Projects / Programmes
Sonogenetics - ultrasound modulation of target gene expression
| Code |
Science |
Field |
Subfield |
| 4.06.00 |
Biotechnical sciences |
Biotechnology |
|
| Code |
Science |
Field |
| T490 |
Technological sciences |
Biotechnology |
| Code |
Science |
Field |
| 3.04 |
Medical and Health Sciences |
Medical biotechnology |
sonogenetics, ultrasound, protein based gas vesicles, synthetic calcium-dependent transcription factor based on NFAT
Researchers (16)
Organisations (1)
| no. |
Code |
Research organisation |
City |
Registration number |
No. of publicationsNo. of publications |
| 1. |
0104 |
National Institute of Chemistry |
Ljubljana |
5051592000 |
10 |
Abstract
Optogenetics, based on light as a spatially and temporally defined signal, made a powerful impact on life sciences and has enabled precise control of transcriptional responses of cells, study of cellular functions in living organism and use of light to trigger therapy of different types of disorders, from metabolic to neurodegenerative to biotechnological applications. The weak penetration of light through tissue limits its application, ultrasound can, on the other hand, also be programmed and precisely focused, and can penetrate deep into tissues, potentially enabling remote control of cells with exciting possibilities for medical and biotechnological applications and for understanding of the mechanisms of mechanosensing. Despite technological advancement and the potential of ultrasound stimulation in preclinical studies, current ultrasound manipulation of cells lacks the targeting specificity of optogenetics and coupling of cell activation to the transcriptional regulation of selected genes.
Objective of the proposal is to design the biotechnological system for the application of the ultrasound as a noninvasive deep tissue penetrating stimulus to induce the expression of target mammalian cells within the defined spatial location and trigger the transcriptional regulation of selected gene(s) within the selected region stimulated by ultrasound. We aim to construct and upgrade the innovative sonogentic tools for ultrasound stimulation, in order to introduce sonogenetics as an alternative to optogenetics.
Specifically, we plan to augment the sensitivity of mammalian cells to ultrasound by the acoustic responsive protein-based gas vesicles (GVs). The genetically encoded self-assembled protein-based gas nanostructures with acoustic and targeting properties for cell stimulation by ultrasound will be added to target cells or expressed within mammalian cells to enhance sensitivity of cells to ultrasound. Moreover, we aim to improve the ultrasound triggered transcriptional activation of mammalian cells through the engineered calcium-responsive NFAT-based transcription factor (Ca-TF). In order to enhance signal-to-background response to ultrasound, the engineered transcription activator will be anchored to the plasma membrane with the signal peptide that the anchorage depends on the membrane electrostatic potential and furthermore the nuclear transport signals (NES and NLS) within transcription factor will be altered. Finally, we plan to demonstrate in vivo ultrasound activation and implementation in medically relevant setting.
Inspired by the paradigm of optogenetics, which in many ways revolutionized synthetic biology and several other branches of life sciences, we propose to introduce the innovative enhancement of sonogenetics, leveraging ultrasound’s deep tissue penetration and spatial patterning. The main novelty of our proposal is enhancement of the sensitivity of cells to the ultrasound by repurposing the self-assembling acoustic protein gas nanostructures and engineering the calcium-dependent signaling pathway based on NFAT protein. Besides direct applications, the project will also provide an insight into the mechanisms of mechanosensing of cells. The project will provide the proof-of-concept results, which could be effectively embraced for various specific applications, for example for local in situ induction of specific growth factors by targeting the selected regions of the parkinsonian brain or to suppress chronic inflammation in neurodegenerative diseases.
Significance for science
The ability to trigger production of bioactive compounds by cells in a noninvasive manner has significant technological, therapeutic and diagnostic application. Sonogenetics has even greater translation potential than optogenetics due to the demonstrated lack of negative adverse effects in diagnostic ultrasound and primarily due to its noninvasiveness, which is probably the most important obstacle to wider therapeutic applications of optogenetics in neurologic diseases.
The project will provide proof-of-concept results, which involve tools for fast spatiotemporal stimulation of cells and local production of bioactive compounds. We plan to engineer the bacterially encoded protein-based gas vesicles to target mammalian cells and enhance ultrasound stimulation. Furthermore, to improve signal-to-background response we will develop a calcium-responsive transducing signaling pathway that couples increased cytosolic calcium ion concentration to the engineered NFAT-based transcription activation. Our interdisciplinary approach combining the expertise from synthetic and cell biology, and immunology represents an innovative and original contribution in these very competitive disciplines and might open new therapeutic possibilities in a variety of diseases; e.g. immunotherapy, neurologic and metabolic diseases.
The interdisciplinary nature of the project opens new possibilities for international cooperation with researchers e.g. within EU programs and bilateral projects (some listed in Section 9.2) and will recruit new PhD and undergraduate students. The developed IP will be protected in form of patent applications and the formation of spin offs specialized in the sonogenetics will be considered. Furthermore, members of the research group will pass their knowledge to the scientific community through training. Results will be shared with academics (international workshops and conferences) and industrial partners and communication activities will be carried out through the project website, promotional materials, and publication of scientific papers in peer-reviewed journals (open access).
Significance for the country
The ability to trigger production of bioactive compounds by cells in a noninvasive manner has significant technological, therapeutic and diagnostic application. Sonogenetics has even greater translation potential than optogenetics due to the demonstrated lack of negative adverse effects in diagnostic ultrasound and primarily due to its noninvasiveness, which is probably the most important obstacle to wider therapeutic applications of optogenetics in neurologic diseases.
The project will provide proof-of-concept results, which involve tools for fast spatiotemporal stimulation of cells and local production of bioactive compounds. We plan to engineer the bacterially encoded protein-based gas vesicles to target mammalian cells and enhance ultrasound stimulation. Furthermore, to improve signal-to-background response we will develop a calcium-responsive transducing signaling pathway that couples increased cytosolic calcium ion concentration to the engineered NFAT-based transcription activation. Our interdisciplinary approach combining the expertise from synthetic and cell biology, and immunology represents an innovative and original contribution in these very competitive disciplines and might open new therapeutic possibilities in a variety of diseases; e.g. immunotherapy, neurologic and metabolic diseases.
The interdisciplinary nature of the project opens new possibilities for international cooperation with researchers e.g. within EU programs and bilateral projects (some listed in Section 9.2) and will recruit new PhD and undergraduate students. The developed IP will be protected in form of patent applications and the formation of spin offs specialized in the sonogenetics will be considered. Furthermore, members of the research group will pass their knowledge to the scientific community through training. Results will be shared with academics (international workshops and conferences) and industrial partners and communication activities will be carried out through the project website, promotional materials, and publication of scientific papers in peer-reviewed journals (open access).
Most important scientific results
Interim report
Most important socioeconomically and culturally relevant results
