Building de novo signaling pathways for multi-stage smart delivery systems in cancer

It has now been established that relapse of prostate cancers can be significantly mitigated by
targeting and inhibiting the androgen receptor in tumours. Critical to this process is the delivery of
corresponding inhibitory peptides but to date this has proved problematic. This is because direct
exposure of protein to the carcinoma environment leads to rapid breakdown through protease
driven action. Here, we propose to develop a unique multi-stage vesicle delivery system, which
differs substantially from previous philosophies as it does not rely on disassembly of the vesicle for
release of its cargo but instead capitalizes upon the construction of the first de novo
mechanosensitive signalling pathway inside artificial cells (ACs) and communication
between real cells and artificial cells. This will be achieved by constructing multi-layered vesicle
machines that are capable of transducing signals from the environment (prostate cancer cells) to
the surface of an AC and then onto the inner lumen of the AC inside through a series of user-
defined protein-protein interactions. These do not require direct protein-protein contacts and are
instead mediated through the recently discovered phenomenon of membrane mediated protein-
protein interactions. As the signal propagates into the AC from the cancer it triggers the opening of
embedded large-pore channels in different layers of the AC, allowing onboard therapeutic agents to
be released in bursts or different agents to be released in series. The channels are based around
the mechanically sensitive channels of large conductance (MscL). MscL has shown a robust and
readily overexpressed (~mg) membrane channel protein that exhibits a wide range of large pores
(up to 40Å) whose size range can be carefully controlled. Once it is embedded in a lipid bilayer, it
has been shown to be sensitive (and opens) to membrane asymmetry (figure 1). One method of
generating asymmetry is to use phospholipase A1 or A2 (PLA). PLA rapidly alters phospholipid
structure by converting them into lysophospholipid (single chained) and the concomitant free fatty
acid, which generates asymmetry when only acting from one side of the bilayer. We have previously
demonstrated that through this asymmetry generation, it is possible to engineer MscL-PLA
communication in analogue and digital format (Charalambous et al. JACS, 2012), a phenomenon
we aim to exploit in this project as it is now well understood that PLA is aberrantly expressed by a
wide variety of tumour cells including prostate cancer cells. The function of the resultant ACs will
depend on the delicate interplay of lipid-lipid, lipid-protein and membrane mediated protein-protein
interactions and new platform technologies for fabricating multi-layered vesicle constructs.

Addressing UK skills demand: The most important impact of the CDT will be to train a new generation of Chemical Biology PhD graduates (~80) to be future leaders of enterprise, molecular technology innovation and translation for academia and industry. They will be able to embrace the life science's industrialisation thereby filling a vital skills gap in UK industry. These students will be able to bridge the divide between academia/industry and development/application across the physical/mathematical sciences and life sciences, as well as the human-machine interfaces. The technology programme of the CDT will empower our students as serial inventors, not reliant on commercial solutions.
CDT Network-Communication & Engagement: The CDT will shape the landscape by bringing together >160 research groups with leading players from industry, government, tech accelerators, SMEs and CDT affiliates. The CDT is pioneering new collaboration models, from co-located prototyping warehouses through to hackathons-these will redefine industry-academic collaborations and drive technology transfer.
UK plc: The technologies generated by the CDT will produce IP with potential for direct commercial exploitation and will also provide valuable information for healthcare and industry. They will redefine the state of the art with respect to the ability to make, measure, model and manipulate molecular interactions in biological systems across multiple length scales. Coupled with industry 4.0 approaches this will reduce the massive, spiralling cost of product development pipelines. These advances will help establish the molecular engineering rules underlying challenging scientific problems in the life sciences that are currently intractable. The technology advances and the corresponding insight in biology generated will be exploitable in industrial and medical applications, resulting in enhanced capabilities for end-users in biological research, biomarker discovery, diagnostics and drug discovery.
These advances will make a significant contribution to innovation in UK industry, with a 5-10 year timeframe for commercial realisation. e.g. These tools will facilitate the identification of illness in its early stages, minimising permanent damage (10 yrs) and reducing associated healthcare costs. In the context of drug discovery, the ability to fuse the power of AI with molecular technologies that provide insight into the molecular mechanisms of disease, target and biomarker validation and testing for side effects of candidates will radically transform productivity (5-10 yrs). Developments in automation and rapid prototyping will reduce the barrier to entry for new start-ups and turn biology into an information technology driven by data, computation and high-throughput robotics. Technologies such as integrated single cell analysis and label free molecular tracking will be exploitable for clinical diagnostics and drug discovery on shorter time scales (ca.3-5 yrs).
Entrepreneurship & Exploitation: Embedded within the CDT, the DISRUPT tech-accelerator programme will drive and support the creation of a new wave of student-led spin-out vehicles based on student-owned IP.
Wider Community: The outreach, responsible research and communication skill-set of our graduates will strengthen end-user engagement outside their PhD research fields and with the general public. Many technologies developed in the CDT will address societal challenges, and thus will generate significant public interest. Through new initiatives such as the Makerspace the CDT will spearhead new citizen science approaches where the public engage directly in CDT led research by taking part in e.g hackathons. Students will also engage with a wide spectrum of stakeholders, including policy makers, regulatory bodies and end-users. e.g. the Molecular Quarter will ensure the CDT can promote new regulatory frameworks that will promote quick customer and patient access to CDT led breakthroughs.