Developing an integrated sorting and single cell analysis tool for the analysis of rare T Cells subsets in Autoimmunity

Autoimmune disease AD affects up to million Americans which costs approximately $ billion on health care annually and accounts for one of the leading causes of death in women T cells and cytokine function play an important role in autoimmune diseases such as Type Diabetes Multiple Sclerosis Rheumatoid Arthritis RA and Systemic Lupus Erythematosus SLE The most successful auto immune therapies already target the factors secreting from or binding to T cells e g TNF IL IL and IL Diagnosis of autoimmune diseases is typically made post presentation of symptoms enabling earlier diagnosis through detecting the T cells that drive disease progression pathogenic T cells could shift paradigms towards earlier and more successful treatment IsoPlexis is developing a system to target and measure single T cells integral to this early diagnosis In this Phase I project we will use SLE as a preliminary disease with which to evaluate the strength of our technology SLE is an autoimmune disease mediated by autoreactive antibodies yet a subset of T cells called follicular helper T Tfh cells when dysregulated or pathogenic promote the induction of B cell autoantibody production Identifying these rare disease causing T cells also present in circulation has been proposed as a critical early diagnostic and a pressing need in the clinic However there are major challenges that need to be addressed pathogenic circulating Tfh cells are present at low frequency in blood these cells are highly heterogeneous with diverse combinations of effector cytokine functions requiring single cell analysis and the range of cytokine functional groups that matters numbers The IsoPlexis technology addresses these unmet challenges by simultaneously assaying cells analyzing cell functionality at the single cell level and co detecting effector proteins per cell This SBIR application aims to develop a suitable solution and an alpha test prototype device by incorporating an on chip upstream cell sorting method for capturing low quantities of circulating Tfh cells followed by a validated single cell cytokine function profiling platform for identifying pathogenic T cells reproducibly This NIH Phase I SBIR project will focus on developing this assay and device for lupus and autoimmune research We propose the following specific aims develop an NACS based cell capture device for profiling circulating Tfh cells with SCBC technology to eliminate flow sorting and establish a single cell plex cytokine and chemokine panel specific to profiling Tfh cells and develop the first components of an integrated automation workflow system for capturing and analyzing single Tfh cells with required reproducibility With a validated test system we would in Phase II apply this test to a large cohort clinical study and fully validate this for further commercialization in a critical area for SLE patients and physicians If successful this will open new opportunities for autoimmune disease diagnosis and monitoring of other T cell cytokine mediated diseases Pathogenic T cells presenting in circulation play an important role in autoimmune diseases We propose to create an automated prototype microdevice that allows for capturing specific T helper cells from patient blood samples i e SLE patient and analyzing the cytokine polyfunctionality of these cells at the single cell level which will be used as an early stage biomarker to detect SLE and monitor SLE patient upon treatment This technique has the potential to be scaled up and delivered to a broader range of autoimmune diseases and applicable to hospital disease diagnosis pharmaceutical drug development and clinical treatment monitoring