Professional Documents
Culture Documents
Neuropsychiatric Genetics
Prevention and Treatment Center for Psychological Diseases, No.102 Hospital of Chinese Peoples Liberation Army,
Changzhou, Jiangsu, P.R. China
Department of Rehabilitation, No.102 Hospital of Chinese Peoples Liberation Army, Changzhou, Jiangsu, P.R. China
Administration office, No.102 Hospital of Chinese Peoples Liberation Army, Changzhou, Jiangsu, P.R. China
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Department of Psychiatric Medicine, Bengbu Medical College, Bengbu, Anhui, P.R. China
Department of Burn Surgery, Changhai Hospital of Second Military Medical University, Shanghai, P.R. China
Department of Psychology and Psychiatry, Second Military Medical University, Shanghai, P.R. China
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Department of Neurology and Psychiatry, Postgraduate school, Xuzhou Medical College, Xuzhou, Jiangsu, P.R. China
Department of Laboratory, No.102 Hospital of Chinese Peoples Liberation Army, Changzhou, Jiangsu, P.R. China
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Xin-yang Sun, Jin Zhang, and Wei Niu have contributed equally to this paper and agreed to share the first authorship position together.
Conflict of interest: none.
Correspondence to: Dr. Li-yi Zhang, MM, Prevention and Treatment Center for Psychological Diseases, No.102 Hospital of Chinese Peoples
Liberation Army, North Peace Road 55, Changzhou213003, Jiangsu, Peoples Republic of China.
E-mail: zly102@126.com
Correspondence to: Jim Lu, MD, GoPath Laboratories LLC, 1351 Barclay Blvd, Buffalo Grove, IL 60089, United States of America.
E-mail: JLu@gopathlabs.com
Article first published online in Wiley Online Library
(wileyonlinelibrary.com): 00 Month 2015
DOI 10.1002/ajmg.b.32292
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treatment, expression levels of miR-132, miR-181b, miR-432 and
miR-30e were significantly decreased. In addition, the improvement of clinical symptomatology was significantly correlated
with the changes of miR-132, miR-181b, miR-212 and miR-30e
expression levels. Furthermore, the decreases of plasma levels of
miR-132 and miR-432 were significantly greater in high-effect
subgroup than those in low-effect subgroup after six-week
treatment course. We conclude that miR-30e, miR-181b, miR34a, miR-346 and miR-7 combined as a panel are potentially
useful non-invasive biomarkers for schizophrenia diagnosis.
Markers miR-132, miR-181b, miR-30e and miR-432 are potential
indicators for symptomatology improvements, treatment
responses and prognosis for schizophrenia patients.
2015 Wiley Periodicals, Inc.
Key words: schizophrenia; microRNA; biomarker; antipsychotics; qPCR; PANSS; GAS; CGI
INTRODUCTION
Reliable diagnosis and selective choice of more effective therapeutic
regimens remain challenge for clinical management of schizophrenia (SZ) patients [Kawanishi et al., 2000];[Killackey and Yung,
2007];[Schwarz and Bahn, 2008a];[Schwarz and Bahn, 2008b];
[Lakhan and Kramer, 2009]. Currently, diagnoses of SZ patients
largely rely on self-reported experience and abnormalities in behavior, followed by a clinical assessment of psychiatrists. There is no
objective test available for assisting clinical diagnosis of SZ patients.
Furthermore, there is an urgent need for laboratory tests which
guide the selection of more effective antipsychotic drugs for individual SZ patients. Therefore, biomarkers for differential diagnoses
and prediction of responses to antipsychotic therapy will be essential in developing novel strategies for successful treatment of SZ
patients.
