Neuroprotective Effects of σ₂R/TMEM97 Receptor Modulators in the Neuronal Model of Huntington's Disease

Huntington's disease (HD) is a genetic neurodegenerative disease caused by an expanded CAG repeat in the Huntingtin (HTT) gene that encodes for an expanded polyglutamine (polyQ) repeat in exon-1 of the human mutant huntingtin (mHTT) protein. The presence of this polyQ repeat results in neuronal degeneration, for which there is no cure or treatment that modifies disease progression. In previous studies, we have shown that small molecules that bind selectively to σ₂R/TMEM97 can have significant neuroprotective effects in models of Alzheimer's disease, traumatic brain injury, and several other neurodegenerative diseases. In the present work, we extend these investigations and show that certain σ₂R/TMEM97-selective ligands decrease mHTT-induced neuronal toxicity. We first synthesized a set of compounds designed to bind to σ₂R/TMEM97 and determined their binding profiles (K_i values) for σ₂R/TMEM97 and other proteins in the central nervous system. Modulators with high affinity and selectivity for σ₂R/TMEM97 were then tested in our HD cell model. Primary cortical neurons were cultured in vitro for 7 days and then co-transfected with either a normal HTT construct (Htt N-586-22Q/GFP) or the mHTT construct Htt-N586-82Q/GFP. Transfected neurons were treated with either σ₂R/TMEM97 or σ₁R modulators for 48 h. After treatment, neurons were fixed and stained with Hoechst, and condensed nuclei were quantified to assess cell death in the transfected neurons. Significantly, σ₂R/TMEM97 modulators reduce the neuronal toxicity induced by mHTT, and their neuroprotective effects are not blocked by NE-100, a selective σ₁R antagonist known to block neuroprotection by σ₁R ligands. These results indicate for the first time that σ₂R/TMEM97 modulators can protect neurons from mHTT-induced neuronal toxicity, suggesting that targeting σ₂R/TMEM97 may lead to a novel therapeutic approach to treat patients with HD.