Whole Exome Sequencing of Familial ALS and Trios of Sporadic ALS to Identify New Genes Associated with ALS (P5.047)

Objective: Identify novel mutations in FALS and SALS by whole exome sequencing. Background: 90[percnt] of ALS cases are defined as sporadic and 10[percnt] are defined as familial. Our goal is to classify ALS patients by their genotypes and to discover common functional pathways and pathologies which could be used to formulate strategies to stop or slow down the progression of disease. Exome sequencing was used to identify novel mutations. This technique led to discovery of novel mutations in ALS. Methods: This study was approved by the IRB. Clinical data were collected by qualified specialists and by examination of medical records. DNA and other samples were taken after obtaining written informed consent. DNA samples from 300 subjects were used for Exome Sequencing. Samples were prepared using we used Agilent Sure-Select Kit. Illumina HiSeq 2500 was used for sequencing. The average read per targeted base was >65X with the quality score of ≥30. Open source bioinformatics tools and in house written scripts were used to perform Bioinformatics. Validation of common SNPs was conducted using NCBI dbSNP and Exome Variant Server. Results: The data were separated into three categories, novel mutations within the coding regions, novel mutations outside of the coding regions and mutations in dbSNP. The average number of variants per sample was between 350,000 and 420,000. All of the samples were Sanger sequenced for the variance verified as mutations. Conclusions: Using NGS we have identified novel mutations in CHCHD10, R15L, HNRNPA1 (P288S), PFN1 (C71G) was identified in several families. A correlation between the depth and the number of SNPs has been found using linear regression. No correlation between the onset, disease duration and gender has been identified using Ordinal Linear Regression. Using NGS we have identified mutations in three other novel genes in ALS which are currently being tested for functionality. Disclosure: Dr. ahmeti has nothing to disclose. Dr. Ahmeti has nothing to disclose. Dr. Yan has nothing to disclose. Dr. Fecto has nothing to disclose. Dr. Siddique has nothing to disclose. Dr. Pericak-Vance has received personal compensation for activities with the University of Alaska as a scientific advisory board member. Dr. Deng has nothing to disclose. Dr. Siddique has nothing to disclose.