Altered expressions of miRNAs have been demonstrated to
contribute to the genetic and biological basis of neuropsychiatric
disorders [Abelson et al., 2005; Kim et al., 2007; Schipper et al., 2007;
Feng et al., 2009; Cox et al., 2010] and specific miRNA expression
profiling has been also observed in postmortem gray matter including the prefrontal cortex, parietal and temporal cortices of SZ
patients [Thomson et al., 2004; Perkins et al., 2007; Beveridge et al.,
2008; Mellios et al., 2009]. As exemplified by the parallel changes in
the levels of certain mRNAs between CNS and peripheral blood
[Vawter et al., 2001; Marazziti et al., 2010], it is plausible to suggest
that disease associated changes in miRNA expression might also be
detectable in peripheral tissues such as plasma and mononuclear
leucocytes. Indeed, blood-based miRNA profiling has become an
increasingly important approach to identify clinically applicable
biomarkers for SZ since brain tissue is not readily accessible.
Hansen et al. studied the association between SZ and genetic
variants of miRNA genes dominantly expressed in brain tissue
using peripheral blood samples, and found that rs17578796 and
rs1700 in miR-206 and miR-198 showed nominal significant allelic
association with SZ[Hansen et al., 2007]. Gardiner et al. explored
miRNA expression profiling in peripheral blood from 112 patients
with SZ and 76 non-psychiatric controls, and revealed 83 signifi-
METHODS
Patients
Sixty-one SZ patients fulfilling the criteria as defined by the
Diagnostic and Statistical Manual 4th edition (DSM-IV) were
prospectively recruited from No.102 Hospital of the Chinese
Peoples Liberation Army from July 2012 to May 2013. Clinical
diagnoses of the patients were made by at least two consultant
psychiatrists, and the diagnoses were further confirmed by an
additional experienced clinical psychiatrist. All patients were first
visitors to the clinics and prior to any clinical treatment for SZ, or in
the absence of psychotropic medication within at least three
months. No patient had history of severe medical diseases, other
psychiatric disorders, structural brain disorders, mental retardation, unstable psychiatric features and movement disorders. Also,
patients who had brain injury causing traumatic amnesia longer
than 24 hr and who received blood transfusion within a month or
electroconvulsive therapy within 6 months, were excluded from the
study. In addition, 62 healthy controls without any family history of
major psychiatric disorders (SZ, bipolar disorder and major depression) within the last three generations were recruited. Similarly,
all healthy controls were without any history of blood transfusion or
severe traumatic event within a month. Patients and healthy controls were matched in gender, age and ethnicity. All individuals
recruited in the study were provided with written informed consent. The study was approved by local Institutional Review Boards.
Medication Intervention
To examine effects of clinical medications on plasma miRNA levels
in SZ patients, 25 out of the 61 SZ patients whose PANSS score were
higher than 60 were particularly selected for receiving six-week
medication intervention. Among 25 patients, seven were treated
with olanzapine (initial dosage 5 mg, average dosage 13 mg, and
SUN ET AL.
dosage range 520mg); eight with quetiapine (initial dosage
100 mg, average dosage 550 mg, and dosage range 100800mg);
six with ziprasidone (initial dosage 40 mg, average dosage 135 mg,
and dosage range 40140mg); additional four with risperidone
(initial dosage 2 mg, average dosage 4.7 mg, and dosage range 2
6mg). All four medications were atypical antipsychotics, treating
both positive and negative symptoms, rarely inducing extrapyramidal symptoms under administration of therapeutic dose.
miRNA Selection
The targeted 10 miRNAs(miR-132, miR-181b, miR-195, miR-212,
miR-30e, miR-346, miR-34a, miR-432, miR-7 and miR-137) in this
study were selected by thoroughly and extensively surveying current
literatures on Pubmed database, Springer database and the University of British Columbia library database. All published studies
about miRNAs in schizophrenia and antipsychotics from year 2000
to May 2013 were searched and analyzed. The search showed more
than 30 articles, which were further classified in terms of article type.
These 10 miRNA were reported to be associated with SZ in more
than three independent original research articles (Table S1).
miRNA Preparation
Whole blood (5 ml) was collected from all subjects (SZ patients and
healthy controls) using EDTA anticoagulant tube. The plasma was
separated by centrifugation and transferred into a fresh RNase/
DNase-free 2 ml microcentrifuge tube, and stored at 80C until
use. Exact 2 ml of plasma were used for miRNA preparation for all
plasma samples using the miRNeasy Serum/Plasma Kit (Qiagen,
CA) according to the manufacturers protocol. The total miRNA
samples were stored at 80C. C. elegans synthetic miR-39 was used
as a spiked-in and normalization control (Qiagen) as suggested by
the manufacturer[Thomson et al., 2004; Kroh et al., 2010; Wang
et al., 2011; Song et al., 2014].
