Whole Blood Filter Paper Assay for Alzheimers Disease

Summary In this Phase I program we will establish the feasibility of a novel filter paper whole blood sample based targeted sequencing assay based on the TempO Seqplatform to measure DNA and RNA profiles as the basis for the development of a minimally invasive prognostic diagnostic assay of Alzheimer s DiseaseADWe will use our already functional whole transcriptome targeted gene expression assay to evaluate a set of whole blood samples from AD and normal donors provided by our collaboratorsDrAllan LeveyDirectorAlzheimer s Disease ResCenter at Emory UnivChairDeptof Neurologyand DrAaron RitterCleveland ClinicLou Ruvo Center for Brain HealthWe will utilize the resulting data to define a smaller targeted AD specific gene subsetthen redesign the probes for those genes to be compatible with the TempO Seq DNA assayThis will allow us to generate a combined DNA RNA assay capable of detecting both critical DNA genotypese gAPOE allelesand the diagnostic prognostic gene expression RNA profile from dried blood spot samplesall in a single welladdition only assaywith no extractionreverse transcriptionor wash stepsFinallywe will combine the assay with a fill in elementallowing mapping of significant stretches of genes which can harbor multiple unpredictable mutationssuch as PSENor APPto create aone shotassay capable of detecting all relevant genetic and transcriptomic biomarkers in one single stepall from smallmmareas of blood spots on filter paperThe functionality of the completed assay will be confirmed on the same set of normal AD samplesallowing us to proceed to Phase IIwhere we will commercialize the assay and prove its predictive and diagnostic valueThis novel assay will address the unmet need for a cost effectivesimple way to broadly screen at risk populationsespecially in hard to reach or otherwise underserved areasAbility to derive data of such unprecedented depth from samples that are as non invasive and simple to collect and ship holds promise to reshape AD detection and careThe proposed research program is responsive to the NIA DBSR Special Interest Topic DtoDevelop multiple and reliable assays for limited blood spot specimens for large surveysand Division of Aging Biology Topic JDevelopment of biomarkers for prognosisdiagnosisor treatment monitoring of aging or aging related diseasesand DN Topic Adevelopment of minimally invasive biomarkers that can be used for screening in the general populationsbiomarkers that could serve as surrogate measures for disease progression in MCIAD and ADRDThe outcome will be an assay that can be taken into Phase II to begin the development and validation of a prognostic diagnostic screening assay for AD Narrative Blood spots collected on filter paper represent an easy to collecteasy to storeand easy to ship sample typebut one that also presents significant technical challenges for transcriptomicsas extraction from filter paper tends to produce low RNA yield and qualityHaving already developed a gene expression assay capable of detecting RNA levels directly on small samples of blood on filter paperwe now propose development of a targeted sequencing assay which is performed directly on the filter paper sample itselfand which will produce a full readout of gene expression biomarkers correlated with Alzheimer s Disease detection and progressionFurthermorewe propose combining this assay with a readout of relevant DNA genotypessuch ApoEor presenilin mutantsfor a fully one shotlow cost assay amenable to high throughput processing