Biology subjects

Darnell, M.

Publications and source records attributed to Darnell, M..

4 recordsLinked to original sources

bioRxiv2023

Switching circuits: Modulating NK cell response to TGF-β using novel receptors

Engineered natural killer (NK) cells offer a promising treatment strategy for multiple therapeutic areas, yet tuning cell function to diverse environmental and inhibitory contexts remains a significant challenge. An attractive strategy for functional rescue in the presence of inhibitory ligands is the use of switch receptors, surface-expressed chimeric receptors that convert an external inhibitory cue into an intracellular activation signal. Here, we discover novel engineered switch receptors in NK cells that are responsive to TGF-{beta}, a soluble inhibitory factor present in many therapeutic contexts. Through a pooled screen of an 11,131 member library, we identified multiple novel signaling endodomains and endodomain combinations that improve both the acute cytotoxicity and persistence of NK cells in the presence of TGF-{beta}, demonstrating a novel and flexible approach to switch receptor discovery.

immunology↗

bioRxiv2023

TRAFfic signals: High-throughput CAR discovery in NK cells reveals novel TRAF-binding endodomains that drive enhanced persistence and cytotoxicity

Natural killer (NK) cells are a promising alternative therapeutic platform to CAR T cells given their favorable safety profile and potent killing ability. However, CAR NK cells suffer from limited persistence in vivo, which is, in part, thought to be the consequence of limited cytokine signaling. To address this challenge, we developed an innovative high-throughput screening strategy to identify CAR endodomains that could drive enhanced persistence while maintaining potent cytotoxicity. We uncovered a family of TRAF-binding endodomains that outperform benchmarks in primary NK cells along dimensions of persistence and cytotoxicity, even in low IL-2 conditions. This work highlights the importance of cell-type-specific cell therapy engineering and unlocks a wide range of high-throughput molecular engineering avenues in NK cells.

immunology↗

bioRxiv2022

Matrix viscoelasticity controls spatio-temporal tissue organization

The spatio-temporal patterning of multicellular tissues is driven by the collective dynamics of cell proliferation and active movement. These processes are mediated by the extracellular matrix environment via a combination of biomolecular and physical cues. Here we show that the passive viscoelastic properties of the matrix that encapsulate a proliferating ball of cells (e.g. a developing organoid) play a critical role in guiding tissue organization in space and time. By varying the viscoelasticity of well-defined model matrices, we show how a spheroidal tissue of breast epithelial cells breaks symmetry and forms finger-like protrusions that invade the matrix. A computational model allows us to recapitulate these observations and leads to a phase diagram that demarcates the regions of morphological stability and instability as a function of matrix viscoelasticity, tissue viscosity, cell motility and cell division rate. Experiments that use biomolecular manipulations to independently vary these parameters confirm our predictions. To further test our theory, we also study the self-organization of an in-vitro intestinal organoid and show that the morphological changes of this system also fits within our paradigm. Altogether, our studies demonstrate the role of stress relaxation mechanisms in determining the dynamics of tissue growth and the symmetry breaking instabilities associated with branching, a fundamental process in morphogenesis and oncogenesis, and suggest ways of controlling tissue form using the extracellular matrix.

biophysics↗

bioRxiv2020

Spatial cueing effects do not always index attentional capture: Evidence for a Priority Accumulation Framework

In visual search, improved performance when a target appears at a recently cued location is taken as strong evidence that attention was shifted to this cue. Here, we provide evidence challenging the canonical interpretation of spatial-cueing (or cue-validity) effects and supporting the Priority Accumulation Framework (PAF). According to PAF, attentional priority accumulates over time at each location until the search context triggers selection of the highest-priority location. Spatial-cueing effects reflect how long it takes to resolve the competition and can thus be observed even when attention was never shifted to the cue. Here, we used a spatial-cueing paradigm with abruptly onset cues and search displays varying in target-distractor similarity. We show search performance on valid-cue trials deteriorated the more difficult the search, a finding that is incompatible with the standard interpretation of spatial-cueing effects. By using brief displays (Experiment 1) and by examining the effect of search difficulty on the fastest trials (Experiment 2), we invalidate alternative accounts invoking post-perceptual verification processes (Experiment 1) or occasional failures of the onset cue to capture attention (Experiment 2). In Experiment 3, we used a combination of the spatial-cueing and dot-probe paradigms. We show that the events that occurred in both the cue and search displays affected attentional distribution, and that the relative attentional priority weight that accumulated at the target location determined how easily the competition was resolved. These findings fully support PAFs predictions. Public significance statementMany studies aim at establishing whether certain objects mandatorily capture our attention. Here, we show that there is no "yes-or-no" answer to this question because the context in which an object appears determines whether this object captures attention. We show that our attention is not shifted to the highest-priority object at any given time: instead, information about priority is collected across time until some signal indicates that the appropriate moment for deploying our attention has arrived. Striking failures to notice conspicuous events routinely illustrate how limited our attentional system is: we can attend to very few objects at any given time, and probably to just one. In natural conditions, when we move the focus of our attention from one object to another, we also shift our gaze towards the attended location: this allows us to place the object of most interest in the center of our fovea, which maximizes the quality of its perceptual processing. Tracking the locus of such overt attention is easily achieved by using eye-tracking devices. However, in order to isolate the benefits of attention from the benefits of visual acuity, one must study covert attention - that is, attentional shifts in the absence of eye movements. These shifts are not directly observable and must therefore be inferred using indirect measures of processing.

neuroscience↗