Interplay Between Dox-Induced ICD and STING Pathway: Insights from Gene Expression in dendritic cells

In this RNA sequencing study, the primary objectives were to explore the intricate interplay between the STING pathway and ICD-associated DAMPs in the context of treatment with cGAMP nanoparticle and Dox. The research delved deep into the transcriptomic alterations in BMDCs when exposed to cGAMPnps and conditioned medium from Dox-treated 4T1 breast cancer cells. By analyzing these shifts in gene expression and conducting a gene ontology analysis, the study sought to understand the inflammatory pathways and other gene clusters influenced by these treatments. Ultimately, this insight aims to inform the development of STING-mediated immunotherapies by identifying potential DAMP targets. Overall design: 4T1 breast cancer cells were initially treated with Dox at a concentration of 2 µM. Following this treatment, the medium was replaced and, after a 24-hour interval, the conditioned medium from these cells was harvested. Parallelly, BMDCs were subjected to treatments either with cGAMPnps (1 µM) alone or combined with the aforementioned conditioned medium. Subsequent to these treatments, total RNA was meticulously extracted from the BMDCs employing the TRIzol reagent. This extracted RNA was then used to craft a sequencing library using the SureSelect XT HS2 mRNA Library Preparation Kit from Agilent. The final sequencing was conducted on the Illumina NovaSeq 6000 platform, aiming to produce 150 bp paired-end reads.