Innovating Protein Nanotechnologies for Cancer Analysis and Immune Activation
Co-workers: Michael Brenner, Karl Brune, Can Buldun, Michael Jacobsen, Anthony Keeble, Anselm Schneider, Christopher Schoene, Antonio Serrano, Linda Tchatchouang,
Our research involves manipulating and modifying components of living organisms, to generate molecular tools with desirable activities. Inspired by extraordinary molecular features from the natural world, we are developing new approaches for controlling cell behaviour, vaccine development, and disease diagnosis.
Circulating tumour cells and engineering antibody technologies
Capturing the circulating tumour cells (CTCs) from blood samples is one of the most promising approaches to enable early diagnosis of cancer. CTC capture is already allowing rapid feedback on how a patient is responding to therapy and how the tumour is evolving. Existing magnetic CTC isolation approaches only capture the cells expressing high levels of tumour marker (EpCAM and HER2). We have shown how improving antibody affinity and cholesterol loading make it possible to recover low-expressing cancer cells. To improve further the isolation of CTCs, we are developing antibody technologies with new protein architectures and modes of target binding. This includes antibodies that form covalent bonds to cell-surface targets.
A new generation of protein interactions- Superglue from bacteria
We have harnessed a remarkable feature of adhesion proteins on the pathogenic bacterium Streptococcus pyogenes. This enabled us to achieve an irreversible covalent bond between genetically-encoded protein and peptide partners. This bond is stable over time, at high temperatures, and against the forces in biological systems (blood flow, cell migration, molecular motors). Our latest interaction, SpyTag with SpyCatcher, is the strongest protein interaction yet measured and is finding wide application in basic research and biotechnology. We are extending this family of protein interactions, through protein design, single-molecule force spectroscopy, X-ray crystallography, and advanced Mass Spectrometry, to create new possibilities for synthetic biology:
- protein tentacles for Circulating Tumour Cell capture, built using SpyLigase
- protein dendrimers to broaden the detection of anti-cancer immune responses
- SpyRing cyclised enzymes for robust diagnostic devices.
Vaccine development: Immuno-engineering
We have established an approach to accelerate vaccine development, through our Plug-and-Display technology. A limiting factor in generating vaccines is the time and effort required to turn a promising antigen into a highly immunogenic assembly. We showed rapid and highly efficient decoration of virus-like particles, which elicited a strong immune response even with only a single injection. We have shown the potential towards malaria vaccination. Our future work will explore this route for immunisation against cancer and different infectious diseases. Through further engineering of biological nanoparticles, we will investigate the principles required to surpass the efficacy of current vaccine systems, in terms of long-lasting protection and overcoming immune evasion.
Protein team-building to enhance the control of cell behaviour
Cells integrate multiple signals from their environment. These signals often have precise spatial organisation. Therapeutics usually target a single receptor, but targeting multiple receptors should enhance potency and selectivity. We have developed a modular approach to programme polyprotein teams. This enhanced the apoptosis activation of cancer cells by identifying a specific team combination binding to Death Receptor and Growth Factor receptors. We are recruiting new kinds of ligands into these teams and devising alternative team formations. Polyproteins should provide a new route to discover the spatial requirements for a wide range of signalling events. In particular we are using polyprotein teams to understand and control T cell activation, towards the goal of developing therapeutics to enhance immune-mediated cancer killing.
- Veggiani G, Nakamura T, Brenner MD, Gayet RV, Yan J, Robinson CV, Howarth M. Programmable polyproteams built using twin peptide superglues. Proc Natl Acad Sci U S A. 2016 Feb 2;113(5):1202-7
- Brune KD, Leneghan DB, Brian IJ, Ishizuka AS, Bachmann MF, Draper SJ, Biswas S, Howarth M. Plug-and-Display: decoration of Virus-Like Particles via isopeptide bonds for modular immunization . Sci Rep. 2016 Jan 19;6:19234
- Fierer JO, Veggiani G, Howarth M. SpyLigase peptide-peptide ligation polymerizes affibodies to enhance magnetic cancer cell capture . Proc Natl Acad Sci U S A. 2014 111(13):E1176-81
- Jain J, Veggiani G, Howarth M. (2013) Cholesterol loading and ultrastable protein interactions determine the level of tumor marker required for optimal isolation of cancer cells . Cancer Research 73(7):2310-21
- Aleksic T, Chitnis M, Perestenko O, Gao S, Thomas P, Turner G, Protheroe A, Howarth M, Macaulay V. (2010) The type I IGF receptor translocates to the nucleus of human tumor cells. Cancer Research 70(16):6412-9
Figure 1: Magnetic isolation of cancer cells
Figure 2: Enhancing one of the strongest non-covalent interactions in nature- crystal structure of traptavidin
Figure 3: Surviving a molecular car-crash. An ultra-stable variant of streptavidin (red and blue) survives collision by the motor protein FtsK (yellow and black) moving along biotinylated DNA at 5 kilobases per second
Figure 4: Schematic of our approach to accelerate vaccine development using SpyTag/SpyCatcher technology.
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