The single projects of the CRC 1066 are distributed into the subareas A (Chemistry) and B (Life Science), complemented by the cross-sectional Q-projects and central Z-projects.
The subarea A provides the CRC 1066 with carrier structures, which are made of various chemical structures including covalently linked polymer architectures as well as self-organized structures.
A2: Synthesis of carbohydrate based nanocapsules and their functionalization with glycomimetics for cell-specific targeting
T. Opatz, K. Landfester
This project combines the use of carbohydrates, both as structural material for the preparation of nanoparticles and nanocapsules and as targeting moieties to achieve selective binding to certain immune cells. By working with polysaccharide based nanocapsules with high loading efficiencies it aims to prepare stimulus responsive carriers, which bind selectively to certain subpopulations of immune cells to release their cargo.
A6: Synthetic control of morphology and function in peptide based carrier systems
M. Barz, T. Weil
This project aims to control morphology and function of brush-like peptidic carriers, which are either derived from living polymerization or result from (further processed) natural monodisperse peptides. In addition peptide based targeting units are modified to allow selective ligation. In combination this project aims at the preparation of systems to study, how morphology (rod or sphere like structures) and/or the ligation of antibodies modify the interaction of the carriers with immune cells.
A7: Liposomes and polymersomes with multifunctional endgroups: targeting and controlled release
H. Frey, M. Helm, R. Zentel
Within this project the processing and loading (formulation) of polymersomes and liposomes as carriers for peptides, RNA and/or low molar mass pharmaca will be studied. It is thereby the speciality of these polymersomes that they can be coated with hyperbranched PEG and decorated with targeting structures. To modify the release as recondition for biomedical activity the stability of the basic polymersomal structure will be modified to allow stabilization (e.g. crosslinking) or destabilization by pH dependent degradation.
The cross-section projects (Q) form a natural bridge between the two subareas A and B. They are of central importance to most A- and B-projects, providing research of high specific value to the CRC.
Q1: Complexation and adsorption of polymeric nanocarriers with biological components like RNA, serum proteins and further components of blood and its influence on their stability and cell uptake
K. Landfester, M. Maskos, F. Schmid
This project aims to study the interaction of the nanoparticular carries with different components of blood serum in vitro. This includes the use of AFFF for sample purification. Then dynamic light scattering, micro-calorimetry and mass spectrometry will be used to identify the corona formation. The results will be compared with theoretical simulations of the interaction of synthetic polymers with proteins.
Q2: Optical methods to study endocytosis, intracellular trafficking and cargo release
K. Koynov, I. Lieberwirth, V. Mailänder
This project uses fluorescence correlation spectroscopy to investigate size, loading, stability and cargo-release of the carrier systems. In addition confocal microscopy will be used to characterize (in combination with electron microscopy) cellular uptake and intracellular trafficking, also in dependence of corona formation. The project finally aims at a correlation between intracellular trafficking, release and/or degradation of the pharmacological cargo.
Q3: Multimodale and multiscalar in vivo imaging of biodistribution and the efficiency of immunotherapy
T. Lammers, M. Miederer, F. Rösch
This project aims at in vivo imaging of the biodistribution of nanodimensional carriers on different length scales (from organ to cellular assembly) by PET- and optical methods. It is thereby the intention to validate fluorescence imaging against the quantitative PET method. In addition methods to visualize the activation of the immune system (immuno-imaging) as result of the immunotherapy will be developed.
Q4: Identification and immunotherapeutic characterization of tumor associated antigens and neoantigenes in genetically and carcinogenically induced melanoma models
M. Diken, H. Schild
This project has two targets. At first it will provide different spontaneous or carcinogen-induced mouse tumor models. Secondly it will characterize them with regard to the antigens and neoantigenes presented, so that these tumor models can be used by the different subprojects to evaluate nanoparticle based tumor vaccines.
Subarea B contains all projects which are primarily exploring the use of nanoparticles and polymers for tumor immunotherapie.
