High-content microscopy reveals a morphological signature of bortezomib resistance.

Drug resistance is a challenge in anticancer therapy, particularly with targeted therapeutics and cytotoxic compounds. In many cases, cancers can be resistant to the drug prior to exposure, i.e., possess intrinsic drug resistance. However, we lack target-independent methods to anticipate resistance in cancer cell lines or characterize intrinsic drug resistance without a priori knowledge of its cause. We hypothesized that cell morphology could provide an unbiased readout of drug sensitivity prior to treatment. We therefore isolated clonal cell lines that were either sensitive or resistant to bortezomib, a well-characterized proteasome inhibitor and anticancer drug to which many cancer cells possess intrinsic resistance. We then measured high-dimensional single-cell morphology profiles using Cell Painting, a high-content microscopy assay. Our imaging- and computation-based profiling pipeline identified morphological features typically different between resistant and sensitive clones. These features were compiled to generate a morphological signature of bortezomib resistance, which correctly predicted the bortezomib treatment response in seven of ten cell lines not included in the training dataset. This signature of resistance was specific to bortezomib over other drugs targeting the ubiquitin-proteasome system. Our results provide evidence that intrinsic morphological features of drug resistance exist and establish a framework for their identification.

Regulation of mRNA expression encoding chaperone and co-chaperone proteins of the glucocorticoid receptor in peripheral blood: association with depressive symptoms during pregnancy.

BACKGROUND: Major depressive disorder during pregnancy associates with potentially detrimental consequences for mother and child. The current study examined peripheral blood gene expression as a potential biomarker for prenatal depressive symptoms. METHOD: Maternal RNA from whole blood, plasma and the Beck Depression Inventory were collected longitudinally from preconception through the third trimester of pregnancy in 106 women with a lifetime history of mood or anxiety disorders. The expression of 16 genes in whole blood involved in glucorticoid receptor (GR) signaling was assessed using real-time polymerase chain reaction. In parallel, plasma concentrations of progesterone, estradiol and cortisol were measured. Finally, we assessed ex vivo GR sensitivity in peripheral blood cells from a subset of 29 women. RESULTS: mRNA expression of a number of GR-complex regulating genes was up-regulated over pregnancy. Women with depressive symptoms showed significantly smaller increases in mRNA expression of four of these genes - FKBP5, BAG1, NCOA1 and PPID. Ex vivo stimulation assays showed that GR sensitivity diminished with progression of pregnancy and increasing maternal depressive symptoms. Plasma concentrations of gonadal steroids and cortisol did not differ over pregnancy between women with and without clinically relevant depressive symptoms. CONCLUSIONS: The presence of prenatal depressive symptoms appears to be associated with altered regulation of GR sensitivity. Peripheral expression of GR co-chaperone genes may serve as a biomarker for risk of developing depressive symptoms during pregnancy. The presence of such biomarkers, if confirmed, could be utilized in treatment planning for women with a psychiatric history.

Sleep in schizophrenic patients on and off haloperidol therapy. Clinically stable vs relapsed patients.

We examined the state-dependent contribution of neuroleptic withdrawal and psychotic relapse in influencing sleep measures. Eighteen clinically stable male schizophrenic patients taking haloperidol were studied with 3 nights of polysomnography for baseline measures and again after neuroleptic withdrawal. Sleep measures were also obtained at the point of relapse (n = 9) or after a 6-week drug-free period if the patient remained clinically stable (n = 9). Neuroleptic withdrawal led to a global deterioration of rapid eye movement and non-rapid eye movement sleep and a reduction of rapid eye movement latency in both groups. Relapsers differed from nonrelapsers in that they had a larger decrease in total sleep time, sleep efficiency, total non-rapid eye movement sleep, and stage 2 sleep. The level of psychosis was inversely correlated with sleep efficiency, total sleep time, and stage 4 sleep in the drug-free patients. Our data suggest that clinical state needs to be identified in sleep studies of drug-free patients.

Behavioral vs biochemical prediction of clinical stability following haloperidol withdrawal in schizophrenia.

BACKGROUND: We sought to identify haloperidol-treated subjects who relapsed within 6 weeks of placebo replacement and those who did not, using multivariate analysis. METHODS: In the week prior to discontinuation of haloperidol treatment, global behavioral ratings and a lumbar puncture for cerebrospinal fluid monoamine metabolities were obtained in 88 patients with chronic schizophrenia. Logistic regression analyses were used to evaluate two competing models of relapse prediction. The models were then compared using receiver operating characteristic analysis and a final combined model was derived. RESULTS: The behavioral model was less variable in its prediction than the cerebrospinal fluid monoamine model. The final model consisted of increased psychosis, decreased anxiety, higher cerebrospinal fluid homovanillic acid levels, and lower cerebrospinal fluid 5-hydroxyindoleacetic acid levels. CONCLUSIONS: Several monoamine systems are involved in psychotic relapse within 6 weeks of haloperidol withdrawal. Future studies of relapse prediction should include both clinical and biological measures to fully assess relapse risk.