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reactions using TaqMan MicroRNA Reverse Transcription Kit
(Applied Biosystems, Inc., Grand Island, NY) and miRNA-specific stem-loop primers were supplied by the TaqMan MicroRNA
Assays(Applied Biosystems, Inc.). The primer sequences of the
tested miRNAsare shown in Table S2. The RT reactions was
mixed according to the manufacturers protocol and performed
in an Applied Biosystems 9700 PCR instrument using the following conditions: 16C for 30 min, 42C for 30 min, 85C for
5 min, hold at 4C. Real-time PCR reactions were performed in a
5-mL reaction mixture containing 2.5 mL TaqMan Universal PCR
Master Mix II(Applied Biosystems, Inc.), 0.25mL miRNA-specific
primer/probe mix(Applied Biosystems, Inc.), and 2.25 mL diluted
RT cDNA product. PCR reactions in a 384 well reaction plate
were run in a 7900HT fast real-time PCR system (Applied
Biosystems, Inc.) using following cycling parameters: 95C for
10 min, followed by 40 cycles of 95C for 15 sec and 60 C for
1 min. Each reaction was performed in triplicate. Realtime PCR
data were collected by SDS software. For analysis of relative levels
of the tested miRNAs, we used C. elegans synthetic miR-39 as a
normalization control since exactly the same amount of plasma
was applied for miRNA preparation for all plasma specimens.
This synthetic miRNA as a normalization was suggested by the
miRNeasy Serum/Plasma Kit(Qiagen) and has been commonly
used as a normalization for analysis miRNA levels in peripheral
blood in previous publications[Kroh et al., 2010; Song et al.,
2014; Thomson et al., 2004; Wang et al., 2011]. The relative levels
of the tested miRNA in plasma were presented as DDCt which
was calculated using the following formula: 2-(CtmiRNA CtmiR)
NA39 [Thomson et al., 2004; Kroh et al., 2010; Wang et al., 2011;
Song et al., 2014].
Statistical Analysis
Wilcoxon rank sum test was used to test the age difference between
SZ and healthy controls, and expression levels of the individual
tested miRNA between different gender(male or female), different
age(above P75 or below P25), different sibling status(with or
without siblings), different residence(urban or rural), different
educational levels(above college or below high school), different
income levels(above 10,000 for annual income or below 10,000 for
annual income), different marital status(married or unmarried)
and different family history(family patients or sporadic patients) in
SZ group. Wilcoxon rank sum test was also used to compare the
expression levels of miRNA between SZ and healthy controls.
Univariate repeated measures analysis of variance and LSD test
were performed to compare the plasma levels of miRNA before and
after medication. KruscalWallis H test and MannWhitney test
were employed to test the changes of miRNA expression levels
among the groups receiving different medication. Spearman correlation test was carried out for testing the correlation of miRNA
expression levels with scale scores (total PANSS score, GAS score
and CGI score). Receiver-operating characteristics (ROC) curves
were established to evaluate the diagnostic performance of miRNAs
for differentiating between SZ and healthy controls. Conditional
logistic regression analyses were then performed to select the most
significant miRNA with the maximum acceptable limit for adding a
variable being 0.1 and the minimum acceptable limit for removing a
variable being 0.15. Mann-Whitney U test was carried out to test the
expression levels changes between different treating effect subgroups. All statistical analyses were carried out using SPSS version
17.0 software (Chicago, IL). P < 0.05 was considered statistically
significant.
RESULTS
Clinical Characteristics of the Patients
The demographic information of all the 123 subjects (both the SZ
patients and healthy controls) is shown in Table I. No significant
differences (P > 0.05) were found in age and genders between 61
SZ patients and 62 healthy controls. Among patients from SZ
group, no significant differences (P > 0.05) in miRNA expression
levels were found between genders, sibling status, residential
locations, educational levels, marital statuses and family history.
