Control of gene expression in engineered mammalian cells with a programmable shear-stress inducer
© 2021 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC..
In humans, cellular mechanoperception serves as the basis of touch sensation and proprioception, contributes to the proper programming of cell fate during embryonic development, and plays a pivotal role in the development of mechanosensitive tissues. Molecular mechanoreceptors can respond to their environment by mediating transient adjustments of ion homeostasis, which subsequently trigger calcium-dependent alteration of gene expression via specific signaling pathways such as the nuclear factor of the activated T-cells pathway. Although, mechanoreceptors are potential drug targets for various diseases, current techniques to study mechanically gated processes are often based on custom-tailored microfluidic systems, which require special setups or have limited throughput. Here, we present a platform to characterize shear-stress-triggered, calcium-mediated gene expression, which employs a programmable, 96-well-format, shear-stress induction device to examine the effects of imposing various mechanical loads on mammalian adherent cell lines. The presented method is suitable for high-throughput experiments and provides a large tunable parameter space to optimize conditions for different cell types. Our findings indicate that the device is an effective tool to explore conditions in terms of frequency, intensity, intervals as well as extracellular matrix composition alongside the evaluation of different combinations of mechanosensitive proteins for mechanically activated gene expression. We believe our results can serve as a platform for further investigations into shear stress-controlled gene expression in basic research and drug screening.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2021 |
|---|---|
| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:118 |
|---|---|
| Enthalten in: |
Biotechnology and bioengineering - 118(2021), 12 vom: 15. Dez., Seite 4751-4759 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Strittmatter, Tobias [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
Calcium |
|---|
| Anmerkungen: |
Date Completed 28.02.2022 Date Revised 21.07.2022 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1002/bit.27939 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM330488643 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM330488643 | ||
| 003 | DE-627 | ||
| 005 | 20231225211406.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231225s2021 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1002/bit.27939 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1101.xml |
| 035 | |a (DE-627)NLM330488643 | ||
| 035 | |a (NLM)34506645 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Strittmatter, Tobias |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Control of gene expression in engineered mammalian cells with a programmable shear-stress inducer |
| 264 | 1 | |c 2021 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 28.02.2022 | ||
| 500 | |a Date Revised 21.07.2022 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2021 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC. | ||
| 520 | |a In humans, cellular mechanoperception serves as the basis of touch sensation and proprioception, contributes to the proper programming of cell fate during embryonic development, and plays a pivotal role in the development of mechanosensitive tissues. Molecular mechanoreceptors can respond to their environment by mediating transient adjustments of ion homeostasis, which subsequently trigger calcium-dependent alteration of gene expression via specific signaling pathways such as the nuclear factor of the activated T-cells pathway. Although, mechanoreceptors are potential drug targets for various diseases, current techniques to study mechanically gated processes are often based on custom-tailored microfluidic systems, which require special setups or have limited throughput. Here, we present a platform to characterize shear-stress-triggered, calcium-mediated gene expression, which employs a programmable, 96-well-format, shear-stress induction device to examine the effects of imposing various mechanical loads on mammalian adherent cell lines. The presented method is suitable for high-throughput experiments and provides a large tunable parameter space to optimize conditions for different cell types. Our findings indicate that the device is an effective tool to explore conditions in terms of frequency, intensity, intervals as well as extracellular matrix composition alongside the evaluation of different combinations of mechanosensitive proteins for mechanically activated gene expression. We believe our results can serve as a platform for further investigations into shear stress-controlled gene expression in basic research and drug screening | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a MscL | |
| 650 | 4 | |a NFAT | |
| 650 | 4 | |a Piezo1 | |
| 650 | 4 | |a mechanoperception | |
| 650 | 7 | |a Calcium |2 NLM | |
| 650 | 7 | |a SY7Q814VUP |2 NLM | |
| 700 | 1 | |a Argast, Paul |e verfasserin |4 aut | |
| 700 | 1 | |a Buchman, Peter |e verfasserin |4 aut | |
| 700 | 1 | |a Krawczyk, Krzysztof |e verfasserin |4 aut | |
| 700 | 1 | |a Fussenegger, Martin |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Biotechnology and bioengineering |d 1976 |g 118(2021), 12 vom: 15. Dez., Seite 4751-4759 |w (DE-627)NLM000014702 |x 1097-0290 |7 nnns |
| 773 | 1 | 8 | |g volume:118 |g year:2021 |g number:12 |g day:15 |g month:12 |g pages:4751-4759 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1002/bit.27939 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 118 |j 2021 |e 12 |b 15 |c 12 |h 4751-4759 | ||