C9orf72 Protein study with RNA-Seq data on dbGaP (193 participants)

phs003065 — Uncovering RNA Signatures of C9orf72-Linked ALS and Related Disorders

Substantial clinical and pathological variability has been reported in patients carrying an expanded repeat in <a href="https://www.ncbi.nlm.nih.gov/gene/203228">C9orf72</a>, who are generally diagnosed with amyotrophic lateral sclerosis (ALS) and/or frontotemporal dementia (FTD). Emerging evidence suggests that <a href="https://www.ncbi.nlm.nih.gov/gene/203228">C9orf72</a> expression, RNA foci, and dipeptide-repeat proteins contribute to C9orf72-related diseases; however, the majority of the clinicopathological variability remains unexplained. We hypothesize that RNA sequencing (RNA-Seq) might reveal individual genes or groups of highly correlated genes (modules) that could account for this variability, which may help to identify much-needed druggable targets and/or biomarkers. Our preliminary studies performed in the frontal cortex, where we compared <a href="https://www.ncbi.nlm.nih.gov/gene/203228">C9orf72</a> expansion carriers to (disease) controls, demonstrated that the top differentially expressed gene was <a href="https://www.ncbi.nlm.nih.gov/gene/203228">C9orf72</a> itself. We also noticed that differentially expressed genes were enriched for the endocytosis pathway, which is consistent with the reported function of the C9orf72 protein in vesicle trafficking. Furthermore, we detected a module enriched for vesicular pathways, and additionally, we discovered an association with survival after onset for several promising candidates involved in vesicular transport. To find new therapeutic targets and biomarkers, here we have completed RNA-Seq on an increased number of samples to examine another relevant region: the cerebellum. As such, we performed RNA-Seq on a well-characterized cohort of <a href="https://www.ncbi.nlm.nih.gov/gene/203228">C9orf72</a> expansion carriers, patients without this expansion, and control subjects without neurological diseases. We used multivariable linear regression models for gene-level analysis, weighted correlation network analysis for module-level analysis, and leafcutter for splicing analysis. Using the cerebellar RNA-Seq data, we found a large number (>6,000) of differentially expressed genes, with a notable increase in expression of homeobox genes in <a href="https://www.ncbi.nlm.nih.gov/gene/203228">C9orf72</a> expansion carriers, as well as many gene modules that were associated with <a href="https://www.ncbi.nlm.nih.gov/gene/203228">C9orf72</a> expansion status. Additionally, using leafcutter, we detected over 1,000 differential splicing events and an increase in the number of cryptic splicing events in C9orf72 expansion carriers.