Missense variants in GABA receptor beta2 subunit disrupt receptor biogenesis and cause loss of function.

Gamma-aminobutyric acid type A receptors (GABARs) are the major inhibitory neurotransmitter-gated channel in the mammalian central nervous system. GABARs function as heteropentamers, typically composed of two α1, two β2, and one γ2 subunits. Protein homeostasis between GABAR folding, trafficking, assembly, and degradation is critical to ensure normal physiological functions. Variants in genes encoded for GABARs lead to numerous neurological disorders, such as genetic epilepsy with or without neurodevelopmental delay. While these variants are associated with severe clinical presentations of epilepsy, the molecular mechanisms driving the disease remain to be elucidated. In this study, we focused on four missense epilepsy-associated variants (EAVs) in the gene: Q209F210delinsH (c. 627_629del), R240T (c. 719G>C), I246T (c. 737T>C), and I299S (c. 896T>G). HEK293T cells exogenously expressing these β2 variants exhibited significantly reduced GABA-induced peak chloride current, indicating their loss of function. However, the four β2 EAVs differed in the degree of proteostasis deficiencies, including increased ER retention, compromised assembly, decreased protein stability, and reduced trafficking and surface expression, with Q209F210delinsH and R240T variants leading to the most severe degradation. Collectively, these results indicate that these epilepsy-linked variants have debilitating effects on the early biogenesis of the β2 subunit, causing misfolding, aggregation, and rapid degradation before it can be assembled with other subunits and transported to the plasma membrane. Overall, our work offers crucial mechanistic insight into how specific β2 missense variants impact the proteostasis maintenance of GABARs, which could facilitate the development of effective therapeutics for genetic epilepsy by targeting trafficking-deficient GABAR variants.