Glutamatergic neurotransmission involves structural and clinical deficits of schizophrenia.

BACKGROUND: Phencyclidine and ketamine induce a syndrome closely resembling schizophrenia due to their blockade of N-methyl-D-aspartate (NMDA) receptor. These findings suggested that some aspects of schizophrenia are associated with decreased NMDA--glutamatergic function. We hypothesized that structural and symptomatic deficits in schizophrenia are related to glutamatergic neurotransmission. METHODS: We studied the relationships among cerebrospinal fluid (CSF) glutamatergic markers, clinical presentation of schizophrenia, and CT parameters of brain structure in drug-free schizophrenics. RESULTS: We found no significant differences between patients with schizophrenia and controls in CSF glutamatergic markers. When patients with schizophrenia were considered as a group, significant negative correlations between glutamatergic markers and brain structural measures as well as clinical measures were observed. Cluster analysis reveals a group of lower indices of glutamatergic neurotransmission, and more prominent thought disorder as well as ventricular enlargement, and a group with increased glutamate level. CONCLUSIONS: The findings support the hypothesis that altered glutamatergic neurotransmission plays a role in the brain structure and the clinical symptoms of schizophrenia.

CSF diazepam binding inhibitor and schizophrenia: clinical and biochemical relationships.

Diazepam-binding inhibitor (DBI) is a 9-kD neuropeptide that interacts with the benzodiazepine (BZD) binding sites of the neuronal gamma-aminobutyric acid type A (GABAA) receptor and with the glial mitochondrial BZD receptor (MBR). We explored the involvement of CSF DBI-LI in schizophrenia, based on the potential role of GABA in the negative symptoms associated with schizophrenia, the relationship of its receptors with dopamine and norepinephrine release, and the proposed therapeutic efficacy of BZDs in schizophrenia. Clinical data, CSF DBI-LI and CSF monoamine measures were obtained in 65 drug-free male chronic (DSM-IIIR) schizophrenic patients, 53 of whom were also tested prior to haloperidol withdrawal. Following haloperidol withdrawal, CSF DBI-LI increased significantly. Drug-free CSF DBI-LI did not correlate with CSF monoamines. CSF DBI-LI was significantly higher in paranoid compared to chronic undifferentiated schizophrenic patients. The data suggest that DBI may have a symptom modulatory rather than an etiological role in schizophrenia.

Further studies of elevated cerebrospinal fluid neuronal cell adhesion molecule in schizophrenia.

BACKGROUND: The purposes of the present study were to attempt to replicate a previous finding of increased cerebrospinal fluid (CSF) neuronal cell adhesion molecule (N-CAM) in schizophrenia, and to assess whether the increases could be related to medication, clinical state effects, or brain structural measures. METHODS: CSF N-CAM was measured by the Western blot technique in 45 DSM-III-R diagnosed male schizophrenic patients both on and off haloperidol treatment and in 20 healthy male control subjects. RESULTS: CSF N-CAM was significantly increased in schizophrenic patients, with no overlap in the ranges, when compared to controls. There were no significant effects of medication or exacerbation on CSF N-CAM. No associations with measures of brain structure were found. CONCLUSIONS: Because N-CAM levels were not shown to be different on and off treatment or in exacerbated versus nonexacerbated patients, the higher levels seen in schizophrenic patients may be inherent to the disorder and possibly related to neurodevelopment.

Deterioration in premorbid functioning in schizophrenia: a developmental model of negative symptoms in drug-free patients.

OBJECTIVE: The authors examined the relationship between negative symptoms and premorbid variables in drug-free schizophrenic patients. METHOD: The authors studied 63 clinically stable male schizophrenic inpatients who were not receiving any psychoactive medication. The patients were classified as having negative, positive, or mixed symptoms, and their premorbid functioning during childhood, early adolescence, and late adolescence was assessed by using the Premorbid Adjustment Scale. Correlational analyses were applied to the classification and developmental models. RESULTS: Patients with negative symptoms had significantly lower levels of premorbid functioning during late adolescence and significantly greater premorbid deterioration between childhood and early adolescence. Correlational analysis revealed significant positive relationships between premorbid variables and negative symptoms. CONCLUSIONS: The data suggest that a deterioration in social and intellectual functioning between childhood and adolescence is associated with the development of a negative symptom syndrome in schizophrenia. The premorbid deterioration appears to be an early prodrome of the disorder. Whether this residual negative symptom syndrome is in some way related to the deficit syndrome of schizophrenia awaits a prospective study.

