The recent rise of adoptive T cell
therapy (ATCT) as a promising
cancer immunotherapy has triggered increased interest in therapeutic
T cell bioprocessing. T cell activation is a critical processing step
and is known to be modulated by physical parameters, such as substrate
stiffness. Nevertheless, relatively little is known about how biophysical
factors regulate immune cells, such as T cells. Understanding how
T cell activation is modulated by physical and biochemical cues may
offer novel methods to control cell behavior for therapeutic cell
processing. Inspired by T cell mechanosensitivity, we developed a
multiwell, reusable, customizable, two-dimensional (2D) polyacrylamide
(PA) hydrogel-integrated culture device to study the physicochemical
stimulation of Jurkat T cells. Substrate stiffness and ligand density
were tuned by concentrations of the hydrogel cross-linker and antibody
in the coating solution, respectively. We cultured Jurkat T cells
on 2D hydrogels of different stiffnesses that presented surface-immobilized
stimulatory antibodies against CD3 and CD28 and demonstrated that
Jurkat T cells stimulated by stiff hydrogels (50.6 ± 15.1 kPa)
exhibited significantly higher interleukin-2 (IL-2) secretion, but
lower proliferation, than those stimulated by softer hydrogels (7.1
± 0.4 kPa). In addition, we found that increasing anti-CD3 concentration
from 10 to 30 μg/mL led to a significant increase in IL-2 secretion
from cells stimulated on 7.1 ± 0.4 and 9.3 ± 2.4 kPa gels.
Simultaneous tuning of substrate stiffness and stimulatory ligand
density showed that the two parameters synergize (two-way ANOVA interaction
effect:
p
< 0.001) to enhance IL-2 secretion.
Our results demonstrate the importance of physical parameters in immune
cell stimulation and highlight the potential of designing future immunostimulatory
biomaterials that are mechanically tailored to balance stimulatory
strength and downstream proliferative capacity of therapeutic T cells.
Biopolymers, such as poly-3-hydroxybutyrate (P(3HB)) are produced as a carbon store in an array of organisms and exhibit characteristics which are similar to oil-derived plastics, yet have the added advantages of biodegradability and biocompatibility. Despite these advantages, P(3HB) production is currently more expensive than the production of oil-derived plastics, and therefore, more efficient P(3HB) production processes would be desirable. In this study, we describe the model-guided design and experimental validation of several engineered P(3HB) producing operons. In particular, we describe the characterization of a hybrid phaCAB operon that consists of a dual promoter (native and J23104) and RBS (native and B0034) design. P(3HB) production at 24 h was around six-fold higher in hybrid phaCAB engineered Escherichia coli in comparison to E. coli engineered with the native phaCAB operon from Ralstonia eutropha H16. Additionally, we describe the utilization of non-recyclable waste as a low-cost carbon source for the production of P(3HB).
The meteoric rise of cancer immunotherapy in the last decade has led to promising treatments for a number of hard-to-treat malignancies. In particular, adoptive T cell therapy has recently reached a major milestone with two products approved by the FDA. However, the inherent complexity of cell-based immunotherapies means that their manufacturing time, cost, and controllability limit their effectiveness and geographic reach. One way to address these issues may lie in complementing the dominant, reductionistic mentality in modern medicine with complex systems thinking. In this Opinion, we identify key concepts from complexity theory to address manufacturing challenges in cell-based immunotherapies and raise the possibility of a unifying framework upon which future bioprocessing strategies may be designed.
HighlightsComplexity theory provides a conceptual framework in which biological and artificial networks may be designed or manipulated to intensify cell bioprocessing in cancer immunotherapies.Studies on T cell mechanobiology have revealed how physical parameters may be exploited to perturb intracellular networks as an effective way of controlling T cell fates for immunotherapeutic applications.Systems biology-based computational models open up the potential to predict cues needed to guide T cell differentiation and reprogramming.Advances in immunomodulatory biomaterials, microfabrication and wearable medical technologies raise the possibility of scaling up point-of-care deployment of immunotherapies.
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