Research Paper Volume 16, Issue 17 pp 12168—12190

Figure 1. (A) Overview of the SINGULAR project and its initial motivation. Publicly available datasets for several rejuvenation interventions were analyzed in this study. With the exception of Parabiosis, analyzed from three datasets, Reprogramming, which was analyzed from two datasets, and Metformin, also used in the Rapamycin experiment condition, every condition had data from one study. SINGULAR combines a unified processing pipeline for all the datasets with three main tools to explore transcriptional regulatory networks, signaling pathways, and cell-to-cell ligand receptor interactions. (B) Example comparison of the technical heterogeneity motivating this study. UMI counts across organ systems and studies, as well as the organs sample in each of the different studies, both vary greatly. (C) Comparison of counts of unique upregulated and downregulated genes from different studies grouped at the organ level, derived using the Delegate method. Even after applying SINGULAR’s unified preprocessing pipeline, substantial heterogeneity by organ and study in the transcriptional response to rejuvenation remains.