PUBLICATIONS
LIVING THERAPEUTIC HYDROGELS THAT SAFELY ENCAPSULATE DRUG-PRODUCING BIOFACTORIES AND SUSTAIN THEIR FUNCTION LONG-TERM
A fundamental question for LTM development is how to control the growth and activity of encapsulated biofactories in a nutrient-rich environment. In the LSC we demonstrated that this can be achieved by tailoring the spatial confinement, mechanical and diffusion properties of the encapsulating materials. We identified hydrogel composition windows and physical requirements needed to maximize drug production and maintain a steady-state bacteria number with a constant drug release profile from the LTM.
Examples of technologies used in the LSC and images of LTM laboratory prototypes containing functional biofactories.
Related publications:
- Bhusari et al., Rheological behavior of Pluronic/Pluronic diacrylate hydrogels used for bacteria encapsulation in living materials. bioRxiv 10.1101/2023.03.29.534703, 2023.2003.2029.534703 (2023).
- Bhusari, S. Sankaran, A. del Campo, Regulating Bacterial Behavior within Hydrogels of Tunable Viscoelasticity. Advanced Science 9, 2106026 (2022).
- Bhusari, J. Kim, K. Polizzi, S. Sankaran, A. del Campo, Encapsulation of bacteria in bilayer Pluronic thin film hydrogels: A safe format for engineered living materials. Biomaterials Advances 145, 213240 (2023).
- Bhusari, M. Hoffmann, P. Herbeck-Engel, S. Sankaran, M. Wilhelm, and A. del Campo, Rheological Behavior of Pluronic/Pluronic Diacrylate Hydrogels Used for Bacteria Encapsulation in Engineered Living Materials, Soft Matter 20, 1320 (2024).
- E. Seyfert et al., Darobactins Exhibiting Superior Antibiotic Activity by Cryo-EM Structure Guided Biosynthetic Engineering. Angewandte Chemie International Edition 62, e202214094 (2023).
- Groß et al., Improved broad-spectrum antibiotics against Gram-negative pathogens via darobactin biosynthetic pathway engineering. Chem. Sci. 12, 11882-11893 (2021).
- Ahmed et al., Engineering of Streptomyces lividans for heterologous expression of secondary metabolite gene clusters. Microbial Cell Factories 19, 5 (2020).
LTMS CAPABLE OF ON-DEMAND DRUG DELIVERY
A unique advantage of biofactories is the possibility to integrate sensors and switches for on-demand therapeutic release. The synthetic biology expertise in this topic is provided to the LSC by the Independent Junior Group of Dr. Shrikrishnan Sankaran. The group has engineered switches for light- and heat-responsive bacterial activity in ClearColi and probiotic E. coli Nissle 1917 strains. Controlled release of the antibiotic darobactin, a pro-angiogenic vascular endothelial growth factor (VEGF)-mimetic protein and the flavonoid pinocembrin has been achieved in living hydrogels. Novel genetic parts for the probiotic L. plantarum, for which the genetic toolbox is underdeveloped, have also been engineered, including heat- and sugar-responsive gene expression systems. These results have led to a patent application. Thermoplasmonic hydrogel composites were used to encapsulate E. coli and demonstrate NIR-regulated protein production. These examples demonstrate the flexibility of LTMs for controlled delivery using different external triggers.
Demonstration of a light-regulated LTM being developed for promoting blood vessel regeneration in chronic wounds
Related publications:
- Dhakane, V. S. Tadimarri, S. Sankaran, Light-Regulated Pro-Angiogenic Engineered Living Materials. Advanced Functional Materials 33, 2212695, (2023).
- Basaran, S. Dey, S. Bhusari, S. Sankaran, T. Kraus, Plasmonic stimulation of gold nanorods for the photothermal control of engineered living materials. Biomaterials Advances 147, 213332 (2023).
- Riedel, M. P. Bartolomé, L. L. T. Enrico, C. Fink-Straube, C. N. Duong, F. Gherlone, Y. Huang, V. Valiante, A. Del Campo, S. Sankaran. Engineered Living Materials for the Conversion of a Low-Cost Food-Grade Precursor to a High-Value Flavonoid. Frontiers in Bioengineering and Biotechnology 11 (2023).
