Method for the in vitro differentiation of human dendritic cells from hematopoietic stem cells for research and immunotherapy development
Unmet Need
Dendritic cells (DCs) play a critical role in the adaptive immune response by capturing and presenting antigens to activate cytotoxic T-cells. As such, naturally occurring DCs are thought to have crucial antitumor activity and are an active area of exploration in cancer immunotherapy and vaccine development, including in osteosarcoma, lung, and breast cancers. Since the first DC vaccine entered the market in 2010, there have been increased interest and investment in these interventions, with a global market size of $593.30 million USD in 2022. In contrast to cancer treatments like chemotherapy and radiation therapy, there are limited side effects and increased survival rates with DC-based immunotherapies. Monocyte-derived DCs (MoDCs) make up the current standard for DC-based immunotherapies because monocytes have a high prevalence in the blood compared to naturally occurring DCs, and MoDCs can be readily expanded for use in therapeutics. However, despite their improvement on other, non-DC cancer treatments, MoDCs are distinct from naturally occurring DCs and have demonstrated suboptimal results in T cell proliferation and migration to lymph organs. Alternatively, conventional type-1 DCs (cDC1) represent a more desirable DC subset given their superiority in CD8+ T-cell activation and ability to promote an inflamed tumor phenotype (i.e., “hot” tumor), which are more responsive to immune-based therapies. Given the low prevalence of naturally occurring cDC1s in the blood, there is a clear and urgent need for a method to generate clinically tractable cDC1.
Technology
Duke inventors have developed a novel procedure to produce conventional type-1 dendritic-like (cDC1-like) cells in vitro. These cells may be used as a research tool to better understand the role of cDC1s in cancer immunology and develop improved immunotherapies. Since cDC1 cells are not prevalent in high numbers in the blood naturally, this protocol would enable researchers to generate large quantities of these cells for study and delivery as therapeutics to patients. Specifically, this procedure differentiates a blood-derived CD34+ hematopoietic stem cells into cDC1-like cells using only the cytokines GM-CSF and FLT3L. This is an improvement over alternative protocols that use immortalized xenogeneic feeder cells to generate large numbers of cDC1-like cells. This protocol produces high yields of cDC1-like cells that express high levels of cDC1-specific markers, are responsive to TLR3 agonism, and have enhanced cross-presentation capabilities over Mo-DCs – all of which are hallmark features of cDC1. This has been demonstrated through in vitro tests of the protocol.
Other Applications
This technology could also be used in vaccine development and to study and develop cDC1-based cell therapies.
Advantages
- Generates high yield of cDC1-like cells
- Produces cells that are responsive to TLR3 agonism and have superior antigen cross-presentation compared to MoDC cells
- Amenable to antigen loading/mRNA transfection
- Protocol requires only cytokines, making these cDC1s suitable for clinical application.