Murtada Alsaif

The application of selective reaction monitoring confirms dysregulation of glycolysis in a preclinical model of schizophrenia

BackgroundEstablishing preclinical models is essential for novel drug discovery in schizophrenia. Most existing models are characterized by abnormalities in behavioral readouts, which are informative, but do not necessarily translate to the symptoms of the human disease. Therefore, there is a necessity of characterizing the preclinical models from a molecular point of view. Selective reaction monitoring (SRM) has already shown promise in preclinical and clinical studies for multiplex measurement of diagnostic, prognostic and treatment-related biomarkers.MethodsWe have established an SRM assay for multiplex analysis of 7 enzymes of the glycolysis pathway which is already known to be affected in human schizophrenia and in the widely-used acute PCP rat model of schizophrenia. The selected enzymes were hexokinase 1 (Hk1), aldolase C (Aldoc), triosephosphate isomerase (Tpi1), glyceraldehyde-3-phosphate dehydrogenase (Gapdh), phosphoglycerate mutase 1 (Pgam1), phosphoglycerate kinase 1 (Pgk1) and enolase 2 (Eno2). The levels of these enzymes were analyzed using SRM in frontal cortex from brain tissue of PCP treated rats.ResultsUnivariate analyses showed statistically significant altered levels of Tpi1 and alteration of Hk1, Aldoc, Pgam1 and Gapdh with borderline significance in PCP rats compared to controls. Most interestingly, multivariate analysis which considered the levels of all 7 enzymes simultaneously resulted in generation of a bi-dimensional chart that can distinguish the PCP rats from the controls.ConclusionsThis study not only supports PCP treated rats as a useful preclinical model of schizophrenia, but it also establishes that SRM mass spectrometry could be used in the development of multiplex classification tools for complex psychiatric disorders such as schizophrenia.

Analysis of serum and plasma identifies differences in molecular coverage, measurement variability, and candidate biomarker selection

To compare the use of serum and plasma in multiplex immunoassay analyses of 190 proteins and small molecules, and associated molecular pathways. We also tested whether differences between these biofluids can influence the identification of potential biomarkers in a preliminary study comparing bipolar disorder patients with controls.

Behavioral and Molecular Biomarkers in Translational Animal Models for Neuropsychiatric Disorders

Modeling neuropsychiatric disorders in animals poses a significant challenge due to the subjective nature of diverse often overlapping symptoms, lack of objective biomarkers and diagnostics, and the rudimentary understanding of the pathophysiology. Successful translational research requires animal models that can inform about disease mechanisms and therapeutic targets. Here, we review behavioral and neurobiological findings from selected animal models, based on presumed etiology and risk factors, for schizophrenia, bipolar disorder, and major depressive disorder. We focus on the use of appropriate statistical tools and newly developed Research Domain Criteria (RDoC) to link biomarkers from animal models with the human disease. We argue that this approach will lead to development of only the most robust animal models for specific psychiatric disorders and may ultimately lead to better understanding of the pathophysiology and identification of novel biomarkers and therapeutic targets.

Differential expression of HINT1 in schizophrenia brain tissue

Recent findings in the literature suggest a relation between histidine triad nucleotide-binding protein-1 (HINT1) and psychiatric disorders such as major depression, anxiety, and schizophrenia, although its physiological roles are not completely comprehended. Using Western blot, we compared HINT1 protein expression in the postmortem dorsolateral prefrontal cortex and thalamus of schizophrenia patients and healthy controls for contributing to elucidate the role of HINT1 in schizophrenia pathophysiology. HINT1 was found to be downregulated in the dorsolateral prefrontal cortex and upregulated in the thalamus. Our results combined to previous studies in human samples and preclinical models support the notion that HINT1 must be more explored as a potential target for psychiatric disorders.

Multiplex immunoassay analysis of plasma shows prominent upregulation of growth factor activity pathways linked to GSK3β signaling in bipolar patients

BACKGROUND Our understanding of bipolar disorder (BD) aetiology has advanced in recent years but our ability to translate this to improve patient care in the clinic is still limited. METHODS In this study, we have measured the concentrations of 190 different molecules using sensitive multiplex immunoassays in plasma of 17 BD patients compared to 46 matched control subjects. RESULTS The analyses led to the identification of 26 dysregulated proteins in BD patients compared to controls. These molecules were comprised mostly of growth factors, hormones, lipid transport and inflammatory proteins. Decreased apolipoprotein A1 has previously been associated with BD patients and this was confirmed in our study. LIMITATIONS The present pilot study was limited by its small sample size, use of multiple drug treatments and the lack of dietary restrictions at the time of sampling. CONCLUSIONS Future studies may increase our understanding of BD which will help to pave the way for much-needed patient stratification for better treatment outcomes.

Proteomic profiling in schizophrenia: enabling stratification for more effective treatment

Schizophrenia is a heterogeneous psychiatric disorder characterized by an array of clinical manifestations. Although the best known manifestations include serious effects on mood and behavior, patients can also display co-morbidities, including immune system or metabolic abnormalities. Thorough characterization of these conditions using proteomic profiling methods has increased our knowledge of these molecular differences and has helped to unravel the complexity and heterogeneity of this debilitating condition. This could lead to patient stratification through characterization of biochemically different subtypes of the disease. In addition, proteomic methods have recently been used for molecular characterization of the mechanism of action of antipsychotic medications in both preclinical models and patients. This has resulted in identification of molecular panels that show some promise for prediction of response or for monitoring treatment outcome. This review describes how proteomic profiling methods can impact the future of schizophrenia diagnosis and therapeutics, and facilitate personalized medicine approaches for more effective treatment management of schizophrenia patients.