B3: Therapy of metastatic melanoma by addressing tolerance inducing M2-macrophages with therapeutic nanoparticles
D. Schuppan, A. Tüttenberg, R. Zentel
This project aims at the elimination of immune-suppression by tumor associated M2 macrophages with the help of nanoparticular carriers loaded with siRNA and low molar mass compounds to reprogram them. For this purpose R. Zentel will adjust his siRNA carrier system (cationic nanohydrogels), which have already proven its potential to deliver siRNA in vivo into macrophages in the liver. This carrier system will be optimized to the needs of this projects.
B4: Polymer induced tumor-immunotherapy by in situ activation of antigen presenting cells
S. Grabbe, L. Nuhn, H. Schild
This project will use nanoparticles for selective loading (antigen) and activation of dendritic cells in situ. Together with a PI (L. Nuhn) from chemistry, the PIs aim to establish a modular system, which allows a rather variable combination of antigen, immune activator and targeting moiety. Also checkpoint inhibitors will be tested with regard to local delivery.
B5: Transcriptional addressing of dendritic cells (DCs) with the help of an hPFscn1 derived promotor using DC-addressing polymers for tumor therapy
M. Bros, E. Wagner, R. Zentel
This project will develop DNA-vaccines, which are under the translational control of the Fascin-promotor and thus only active in dendritic cells. Since dendritic cells as –non dividing cells– are rather difficult to transfect, the expertise of E. Wagner and R. Zentel will be combined to develop new, more efficient stimulus responsive transfection systems from block copolymers and cationic oligomers.
B6: Modulation of IL-10- and/or STAT3-mediated tumor-associated tolerance-mechanisms with the help of functionalized nanoparticles
K. Landfester, K. Steinbrink
The applicants will use functionalized hydroxyethyl-starch based nanocapsules for cell specific targeting of immune cells. For this purpose production line of these capsules has been developed. During the next funding period these capsules will be loaded with IL-10-/STAT3-inhibitors to evaluate their therapeutic use in melanoma-models.
B8: Development and testing of nanoparticles to eliminate the cAMP-mediated immune-suppression in malignant melanoma
M. Barz, C. Becker, T. Bopp
This project aims at the elimination of the immune-tolerance in melanoma by modulating the cAMP concentration locally. This will be done with micellar formulations of adenylatcyclase-, protein kinase A- or casein kinase inhibitors, which have –in parts– already shown to work successfully in mouse models. For this purpose the delivery system will be improved with regard to targeting and local release to enable the therapeutic use of this concept.
B11: Proteins on nanocarriers: from the stealth effect to active targeting in vivo
V. Mailänder, S. Tenzer, T. Weil
Starting with the observation that some nanocarriers build up a protein corona nearly instantaneously in context with serum proteins, which has serious results for cellular uptake and intra-cellular trafficking, this project aims at three different topics. At first the possible corona formation will be studied on more carrier systems to determine the generality of this phenomenon. Then the applicants want to look for ways to pre-organize desired proteins in the corona to modify biodistribution. At last it is the aim to bind desired proteins like clusterin and/or antibodies chemically to obtain –in this way– a stealth like behavior.
B12: Nano-particular mRNA-carrier systems to reprogram different immune-cell populations
M. Barz, P. Langguth, U. Sahin
Within this project nano-particular mRNA carriers, which are able to reach different immune-cell population in different parts of the body and reprogram them, shall be developed. This shall be done in a preclinical setting with the aim to establish new mRNA based therapies. As carrier systems both lipo- and polyplexes will be studied, which allow an optimization via high-throughput-screening.
B13: Multifunctional nanosized peptid/glykopeptid conjugates as fully-synthetic vaccine for tumor-immunotherapy
P. Besenius, E. Schmitt
This project aims to use form-anisotropic, self-organized nanorods to induce a humorale anti-tumor response. For this purpose nanorods will be decorated in a multivalent manner with (i) B-cell epitopes derived from surface proteins typical for melanoma or breast cancer cells and (ii) T-helper cells. In addition mannose will be used as targeting moiety and TLR-ligands for immune cell activation.