Periodic Reporting for period 2 - CHIPIN (Tissue-engineering the tumour microenvironment to improve treatment of pancreatic cancer)
Berichtszeitraum: 2022-10-01 bis 2024-03-31
The project addresses a clinical problem with a transformative approach to change our current paradigm in preclinical models. Traditional cell culture models utilising cells grown as monolayers have poor biological relevance, particularly for tumour biology. Our biologically relevant approach provides a tissue-like microenvironment to advance preclinical studies. We use a biomaterial-based 3D approach that allows cells to crosstalk with their microenvironment, which initiates critical events at cellular and molecular levels, changes in cell functions, biomechanics and gene expression. In a 3D structure, cells can recreate their original architecture similar to human tissues providing data of clinical relevance for people diagnosed with cancer. Bioengineered 3D models also minimize the use of research animals, and our biomaterials for the 3D cell cultures are ethically sustainable.
This is important for society because more than 90% of anti-cancer drugs brought to the clinic fail. The gap between preclinical evidence and clinical reality arises, partially, from the inability of traditional experimental models to truly and reproducibly recreate the original tissue composition and biomechanical properties of real tumours. Improvements in preclinical research can enable higher success rates and better clinical outcomes for people diagnosed with cancer. To achieve these goals, we need patient-specific and clinically predictive models using patient-derived samples to observe responses to drugs and optimise therapies like precision medicines. For pancreatic cancer, targeting only the tumour cells has failed. Pancreatic cancer is on its way to become the second deadliest cancer. The tumour microenvironment is a crucial untapped therapeutic target but a better understanding of the mechanisms of cell-cell and cell-matrix interactions within the cancer ecosystem is crucial to progress drug discovery and development.
The overall objectives are to combine tumour biology and tissue engineering to design a new platform and to find and test better treatments for pancreatic cancer. We aim to develop a controllable and reproducible technology platform for modelling the human tumour microenvironment of pancreatic cancer in 3D culture. In a complementary strategy, we are using hydrogel-enabled models and patient-derived cells to recreate the matrix composition, architecture and dynamics of pancreatic cancer and metastasis to decipher its tumour biology and improve its treatment.
This is important for society because more than 90% of anti-cancer drugs brought to the clinic fail. The gap between preclinical evidence and clinical reality arises, partially, from the inability of traditional experimental models to truly and reproducibly recreate the original tissue composition and biomechanical properties of real tumours. Improvements in preclinical research can enable higher success rates and better clinical outcomes for people diagnosed with cancer. To achieve these goals, we need patient-specific and clinically predictive models using patient-derived samples to observe responses to drugs and optimise therapies like precision medicines. For pancreatic cancer, targeting only the tumour cells has failed. Pancreatic cancer is on its way to become the second deadliest cancer. The tumour microenvironment is a crucial untapped therapeutic target but a better understanding of the mechanisms of cell-cell and cell-matrix interactions within the cancer ecosystem is crucial to progress drug discovery and development.
The overall objectives are to combine tumour biology and tissue engineering to design a new platform and to find and test better treatments for pancreatic cancer. We aim to develop a controllable and reproducible technology platform for modelling the human tumour microenvironment of pancreatic cancer in 3D culture. In a complementary strategy, we are using hydrogel-enabled models and patient-derived cells to recreate the matrix composition, architecture and dynamics of pancreatic cancer and metastasis to decipher its tumour biology and improve its treatment.
The PhD student investigated the role of the cancer-associated protease KLK6 as a potential therapeutic target for pancreatic cancer. Harnessing the CRISPR/Cas9 and RNAseq technologies, the impact of KLK6 knockout on cancer cell functions and changes in drug responses in 3D cell culture and xenograft models was explored. Loss of KLK6 gene expression significantly slowed down cancer cell growth and was related to signalling pathways involved in cell survival, proliferation, invasion, epithelial-mesenchymal transition and immunosuppression.
PostDoc #1 (biologist) developed a multicellular pancreatic cancer model and used it to assess the effects of immunotherapy in combination with chemotherapy. Human cancer and cancer-supporting cells were grown encapsulated in hydrogels to mimic key components of tumour tissues. Combining the CD11b agonist ADH-503 with immunotherapy and chemotherapy led to a significant reduction in tumour cell viability, proliferation, metabolic activity, immunomodulation, and secretion of immunosuppressive and tumour growth-promoting cytokines.
PostDoc #2 (engineer) evaluated the dynamics of matrix remodeling during the growth and autonomous organization of pancreatic cancer cells in hydrogel with the adequate initial stiffness according to published datasets from patient-derived tissues. Incorporation of protease-sensitive peptides provided the dynamic degradability of hydrogels and integrin-binding peptides enabled integrin-mediated cell functions. Oscillatory shear measurements and atomic force microscopy were used to evaluate the mechanical properties of hydrogels. The dynamics of matrix remodeling in 3D cell cultures using cancer and cancer-supporting cells were evaluated. While 3D cell cultures had comparable stiffness at the beginning, the cancer-supporting cells contributed to a progressive and significant stiffness reduction. The autonomous organization of cells in hydrogels resulted in the formation of spheroids, followed by degradation of the hydrogel matrix. The size and number of spheroids and matrix remodeling were significantly increased in the presence of cancer-supporting cells.