Empirical validation of primary negative symptoms: independence from effects of medication and psychosis.

OBJECTIVE: Recent studies of negative symptoms in schizophrenia-specifically, those involving the deficit syndrome-have focused on uncovering the symptoms that are primary to the disease rather than secondary to the psychotic process. One of the foremost concerns in this effort is establishing whether the negative symptoms observed are the result of medication effects. METHOD: This study used negative symptom ratings obtained in a drug withdrawal paradigm to compare symptom profiles in the same schizophrenic patients when they were on and off antipsychotic drug treatment. The study group consisted of 93 physically healthy male patients with DSM-III-R-defined schizophrenia. Principal components analysis was performed on negative symptom data obtained separately during haloperidol treatment and again when the patients were drug free to determine whether there were meaningful factor scores that were consistent across medication conditions. Drug withdrawal effects on negative symptom factors were then tested for associations with secondary sources of variance including extrapyramidal side effects, anxiety/depression, and psychosis. RESULTS: Two factors, termed affective flattening and diminished motivation, exhibited similar loadings when the patients were both on and off medication. Changes in motivation were associated with changes in anxiety/depression and psychosis, while changes in affective flattening were associated with changes in extrapyramidal side effects. CONCLUSIONS: The documented secondary sources of negative symptoms are related to different and distinct aspects of negative symptoms; this finding will aid in the identification of primary negative symptoms.

CSF dopamine beta-hydroxylase in schizophrenia: associations with premorbid functioning and brain computerized tomography scan measures.

OBJECTIVE: The authors attempted to replicate previous findings of relationships of CSF dopamine beta-hydroxylase with premorbid functioning and computerized tomography (CT) scan measures in a new cohort of schizophrenic patients. METHOD: Data on CSF dopamine beta-hydroxylase-like immunoreactivity and premorbid functioning, as well as CT scans, were obtained in 60 drug-free, male schizophrenic patients and two groups of normal comparison subjects. RESULTS: CSF dopamine beta-hydroxylase did not differ between the comparison subjects and schizophrenic patients. Lower CSF dopamine beta-hydroxylase was associated with better premorbid social functioning and with less prefrontal sulcal widening. Better premorbid school functioning and more years of education, however, were associated with higher CSF dopamine beta-hydroxylase. CONCLUSIONS: The association between low CSF dopamine beta-hydroxylase and premorbid functioning was confirmed for social functioning, while the opposite was observed for scholastic performance, suggesting that these are different dimensions. Dopamine beta-hydroxylase modulates the prognosis and potentially the course of schizophrenia without necessarily causing the disorder.

Increases in CSF levels of interleukin-2 in schizophrenia: effects of recurrence of psychosis and medication status.

OBJECTIVE: Interleukin-2, traditionally viewed as solely involved in immunological events, has recently been shown to exert profound effects on the development and regulation of the central nervous system. This study examined the relationships between interleukin-2 in the CSF and plasma of schizophrenic patients and clinical measures, including relapse and medication status. Plasma and CSF interleukin-1 alpha levels were also measured to ascertain the specificity of changes in cytokine levels. METHODS: Seventy-nine physically healthy male patients with schizophrenia (DSM-III-R) received diagnostic evaluation and behavioral ratings. Haloperidol treatment was withdrawn for up to 6 weeks and patients were evaluated for symptom recurrence. CSF and plasma were obtained by established procedures before haloperidol withdrawal (N = 79) and after (N = 64). RESULTS: CSF levels of interleukin-1 alpha decreased significantly after haloperidol withdrawal but showed no relation to clinical status. In contrast, levels of CSF interleukin-2 were associated with recurrence of psychotic symptoms. Relapse-prone patients, examined both while medicated and after drug withdrawal, had significantly higher levels of CSF interleukin-2 than patients who did not relapse. CSF interleukin-2 level during haloperidol treatment was a significant predictor of worsening in psychosis. CONCLUSIONS: Levels of interleukin-2, a molecule that plays both neurodevelopmental and neuroregulatory roles, may have a role in relapse in schizophrenia. Levels of CSF interleukin-2 appear to be affected by relapse mechanisms, while peripheral blood levels are not. These changes are specific to interleukin-2, since levels of interleukin-1 alpha were affected by medication withdrawal but not by change in clinical state.

Dopamine turnover in schizophrenia before and after haloperidol withdrawal. CSF, plasma, and urine studies.

