Protease-controlled Secretion and Display of Intercellular Signals

Overview

Journal Nat Commun

Specialty Biology

Date 2022 Feb 18

PMID 35177637

Authors

Alexander E Vlahos

Jeewoo Kang

Carlos A Aldrete

Ronghui Zhu

Lucy S Chong

Michael B Elowitz

Xiaojing J Gao

Affiliations

Soon will be listed here.

Abstract

To program intercellular communication for biomedicine, it is crucial to regulate the secretion and surface display of signaling proteins. If such regulations are at the protein level, there are additional advantages, including compact delivery and direct interactions with endogenous signaling pathways. Here we create a modular, generalizable design called Retained Endoplasmic Cleavable Secretion (RELEASE), with engineered proteins retained in the endoplasmic reticulum and displayed/secreted in response to specific proteases. The design allows functional regulation of multiple synthetic and natural proteins by synthetic protease circuits to realize diverse signal processing capabilities, including logic operation and threshold tuning. By linking RELEASE to additional sensing and processing circuits, we can achieve elevated protein secretion in response to "undruggable" oncogene KRAS mutants. RELEASE should enable the local, programmable delivery of intercellular cues for a broad variety of fields such as neurobiology, cancer immunotherapy and cell transplantation.

Citing Articles

Orthogonalized human protease control of secreted signals.

Aldrete C, Call C, SantAnna L, Vlahos A, Pei J, Cong Q Nat Chem Biol. 2025; .

PMID: 39814991 DOI: 10.1038/s41589-024-01831-x.

AND-gated protease-activated nanosensors for programmable detection of anti-tumour immunity.

Sivakumar A, Phuengkham H, Rajesh H, Mac Q, Rogers L, Silva Trenkle A Nat Nanotechnol. 2025; .

PMID: 39753733 DOI: 10.1038/s41565-024-01834-8.

Orthogonalized human protease control of secreted signals.

Aldrete C, Call C, SantAnna L, Vlahos A, Pei J, Cong Q bioRxiv. 2024; .

PMID: 39484520 PMC: 11526856. DOI: 10.1101/2024.01.18.576308.

Perspectives on Synthetic Protein Circuits in Mammalian Cells.

Aldrete C, An C, Call C, Gao X, Vlahos A Curr Opin Biomed Eng. 2024; 32.

PMID: 39372446 PMC: 11448451. DOI: 10.1016/j.cobme.2024.100555.

Light-triggered protease-mediated release of actin-bound cargo from synthetic cells.

Akter M, Moghimianavval H, Luker G, Liu A bioRxiv. 2024; .

PMID: 39314483 PMC: 11419145. DOI: 10.1101/2024.09.15.613133.

References

Cheng A, Lu T . Synthetic biology: an emerging engineering discipline. Annu Rev Biomed Eng. 2012; 14:155-78. DOI: 10.1146/annurev-bioeng-071811-150118. View

Lim W . Designing customized cell signalling circuits. Nat Rev Mol Cell Biol. 2010; 11(6):393-403. PMC: 2975372. DOI: 10.1038/nrm2904. View

Siciliano V, DiAndreth B, Monel B, Beal J, Huh J, Clayton K . Engineering modular intracellular protein sensor-actuator devices. Nat Commun. 2018; 9(1):1881. PMC: 5951936. DOI: 10.1038/s41467-018-03984-5. View

Chen Z, Elowitz M . Programmable protein circuit design. Cell. 2021; 184(9):2284-2301. PMC: 8087657. DOI: 10.1016/j.cell.2021.03.007. View

Chen Z, Boyken S, Jia M, Busch F, Flores-Solis D, Bick M . Programmable design of orthogonal protein heterodimers. Nature. 2018; 565(7737):106-111. PMC: 6537907. DOI: 10.1038/s41586-018-0802-y. View

Fink T, Lonzaric J, Praznik A, Plaper T, Merljak E, Leben K . Design of fast proteolysis-based signaling and logic circuits in mammalian cells. Nat Chem Biol. 2018; 15(2):115-122. PMC: 7069760. DOI: 10.1038/s41589-018-0181-6. View

Gao X, Chong L, Kim M, Elowitz M . Programmable protein circuits in living cells. Science. 2018; 361(6408):1252-1258. PMC: 7176481. DOI: 10.1126/science.aat5062. View

Kojima R, Fussenegger M . Synthetic Biology: Engineering Mammalian Cells To Control Cell-to-Cell Communication at Will. Chembiochem. 2018; 20(8):994-1002. DOI: 10.1002/cbic.201800682. View

Buddingh B, Elzinga J, van Hest J . Intercellular communication between artificial cells by allosteric amplification of a molecular signal. Nat Commun. 2020; 11(1):1652. PMC: 7125153. DOI: 10.1038/s41467-020-15482-8. View

10.

Thurley K, Wu L, Altschuler S . Modeling Cell-to-Cell Communication Networks Using Response-Time Distributions. Cell Syst. 2018; 6(3):355-367.e5. PMC: 5913757. DOI: 10.1016/j.cels.2018.01.016. View

11.

Thurley K, Gerecht D, Friedmann E, Hofer T . Three-Dimensional Gradients of Cytokine Signaling between T Cells. PLoS Comput Biol. 2015; 11(4):e1004206. PMC: 4414419. DOI: 10.1371/journal.pcbi.1004206. View

12.

Francisco L, Sage P, Sharpe A . The PD-1 pathway in tolerance and autoimmunity. Immunol Rev. 2010; 236:219-42. PMC: 2919275. DOI: 10.1111/j.1600-065X.2010.00923.x. View

13.

Glen C, McDevitt T, Kemp M . Dynamic intercellular transport modulates the spatial patterning of differentiation during early neural commitment. Nat Commun. 2018; 9(1):4111. PMC: 6173785. DOI: 10.1038/s41467-018-06693-1. View

14.

Lander A . How cells know where they are. Science. 2013; 339(6122):923-7. PMC: 3932337. DOI: 10.1126/science.1224186. View

15.

Li P, Markson J, Wang S, Chen S, Vachharajani V, Elowitz M . Morphogen gradient reconstitution reveals Hedgehog pathway design principles. Science. 2018; 360(6388):543-548. PMC: 6516753. DOI: 10.1126/science.aao0645. View

16.

Roder P, Wu B, Liu Y, Han W . Pancreatic regulation of glucose homeostasis. Exp Mol Med. 2016; 48:e219. PMC: 4892884. DOI: 10.1038/emm.2016.6. View

17.

Barcellos-Hoff M, Lyden D, Wang T . The evolution of the cancer niche during multistage carcinogenesis. Nat Rev Cancer. 2013; 13(7):511-8. DOI: 10.1038/nrc3536. View

18.

Nissim L, Wu M, Pery E, Binder-Nissim A, Suzuki H, Stupp D . Synthetic RNA-Based Immunomodulatory Gene Circuits for Cancer Immunotherapy. Cell. 2017; 171(5):1138-1150.e15. PMC: 5986174. DOI: 10.1016/j.cell.2017.09.049. View

19.

Frangogiannis N . Transforming growth factor-β in tissue fibrosis. J Exp Med. 2020; 217(3):e20190103. PMC: 7062524. DOI: 10.1084/jem.20190103. View

20.

Subramanian N, Torabi-Parizi P, Gottschalk R, Germain R, Dutta B . Network representations of immune system complexity. Wiley Interdiscip Rev Syst Biol Med. 2015; 7(1):13-38. PMC: 4339634. DOI: 10.1002/wsbm.1288. View