Integrated genetic and epigenetic approach to developmental psychiatric disorders: analysis of human blood and brain

Environmental stimuli influence the developmental trajectories of neural circuits from birth through adolescence. Exposure to harmful environmental stimuli during these developmental stages may result in increased vulnerability to psychiatric disorders. These effects are suggested to be partly dependent on genotype and mediated by epigenetic mechanisms. We aim to identify biomarkers and new therapeutic targets for the treatment of developmental psychiatric disorders, primarily alcohol dependence. We will perform genome-wide analysis of DNA methylation in blood from 2000 adolescents part of the IMAGEN study on factors that influence mental health in adolescents (, and of genotype, DNA methylation and gene expression in brain from circa 400 controls at different developmental stages and adult alcohol dependents. Loci associated with phenotypic traits relevant to alcohol dependence, DNA methylation and gene expression in the IMAGEN sample and diagnosis, DNA methylation and gene expression in the brain sample will be considered candidate biomarkers for alcohol dependence. Mechanisms underlying these associations will be considered candidate therapeutic targets for the treatment of alcohol dependence.

Alcohol binge drinking – induced cognitive impairment in health and working life: genetic vulnerability and novel pharmacotherapy

Alcohol binge drinking patterns and excessive alcohol consumption have severe effects on public health and working life in Sweden and its Nordic neighbors. Delayed impairment of cognitive capabilities and executive functions are the most deteriorating consequence of heavy drinking. We examine the novel pathophysiological mechanism of this impairment, and develop clinically manageable strategies to identify subpopulations at risk and effective pharmacotherapy to treat alcohol-produced cognitive impairment. According to the mechanism, cycles of alcohol intoxication and withdrawal impair cognition through dysregulations in neurotransmission that is controlled by the endogenous opioid systems (EOS) including kappa-opioid receptor (KOPr) and its ligands dynorphins, prodynorphin (PDYN) products. 1) We examine whether genetic variants of the KOPr/OPRK1 and PDYN genes contribute to alcohol use, abuse and dependence, and to alcohol-induced cognitive impairments. Population-based cohorts of young adult monozygotic and dizygotic twin pairs and office workers/social drinkers are analysed. 2) By conducting human post-mortem studies, we are identifing and characterizing dysregulations in the opioid and glutamate neurotransmitter systems, potential targets for pharmacological interference, in human heavy alcohol drinkers; and analyze underlying epigenetic mechanism. We focus on epigenetic modifications that may cause long-lasting behavioral alterations. 3) We plan to use animals models to mimic binge alcohol - induced impairment of cognitive functions and evaluate if these detrimental effects are reversed by available and novel pharmacological means. Characterization of the novel mechanism of alcohol binge drinking - induced cognitive impairment will open new possibilities for identification of subpopulations at risk, and for therapeutic intervention with opioid or glutamate antagonists.

Pathogenic mechanisms of human neuropeptide mutations: implications for regulation of the anti-reward system

Much of our knowledge about functions of genes and proteins in human brain has come from analysis of rare human and mouse mutations. Such mutations represent entry points into novel signaling pathways and molecular mechanisms. In collaboration with Dr. Dineke Verbeek, Dept. Genetics, Univ. Med. Center Groningen, The Netherlands, we have identified missense mutations in the human prodynorphin gene (PDYN) coding for a precursor to opioid peptides dynorphins (Bakalkin et al., 2010). These mutations cause profound neurodegeneration in the cerebrum and cerebellum underlying the development of spinocerebellar ataxia type 23, a dominantly inherited neurodegenerative disorder. Remarkably, seven out of eight mutations are located in dynorphins, which also have non-opioid neurodegenerative activities. This is the first finding on neuropeptide mutations underlying human neuropathology. The best-established function of the dynorphins in the brain is regulation of the anti-reward system by inducing dysphoria that counterbalances positive hedonic effects of endorphins and enkephalins, and addictive substances. Analysis of PDYN mutations could uncover novel molecular and cellular mechanisms of anti-reward - reward regulation that are generally unknown. We focus on two mechanisms. First, the mutations may impair correct folding of PDYN molecules in the endoplasmic reticulum, resulting in PDYN aggregation, cellular stress and the unfolded protein response. Long-lasting activation of the unfolded protein response by mutant PDYNs, or by wild-type - PDYN excessively produced under pathological conditions such as substance addiction, may lead to neuronal dysfunction, atrophy and neurodegeneration. Second, PDYN mutations may enhance non-opioid pathogenic activities of dynorphins. Our previous analysis suggests that dynorphins may mediate communications between neurons through non-receptor excitatory mechanism. This mechanism may be engaged in pathogenic effects of upregulated wild-type - dynorphins and mutant peptides. Our general goal is to understand pathogenic mechanisms of PDYN mutations and, in the following studies to evaluate whether wild-type - PDYN and dynorphins engage these mechanisms to regulate reward circuits under normal conditions and in substance addiction when these neuropeptides are excessively produced. We explore pathogenic mechanisms underlying actions of wild-type- and mutant-PDYN in cellular and in vitro biochemical/ biophysical studies, and are planning to use in vivo transgenic mice expressing human wild-type- or pathogenic mutant-PDYN that have been produced by Dr. Verbeek. Generalized pathological changes including cerebral cortical and subcortical atrophy, and agenesis of corpus callosum in patients caring PDYN mutations emphasize the fundamental role of neuropeptides in brain functions, and propose that the essential molecular mechanisms are affected; the phenomenon that requires detailed investigation. Dissection of these mechanisms is critically important for understanding of reward – anti-reward, substance addiction, depression and chronic pain, all neuropathological conditions in which dynorphins play a critical role, as well as for the neuropeptide and neurodegeneration fields in general because of the novelty of mechanisms identified.