- Dey, M. Blanch-Asensio, S. Balaji Kuttae, and S. Sankaran, Novel Genetic Modules Encoding High-Level Antibiotic-Free Protein Expression in Probiotic Lactobacilli, Microbial Biotechnology 16, 1264 (2023).
- Blanch-Asensio, V. S. Tadimarri, A. Wilk, and S. Sankaran, Discovery of a High-Performance Phage-Derived Promoter/Repressor System for Probiotic Lactobacillus Engineering, Microbial Cell Factories 23, 42 (2024).
- Blanch-Asensio, S. Dey, V. S. Tadimarri, and S. Sankaran, Expanding the Genetic Programmability of Lactiplantibacillus Plantarum, Microbial Biotechnology 00, 1 (2023).
- Rodrigo-Navarro, S. Sankaran, M. J. Dalby, A. del Campo, and M. Salmeron-Sanchez, Engineered Living Biomaterials, Nat Rev Mater 6, 1175 (2021).
- Pearson, J. Feng, A. del Campo. Lighting the Path: Light Delivery Strategies to Activate Photoresponsive Biomaterials In Vivo, Advanced Functional Materials, 31, (50), 2105989 (2021).
EVALUATION OF HOST-LTMS INTERACTIONS AND DEVELOPMENT OF ASSAYS FOR RISK ASSESSMENT
The evaluation of the immunogenic potential of LTMs is a fundamental step to assess their clinical transferability. Although LTMs prevent the physical contact between biofactories and host cells, the high permeability of the matrix allows exchange of their secretomes. Interestingly, the first studies of the immune response to LTM prototypes have provided encouraging results. Human peripheral blood mononuclear cells from healthy donors showed no significant activation when exposed to LTMs containing engineered ClearColi.
Thin film bacterial encapsulation format for in vitro testing of immune responses using human-derived peripheral blood mononuclear cells (PBMCs).
Related publications:
- A. K. Yanamandra, S. Bhusari, A. del Campo, S. Sankaran, B. Qu, In vitro evaluation of immune responses to bacterial hydrogels for the development of living therapeutic materials. Biomaterials Advances 153, 213554 (2023).
JOURNAL PUBLICATIONS
A Rodrigo-Navarro, S Sankaran, M Dalby, A del Campo, M Salmeron-Sanchez
Engineered Living Biomaterials
Nat Rev Mater 2021, 6, 1175–1190
S Groß, F Panter, D Pogorevc, CE Seyfert, S Deckarm, CD Bader, J Herrmann, R Mueller
Improved broad-spectrum antibiotics against Gram-negative pathogens via darobactin biosynthetic pathway engineering
Chem. Sci., 2021, 12, 11882-11893
S Bhusari, S Sankaran, A del Campo
Regulating Bacterial Behavior within Hydrogels of Tunable Viscoelasticity
Advanced Science 2022, 9 (17), 2106026
G Toth, B Berko-Gottel, B Seitz, A Langenbucher, T Stachon, MT Pluzsik, ZZ Nagy, S Smola, N Szentmary
Herpes simplex virus PCR in 2230 explanted corneal buttons
Acta Ophthalmol. 2022;100(1):e77-e82
J Rehner, GP Schmartz, L Groeger, J Dastbaz, N Ludwig, M Hannig, S Rupf, B Seitz, E Flockerzi, T Berger, MC Reichert, M Krawczyk, E Meese, C Herr, R Bals, SL Becker, A Keller, R Mueller
Systematic cross-biospecimen evaluation of DNA extraction kits for long- and short-read multi-metagenomic sequencing studies / Genomics
Proteomics & Bioinformatics, 20, 2, 2022, 405 – 417
M Blanch Asensio, S Dey, S Sankaran
Gibson Assembly-based direct cloning of plasmid DNA in Lactiplantibacillus plantarum WCSF1
BioRxiv, 2022 / https://www.biorxiv.org/content/10.1101/2022.09.09.507304v1
S Dey, M Blanch Asensio, S Balaji Kuttae, S Sankaran.
