Community Research and Development Information Service - CORDIS

FP7

Light2NanoGene Report Summary

Project ID: 624888
Funded under: FP7-PEOPLE
Country: United Kingdom

Periodic Report Summary 1 - LIGHT2NANOGENE (Cellular bioengineering by plasmonic enhanced laser nanosurgery)

The use of nanoscience technologies to either perform therapy or diagnosis at the cellular level is expected to revolutionize 21st Century medicine by opening new approaches to cure various illnesses. However, cellular bioengineering is technologically challenging and becomes feasible only when different scientific disciplines are combined together to provide advanced cellular level surgery tools. To this aim, nanosurgery (i.e., surgery on the nanoscale) employs ultrafast laser technology and/or nanoscience emerging technologies to perform cell or even nucleus surgery. The major advantage of the nanosurgery approach is the prospect to disrupt submicrometer-sized organelles within living cells or tissue without affecting the surrounding material or compromising viability of the cell or organism.
The Light2NanoGene project aims to the development and optimization of a novel femtosecond laser technique for nanosurgery of cancer cells. The technique, named plasmonic enhanced laser nanosurgery, combines the advantages of two rapidly expanding research and technological fields, namely plasmonics and ultrafast lasers, to build a versatile tool capable of performing high throughput cell nanosurgery. The main innovative goal of the project is the optical fiber integration of the plasmonic nanosurgery tool towards in-vivo (i.e., living subject) applications. In-vitro cell transfection (i.e., introduction of siRNA through the membrane of breast cancer stem cells (CSCs)) is the specific nanosurgery application of the Light2NanoGene project. The latter, is driven by the remarkable ability of these undifferentiated cells within a tumour to self-renew and promote metastases. The successful transfection of the CSCs with siRNA will silence the expression of key genes involved in their aggressive behaviour. We expect proof-of-concept elimination of their capacity for self-regeneration and induction of metastases.
The Light2NanoGene work plan is organized in 5 work packages, aiming to the development and optimization of the plasmonic nanoparticles assisted laser nanosurgery technology. Briefly, the research methodology is organized towards the accomplishment of three main objectives: (a) Selection of the most efficient plasmonic material for laser nanosurgery by employing theoretical modelling and experimental investigations of their interactions with laser pulses. (b) Intracellular delivery of exogenous material (fluorescence molecules, siRNA) into living cancer cells using optimal nanomaterials. (c) Integration of the developed technology towards a fibre-based, portable and low cost system.
So far the following results have been achieved (only published results are included):
We demonstrated with single-particle monitoring that 100 nm gold nanoparticles (AuNPs) irradiated by off-resonance femtosecond (fs) laser in the tissue therapeutic optical window (λ = 800 nm), can act as a durable nanolenses in liquid and provoke nanocavitation while remaining intact (Boutopoulos, C.; Hatef, A.; Fortin-Deschênes, M.; Meunier, M. Nanoscale 2015, 7 (27), 11758–11765). We have employed combined time-resolved shadowgraphic imaging, in situ dark field imaging, and dynamic tracking of AuNP Brownian motion to ensure the study of individual AuNPs/nanolenses under multiple fs laser pulses. We showed that 100 nm AuNPs can generate multiple, highly confined (radius down to 550 nm) and transient (life time < 50 ns) nanobubbles. The latter is of significant importance towards in vivo application of the AuNP-assisted laser nanosurgery, where AuNP fragmentation should be avoided to prevent side effects, such as cytotoxicity and immune system’s response. The experimental results have been correlated with theoretical modeling to provide an insight to the AuNP-safe cavitation mechanism as well as to investigate the deformation mechanism of the AuNPs at high laser fluence.
For the AuNP assisted laser induced intracellular delivery optimization process, we used a dye as indicator for the perforation of the cancer cells (Boutopoulos, C.; Bergeron, E.; Meunier, M. J. Biophotonics 2016, 9 (1-2), 26–31). We performed transient membrane perforation of living cancer cells using AuNPs enhanced single near infrared (NIR) femtosecond (fs) laser pulse. Under optimized laser energy fluence, single pulse treatment (τ = 45 fs, λ = 800 nm) resulted in 77% cell perforation efficiency and 90% cell viability. Using dark field and ultrafast imaging, we demonstrated that the generation of submicron bubbles around the AuNPs is the necessary condition for the cell membrane perforation. AuNP clustering increased drastically the bubble generation efficiency, thus enabling an effective laser treatment using low energy dose in the NIR optical therapeutical window. The latter can be the basis for further development of AuNP enhanced cell perforation and transfection methods towards the implementation of an effective laser therapeutic tool for safe in vivo treatments.
The reader is referred to the Light2NanoGene web site for latest news and publications related to the project. (http://cboutop.com/light2nanogene/)

Contact

Trish Starrs, (Business Development Manager)
Tel.: +44 1334 467286
Fax: +44 1334 462217
E-mail

Subjects

Life Sciences
Record Number: 187537 / Last updated on: 2016-08-22