Genetic and Epigenetic Dysregulation of CR1 is Associated with Catastrophic Antiphospholipid Syndrome (CAPS)
Listed in
This article is not in any list yet, why not save it to one of your lists.Abstract
Objective
Catastrophic antiphospholipid syndrome (CAPS), characterized by widespread thrombosis and multi-organ failure, is associated with high morbidity and mortality. We previously established complement activation as a pathogenic driver of CAPS and identified rare germline variants in complement-regulatory genes including Complement Receptor 1 ( CR1 ) in 50% of CAPS.
Methods
We quantified CR1 expression by flow cytometry across hematopoietic cell types. CRISPR/Cas9 genome editing of TF-1 (erythroleukemia) cells was performed to generate CR1 “knock-out” and “knock-in” lines with patient-specific CR1 variants. Multiomics analysis was performed to investigate the role of methylation in CR1 expression in patients with reduced CR1 expression. Functional impact of low CR1 expression was assessed by complement-mediated cell killing using modified Ham (mHam) assay, cell-bound complement degradation products through flow cytometry and circulatory immune complexes (CIC) in serum samples through ELISA.
Results
CR1 expression in erythrocytes was markedly reduced on CAPS erythrocytes (n=9, 21.80%) compared to healthy controls (HC; n=32, 82.40%), with promoter hypermethylation emerging as a plausible epigenetic mechanism for CR1 downregulation. A novel germline variant ( CR1- V2125L; rs202148801 ) mitigated CR1 expression and increased complement-mediated cell death of knock-in cell lines. Erythrocytes from the patient with the CR1- V2125L variant had low CR1 expression. Levels of CIC, which are bound and cleared by CR1 on erythrocytes, were higher in acute CAPS (n=3, 25.55 µg Eq/ml) than healthy controls (n=3, 7.445 µg Eq/ml). Five patients were treated with C5 inhibition which mitigated thrombosis.
Conclusion
Genetic or epigenetic-mediated CR1 deficiency is a potential hallmark of CAPS and predicts response to C5 inhibition.