The 61 SZ patients were further classified into higher and lower
age subgroups according to age percentile. Patients with ages
above 37.5 were classified into the higher age subgroup (P75,
n 16) and those with ages below 19 into the lower age subgroup
(P25, n 13). Statistical analysis revealed that only miR-30e
expression level was significantly elevated in the lower age
subgroup as compared to the higher age subgroup(P < 0.01,
Table S3). In addition, 25 out of all the 61 patients who had
PANSS score more than 60 were selected to undergo six-week
antipsychotics treatment. These 25 patients who underwent
three-week treatment and 18 patients who underwent six-week
treatment with complete experimental and clinical data were
included for the final analysis.
SZ(n 61)
Control(n 62)
27.84 (10.64)
15-56
28.08(10.98)
16-56
39(63.93%)
22(36.07%)
40(64.52%)
22(35.48%)
44
17
46
15
31
30
21
40
11
50
SUN ET AL.
Fig. 1. Comparision of aberrant miRNAs expression levels between SZ and healthy controls. Plasma levels of selected MiRNAs were analyzed
in 61 SZ patients and 62 matched healthy controls using quantitative real-time PCR. The figure shows DDCT of 5 miRNAs significantly
different between SZ patients and healthy controls. The plots were constructed using GraphPad Prism 5 software and statistical difference
was analyzed by Wilcoxon sum test.
DISCUSSION
212, and miR-30e) highly correlated with the clinical scores changes
of the patients after medication (Table III). To verify the role of
plasma miRNAs as biomarkers predicting therapeutic effects in SZ
patients, we reclassified all patients into high-effect and low-effect
subgroup based on the PANSS score reduction rate((Spre-Spost)/
Spre, Spre represents pre-medication score, Spost represents postmedication score) and the degree of improvement of the symptoms
TABLE II. Comparison of miRNA Expression in SZ Patients Between Pre-Medication, Three-Week and Six-Week Post-Medication(DDCt)
(n 18)
miRNA
miRNA-132
miRNA-181b
miRNA-195
miRNA-212
miRNA-30e
miRNA-346
miRNA-34a
miRNA-432
miRNA-7
Pre-medication
5.14 0.33
2.06 0.26
3.64 0.43
5.66 0.27
1.95 0.42
4.95 0.43
9.21 0.43
4.79 0.39
9.78 0.32
Three-week post-medication
5.18 0.33*
1.95 0.27
3.67 0.28
5.85 0.32
2.40 0.40
5.42 0.22
9.31 0.28
4.88 0.38
11.13 0.56
Note: Compared with pre-medication,* represents P < 0.05, ** represents P < 0.01; compared with three-week post-medication, D represents P < 0.05.
Six-week post-medication
5.95 0.28*D
2.80 0.24**
4.27 0.36
6.13 0.24
3.07 0.34*
5.30 0.21
9.97 0.59
5.91 0.44D
10.76 0.42
SUN ET AL.
TABLE III. Correlation Between PANSS and GAS Scores Changes and miRNA DDCt Value Changes in SZ Patients Before, Three-Week and
Six-Week Post-Medication(r)
PANSS total score changes
DmiRNA
miRNA-132
miRNA-181b
miRNA-212
miRNA-30e
D1
-0.129
-0.270
0.103
0.024
D2
-0.282
-0.191
-0.557*
-0.204
D1
-0.119
0.574*
-0.106
-0.108
D2
-0.383
-0.162
-0.336
-0.320
D3
-0.202
0.021
-0.043
-0.277
Note: D1represents the differences of three-week post-medication and pre-medication;D2 represents the differences of six-week post-medication and three-week post-medication;D3represents the
differences of six-week post-medication and pre-medication;* represents P < 0.05.
