Clinical translation and regulatory aspects of CAR/TCR-based adoptive cell therapies-the German Cancer Consortium approach.

Adoptive transfer of T cells genetically modified by TCRs or CARs represents a highly attractive novel therapeutic strategy to treat malignant diseases. Various approaches for the development of such gene therapy medicinal products (GTMPs) have been initiated by scientists in recent years. To date, however, the number of clinical trials commenced in Germany and Europe is still low.

Aggregation-induced changes in the chemical exchange saturation transfer (CEST) signals of proteins.

Chemical exchange saturation transfer (CEST) is an MRI technique that allows mapping of biomolecules (small metabolites, proteins) with nearly the sensitivity of conventional water proton MRI. In living organisms, several tissue-specific CEST effects have been observed and successfully applied to diagnostic imaging. In these studies, particularly the signals of proteins showed a distinct correlation with pathological changes.

Integrin α5β1 and p53 convergent pathways in the control of anti-apoptotic proteins PEA-15 and survivin in high-grade glioma.

Integrin α5β1 expression is correlated with a worse prognosis in high-grade glioma. We previously unraveled a negative crosstalk between integrin α5β1 and p53 pathway, which was proposed to be part of the resistance of glioblastoma to chemotherapies. The restoration of p53 tumor-suppressor function is under intensive investigations for cancer therapy.

Signature of protein unfolding in chemical exchange saturation transfer imaging.

Chemical exchange saturation transfer (CEST) allows the detection of metabolites of low concentration in tissue with nearly the sensitivity of MRI with water protons. With this spectroscopic imaging approach, several tissue-specific CEST effects have been observed in vivo. Some of these originate from exchanging sites of proteins, such as backbone amide protons, or from aliphatic protons within the hydrophobic protein core.

Tumor suppressing properties of rodent parvovirus NS1 proteins and their derivatives.

Cancer chemotherapy with monospecific agents is often hampered by the rapid development of tumor resistance to the drug used. Therefore, combination treatments aiming at several different targets are sought. Viral regulatory proteins, modified or not, appear ideal for this purpose because of their multimodal killing action against neoplastically transformed cells.

Characterization of creatine guanidinium proton exchange by water-exchange (WEX) spectroscopy for absolute-pH CEST imaging in vitro.

Chemical exchange saturation transfer (CEST) enables indirect detection of small metabolites in tissue by MR imaging. To optimize and interpret creatine-CEST imaging we characterized the dependence of the exchange-rate constant k(sw) of creatine guanidinium protons in aqueous creatine solutions as a function of pH and temperature T in vitro. Model solutions in the low pH range (pH = 5-6.

Further studies with a cell immortalization assay to investigate the mutation signature of aristolochic acid in human p53 sequences.

To test hypotheses on the origins of p53 mutations in human tumors, novel strategies are needed for generating mutation spectra experimentally. To this end we developed an assay employing Hupki (Human p53 knock-in) mouse embryonic fibroblasts (HUFs). Here we examine p53 mutations induced by aristolochic acid I (AAI)), the carcinogen probably responsible for Chinese herbal nephropathy.

Multimodal imaging and computer assisted diagnosis for functional tumour characterisation.

Radiological modalities, especially CT, mainly provide morphological and structural information with high spatial resolution covering large volumes. Novel developments, which are predominantly MR-based, also deliver 'functional' information, which can be used for individual characterisation of tumour biology. Both aspects and modalities, additionally complemented by ultrasound, have to be combined in the radiological workflow of cancer patients including volumetric visualisation, information extraction from multimodal imaging, quantitative surrogates, intelligent interpretation assistance and image-guided procedures.

Analysis of radon-associated squamous cell carcinomas of the lung for a p53 gene hotspot mutation.

Squamous cell lung carcinomas (SCC) from former employees of the Wismut uranium mining company (Saxony, Germany) were obtained from the Stollberg Archive in order to screen for p53 tumour suppressor gene codon 249 arg-->met hotspot mutations, a putative molecular bio-dosimeter of alpha-particle (radon) exposure (Taylor et al (1994) Lancet 343: 86-87; McDonald et al (1995) Cancer Epidemiol Biomarkers Prevent 4: 791-793). Of the 29 archived samples of SCC meeting quality criteria for DNA analysis by polymerase chain reaction (PCR) and Haelll restriction enzyme digestion, two tumours were found that harboured this mutation. DNA sequencing confirmed the presence of a G to T base substitution within the Haelll site spanning codons 249 and 250 of the p53 gene that results in replacement of arginine (wild-type) by methionine at residue 249.

