Light INduced Cell control by Exogenous organic semiconductors

Project Information

LINCE

Grant agreement ID: 803621

DOI

10.3030/803621

Project closed

EC signature date 12 December 2018

Start date 1 March 2019

End date 31 August 2025

Funded under

• EXCELLENT SCIENCE - European Research Council (ERC)

Total cost

€ 1 866 250,00

EU contribution

€ 1 866 250,00

Coordinated by

FONDAZIONE ISTITUTO ITALIANO DI TECNOLOGIA
Italy

Periodic Reporting for period 4 - LINCE (Light INduced Cell control by Exogenous organic semiconductors)

Reporting period: 2023-09-01 to 2025-08-31

Summary of the context and overall objectives of the project

LINCE developed light-sensitive devices based on novel functional materials for optical regulation of living cells functions.
The possibility to control the activity of biological systems is a timeless mission for neuroscientists, since it allows both to understand specific functions and to manage dysfunctions. Optical modulation provides, respect to traditional electrical methods, unprecedented spatio-temporal resolution, lower invasiveness, and higher selectivity. However, the vast majority of animal cells does not bear specific sensitivity to light. Search for new materials capable to optically regulate cell activity is thus an extremely hot topic. LINCE focused on organic semiconductors as ideal candidates, since they are inherently sensitive to visible light and highly biocompatible, sustain both ionic and electronic conduction, can be functionalized with biomolecules and drugs. The application of organic semiconductors in biotechnology was significantly broadened within LINCE lifespan, and open issues of high biological relevance, in both neuroscience and regenerative medicine, could be successfully addressed by taking advantage of the proposed approach.
In particular, LINCE demonstrated new devices for: (i) regulation of astrocytes functions, active in many fundamental processes of the central nervous system and in pathological disorders; (ii) effective control of cell migration and maturation processes, by acting on intracellular redox metabolism in a non-detrimental manner, in several cell models, relevant form a therapeutic point of view, including endothelial progenitors, epithelial cells, adipose stem cells and neural progenitors; (iii) control of tissue regeneration and animal behavior; device biocompatibility and efficacy in therapeutically relevant 3D systems and in vivo was successfully reported.
LINCE tools are sensitive to visible and NIR light, flexible, biocompatible, and easily integrated with any standard physiology set-up. They combine electrical, chemical and thermal stimuli, offering high spatio-temporal resolution, reversibility, specificity and yield. The combination of all these features is still not achievable by currently available technologies, and sizable impact is expected in the application to cell-based therapies, implantable organoids, and overall in the field of regenerative medicine.
In brief, LINCE made available to neuroscientists and medical doctors unprecedented tools for both in vitro and in vivo investigation, based on optical stimulation of biocompatible materials while avoiding the need for genetic manipulation.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

LINCE specifically targeted optical modulation of cells belonging to the huge class of non-excitable cells, because in this field the need for spatio-temporally selective, high throughput and highly effective methods to gain reversible cell activity modulation is extremely urgent. By definition, non-excitable cells are characterized by the inability to generate an all-or-none action potential in response to external stimuli. With the exception of neuron, muscle, and some endocrine cells, all other cells in the body are non-excitable.
During LINCE lifespan, all targeted objectives were successfully achieved, and in particular:
1. Phototransduction mechanisms governing the interface between photoactive materials and in vitro cells were elucidated and parametrized; different contributing processes were identified over relevant timescales from a biological point of view and modeled;
2. Polymer beads and nanostructured architectures able to maximize phototransduction efficiency were implemented and ad hoc engineered, according to specific needs/requirements of the targeted biological models;
3. Microstructured devices were engineered, used as highly biocompatible and optically active substrates for modulation of primary astrocytes activity and neurospheres development (in collaboration with Dr. Valentina Benfenati at CNR, Bologna, Italy);
4. LINCE tools were proven to be highly reliable and effective in the optical modulation of the cell fate. In more detail, (i) bimodal modulation of angiogenesis was achieved in endothelial cell models (in collaboration with Prof. F. Moccia at University of Pavia, Italy); (ii) epithelial and endothelial cell migration was enhanced in both in vitro and in vivo pilot experiments; (iii) modulation of adipose stem cells physiological behaviour was obtained; (iv) modulation of specific ion channels (TRP and PIEZO1) was achieved; (v) optical control of cell maturation processes was demonstrated;
3. Optical excitation of active materials lead to effective tissue regeneration of Hydra Vulgaris invertebrate models (in collaboration with Dr. Claudia Tortiglione at CNR, Pozzuoli, Italy). This result fostered furtherstudies in the field of skin repair and wound healing applications, currently ongoing (In collaboration with Prof. P. Netti at IIT, Napoli, Italy).

Overall, the ensemble of results obtained in a variety of different biological models fully confirmed the reliability of the LINCE paradigm, and lead to the realization of several tools for optical modulation of the cell fate. In all cases, the active materials were engineered and the photostimulation protocol was adapted to the targeted application, and the phototransduction process was elucidated by taking into account not only the properties of the material but also the biological response of the model, which is highly peculiar and can vary a lot from case to case. Thanks to this cross-disciplinary approach, LINCE could develop unprecedented optoceutic tools and make them available to biologists and clinicians for further refinement, in view of therapeutic applications and fundamental studies.

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)

There is an increasing need of tools capable to selectively modulate, in a spatially and temporally selective manner, the specific functions of diverse, non-excitable cells. Ideally, new technologies should be also minimally invasive and reversible, qualities which cannot be achieved by currently available chemical or electrical cues. Even optogenetic tools, widely employed to modulate the neural activity, have been scarcely applied to non-excitable tissues, being limited by the lack of ion selectivity and inability to extend their spectral sensitivity into the NIR range.
In this framework, LINCE program coupled three main ground-breaking elements: the choice of new materials for functional bio-interfaces; the choice of exogenous optical stimulation, as the most suitable technique to provide neuroscientists, biologists and bioengineers with not invasive tools and high spatio-temporal resolution; the selection of specific open issues of high biological relevance and the choice to focus on non-excitable cells, whose precise and selective modulation is currently challenging.
Light addressable interfaces based on organic semiconductors realized by LINCE represent valuable candidates for the toolbox of the neuroscientist and the medical doctor, allowing direct, minimally invasive, selective and rapid control over in vitro cells, during their whole life-span, from genesis to differentiation, to specific function. LINCE represents a breakthrough optical strategy, providing high resolution, selectivity and yield, easily coupled to any standard physiology set-up, without making recourse to highly expensive instrumentation, and avoiding the need for viral transfer. These features, widely demonstrated and implemented along the project lifespan, can find useful exploitation in a number of different applications. Some examples include: modulation of tubulogenesis and implementation of better vascularization in a non-invasive, drug-free manner; fine modulation of the endothelial and epithelial barriers, with interesting perspectives in the study of the blood-brain barrier as well as in drug delivery; significant enhancement in cell migration and cell maturation processes, with possible outcomes in cell-based therapies and functional organoids development; sizable promotion and fastening of wound healing and tissue regeneration, with potential spin off in clinics, as well as in cosmetic and dermatology industrial sectors.
In brief, LINCE final outcomes are expected to open completely new perspectives for the use of optical stimulation not only in fundamental physiology, but also in the field of regenerative medicine and in clinics.