Optogenetic light-tunable extracellular matrix
Maximilian
Hörner1, 2, 3,
Natascha
Hotz1, 2, 3,
Raphael
J.
Gübeli1, 2, 3,
Erik
H.
Christen1, 3,
Katrin
Raute1, 2, 3,
Barbara
Hummel4,
Josef
Madl1, 3,
Susana
Minguet1, 3,
Winfried
Römer1, 2, 3,
Jan
Pruszak2, 3, 5,
Ritwick
Sawarkar4,
Wolfgang
W.
Schamel1, 2, 3,
Gerald
Radziwill1, 3,
Matias
D.
Zurbriggen1, 3 and
Wilfried
Weber1, 2, 3
-
1
University of Freiburg, Faculty of Biology, Germany
-
2
University of Freiburg, Spemann Graduate School of Biology and Medicine (SGBM), Germany
-
3
University of Freiburg, Centre for Biological Signalling Studies (BIOSS), Germany
-
4
Max Planck Institute of Immunobiology and Epigenetics, Germany
-
5
University of Freiburg, Emmy Noether-Group for Stem Cell Biology, Institute of Anatomy and Cell Biology, Germany
Introduction: Since stiffness of the cellular microenvironment plays an important role in various biological processes such as development, cell differentiation or tumor progression, a deeper understanding of the matrix cell interactions remains crucial. While several studies have investigated the influence of static matrix stiffness on diverse cellular processes, only few studies have addressed the impact of dynamic changes in substrate elasticity on cells. Therefore our aim was to engineer a synthetic cell matrix the stiffness of which can be fully reversibly adjusted in a biologically relevant range by light illumination.
Material and Methods: This was realized by combining light-gated molecular switches from the field of optogenetics with cell-compatible polymers from material sciences. To this end, we covalently coupled the cyanobacterial phytochrome Cph1 to an eight-arm polyethylene glycol thus forming a biohybrid hydrogel. Cph1 was produced together with its chromophore phycocyanobillin in E. coli.
Results: Illumination with red light triggers dimerization of Cph1, thereby increasing the number of crosslinks within the hydrogel and enhancing its stiffness. Vice versa, far-red light illumination induces Cph1 monomerization and decreases hydrogel stiffness. Due to the switching properties of Cph1 and the use of light as a trigger, the stiffness can be adjusted in a wavelength-specific, dose-dependent, spatially controlled and fully reversibly manner within seconds. By incorporation of RGD cell adhesion motifs our hydrogel serves as suitable matrix for primary cells as well as cell lines. We show that the light-induced stiffness and crosslink modifications can be used to control the cellular localization of the transcriptional co-activator YAP and the migration of primary T cells. Utilizing phospho-antibody microarrays and RNA-Seq technologies we are currently investigating the influence of dynamic stiffness changes on cellular signaling.
Conclusion: For the first time we engineered a synthetic extracellular matrix the stiffness of which can be fully reversibly modulated by light in order to control cellular signaling or migration. We propose that this system is a unique tool to study matrix cell interactions for obtaining deeper insights into related physiological and pathological processes.
Keywords:
Extracellular Matrix,
Hydrogel,
Light,
mechanical property
Conference:
10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.
Presentation Type:
New Frontier Oral
Topic:
Biomaterials for cellular programming
Citation:
Hörner
M,
Hotz
N,
Gübeli
RJ,
Christen
EH,
Raute
K,
Hummel
B,
Madl
J,
Minguet
S,
Römer
W,
Pruszak
J,
Sawarkar
R,
Schamel
WW,
Radziwill
G,
Zurbriggen
MD and
Weber
W
(2016). Optogenetic light-tunable extracellular matrix.
Front. Bioeng. Biotechnol.
Conference Abstract:
10th World Biomaterials Congress.
doi: 10.3389/conf.FBIOE.2016.01.00357
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Received:
27 Mar 2016;
Published Online:
30 Mar 2016.