RT Journal Article
SR Electronic
T1 Multiscale profiling of enzyme activity in cancer
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.11.11.468288
DO 10.1101/2021.11.11.468288
A1 Ava P. Soleimany
A1 Jesse D. Kirkpatrick
A1 Cathy S. Wang
A1 Alex M. Jaeger
A1 Susan Su
A1 Santiago Naranjo
A1 Qian Zhong
A1 Christina M. Cabana
A1 Tyler Jacks
A1 Sangeeta N. Bhatia
YR 2021
UL http://biorxiv.org/content/early/2021/11/12/2021.11.11.468288.abstract
AB Diverse processes in cancer are mediated by enzymes, which most proximally exert their function through their activity. Methods to quantify enzyme activity, rather than just expression, are therefore critical to our ability to understand the pathological roles of enzymes in cancer and to harness this class of biomolecules as diagnostic and therapeutic targets. Here we present an integrated set of methods for measuring specific enzyme activities across the organism, tissue, and cellular levels, which we unify into a methodological hierarchy to facilitate biological discovery. We focus on proteases for method development and validate our approach through the study of tumor progression and treatment response in an autochthonous model of Alk-mutant lung cancer. To quantitatively measure activity dynamics over time, we engineered multiplexed, peptide-based nanosensors to query protease activity in vivo. Machine learning analysis of sensor measurements revealed dramatic protease dysregulation in lung cancer, including significantly enhanced proteolytic cleavage of one peptide, S1 (Padj < 0.0001), which returned to healthy levels within three days after initiation of targeted therapy. Next, to link these organism-level observations to the in situ context, we established a multiplexed assay for on-tissue localization of enzyme activity and pinpointed S1 cleavage to endothelial cells and pericytes of the tumor vasculature. Lastly, to directly link enzyme activity measurements to cellular phenotype, we designed a high-throughput method to isolate and characterize proteolytically active cells, uncovering profound upregulation of pro-angiogenic transcriptional programs in S1-positive cells. Together, these methods allowed us to discover that protease production by angiogenic vasculature responds rapidly to targeted therapy against oncogene-addicted tumor cells, identifying a highly dynamic interplay between tumor cells and their microenvironment. This work provides a generalizable framework to functionally characterize enzyme activity in cancer.Competing Interest StatementT.J. is a member of the Board of Directors of Amgen and Thermo Fisher Scientific and a co-founder of Dragonfly Therapeutics and T2 Biosystems; serves on the Scientific Advisory Board of Dragonfly Therapeutics, SQZ Biotech, and Skyhawk Therapeutics; and is President of Break Through Cancer. T.J.'s laboratory currently receives funding from Johnson & Johnson and The Lustgarten Foundation, but these funds did not support the research described in this manuscript. S.N.B. is a director at Vertex; is a co-founder of and consultant for Glympse Bio, Satellite Bio, and Cend Therapeutics; holds equity in Glympse Bio, Satellite Bio, Cend Therapeutics, and Catalio Capital; consults for Moderna; and receives sponsored research funding from Johnson & Johnson. Funds from Johnson & Johnson to S.N.B.'s laboratory supported the research described in this manuscript.