Poly(GP) proteins are a useful pharmacodynamic marker for <i>C9ORF72</i> -associated amyotrophic lateral sclerosis

There is no effective treatment for amyotrophic lateral sclerosis (ALS), a devastating motor neuron disease. However, discovery of a G₄C₂ repeat expansion in the <i>C9ORF72</i> gene as the most common genetic cause of ALS has opened up new avenues for therapeutic intervention for this form of ALS. G₄C₂ repeat expansion RNAs and proteins of repeating dipeptides synthesized from these transcripts are believed to play a key role in <i>C9ORF72</i>-associated ALS (c9ALS). Therapeutics that target G₄C₂ RNA, such as antisense oligonucleotides (ASOs) and small molecules, are thus being actively investigated. A limitation in moving such treatments from bench to bedside is a lack of pharmacodynamic markers for use in clinical trials. We explored whether poly(GP) proteins translated from G₄C₂ RNA could serve such a purpose. Poly(GP) proteins were detected in cerebrospinal fluid (CSF) and in peripheral blood mononuclear cells from c9ALS patients and, notably, from asymptomatic <i>C9ORF72</i> mutation carriers. Moreover, CSF poly(GP) proteins remained relatively constant over time, boding well for their use in gauging biochemical responses to potential treatments. Treating c9ALS patient cells or a mouse model of c9ALS with ASOs that target G₄C₂ RNA resulted in decreased intracellular and extracellular poly(GP) proteins. This decrease paralleled reductions in G₄C₂ RNA and downstream G₄C₂ RNA-mediated events. These findings indicate that tracking poly(GP) proteins in CSF could provide a means to assess target engagement of G₄C₂ RNA-based therapies in symptomatic <i>C9ORF72</i> repeat expansion carriers and presymptomatic individuals who are expected to benefit from early therapeutic intervention.