Autoantibody- and microRNA-based Next-Gen Multiplex Test for Type 1 Diabetes

DESCRIPTION provided by applicant Diabetes is a major cause of morbidity and mortality in the U S Given that the diabetes epidemic continues to grow worldwide there is a clear need for improvements in the management of the disease and its complications Early diagnosis and intervention can significantly improve long term prognosis Autoantibodies against islet antigens are a serologic hallmark of patients with Type Diabetes T D In addition circulating microRNAs miRNAs have been shown recently to be systematically altered indicative of miRNA signatures with diagnostic utility Growing evidence suggests that a multi marker strategy containing a combination of biomarkers with high clinical sensitivity and specificity may enhance diagnostic and prognostic accuracy in the future compared to single marker tests To support efforts of identifying the most informative biomarker panels reliable next generation platform technologies are needed that permit multiplexed detection of markers in small samples and are suitable for automation and integration into the clinical laboratory work flow Nesher Technologies Inc NTI has exclusively licensed the intellectual property for an ultrasensitive and specific biodetection technology developed at the UCLA Single Molecule Biophysics Lab headed by Prof Shimon Weiss with high single well multiplexing potential minimal sample requirements and simplified handling procedures no separation washing and amplification steps It is based on alternating laser excitation ALEX single molecule fluorescence spectroscopy whereby target recognition molecules are tagged with different color fluorescent dyes and quenchers NTI recently achieved extension from color to color ALEX substantially expanding its multiplexing power and demonstrated diagnostic utility for direct protein as well as miRNA quantification Furthermore recent work by Profs Steve Quake and Shimon Weiss shows i combination of microfluidics based sample handling with ALEX spectroscopy termed andquot single molecule optofluidicsandquot and ii enhanced throughput using a multifoci excitation detection geometry NTIandapos s long term goal is to develop rapid highly multiplexed ultrasensitive and specific as well as fully automated nucleic acid and protein based diagnostic tests that require minimal sample sizes Here we propose assay development and clinical validation of a next generation test with significantly improved diagnostic prognostic and treatment guiding properties implementing a panel of autoantibody and miRNA biomarkers and overcoming limitations of current T D testing Our Specific Aims are Initial reagent development for a multiplex autoantibody and miRNA based next generation test for T D Separate as well as multiplexed biomarker detection and quantification using spiked samples ALEX based analysis of archived clinical samples and cross validation to ELISA and qPCR methods SBIR Phase II will propose assay expansion to include more markers miniaturization and development of a user friendly andquot sample in answer outandquot diagnostic system offering significant cost and patient sample savings PUBLIC HEALTH RELEVANCE Type Diabetes characterized by a prolonged and variable latent period that culminates in the destruction of pancreatic beta cells and the development of hyperglycemia is a debilitating autoimmune disease and without proper management patients develop serious complications that reduce their quality of life and life expectancy As current single marker tests for T D are inadequate there will be a great need for a versatile next generation platform technology to detect and quantify panels of diagnostic and prognostic protein and nucleic acid biomarkers as they become available in order to better assist in establishing early diagnosis of T D refine prognosis guide management target treatment and finally improve patient outcome Based on patent protected alternating laser excitation ALEX single molecule fluorescence spectroscopy Nesher Technologies Inc intends to make its single molecule detection SMD platform technology robust and easy to use for diagnostic labs as well as the broad research community and proposes to develop a next generation test for T D monitoring a panel of autoantibody and microRNA biomarkers present in very small patient samples thereby translating cutting edge innovations in nanobiotechnology into benefits for the society at large by saving human lives and reducing healthcare costs