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Investigating Target Gene Function in a CD40 Agonistic Antibody-induced Colitis Model using CRISPR/Cas9-based Technologies
Published on: June 2, 2021
Endogenous antisense RNA curbs CD39 expression in Crohn's disease
Rasika P Harshe1, Anyan Xie2, Marta Vuerich1
1Department of Anesthesia, Critical Care & Pain Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, Boston, MA, 02215, USA.
A novel antisense RNA regulates CD39, an enzyme crucial for immune suppression. Increased antisense in Crohn's disease patients suggests a therapeutic target for restoring immune balance.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- CD39 (an ectonucleotidase) converts nucleotides to immunosuppressive adenosine.
- CD39 is expressed on regulatory T (Treg) and T-helper (Th) 17 cells, modulating immune responses.
- Known CD39 regulation involves genetic variations and signaling pathways.
Purpose of the Study:
- To investigate a novel regulatory mechanism of CD39 involving endogenous antisense RNA.
- To assess the role of CD39-specific antisense RNA in Crohn's disease pathogenesis.
- To explore therapeutic potential of targeting this antisense RNA.
Main Methods:
- Analysis of CD39-specific antisense RNA levels in patient immune cells.
- Investigation of antisense RNA localization and interaction with nuclear proteins (nucleolin, hnRNP-A1).
- In vitro antisense silencing and in vivo colitis model in humanized mice.
Main Results:
- A CD39-specific antisense RNA, transcribed from the gene's 3'-end, was identified.
- Increased antisense RNA was observed in Treg and Th17 cells from Crohn's disease patients.
- Antisense silencing led to CD39 upregulation and improved colitis symptoms in mice.
Conclusions:
- Endogenous antisense RNA represents a new mechanism for CD39 regulation.
- CD39-specific antisense RNA is implicated in Crohn's disease.
- Targeting this antisense RNA could restore CD39 function and immune homeostasis in Crohn's disease.
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