Novel genetic modules encoding high-level antibiotic-free protein expression in probiotic lactobacill
BioRxiv, 2022, https://www.biorxiv.org/content/10.1101/2022.08.04. 502766v1
S K. Srikakulam, S Keller, F Dabbaghie, R Bals, O V. Kalinina.
MetaProFi: A protein-based Bloom filter for storing and querying sequence data for accurate identification of functionally relevant genetic variants
BioRxiv, 2022 / https://doi.org/10.1101/2021.08.12.456081
A Yanamandra, S Bhusari, A del Campo, S Sankaran, B Qu. In vitro evaluation of immune responses to bacterial hydrogels for the development of living therapeutic materials. BioRxiv, 2022 / https://www.biorxiv.org/content/10.1101/2022.09.16.508081v1
S Bhusari, J Kim, K Polizzi, S Sankaran, A del Campo.
Encapsulation of bacteria in bilayer Pluronic thin film hydrogels: a safe format for engineered living materials. Biomaterials Advances, doi: 10.1016/J.bioadv.2022.213240
L Jungmann, S L Hoffmann, C Lang, R De Agazio, J Becker, M Kohlstedt, C Wittmann
High-efficiency production of 5-hydroxyectoine using metabolically engineered Corynebacterium glutamicum. Microbial Cell Factories, 2022, 21, 274, https://doi.org/10.1186/s12934-022-02003-z
M Brand, F Ritzmann, D Milasius, Y Yao, R Mueller, D Yildiz, R Bals, C Beisswenger
3D lung organoid platform for pre-clinical testing of active substances targeting senescence. Pneumologie 2023; 77(S 01): S24, DOI: 10.1055/s-0043-1760921
S Basaran, S Dey, S Bhusari, S Sankaran, T Kraus
Plasmonic stimulation of gold nanorods for the photothermal control of engineered living materials
Biomaterials Advances, 147, 2023, 213332, https://doi.org/10.1016/j.bioadv.2023.213332
N Eckert, L Horbal, Y Rebets, J Zapp, A Luzhetskyy
Discovery and isolation of novel highly active pamamycins through transcriptional gene cluster engineering, in preparation
Conference contributions (oral or posters)
S Bhusari, S Shrikrishnan, A del Campo
Design of 3D printed hydrogel biofilm mimics
Cell Physics 2021, 29.09 – 01.10.2021, Saarbruecken, Germany
S Bhusari, S Sankaran, A del Campo
Bacteria Material Relationship – it’s complicated! Bacterial behavior in Ploronic based bioinks for Living Therapeutic Devices
ESB 2021, 31st Annual Conference of the European Society for Biomaterials, 05-09.09.2021
S Bhusari, S Sankaran, A del Campo
Living therapeutic devices – Marrying synthetic biology with materials science / German Conference on Synthetic Biology (GCSB21) – Engineering Living Systems, 13-17.09.2021
S Bhusari, S Sankaran, A del Campo
Living therapeutic devices: bacterial behavior in viscoelastically tunable confinement / 2nd Engineered Living Materials 2021, 04-07.05.2021
S Sankaran, P Dhakane, S Bhusari, S Dey, M Blanch-Asensio, A del Campo
Living therapeutic materials – hydrogel-confined bacteria for smart drug delivery.
