Coiled-Coil Domain Containing 9 is a novel gene located beside C5aR1/R2 on murine chromosome 7. This proximal relationship is mirrored on human chromosome 19, making this research translational and relevant to the medical field. CCDC9-/- mice were created through a contract research organization utilizing CRISPR/Cas9 gene editing. Necropsy, gross examination, hematology, and clinical chemistry analysis were conducted to ensure that the global knockout of the CCDC9 gene did not induce any abnormalities in the development of the mice. Bone Marrow Derived Macrophages and Thioglycolate Elicited Peritoneal Macrophages were utilized from wild type and CCDC9-/- mice to explore the cytokine response from these macrophages when stimulated with various pattern recognition receptor agonists. Poly (I:C) and LPS were the two agonists that led to the most significant difference in cytokine release. Interactions with Poly (I:C), a synthetic double-stranded RNA that activates anti-viral RNA sensing immune responses, were targeted to elucidate how CCDC9 may be interacting with RNA. Meanwhile, interactions with LPS demonstrated that the role of CCDC9 is not specific to RNA-sensing but plays a modulatory role in the TLR4 and LPS pathway. These agonists were then paired with complement anaphylatoxin, C5a, and stimulated TEPM to explore the proximal relationship between CCDC9 and C5aR1/R2. The pairing with Poly (I:C) did not reveal any significant changes, but the pairing with LPS lead to a cytokine decrease that was not mirrored in CCDC9-/- macrophages. In-vivo applications were established in which wild type and CCDC9 -/- mice were injected intraperitoneally with LPS and Poly (I:C) to investigate the cytokine release. CCDC9-/- mice and cells consistently displayed a greater inflammatory cytokine induction when encountering stimulators of viral and bacterial pathogen sensing, suggesting that CCDC9 plays a role in immune function. / 2025-03-07T00:00:00Z
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/48366 |
| Date | 08 March 2024 |
| Creators | Pesta, Melissa M. |
| Contributors | Bosmann, Markus, Duffy, Elizabeth R. |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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