Session Type: Poster Session (Tuesday)
Session Time: 9:00AM-11:00AM
Background/Purpose: Systemic lupus erythematosus is an autoimmune disease characterized by chronic inflammation. We have previously shown that TLR7 and TLR8 are significantly upregulated in PBMCs of Lupus patients. Recent studies have discovered that miR-21, mir-29a, and miR-29b packaged and secreted in extracellular vesicles (EVs) can also bind to these receptors.
Methods: Twenty four lupus patients meeting the revised ACR criteria and 15 healthy controls provided informed consent to participate in this IRB-approved study. Plasma- and urine-derived EVs were isolated from 16 lupus patients including lupus nephritis (LN) and 6 healthy subjects by differential ultracentrifugation and validated by Nanosight and ELISA. Moreover, urine and plasma EVs from an additional 5 LN patients and 9 healthy subjects were subjected to RNA sequencing to identify RNA species including lnRNA, miRNA, and mRNA. A novel human-mouse chimeric model for testing therapeutics in SLE was created by adoptively transferring PBMCs from 3 active lupus patients into immunodeficient mouse recipients. Prior to transfer, PBMCs were incubated with synthetic liposomal EVs containing miR antagonists to miR-21, mir-29a, and miR-29b, or a control. After 21 days, PBMCs were collected for immunophenotyping and ELISA; tissues were collected for histopathological analysis by H&E staining and immunohistochemistry (IHC) for human CD3.
Results: There was a significant upregulation of EVs detected in the plasma of lupus patients relative to healthy controls. Additionally, the lupus nephritis patients that had RNAseq data resulted in a collection of statistically significant small RNA reads, such as miR-142-3p and let-7b-5p. The unique RNA signature observed by volcano plots showed multiple RNA species significantly upregulated and downregulated in lupus nephritis plasma samples. Furthermore, principal component analysis of both plasma and urine derived EVs revealed distinct lupus nephritis and healthy populations. Finally, in the adoptive transfer model, human CD4+, CD8+, B-cells, monocytes, and NK cells were successfully recovered from whole blood of chimeric mice at similar levels, but levels of human IL-2, IL-6, IL-10, and TNF-α were reduced with miR inhibition. Moreover, miR inhibition significantly reduced the inflammatory histopathology in the small intestine, liver, and kidney, demonstrated by H&E and digital quantitation of positive human CD3 IHC staining.
Conclusion: Our data shows elevated levels of EVs in lupus patients and reveals unique EV-derived RNA signatures in both urine and plasma that may be targeted therapeutically or used as diagnostic biomarkers for Lupus.
To cite this abstract in AMA style:Koenig T, Young N, Valiente G, Okafor I, Schwarz E, Harb P, Henry C, Jablonski K, Wu L, Roberson E, Jarjour W. RNA Sequencing of Plasma and Urine-derived Extracellular Vesicles from Lupus Nephritis Patients Identifies Disease-associated Small RNA Signatures and Putative Therapeutic Targets [abstract]. Arthritis Rheumatol. 2019; 71 (suppl 10). https://acrabstracts.org/abstract/rna-sequencing-of-plasma-and-urine-derived-extracellular-vesicles-from-lupus-nephritis-patients-identifies-disease-associated-small-rna-signatures-and-putative-therapeutic-targets/. Accessed August 5, 2020.
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ACR Meeting Abstracts - https://acrabstracts.org/abstract/rna-sequencing-of-plasma-and-urine-derived-extracellular-vesicles-from-lupus-nephritis-patients-identifies-disease-associated-small-rna-signatures-and-putative-therapeutic-targets/