TABLE IV. Comparison of miRNA Expression Changes in SZ Patients Between Different Symptomatology Improvement Subgroups After
Six-Week Medicaiton(x s)
miRNA
miRNA-132
miRNA-181b
miRNA-195
miRNA-212
miRNA-30e
miRNA-346
miRNA-34a
miRNA-432
miRNA-7
*
High-effect subgroup (n 8)
1.59 1.05*
1.35 1.52
0.79 1.86
1.01 1.82
1.71 1.92
0.33 2.80
1.18 3.23
2.04 2.17*
2.06 3.12
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sensitivity of 35.5% sensitivity and specificity of 90.2% respectively.
Among these five miRNA, miR-181b was the most significant
predictor for SZ with OR of 2.438 as demonstrated by multivariate
logistic regression analysis. Taken together, findings from our
current and other previous studies suggest that miR-181b, miR30e, miR-34a, miR-346 and miR-7 can be developed to blood-based
biomarkers for SZ diagnosis.
Currently, dopamine receptors system, 5-HT receptors, and
GABA system, are the major pharmacological treatments for SZ
patients. However, the pharmacological actions of these drugs
remain unclear, making it difficult to select effective treatment
and predict treatment responses. Up to date, very limited numbers
of studies have investigated pharmaceutical impact on miRNA
expression. Our current study observed altered expression levels of
miR-30e, miR-181b, miR-132, and miR-432 upon medication,
suggesting that these four miRNAs might be functionally implicated in clinical responses to psychotropic drugs in SZ patients. We
further explored the effects of different psychotropic medications
on plasma miRNA levels. Among the four medication subgroups,
the expression of miR-30e, miR-195, miR-132, and miR-432
showed significantly different patterns of changes, with the olanzapine showing the strongest effect. The findings imply that these
four miRNA are the most sensitive to psychotropic medication, and
olanzapine has the most effective therapeutic effect on SZ patients.
Association of plasma miRNA changes with therapeutic effects of
drugs was further supported by the observations that symptomatic
improvements of SZ after treatment tightly correlated with plasma
levels changes of four miRNAs, including miR-132, miR-181b,
miR-212 and miR-30e. In addition, we found that the decreases of
plasma levels of miR-132 and miR-432 were significantly greater in
high-effect subgroup than those in the low-effect subgroup at
different periods of treatment course. Recent findings of Miller
et al. suggest that miR-132 dysregulation and subsequent abnormal
expression of miR-132 target genes contribute to the neurodevelopmental and neuromorphological pathologies present in schizophrenia [Miller et al., 2012]. These findings taken together suggest
that miR-30e, miR-181b, miR-132, and miR-432 can be potential
indicators for therapeutic responses of SZ patients.
In conclusion, miR-181b, miR-30e, miR-34a, miR-346 and miR7 combined as a panel might serve as useful biomarkers for
diagnosis of SZ patients, and markers miR-30e, miR-181b, miR132 and miR-432 combined are potential indications for symptomatology improvements, treatment responses and prognosis for
schizophrenia patients.
LIMITATIONS
It would be ideal to have a microarray analysis before the PCR
verification. Another limitation of our study was the sample size,
which may not be powerful enough to detect variants with minor
effect. Besides, observational study results need to be confirmed by
prospective studies. Thus, further epidemiological and functional
studies in a larger cohort are warranted to validate these results.
Since this study did not involve other psychiatric diseases, such as
bipolar disorder and major depressive disorder and so on, the
conclusion of this study needs further differential verification
against other psychiatric diseases.
ACKNOWLEDGMENTS
The authors wish to thank all the subjects who participated in the
study. Authors also acknowledge invaluable assistance by numerous mental health professionals in different clinical departments
within and without No.102 Hospital of PLA. In particular, the
authors wish to convey sincere thanks to Gopath Global LLC.,
Chicago in USA for their professional laboratory services and
No.102 Hospital clinical laboratory of PLA for professional laboratory assistance.
REFERENCES
Abelson JF, Kwan KY, ORoak BJ, Baek DY, Stillman AA, Morgan TM,
Mathews CA, Pauls DL, Rasin M-R, Gunel M. 2005. Sequence variants in
SLITRK1 are associated with Tourettes syndrome. Science 310(5746):317320.