Arachidonic acid metabolism as a reporter of skin irritancy and target of cancer chemoprevention.

Keratinocytes respond to skin irritation and injury by cytokine release and a rapid but transient activation of arachidonic acid metabolism along both the cyclooxygenase and lipoxygenase pathways. In the first part of this article results are reviewed indicating that the release of pro-inflammatory mediators such as eicosanoids and interleukin-1 from keratinocytes provides a suitable in vitro parameter of irritancy. Based on this response an assay system has been established which may partially replace animal tests such as the Draize test.

The German teleradiology system MEDICUS: system description and experiences in a German field test.

MEDICUS is a teleradiology system which has been developed in a joint project of the German Cancer Research Center (Deutsches Krebsforschungszentrum) and the Transfer Center Medical Informatics (Steinbeis-Transferzentrum Medizinische Informatik) in Heidelberg, Germany. The system is designed to work on ISDN lines as well as in a local area network. Special attention has been given to the design of the user interface and data security, integrity, and authentication.

Cognition-based development and evaluation of ergonomic user interfaces for medical image processing and archiving systems.

Whether or not a computerized system enhances the conditions of work in the application domain, very much demands on the user interface. Graphical user interfaces seem to attract the interest of the users but mostly ignore some basic rules of visual information processing thus leading to systems which are difficult to use, lowering productivity and increasing working stress (cognitive and work load). In this work we present some fundamental ergonomic considerations and their application to the medical image processing and archiving domain.

Whole genome amplification of single cells from clinical peripheral blood smears.

Molecular analysis of clinical samples has been hampered by the lack of fresh or frozen specimens and the presence of contaminating background cells within samples obscuring the molecular analysis of the pathological cells of interest. Routine cytology specimens are a ubiquitous and abundant, yet largely untapped, source of clinical samples for molecular analysis. Morphologically defined single cells from peripheral blood smears can be microdissected from contaminating background cells and their whole genome amplified by primer extension preamplification, followed by polymerase chain reaction analysis of the specific DNA of interest.

Differentiative action of K252a on protein kinase C and a calcium-unresponsive, phorbol ester/phospholipid-activated protein kinase.

The Triton X-100 extract of the particulate fraction of porcine spleen contains a protein kinase which can be activated by phospholipid and the phorbol ester TPA but does not respond to phospholipid and calcium. The partially purified kinase has a molecular weight of 76 kDa (p76-kinase) and hence is somewhat smaller than the similarly behaving p82-kinase from mouse epidermis and spleen. The p76-kinase shows strong autophosphorylation.

Cyclosporine A suppresses an early process in phorbol ester action.

Simultaneous application of cyclosporine A (CsA) and the phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA) on mouse skin strongly inhibits various biological responses to TPA. Delayed application of CsA is less effective. CsA given 1-2 h after TPA is unable to suppress TPA effects.

Cell cycle specificity and reversibility of the inhibitory effect of epidermal G1-chalone on DNA synthesis in partially synchronized RTE-2 keratinocytes in vitro.

The specific action of a pig skin fraction enriched in epidermal G1-chalone, a tissue-specific inhibitor of epidermal DNA synthesis, was investigated by means of flow cytofluorometry. The results indicate that G1-chalone inhibits progression of partially synchronized rat tongue epithelial cells (line RTE-2) through the cell cycle at a point 2 h prior to the beginning of the S-phase. Approximately 8 h after chalone addition, the cells can overcome the inhibition and begin to enter the S-phase.

cDNA-cloning, sequencing and expression in glucocorticoid-stimulated quiescent Swiss 3T3 fibroblasts of mouse lipocortin I.

We isolated and sequenced mouse lipocortin I cDNA clones from a lambda gt10 cDNA library prepared from Swiss 3T3 mRNA. The homology with human lipocortin I at the amino acid level is 86%. When confluent layers of Swiss 3T3 cells were stimulated with 10% fetal calf serum, expression of lipocortin I was strongly stimulated.