Transcriptome comparison of monocytes, activated CD4 T cells, and activated CD8 T cells in healthy human peripheral blood

Molecular tether-mediated extracellular targeted protein degradation (ePTD) emerges as a promising drug modality. The existing strategies for ePTD have exploited several membrane degrader proteins. However, the current panel of ePTD degraders clearly needs to be expanded, as a given degrader may present tissue-dependent expression and activity. Based on screening of >50 receptors on endocytic rates and on their tissue distribution, we established a resource of potential endocytic carriers that may mediate effective ePTD. We subsequently introduced an adaptable design to assemble “Selected endocytic carrier-targeting chimera (SecTAC)” based on bispecific antibody constructs. The resultant modular chimeras, by co-opting newly identified degraders, directed efficient internalization of extracellular protein cargoes (or nucleic acids). Importantly, shaped by the availability of the corresponding endocytic carrier, the output of a SecTAC could present cell subset selectivity. We also validated the therapeutic potential of SecTAC agents for targeting several cell membrane proteins in tumor cells and primary T cells. Furthermore, we adapted the platform to establish multi-cargo tethers that triggered simultaneous degradation of two cell surface proteins in primary T cells. Taken together, our SecTAC platform has laid the foundation for development of preclinical drug candidates capable of inducing extracellular and membrane target degradation in desirable population of cells. Monocytes were directly isolated from human whole blood by immunomagnetic negative selection using EasySep™ Direct Human Monocyte Isolation Kit (STEMCELL). CD3+ T cells were isolated from PBMCs and activated and proliferated in vitro for 3 days. Total RNA was isolated and perform RNAseq services.The data were mainly used to compare gene expression differences between different cell populations