PostDoc #3 (HZDR) transferred the multicellular pancreatic cancer model to a xenograft approach to add the dimensions of an intact organism in which applied substances are subject to specific pharmacokinetics and pharmacodynamics. Cell-seeded hydrogels were implanted into immunodeficient NXG mice. Tumour growth was quantified by magnetic resonance imaging and dependent on the initially seeded cell populations. Addition of cancer-supporting cells accelerated tumour growth and resulted in tumours with intratumoral heterogeneity containing diverse cell populations and desmoplastic areas with poor vascularisation. Uptake of a radiolabeled FAP inhibitor by the tumours was measured by positron emission tomography, confirming the presence of functional FAP-expressing cells. Addition of cancer-supporting cells induced a switch in phenotype from differentiated expressing both mesenchymal and epithelial markers towards de-differentiated losing these key markers in tumours containing cancer-supporting cells.
PostDoc #1 (biologist) developed a multicellular pancreatic cancer model and used it to assess the effects of immunotherapy in combination with chemotherapy. Human cancer and cancer-supporting cells were grown encapsulated in hydrogels to mimic key components of tumour tissues. Combining the CD11b agonist ADH-503 with immunotherapy and chemotherapy led to a significant reduction in tumour cell viability, proliferation, metabolic activity, immunomodulation, and secretion of immunosuppressive and tumour growth-promoting cytokines.
PostDoc #2 (engineer) evaluated the dynamics of matrix remodeling during the growth and autonomous organization of pancreatic cancer cells in hydrogel with the adequate initial stiffness according to published datasets from patient-derived tissues. Incorporation of protease-sensitive peptides provided the dynamic degradability of hydrogels and integrin-binding peptides enabled integrin-mediated cell functions. Oscillatory shear measurements and atomic force microscopy were used to evaluate the mechanical properties of hydrogels. The dynamics of matrix remodeling in 3D cell cultures using cancer and cancer-supporting cells were evaluated. While 3D cell cultures had comparable stiffness at the beginning, the cancer-supporting cells contributed to a progressive and significant stiffness reduction. The autonomous organization of cells in hydrogels resulted in the formation of spheroids, followed by degradation of the hydrogel matrix. The size and number of spheroids and matrix remodeling were significantly increased in the presence of cancer-supporting cells.
PostDoc #3 (HZDR) transferred the multicellular pancreatic cancer model to a xenograft approach to add the dimensions of an intact organism in which applied substances are subject to specific pharmacokinetics and pharmacodynamics. Cell-seeded hydrogels were implanted into immunodeficient NXG mice. Tumour growth was quantified by magnetic resonance imaging and dependent on the initially seeded cell populations. Addition of cancer-supporting cells accelerated tumour growth and resulted in tumours with intratumoral heterogeneity containing diverse cell populations and desmoplastic areas with poor vascularisation. Uptake of a radiolabeled FAP inhibitor by the tumours was measured by positron emission tomography, confirming the presence of functional FAP-expressing cells. Addition of cancer-supporting cells induced a switch in phenotype from differentiated expressing both mesenchymal and epithelial markers towards de-differentiated losing these key markers in tumours containing cancer-supporting cells.
Beyond the state of the art [Expected results]
- CRISPR/Cas9 KO KLK6 in 3 different cell lines [find interactive partners in the KLK6 proteolytic network]
- neutrophil isolation and characterization [proof the immunosuppressive role of neutrophils]
- new 3D disease model for pancreatic cancer [identify tumour-immune cell interactions]
- rational design strategy [guide better therapeutic options for patients]
- mimic specific adhesion profile of cancer cells [instruct disease-relevant integrin profile]
- new orthotopic xenograft approach [explore factors driving disease progression and metastasis]
- preclinical evaluation of therapeutic strategies [impact of cancer-supporting cells on drug responses and resistance]
- drug combinations targeting cancer-supporting cells [impact of phenotypical switch on responses to drugs]
- CRISPR/Cas9 KO KLK6 in 3 different cell lines [find interactive partners in the KLK6 proteolytic network]
- neutrophil isolation and characterization [proof the immunosuppressive role of neutrophils]
- new 3D disease model for pancreatic cancer [identify tumour-immune cell interactions]
- rational design strategy [guide better therapeutic options for patients]
- mimic specific adhesion profile of cancer cells [instruct disease-relevant integrin profile]
- new orthotopic xenograft approach [explore factors driving disease progression and metastasis]
- preclinical evaluation of therapeutic strategies [impact of cancer-supporting cells on drug responses and resistance]
- drug combinations targeting cancer-supporting cells [impact of phenotypical switch on responses to drugs]