Periodic Reporting for period 3 - ElectroGene (Electrogenetics – Shaping Electrogenetic Interfaces for Closed-Loop Voltage-Controlled Gene Expression) Reporting period: 2021-11-01 to 2023-04-30 Summary of the context and overall objectives of the project Physical objects equipped with electronic sensors and machine-learning software are increasingly exchanging data over the internet of things to collaborate and shape our future. With their molecular structure and analog metabolic control circuits taking generations to upgrade by evolution, living systems including humans are incompatible with the digital electronic world and the internet of things. Indeed, smart wearable devices profiling our habits and fitness day and night are impacting our behavior but for lack of compatibility, they have no access to control our genes and metabolism. However, despite the differences between the biologic and electronic worlds their information processing and control share some basic principles. Biological systems use ion gradients across insulating cell membranes and protein-receptor interactions to propagate analog signals, while electronic devices utilize electrons flowing through metal wires to transfer digital information. Also, biological systems can use airborne volatile molecules to communicate, while electronic systems can employ wireless broadcasting. Furthermore, single cells in a multicellular organism can simultaneously process an enormous amount of analog metabolic information in parallel at slow speed, whereas central processing units in electronic devices process digital information sequentially at enormous speed, but parallel processing requires a multi-core architecture in combination with hyper-threading technology. With the advent of the internet of things, the complexity of the digital world may be approaching that of biological systems, and electrogenetic interfaces to manage information transfer between the biological and electronic worlds may lead to novel strategies for diagnosis, treatment and prevention of diseases.ElectroGene was designed with a mission to pioneer an interface between the electronic and the genetic worlds, so that eventually electronic devices can communicate with genes and metabolism and that genes and metabolism may communicate with electronic devices, ideally using seamless closed-loop communication. By functionally interconnecting metabolism and electronics, medical conditions could be monitored and therapeutic interventions executed in real time, eventually using closed-loop control algorithms and machine learning-assisted artificial intelligence. As a non-limiting proof-of-concept example which can be extended to any medical condition, cells of the body will be engineered to report the blood-glucose concentration via the electrogenetic interface to an electronic device. This electronic information, can be stored, processed, curated by an AI-assisted algorithm or a physician and used to electrically stimulate the production and release of the therapeutic protein insulin, at the right time, dose and dynamics. In its first reporting period, ElectroGene has set the foundation for a closed-loop electrogenetic interface at several levels. For example, ElectroGene research has pioneered the genetic tools and technologies to design extremely fast molecular communication systems within the cell to come closer in matching the speed of electronic information processing. Also, ElectroGene pioneered protein degradation systems that are extremely rapid, predictable and programmable to control the timing and turnover dynamics of molecular communication systems with unprecedented precision. Most importantly, ElectroGene resulted in the prototypic design of three electrogenetic interfaces by which electronic devices could be used to control genetic activities in the body: (i) The electrothermal device, (ii) the direct electrical current-triggered gene expression and (iii) the electro-genetic diabetes control using the green LED of a commercial smartwatch or smartphone. The electrothermal device enabled therapeutic transgene expression using cooling sensation by menthol as a proxy for electro-cooling to control diabetes and muscular distrophy. The first-of-its-kind direct electro-genetic interface enables direct electrical stimulation of engineered cells via a combination of eningeered membrane channels linked to a tailored molecular signaling cascade to control the production and rapid release of insulin for the treatment of experimental type-1 diabetes. Finally, ElectroGene has set a real-world example for the potential of electrogenetic interfaces by having experimental diabetes controlled by tuning the expression and release of therapeutic proteins by subcutaneously implanted cells using percutaneous illumination by programmed smartwatches or smartphones. Capitalising on this head start, ElectroGene is set to deliver on its promises to pioneer new diagnosis, treatment and prevention strategies by creating an internet of the body and connecting the human metabolism to the internet of things. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far We have designed the first bioelectronic interface that uses wireless-powered electrical stimulation of human cells to promote the release of insulin. The human cells were genetically engineered to respond to electrically induced membrane depolarization by rapidly releasing insulin from intracellular storage vesicles. A bioelectronic device that incorporates the cells can be wirelessly triggered by a field generator. When subcutaneously implanted in diabetics mice, the device could be triggered to restore normal blood glucose levels. Bioelectronic devices with electrogenetic interfaces will create an internet of the body and advance the integration of human metabolism with the internet of things. This will enable unprecedented therapeutic opportunities for currently uncurable disease such as Diabetes that will soon be affecting over 10 percent of the world’s population. Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) ElectroGene will continue to pioneer and put into practice direct interfaces between the electronic and genetic worlds to realise the Internet of the body and connect human metabolism to the internet of things.Therefore a variety of electro-genetic (conversion of electric signals into genetic control) and gene-electric (conversion of genetic into electric signals) interfaces will be pioneered using iterative design cycles and be validated for the treatment of a variety of experimental medical conditions including Diabetes mellitus. Eventually, both interfaces will be combined to pioneer closed-loop metabolic control in which metabolic disturbances are monitored and recorded by electronics, the electronic information will be stored, analysed and processed and used to electronically control and correct the metabolism. Such self-sufficient closed-loop control circuits interfacing with human metabolism will create an internet of the body as well as connect human metabolism to the internet of things. Thus, ElectroGene will provide novel diagnostic, treatment and prevention options - a type of an electronic pill - that has the potential to cure mankind from major metabolic disorders. Electrogenetic Implant (Bottom View) Vesicular Secretion of Electrogenetic Beta Cells Apple Watch-controlled diabetic mouse Electrogenetic Implant (Top View) Smart Watch-Controlled Gene Expression Electrogenetic Gene Switches
