A Combined High-Throughput Single-Cell Transcriptomic and Secretomic Assay for Neurodegenerative Disease Biomarker Discovery

In this Phase I application IsoPlexis proposes to deliver a novel method for targeted profiling of both the transcriptome and secretome from an array ofsingle central and peripheral immune cellsSpecificallywe will deliver a single cellmRNA and secreted protein capture assay with proven sensitivity for detecting the most important predictive biomarkers for neurodegenerative diseasesThere is growing evidence of the immune system acting in a functional or dysfunctional manner in neurodegenerative diseases and their cellularly heterogeneous microenvironments taken on therapeutic significanceThe function of microglia has been implicated in Alzheimerandapos s DiseaseFrontotemporal DementiaALSand MS among many moreTo best utilize the growing scientific understanding of immune response to genetic epigenetic factors contributing to neurodegenerative diseasean integrated tool is needed toimake highly multiplexedquantitative measurements thousands of single cellsiiwhile being able to gather both precise proteomic and transcriptomic data from the same celliiiwith fully automatedhigh throughputWe at IsoPlexis propose to develop an integrated and comprehensive platform to capture and characterize secreted protein response and the mRNA transcriptomefor downstream RNA Seqin a cell specific mannerwith minimal user variabilityAimDevelop an SCBC injection flow cell for the dual capture of secreted proteins and subsequent cell lysis for transcriptome on deviceAimDemonstrate the ability to monitor transcriptome and multiplexed secreted protein biomarkers fromsingle microglia and monocytes from aneurodegenerative disease mouse study and monocytes fromAlzheimerandapos s Disease andFrontotemporal Dementia patient samples in collaboration with UCI and UCSFTransition to Phase IIiWith the fully functional and validated flow cellwe will seek outclinical preclinical collaborations with institutes currently researching genetic correlations and immune system involvement in neurodegenerative diseasesiiWe will demonstrate the mRNA secretion assay on a fully automated SCBC instrumentthe IsoLightfrom cell isolation all the way to the final analysis improving throughput and reducing variability The role the central and peripheral immune systems acting in a functional or dysfunctional manner in neurodegenerative diseases has taken on therapeutic significance in recent yearsElucidating the interplay individual immune cell subsets for both the transcriptomic expression and proteomic signaling will define whether individual cells are predictably responding to treatment or are demonstrating mechanisms of disease progressionBy developing the IsoPlexis automated high throughput platform to make these highly multiplexedsingle cell measurements that simultaneously resolve the transcriptomic and proteomic information will enable researchers and clinicians make precise therapy decisions for a variety of neurodegenerative diseases such as Alzheimer s Disease and Frontotemporal Dementia