Denunded HUVEC/HUASMC spheroids vs. HUASMC spheroids

Here, we investigated changes of the VSMC transcriptome by utilizing 3D human vascular organoids organized as a core of VSMCs enclosed by a monolayer of ECs. Unbiased microarray-based analyses indicated that interaction with ECs for 48 hours down-regulates the VSMC expression of genes controlling rate-limiting steps of the cholesterol biosynthesis such as HMGCR, HMGCS1, DHCR24 and DHCR7. Protein analyses revealed a decrease in the abundance of 24-dehydrocholesterol reductase and lower cholesterol levels in VSMCs co-cultured with ECs. On the functional level, the blockade of the DHCR24 activity impaired spreading, migration and proliferation of VSMCs. Collectively, these findings indicate that ECs have the capacity to instruct VSMCs to shut down the expression of DHCR24 thereby limiting their capacity for cholesterol biosynthesis which may support their functional steady state. 3 samples of denunded HUVEC/HUASMC spheroids (80-90% of the HUVEC were removed) and 3 samples of HUASMCs spheroids were processed for RNA extraction