Death-seq identifies regulators of cell death and senolytic therapies

• Published in: bioRxiv
• Main Authors: Colville, Alex, Jie-Yu, Liu, Thomas, Samantha, Ishak, Heather D, Zhou, Ronghao, Klein, Julian Dd, Morgens, David W, Armon Goshayeshi, Salvi, Jayesh S, Yao, David, Spees, Kaitlyn, Bassik, Michael C, Rando, Thomas A
• Format: Paper
• Language: English
• Published: Cold Spring Harbor Cold Spring Harbor Laboratory Press 03.04.2022; Cold Spring Harbor Laboratory
• Edition: 1.1
• Subjects: Age; Apoptosis; Cell Biology; Cell culture; Cell death; CRISPR; DIABLO protein; Enhancers; Genetic screening; Molecular modelling; Neurodegeneration; Patent applications; Positive selection; Senescence; Synergism; Therapeutic targets; Thrombocytopenia; senescence; CRISPR; genome-wide; Death-seq; cell death; pulmonary fibrosis; synthetic lethality; senolytics; positive selection; Death screen
• ISSN: 2692-8205; 2692-8205
• DOI: 10.1101/2022.04.01.486768

Selectively ablating senescent cells ("senolysis") is an evolving therapeutic approach for age-related diseases. Current senolytics are limited to local administration by potency and side effects. While genetic screens could identify senolytics, current screens are underpowered for identifying genes that regulate cell death due to limitations in screen methodology. Here, we establish Death-seq, a positive selection CRISPR screen optimized to identify enhancers and mechanisms of cell death. Our screens identified synergistic enhancers of cell death induced by the known senolytic ABT-263, a BH3 mimetic. SMAC mimetics, enhancers in our screens, synergize with ABT-199, another BH3 mimetic that is not senolytic alone, clearing senescent cells in models of age-related disease while sparing human platelets, avoiding the thrombocytopenia associated with ABT-263. Death-seq enables the systematic screening of cell death pathways to uncover molecular mechanisms of regulated cell death subroutines and identify drug targets for diverse pathological states such as senescence, cancer, and neurodegeneration. Competing Interest Statement A.C. and T.A.R. have filed a patent application related to the subject matter of this paper. A.C. was formerly a paid consultant for Maze Therapeutics and Rubedo Life Sciences during this work.