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Biology subjects

DeGrandchamp, J. B.

Publications and source records attributed to DeGrandchamp, J. B..

3 recordsLinked to original sources

Morphology-guided convolutional Graph Neural Network decodes optically barcoded nanoparticles for one-pot, purification-,amplification-, and enzyme-free femtomolar nucleic-acid diagnostics

Affordable, accurate, and rapid point-of-care diagnostic tests remain elusive due to inherent trade-offs between performance and cost. Conventional nucleic acid tests offer high sensitivity but require complex, expensive steps such as amplification and purification, whereas lateral flow assays are simple and low-cost but lack the necessary sensitivity for many applications. To bridge this gap, we present a miniaturized and simplified chip-based platform that combines three components into a single diagnostic pipeline: we use (i) spectrally distinct silver and gold nanoparticles that form analyte-dependent clusters with unique spectral fingerprints, (ii) a one-pot, enzyme- and purification-free assay on a chip integrated with a high-throughput automated low-cost microscope, and (iii) a morphology-guided convolutional Graph Neural Network that embeds morphology information into convolutional kernels and performs graph-based relational learning across particle-level features. This integration captures spectral, spatial, and morphological quantification at the particle level, rather than relying on bulk spectral shifts, thereby overcoming the limitations of contemporary nanoparticle assays and image-level deep learning approaches. Processing up to 5000 particles per image using only <5 GB GPU memory, Mc-GNN achieves femtomolar sensitivity with 98.2% recall for synthetic DNA and 94.8% for SARS-CoV-2 RNA from whole virus, despite variations in nanoparticle selection and sample complexity. By embedding morphological information into the biosensing pipeline, our diagnostic platform is computationally efficient, smartphone-compatible and is readily extensible to new analytes and multiplexing, offering a scalable solution for a fieldable diagnostic tool.

bioengineering↗

Grb2 Phosphorylation Antagonizes EGFR-driven Ras Activation by Interfering with Condensate Assembly

Upon ligand binding, the kinase domain of EGFR phosphorylates multiple tyrosine residues on the receptor cytoplasmic tail through a trans-autophosphorylation process. Phosphotyrosine sites on activated receptors recruit Grb2, which further recruits SOS to initiate downstream signaling by activating Ras. Multivalent binding between SOS and Grb2, as well as direct Grb2:Grb2 interactions, contribute to formation of a protein condensate of activated EGFR. The condensed state of EGFR facilitates autoinhibition release in SOS and exerts regulatory control over signal propagation from activated EGFR to Ras. While kinase activity of EGFR is an essential driver of this signaling process, phosphorylation at residue Y160 on Grb2 blocks Grb2:Grb2 binding and can interfere with EGFR condensation. Here, using a reconstituted system, we examine how titrating kinase activity in the EGFR system can both promote and inhibit signal output to Ras. The results reveal how effects of tyrosine kinase inhibition can, under some circumstances, promote Ras activation by inhibiting negative feedback through Grb2 phosphorylation and disruption of a Grb2 SH2/SH3 dimer interface. Statement of SignificanceActivated EGFR forms a biomolecular condensate, via linkage of multiple EGFR through Grb2 and SOS, and the condensation state of EGFR influences signal propagation to Ras. While tyrosine phosphorylation is a critical step in EGFR activation, phosphorylation of Grb2 can have an inhibitory effect on EGFR condensation and subsequent Ras activation. Under some conditions, kinase inhibition can promote signaling from EGFR to Ras.

biophysics↗

Bimodality in Ras signaling originates from processivity of the Ras activator SOS without classic kinetic bistability

Ras is a small GTPase that is central to important functional decisions in diverse cell types. An important aspect of Ras signaling is its ability to exhibit bimodal, or switch-like activity. We describe the total reconstitution of a receptor-mediated Ras activation-deactivation reaction catalyzed by SOS and p120-RasGAP on supported lipid membrane microarrays. The results reveal a bimodal Ras activation response, which is not a result of classic kinetic bistability, but is rather driven by the distinct processivity of the Ras activator, SOS. Furthermore, the bimodal response is controlled by the condensation state of the scaffold protein, LAT, to which SOS is recruited. Processivity-driven bimodality leads to stochastic bursts of Ras activation even under strongly deactivating conditions. This behavior contrasts classic kinetic bistability and is distinctly more resistant to pharmacological inhibition.

biophysics↗