A platform for integratingcell signaling and single cell transcriptomics.

PROJECT SUMMARY Cells communicate through physical interaction of cell surface proteins or through autocrineparacrinejuxtacrine or endocrine mechanismsHowevercell types identified by current large scale single cell sequencing and spatial transcriptomics efforts may mask heterogeneity in functional responses between cell typesOur ultimate vision for a product would be a platform to interrogate multiple cell types in different combinations and under different biochemical conditions to understand heterogenous gene expression responsesIn phase I of this SBIRScribe Biosciences will adapt its proprietary Printed Droplet MicrofluidicsPDMtechnology to study the impact of perturbations on single cell transcriptomicsPDM can precisely deliver reagents and single cells to a microwell array at any timeenabling flexible multi step workflows while maintaining high experimental throughputsPDM will be used to encapsulate single cellsincubate them either with an agonist antagonist or additional cell sof different cell typesand perform scRNA seq to understand induced transcriptional changesA novel lipid barcoding scheme will be used to link cells encapsulated in the same dropletsenabling cells to be recovered with traceable signatures in bulk and sequenced by established singlecell methodsIn Specific Aimwe will demonstrate the ability to perturb and incubate single cells in a test system where single Jurkat cells isolated in droplets are treated with anti CDand anti CDantibodiesthen assayed for IL p expressionIn Specific Aimwe will demonstrate a barcoding method to bioinformatically link cells incubated within the same droplet while allowing them to be sequenced separatelyThe successful execution of this phase I program will demonstrate the technical feasibility of a high throughput functional genomics platform using our PDM technology and will set the stage for a numerical scale up of the platform to analyze up tosingle cells PROJECT NARRATIVE Although single cell genomics have become widespreadthese technologies do not include information the original biological context of an analyzed sampleNext generation technologies will integrate biochemical and cellular contextsleading to a more complete understanding of the relationship between functional perturbations and the transcriptomic changes they engenderIn this projectwe will apply Scribe Biosciencesproprietary Printed Droplet Microfluidics technology to deterministically perturb single cells with agents or other single cellsthen use a new barcoding technology link perturbation conditions to single cell sequencing data