Bajestan SN, Sabouri AH, Nakamura M, Takashima H, Keikhaee MR,
Behdani F, Fayyazi MR, Sargolzaee MR, Bajestan MN, Sabouri Z. 2006.
Association of AKT1 haplotype with the risk of schizophrenia in Iranian
population. Am J Med Genet Part B Neuropsychiatr Genet 141(4):
383386.
Beveridge N, Gardiner E, Carroll A, Tooney P, Cairns M. 2009. Schizophrenia is associated with an increase in cortical microRNA biogenesis.
Mol Psychiatr 15(12):11761189.
Beveridge NJ, Tooney PA, Carroll AP, Gardiner E, Bowden N, Scott RJ,
Tran N, Dedova I, Cairns MJ. 2008. Dysregulation of miRNA 181b in the
temporal cortex in schizophrenia. Hum Mol Genet 17(8):11561168.
Chang TC, Wentzel Ea Fau - Kent OA, Kent Oa Fau - Ramachandran K,
Ramachandran K Fau - Mullendore M, Mullendore M Fau - Lee KH, Lee
Kh Fau - Feldmann G, Feldmann G Fau - Yamakuchi M, Yamakuchi M
Fau - Ferlito M, Ferlito M Fau - Lowenstein CJ, Lowenstein Cj Fau - Arking
DE, Arking De Fau - Beer MA, Beer Ma Fau - Maitra A, Maitra A Fau Mendell JT, Mendell JT. 2007. Transactivation of miR-34a by p53 broadly
influences gene expression and promotes apoptosis 26(5):745752.
Consortium SPG-WAS. 2011. Genome-wide association study identifies
five new schizophrenia loci. Nat Genet 43(10):969976.
Cox MB, Cairns MJ, Gandhi KS, Carroll AP, Moscovis S, Stewart GJ,
Broadley S, Scott RJ, Booth DR, Lechner-Scott J. 2010. MicroRNAs miR17 and miR-20a inhibit T cell activation genes and are under-expressed in
MS whole blood. PLoS One 5(8):e12132.
Emamian ES, Hall D, Birnbaum MJ, Karayiorgou M, Gogos JA. 2004.
Convergent evidence for impaired AKT1-GSK3b signaling in schizophrenia. Nat Genet 36(2):131137.
Fallin MD, Lasseter VK, Avramopoulos D, Nicodemus KK, Wolyniec PS,
McGrath JA, Steel G, Nestadt G, Liang K-Y, Huganir RL. 2005. Bipolar I
disorder and schizophrenia: A 440single-nucleotide polymorphism
screen of 64 candidate genes among Ashkenazi Jewish case-parent trios.
Am J Hum Genet 77(6):918936.
Feng J, Sun G, Yan J, Noltner K, Li W, Buzin CH, Longmate J, Heston LL,
Rossi J, Sommer SS. 2009. Evidence for X-chromosomal schizophrenia
associated with microRNA alterations. PLoS One 4(7):e6121.
Gardiner E, Beveridge N, Wu J, Carr V, Scott R, Tooney P, Cairns M. 2011.
Imprinted DLK1-DIO3 region of 14q32 defines a schizophrenia-associated miRNA signature in peripheral blood mononuclear cells. Mol
Psychiatry 17(8):827840.
Guo S-Z, Huang K, Shi Y-Y, Tang W, Zhou J, Feng G-Y, Zhu S-M, Liu H-J,
Chen Y, Sun X-D. 2007. A Case-control association study between the
GRID1 gene and schizophrenia in the Chinese Northern Han population.
Schizophr Res 93(1):385390.
SUN ET AL.
Hansen T, Olsen L, Lindow M, Jakobsen KD, Ullum H, Jonsson E,
Andreassen OA, Djurovic S, Melle I, Agartz I. 2007. Brain expressed
microRNAs implicated in schizophrenia etiology. PloS one 2(9):e873.
Hunsberger JG, Austin DR, Chen G, Manji HK. 2009. MicroRNAs in
mental health: from biological underpinnings to potential therapies.
Neuromolecular Med 11(3):173182.
