CRISPLD2 regulates adipocyte function

Obesity with accompanying adipose fibrosis is associated with worsening insulin resistance. Weight loss and adipose tissue turnover are conversly associated with improved insulin signaling and overall health. We previously found the secreted protein CRISPLD2 up-regulated in adipocytes during weight loss; CRISPLD has been suggested regulate inflammation and tissue turnover. As loss of CRISPLD2 is embryonic lethal, we developed mice with adipocyte-specific inducible CRISPLD2 overexpression to explore its function. Single-cell RNAseq analysis of the stromal vascular fraction (SVF) during long-term adipocyte CRISPLD2 overexpression demonstrated broad down-regulation of collagen transcription across multiple cell types; we found similar down-regulation of fibrosis via picosirius red staining of adipose tissue overexpressing CRISPLD2, along with a decrease in adipocyte cell size in chow-fed animals. Overall design: scRNAseq of Mouse stromal vascular fraction from control and CRISPLD2 overexpression mice. This was performed on 2 males of each genotype, Tg and Double Tg. A double transgenic mouse was developed by crossing mice carrying a the inducible crispld2 plasmid with C57BL/6-Tg(Adipoq-rtTA)2Zvw/J mice from Jackson lab (RRID:IMSR_JAX:033448) (CLD2AD). Single transgenic mice carrying only the TRE-mCLD2 served as the control group. These mice were fed Teklad Rodent Diet plus doxycycline to induce the crispld2 transgene. Single Cell experiment was done on the stromal vascular fraction of these mice following isolation. Cell ranger and Parktek Software was used to visualize the data,, quality control was performed then cell types were identified and additioanl analsis of gene expression in those populations was analyzed.