German Conference on Synthetic Biology (GCSB21) – Engineering Living Systems, 13.09.-17.09.2021, virtual, Oral presentation
S Sankaran, S Bhusari, A del Campo
Living materials for life – Burglabs Café 2021, 09.09.2021, Burg Giebichenstein University of Art and Design, Oral presentation
S Sankaran, P Dhakane, S Bhusari, A del Camp
Living therapeutic materials -hydrogel-confined bacteria for smart drug deliver – BMT2020 – 54th DGBMT Annual Conference on Biomedical Engineering 29.09.- 01.10.2021, Virtual, Oral presentation
P Dhakane, V Tadimarri, A del Campo, S Sankaran
Living therapeutic materials to release angiogenesis-promoting QK that can bind to ECM collagen – German Conference on Synthetic Biology (GCSB21) – Engineering Living Systems, 13.09-17.09.2021, virtual, Oral presentation
P Dhakane, V Tadimarri, S Bhusari, A del Campo, S Sankaran
Living therapeutic materials for sustained drug release – Designer Biology Conference, 08.09.-10.09.2021, virtual, Oral presentation
M Blanch Asensio, S Sankaran
Light-regulated release of recombinant Nerve Growth Factor from Bacterial Hydrogels. German Conference on Synthetic Biology (GCSB 2021) – Engineering Living Systems, Virtual, 13.09. –17.09.2021, poster presentation
S Dey, S Sankaran
Thermoresponsive living therapeutic materials – Darobactin releasing bacterial hydrogels for treating chronic pathogen infections – German Conference on Synthetic Biology (GCSB 2021) – Engineering Living Systems, 13.09.– 17.09.2021, virtual, poster presentation
S Basaran, S Dey, S Bhusari, A del Campo, S Sankaran, T Kraus
Plasmonic Stimulation of Gold Nanorods for the Control of Living Materials
17th Zsigmondy Conf., Aachen, 6 – 8.4.2022
S Basaran, S Dey, S Sankaran, T Kraus
Thermoplasmonic Stimulation of Living Materials with Gold Nanorod Composites Engineered Engineered Living Materials 2022, Saarbrücken, 21 – 23.06.2022
S Basaran, S Dey, S Bhusari, A del Campo, S Sankaran, T Kraus
Plasmonic Stimulation of Gold Nanorods for the Control of Living Materials
36th Europ. Colloid & Interface Soc. Conf., Crete/ Greece, 4– 9.9.2022
S Basaran, S Dey, S Sankaran, T Kraus
Thermoplasmonic Stimulation of Gold Nanorods for the Control of Living Materials
51st Conf. of the German Colloid Society, Berlin, 28– 30.9.2022
S Beganovic, F Weiland, S Sankaran, A del Campo, S Smola, C Wittmann
Streamlining Coryne-bacterium glutamicum into a living therapeutic for treatment of HPV-induced cervical lesions
Engineered Living Materials, Saarbrücken, 21- 23.6.2022
S Beganovic, L Gläser, W Shu, C Rückert, J Kalinowski, H Sucipto, A Luzhetskyy, C Wittmann
Systems biology of oxytetracycline biosynthesis in industrial Streptomyces rimosus
Metabolic Engineering & Systems Biology, Gothenburg, Sweden, 22- 26.8.2022
S Bhusari, S Sankaran, A del Campo
Giving Life to Materials: Tuning bacterial behavior in Pluronic based bioinks for living therapeutic applications
ESB 2022, the 32nd Annual Conference of the European Society for Biomaterials, 4-9.09 2022, Bordeaux, France
S Bhusari, S Sankaran, A del Campo
Therapeutic Materials with Life: Bacterial behavior in Pluronic based bioinks for smart drug delivery
6th International School and Conference on Biological Materials Science : Bioinspired Materials 2022, 21- 24.03.2022
B Muhovic, A del Campo, S Sankaran, C Wittmann
Towards living therapeutics in ophthalmology: Supply of the cell protectant ectoine using engineered Corynebacterium glutamicum
Engineered Living Materials, Saarbrücken, 21- 23.6.2022
C Porten, S Dey, S Basaran, R Müller, S Sankaran, T Kraus.
