addition to RVCL, mutations in other areas of TREX1 play major roles in a variety of neurovascular and autoimmune-related disorders such as Aicardi-Goutieres syndrome, chilblain lupus, and Cree encephalitis. TREX1 mutations also have been linked to systemic lupus erythematosus (SLE) and Sjögren’s syndrome.
There are two known functions for TREX1:
TREX1 was originally discovered and identified as the “3 prime repair exonuclease 1” that cleans up and digests DNA debris that is generated when tissue is damaged or cells die. It degrades single-stranded DNA ~4-fold more efficiently than double-stranded DNA. TREX1 is also important for handling DNA from viruses that invade and infect cells.
A newly discovered function of TREX1 is in overseeing the ‘sugar polishing’ step (i.e., glycosylation) for newly made proteins. Thus, TREX1 interacts with cellular machinery (the oligosaccharyltransferase or “OST” complex) to add N-glycans to proteins as they are produced. These glycans are important for modulating the function and stability of proteins.
When the sugar polishing function is disrupted, TREX1 no longer can bind to its partner proteins in the OST complex. This leads to the buildup of glycans. The disruption of this important interaction also may diminish blood vessel life-span and integrity and lead to disturbances in the immune system. These studies also identified an anthracycline antibiotic (Aclarubicin) that corrected the defect in both mouse disease models and in human patient cells. Studies are underway to better understand and expand on these exciting discoveries. Additionally, human clinical trials are in development.
Our research utilizes two types of models: human lymphoblast cell lines and mouse disease models. For the former, we prepare immortalized EBV-transformed patient B cells. Secondly, we and others have developed TREX1 mouse models that knock out of mouse TREX1 (TREX1-/-) or knock in human RVCL TREX1 mutation (V235fs-KI).
Parul Kothari, MD, PhD, who studied this disease at Washington University, wrote her doctoral dissertation on these studies.
Our teams have produced recombinant human TREX1 and recombinant RVCL TREX1 as well as antibodies to normal TREX1. These reagents are valuable tools to study the disease in vitro (in the test tube) as well as in vivo (in living systems).
Researchers at the RVCL Research Center continue to seek a better understanding of the disease process and to find an effective treatment to replace, bypass, correct or negate the effects of defective TREX1 protein.
For additional information on RVCL, see our review for the National Organization for Rare Disorders (NORD).