Distinct proteomic profiles in post-mortem pituitary glands from bipolar disorder and major depressive disorder patients

Disturbances of the hypothalamic-pituitary-adrenal axis have been implicated in the pathophysiology of bipolar disorder (BD) and major depressive disorder (MDD). To examine this further, we carried out proteomic profiling of post-mortem pituitaries from 13 BD and 14 MDD patients, in comparison to 15 controls. Liquid chromatography-mass spectrometry (LC-MS(E)) analysis showed that BD patients had significantly increased levels of the major pituitary hormones pro-opiomelanocortin (POMC) and galanin. BD patients also showed changes in proteins associated with gene transcription, stress response, lipid metabolism and growth signalling. In contrast, LC-MS(E) profiling revealed that MDD patients had significantly decreased levels of the prohormone-converting enzyme carboxypeptidease E and follow-up enzymatic analysis showed decreased activity of prolyl-oligopeptidase convertase. This suggested that altered prohormone processing may occur in pituitaries of MDD patients. In addition, MDD patients had significant changes in proteins involved in intracellular transport and cytoskeletal signalling. Finally, we carried out selective reaction monitoring (SRM) mass spectrometry profiling for validation of protein changes in key biological pathways. This confirmed increased POMC levels in BD patients with no change in the levels of this prohormone in MDD. This study demonstrates that proteomic profiling analysis of the pituitary can lead to new insights into the pathophysiology of BD and MDD. Also, given that the pituitary directly releases a variety of bioactive molecules into the bloodstream, many of the proteins identified here could serve as focal points in the search for peripheral biomarkers in clinical or drug treatment studies of BD and MDD patients.

Translating potential biomarker candidates for schizophrenia and depression to animal models of psychiatric disorders

Schizophrenia and major depressive disorder are severe mental illnesses, which are diagnosed based on patient interviews. Despite many years of extensive research, scientists have not yet fully deciphered how genetic and environmental factors interact to cause these illnesses. Biomarker tests that can confirm diagnoses of schizophrenia or depression are only now beginning to emerge, and could result in a paradigm shift in this field. These tests will help to evaluate the validity of animal models of psychiatric disorders, which are currently characterized based on behavioral measures. In this article, we explore the utility of translating both behavioral and molecular phenotypes of such models to the corresponding human disorders. This approach may help to provide construct validity to animal models and could lead to the identification of models corresponding to defined subtypes of neuropsychiatric disorders based on molecular profiles. Here, we review the molecular and biological pathway alterations that have been found in animal models of schizophrenia and depression and focus on those that are mirrored by similar abnormalities in human patients. Such parallels may provide insight into the validity of specific animal models and therefore help to provide more valuable and accurate tools for the discovery and development of improved psychiatric medications.

Multiplex immunoassay analysis of plasma shows differences in biomarkers related to manic or mixed mood states in bipolar disorder patients

BACKGROUND The molecular understanding of bipolar disorder (BD) aetiology has advanced over the last years through the identification of peripheral disease biomarkers. Here, we have attempted to identify plasma biomarkers associated with distinct BD mood states. METHODS Plasma from BD patients with either a current manic (n=29) or mixed (n=17) mood state and healthy controls (n=53) were analysed using a multiplex immunoassay platform. A total of 145 hormones, growth factors, transport proteins and inflammatory factors were measured. RESULTS Plasma levels of the hormones C-peptide, progesterone and insulin, and the inflammatory protein cancer antigen 125 were altered in both mood states. The hormone peptide YY and the growth factor trafficking protein sortilin were changed only in mania patients. Finally, the inflammatory factors haptoglobin, chemokine CC4 and matrix metalloproteinase 7 were altered specifically in mixed mood patients. LIMITATIONS This study was limited by a small sample size, potential confounding effects of multiple drug treatments in the patient groups, and lack of dietary restrictions at sampling. CONCLUSIONS Plasma from mania and mixed mood BD patients revealed similar changes in proteins related to insulin signalling, suggesting that these could be trait biomarkers. However, mania patients showed specific changes in hormonal and growth factor functions and mixed mood patients had a higher number of changes in inflammation-related molecules. Further studies of these and other biomarker candidates will increase our understanding of the systemic biological pathways affected in different BD mood states. This could lead to the identification of differential surrogate readouts and potential new drug targets for improved treatment outcomes.

Challenges in drug target discovery in bipolar disorder

Introduction: Misdiagnosis and subsequent inappropriate treatment of patients with bipolar disorder (BD) can worsen their clinical condition and outcome. Areas covered: This review focuses on the therapeutic targets which have been implicated in BD, including the glycogen synthase kinase 3 (GSK-3) and phosphoinositide signaling pathways. In addition, evidence is presented for potential new molecular strategies which involve targeting neuropeptide-converting endopeptidases, glutamatergic excitotoxicity, insulin signaling and dysfunctions in mitochondrial metabolism. Current limitations in study design, molecular platforms, preclinical and cellular models in the context of BD drug target discovery, suggest that there are many areas for improvement. Expert opinion: For the future outlook, this review outlines the importance of developments such as the use of BD patient-derived cellular models for providing better understanding of the BD etiology and robust translational drug screening tools in combination with developments in the fields of bioinformatics and systems biology.