Kawanishi Y, Tachikawa H, Suzuki T. 2000. Pharmacogenomics and
schizophrenia. Eur J Pharmacol 410(2):227241.
Killackey E, Yung AR. 2007. Effectiveness of early intervention in psychosis.
Curr Opin Psychiatry 20(2):121125.
Kim AH, Reimers M, Maher B, Williamson V, McMichael O, McClay JL,
van den Oord EJ, Riley BP, Kendler KS, Vladimirov VI. 2010. MicroRNA
expression profiling in the prefrontal cortex of individuals affected with
schizophrenia and bipolar disorders. Schizophr Res 124(1):183191.
9
Pepe MS, Thompson ML. 2000. Combining diagnostic test results to
increase accuracy. Biostatistics 1(2):123140.
Perkins DO, Jeffries CD, Jarskog LF, Thomson JM, Woods K, Newman MA,
Parker JS, Jin J, Hammond SM. 2007. MicroRNA expression in the
prefrontal cortex of individuals with schizophrenia and schizoaffective
disorder. Genome Biol 8(2):R27.
Schipper HM, Maes OC, Chertkow HM, Wang E. 2007. MicroRNA
expression in Alzheimer blood mononuclear cells. Gene Regul Sys
Biol 1:263.
Schwarz E, Bahn S. 2008a. Biomarker discovery in psychiatric disorders.
Electrophoresis 29(13):28842890.
Schwarz E, Bahn S. 2008b. The utility of biomarker discovery approaches
for the detection of disease mechanisms in psychiatric disorders. Br J
Pharmacol 153(S1) S133S136.
Kroh EM, Parkin RK, Mitchell PS, Tewari M. 2010. Analysis of circulating
microRNA biomarkers in plasma and serum using quantitative reverse
transcription-PCR (qRT-PCR). Methods 50(4):298301.
Song H-t, Sun X-y, Zhang L, Zhao L, Guo Z-m, Fan H-m, Zhong A-f, Niu
W. 2014. A preliminary analysis of association between the downregulation of microRNA-181b expression and symptomatology improvement in schizophrenia patients before and after antipsychotic
treatment. J Psychiatr Res 54:134140.
Lai C-Y, Yu S-L, Hsieh MH, Chen C-H, Chen H-Y, Wen C-C, Huang Y-H,
Hsiao P-C, Hsiao CK, Liu C-M. 2011. MicroRNA expression aberration
as potential peripheral blood biomarkers for schizophrenia. PloS one
6(6):e21635.
Lakhan SE, Kramer A. 2009. Schizophrenia genomics and proteomics: are
we any closer to biomarker discovery. Behav Brain Funct 5(2):19.
Marazziti D, Catena DellOsso, Baroni M, Masala S, DellOsso I, Consoli B,
Giannaccini G, Betti G, Lucacchini L. 2010. Alterations of the dopamine
transporter in resting lymphocytes of patients with different psychotic
disorders. Psychiatr Res 175(1):5457.
Mellios N, Huang H-S, Baker SP, Galdzicka M, Ginns E, Akbarian S. 2009.
Molecular determinants of dysregulated GABAergic gene expression in
the prefrontal cortex of subjects with schizophrenia. Biol Psychiatry
65(12):10061014.
Miller BH, Zeier Z Fau - Xi L, Xi L Fau - Lanz TA, Lanz Ta Fau - Deng S,
Deng S Fau - Strathmann J, Strathmann J Fau - Willoughby D, Willoughby D Fau - Kenny PJ, Kenny Pj Fau - Elsworth JD, Elsworth Jd Fau Lawrence MS, Lawrence Ms Fau - Roth RH, Roth Rh Fau - Edbauer D,
Edbauer D Fau - Kleiman RJ, Kleiman Rj Fau - Wahlestedt C, Wahlestedt
C. 2012. MicroRNA-132 dysregulation in schizophrenia has implications
for both neurodevelopment and adult brain function. (1091 -6490
(Electronic)) 109(8):31253130.
Pepe MS, Cai T, Longton G. 2006. Combining predictors for classification
using the area under the receiver operating characteristic curve. Biometrics 62(1):221229.
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