Opto-thermally regulated release of novel antibiotics from ELMs
Engineered Living Materials 2022, Saarbrücken, 21 – 23.06.2022
A Yanamandra, S Bhusari, A del Campo, S Sandaran, B Qu
In vitro evaluation of immune responses to bacterial hydrogels for the development of living therapeutic materials
Engineered Living Materials, Saarbrücken, 21 – 23.6.2022
F Weiland, J Becker, M Kuhl, M Kohlstedt, C Wittmann
Metabolic engineering of Corynebacterium glutamicum for the valorization of lignin-derived phenylpropanoids
Gordon Research Conference, Lignin: Realizing Lignin’s Potential in Biorefining by Bridging Plant Biology, Chemistry and Engineering; Easton, USA, 31.7 – 5.8.2022
A del Campo
Mechanopharmacology with living therapeutic hydrogels
Molecular Systems Engineering for Bioapplications (MSEB), 19-21.10.2022, Heidelberg, Germany (invited)
A del Campo
Hydrogels that talk to cells when lighted
ACTIVE 2022 Electrical and Optical Active Molecular Materials for Bio-Applications, 06-07.10.2022, Bellaterra, Spain (keynote)
A del Campo
Regulating bacterial functions by confinement
GBM Fall Conference 2022 Molecular Basis of Life, 14-16.09.2022, Heinrich-Heine University Duesseldorf, Germany (keynote)
A del Campo
Mechanopharmacology with living therapeutic hydrogels, GRC Multiscale Mechanochemistry and Mechanobiology
From Molecular Mechanics to Smart Materials, 31– 05.08.2022, Ventura, USA (invited)
A del Campo
Engineered materials with bioprogrammable functions
International Conference on Programmable Materials 2022, 12-15.07. 2022, Berlin/Germany (plenary)
A del Campo
Engineered living therapeutic materials: new concepts for sustained and sustainable drug delivery
N4M Nanoengineering for Mechanobiology, 27-31.03.2022, Camogli, Italy (invited)
S Sankaran
ELMs for sensing – Nachwuchsakademie ELM workshop, Saarbrucken, Germany, 17.06.–21.06.2022, Oral presentation
S Sankaran, P Dhakane, S Bhusari, S Dey, M Blanch-Asensio, F Riedel, V Tadimarri, A del Campo
Living therapeutic materials – hydrogel-confined bacteria for smart drug delivery. Invited talk, Bharathiar University, Coimbatore, India, 05.05.2022, Oral presentation
S Sankaran, P Dhakane, S Bhusari, S Dey, M Blanch-Asensio, F Riedel, V Tadimarri, A del Campo
Living therapeutic materials – hydrogel-confined bacteria for smart drug delivery. Invited talk, IIT Madras, Chennai, India, 17.05.2022, Oral presentation
S Sankaran, S Bhusari, A del Campo.
Living materials for life. Biophilic Intelligence seminar in the Future of Life summer school -Bio Design Lab at HfG Karlsruhe, Germany, 28.07.2022, Oral presentation
V Tadimarri, S Sankaran
Study of behavioral dynamics of lactic acid bacteria in hydrogels – BioSynsys, symposium on synthetic and systems biology, Paris. 12.09.-14.09.2022, Poster presentation
M Blanch Asensio, S Sankaran
Building an optogenetic circuit in probiotic bacteria as a novel living therapeutic -Third International Conference on Engineered Living Materials (ELMs), Saarbruecken, 21.06. –23.06.2022, Poster presentation
S Dey, CE Seyfert, R Mueller, S Sankaran
Thermoresponsive living therapeutic materials – Darobactin releasing bacterial hydrogels for treating chronic pathogen infections – Third International Conference on Engineered Living Materials (ELMs), Saarbruecken, 21.06.–23.06.2022, Poster presentation
S Dey, S Basaran, C Porten
Opto-thermally regulated release of novel antibiotics from ELMs – Third International Conference on Engineered Living Materials (ELMs), Saarbruecken, 21.06.– 23.06.2022, Oral presentation
M Blanch Asensio, Sourik Dey, S Sankaran
Novel genetic modules encoding high-level antibiotic-free protein expression in probiotic lactobacilli – BioSynsys, symposium on synthetic and systems biology, Paris, 12.09.–14.09. 2022, Oral presentation
S Dey, S Sankaran
LactoBio – Novel genetic tools to unleash probiotics for Live Biotherapeutic Products (LBP) – International Summer School on Technology Transfer in Life Sciences, In person, September 26.09. – 30.09.2022, Oral presentation
S Basaran, S Dey, S Sankaran, T Kraus
Plasmonic Stimulation of Gold Nanorods for the Photothermal Control of Engineered Living Materials
2023 MRS Spring Meeting & Exhibit, San Francisco, USA, 10.04. – 14.04.2023
PATENTS
S Dey, M Blanch Asensio, S Sankaran,
Patent application: “Neuartige genetische Werkzeuge”.
DE 10 2022 119 024.2