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A Novel Feeder-free System for Mass Production of Murine Natural Killer Cells In Vitro
Published on: January 9, 2018
In vitro vascular differentiation system efficiently produces natural killer cells for cancer immunotherapies
Yekaterina Galat1,2,3, Yuchen Du3, Mariana Perepitchka1,2,3
1Developmental Biology Program, Stanley Manne Children's Research Institute, Ann & Robert H. Lurie Children's Hospital, Chicago, IL, USA.
Background:
Immunotherapeutic innovation is crucial for limited operability tumors. CAR T-cell therapy displayed reduced efficiency against glioblastoma (GBM), likely due to mutations underlying disease progression. Natural Killer cells (NKs) detect cancer cells despite said mutations - demonstrating increased tumor elimination potential. We developed an NK differentiation system using human pluripotent stem cells (hPSCs). Via this system, genetic modifications targeting cancer treatment challenges can be introduced during pluripotency - enabling unlimited production of modified "off-the-shelf" hPSC-NKs.
Methods:
hPSCs were differentiated into hematopoietic progenitor cells (HPCs) and NKs using our novel organoid system. These cells were characterized using flow cytometric and bioinformatic analyses. HPC engraftment potential was assessed using NSG mice. NK cytotoxicity was validated using in vitro and in vitro K562 assays and further corroborated on lymphoma, diffuse intrinsic pontine glioma (DIPG), and GBM cell lines in vitro.
Results:
HPCs demonstrated engraftment in peripheral blood samples, and hPSC-NKs showcased morphology and functionality akin to same donor peripheral blood NKs (PB-NKs). The hPSC-NKs also displayed potential advantages regarding checkpoint inhibitor and metabolic gene expression, and demonstrated in vitro and in vivo cytotoxicity against various cancers.
Conclusions:
Our organoid system, designed to replicate in vivo cellular organization (including signaling gradients and shear stress conditions), offers a suitable environment for HPC and NK generation. The engraftable nature of HPCs and potent NK cytotoxicity against leukemia, lymphoma, DIPG, and GBM highlight the potential of this innovative system to serve as a valuable tool that will benefit cancer treatment and research - improving patient survival and quality of life.
Insights
Human pluripotent stem cell-derived Natural Killer (NK) cells offer a promising cancer immunotherapy. This novel organoid system enables the production of "off-the-shelf" NK cells for enhanced glioblastoma treatment.
Area of Science:
- Stem cell biology
- Immunology
- Cancer research
Background:
- Limited operability tumors require immunotherapeutic innovation.
- CAR T-cell therapy shows reduced efficiency against glioblastoma (GBM) due to disease-driving mutations.
- Natural Killer (NK) cells offer potential for tumor elimination, even with mutations.
Purpose of the Study:
- To develop an NK differentiation system using human pluripotent stem cells (hPSCs).
- To enable genetic modifications during pluripotency for enhanced cancer treatment.
- To facilitate unlimited production of modified "off-the-shelf" hPSC-NKs.
Main Methods:
- hPSCs were differentiated into hematopoietic progenitor cells (HPCs) and NKs using a novel organoid system.
- Cell characterization involved flow cytometry and bioinformatic analyses.
- HPC engraftment was assessed in NSG mice, and NK cytotoxicity was validated against various cancer cell lines.
Main Results:
- HPCs demonstrated peripheral blood engraftment.
- hPSC-NKs exhibited morphology and functionality similar to peripheral blood NKs (PB-NKs).
- hPSC-NKs showed enhanced gene expression and potent in vitro and in vivo cytotoxicity against multiple cancer types.
Conclusions:
- The organoid system effectively supports HPC and NK generation.
- Engraftable HPCs and potent NK cytotoxicity highlight the system's potential.
- This innovative approach may improve cancer treatment, research, patient survival, and quality of life.
