Experimental Immuno-Oncology
Diefenbacher Lab
Lung cancer is the leading cause of cancer death globally, claiming 1.8 million lives in 2020. Projections show that lung cancer annual cases are expected to reach 3.8 million in 2050. This demonstrates how important it is to learn more about the development of this disease and to investigate novel therapeutic approaches.
Like all cancer researchers, we want to tear away the cloak of invisibility from tumor cells, we want to see what they do. To do this, we analyze the interaction between tumor and genetics as well as between tumor and environment: What processes take place, which proteins are involved, how are they regulated? Our visions are new, advanced models to read, control and stop or reverse these processes with the help of the immune system.
Lung cancer is the leading cause of cancer death globally, claiming 1.8 million lives in 2020. Projections show that lung cancer annual cases are expected to reach 3.8 million in 2050. This demonstrates how important it is to learn more about the development of this disease and to investigate novel therapeutic approaches.
Like all cancer researchers, we want to tear away the cloak of invisibility from tumor cells, we want to see what they do. To do this, we analyze the interaction between tumor and genetics as well as between tumor and environment: What processes take place, which proteins are involved, how are they regulated? Our visions are new, advanced models to read, control and stop or reverse these processes with the help of the immune system.
Target identification and validation
Our focus is the deregulation of protein turnover as a central driver in tumorigenesis, using state-of-the-art ex vivo and in vivo tools like CRISPR/Cas9 and patient derived Organoids.
One central mechanism that tumor cells alter during oncogenic transformation is the ubiquitin proteasome system (UPS), which is regulating virtually all biological processes, including, but not limited to DNA damage, senescence, cell cycle, DNA replication and the immune system. Despite the prominent involvement of the UPS in cancer, our understanding of how tumor cells alter the UPS system very early in transformation is rather limited.
The targeting of these common essential pathways and exploiting tumor intrinsic vulnerabilities holds the potential to develop preclinical models as well as personalized therapies not only for end- stage but also for early-stage patients.
Methods - CRISPR/Cas9
We utilize and combine novel techniques like CRISPR/Cas9 for tumor onset and develop isogenic transplant models. With the advent of CRISPR-mediated genome editing, gene deletion as well as site-directed integration of point mutations enabled us to model human malignancies in more detail than ever before.
We employ these models to study the early consequences of oncogenic transformation in the lung, and how cells, in combination with expression of different tumor drivers, reshape their niche and respond to standard of care therapy. These new models will provide a foundation for understanding lung cancer formation in molecular detail, from onset to late-stage disease.
Green: Deubiquitylating enzyme overrepresented in tumours
Red: Marker for club cells
Methods - Organoids
Another focus of our Lab is to culture primary murine and human lung epithelial and primary tumor cells as spheroids or ‘so called’ organoids. In the last years, we have established a novel protocol which simplifies the culture conditions and enabled us to cultivate not only primary murine cells derived from the trachea and alveolar compartment, but also patient material derived primary organoids from resected material.
Apart from genetically tailored murine respiratory organoid models, we culture and propagate patient organoids, derived from resection material. Combining genetically modified organoids with patient organoids, together with the corresponding control organoids, allows us the testing of novel therapeutic strategies for the treatment of all lung cancer variations.
Methods - Adeno-associated viruses (AAV)
As vehicle to facilitate gene transfer into recipient animals or patient samples do we use adeno-associated viruses, or short AAVs. These non-integrating, non-pathologic vectors carry as a payload all genetic components for e.g. CRISPR mediated gene editing, including the homology repair template or sgRNA sequences. This allows us to ‘just’ clone our genotypes of interest, in the case of modelling human disease, or express shRNA or cDNA variants of disease-causing genes. We established a simplified production protocol, hence, allowing to perform all steps required form design to production on site, making it the go-to tool for gene delivery in our model systems and beyond.
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