Abstract
Homozygous DNAJC12 c.79-2A>G (p. V27Wfs*14) loss-of-function mutations were first reported as a cause of young-onset Parkinson's disease. However, bi-allelic autosomal recessive pathogenic variants in DNAJC12 may lead to an alternative constellation of neurological features, including infantile dystonia, developmental delay, intellectual disability and neuropsychiatric disorders. DNAJC12 is understood to co-chaperone aromatic amino acid hydroxylases to foster the synthesis of biogenic amines. In vitro, we discover overexpressed DNAJC12 forms a complex with guanine triphosphate cyclohydrolase 1 (GCH1), the rate-limiting enzyme in the synthesis of tetrahydrobiopterin, a cofactor paramount for biogenic amines synthesis. We also confirm DNAJC12's interaction with tyrosine (TH) and tryptophan hydroxylase (TPH), which are rate-limiting enzymes for synthesis of biogenic amines dopamine (DA) and serotonin (5-HT). In-vitro knock-down of DNAJC12 with a siRNA destabilizes the DNAJC12-TH-GCH1 complex, reducing GCH1 levels, whereas reciprocal overexpression of both TH and GCH1 increases endogenous DNAJC12, alluding to the significance of modulating the DNAJC12-TH-GCH1 complex as a therapy for DNAJC12 and other biogenic amine disorders. We extend these investigations to a Cre-conditional knock-out mice (cDKO) in which loxP sites flanking Dnajc12 exon 2 enable its excision by cre-recombinase. With germline Cre expression, we have created a constitutive Dnajc12 knock-out (DKO). DKO mice exhibit reduced locomotion/ exploratory behavior at 3 months in automated open-field testing, accompanied by increased plasma phenylalanine which is a cardinal feature of patients with pathogenic DNAJC12 variants. In striatal tissue, total DA and 5-HT, their metabolites, and electrically-evoked DA release are all reduced. Biochemical alterations in synaptic proteins are also apparent, with enhanced phosphorylation of Th pSer31 and pSer40 reflecting biological compensation. Most immediately, cDKO and DKO mice present models to develop and refine therapeutic approaches for biogenic amines disorders, including dystonia and parkinsonism. They will also enable the pleiotropic functions of biogenic amines (including DA), usually synthesized in the brain or periphery, to be separated.