The dopamine hypotheses of schizophrenia and antipsychotic drug action suggest that the dopamine metabolite homovanillic acid (HVA) should change with drug withdrawal and change in clinical state. We designed a study of cerebrospinal fluid (CSF), plasma, and urinary HVA on and off haloperidol to examine the effects of drug withdrawal. CSF and plasma HVA samples were obtained in 72 healthy schizophrenic (DSM-III-R) males (age: 36 +/- 7.4 years), before and after haloperidol withdrawal, which was after 6 weeks on placebo or sooner if they met specific criteria for relapse. We collected three 24-hour urine samples in 34 of these patients. In addition, CSF HVA was obtained in 24 well-screened age-matched male normal controls. HVA was measured with high-pressure liquid chromatography (HPLC). CSF HVA decreased significantly after drug withdrawal, particularly in those who met relapse criteria; drug-free CSF HVA levels were not significantly different from those of normals. Plasma HVA increased significantly after haloperidol withdrawal in relapsing patients, but not in clinically stable patients. Urinary HVA excretion decreased after withdrawal with decreased HVA clearance. We conclude that haloperidol withdrawal had a strong effect on dopamine turnover, whereas the patient's clinical state had only a weak central effect, without affecting total body production of HVA. Conceivably, dopamine involvement in schizophrenia reflects the failure of the homeostatic mechanisms that allow for integration of different functional brain components as needed.

Plasma catecholamine metabolites as markers for psychosis and antipsychotic response in schizophrenia.

The objective of this study was to determine the association between the patterns of change in the dopaminergic metabolite plasma homovanillic acid (HVA), the noradrenergic metabolite 3-methoxy-4-hydroxyphenylglycol (MHPG), and psychosis following haloperidol withdrawal in schizophrenic patients. Weekly plasma measurements were obtained in 107 subjects with schizophrenia or schizoaffective disorder. Random regression was used to control for individual variance while modeling metabolite changes over time and relationships with psychosis. Changes in plasma MHPG were not significantly associated with relapse or psychosis, while increased plasma HVA was found to be associated with relapse. Psychosis was correlated negatively with plasma HVA levels. The current analysis, controlling for individual variance, indicates that there is evidence for pharmacological effects on plasma HVA, but not plasma MHPG. In addition, these metabolites do not appear to be direct markers of psychosis, but may be associated with a compensatory response by the system to return to the steady state.

Predicting duration of clinical stability following haloperidol withdrawal in schizophrenic patients.

Although chronic maintenance antipsychotic drug treatment is the most effective way of preventing relapse in schizophrenic patients, it is not very successful. A considerable number of patients relapse on medication, and many others do not take their medications as prescribed after leaving the hospital. Unfortunately, clinicians are not able to identify how long patients will remain clinically stable after drug discontinuation. To develop a model consisting of behavioral and monoaminergic variables to identify the risk of symptom exacerbation, we obtained in the week prior to haloperidol discontinuation global behavioral ratings and cerebrospinal fluid (CSF) values for monoamine metabolites in a sample of 109 DSM-III-R schizophrenic patients. Patients were followed until specific criteria for increases in psychosis were met for up to 1 year and then returned to antipsychotic drug treatment. Cox regression analysis identified predictors of the survival function, or the probability of relapse at a given time drug free. The best model indicated that increased psychosis, decreased anxiety, an increased CSF homovanillic acid (HVA) to 5-hydroxyindoleacetic acid (5-HIAA) ratio, and decreased CSF 3-methoxy-4-hydroxyphenylglycol (MHPG) prior to haloperidol withdrawal were associated with early increases in psychosis. Our study indicates that it is possible to identify those patients who are more likely to remain clinically stable without medication. When the model is validated, it will help clinicians assess the relapse risk over time, lower doses in treatment-resistant patients, and possibly determine the optimal time for aftercare visits following hospital discharge.

Delta sleep-inducing-peptide-like immunoreactivity (DSIP-LI) and delta sleep in schizophrenic volunteers.

Delta sleep-inducing-peptide (DSIP) has been reported to increase sleep in subjects with insomnia. The authors studied cerebrospinal fluid (CSF) DSIP-like immunoreactivity (DSIP-LI) in 15 drug-free male subjects with a DSM-IIIR diagnosis of schizophrenia. The subjects underwent a lumbar puncture and three nights of polysomnography. CSF DSIP-LI was significantly correlated with polysomnography the night before the LP: with stage 3 sleep (p = 0.05), stage 3 and delta (stages 3 + 4) sleep during the first nonrapid eye movement NREM period (p = 0.02 and p = 0.05, respectively) and the ratio of the first and second NREM period (p < 0.05), and negatively with stage 2% sleep (p < 0.05). Whether this first report of a potential relationship between CSF DSIP-LI and slow-wave sleep in man might be generalized to sleep in nonpsychiatric subjects awaits further study.

Provider attitudes and self-reported behaviors related to hormone replacement therapy.

OBJECTIVE: The purpose of this study was to survey providers within a large health maintenance organization regarding their attitudes and practice patterns related to counseling women about hormone replacement therapy (HRT). DESIGN: A total of 260 providers from gynecology (n = 81), family practice (n = 96), and internal medicine (n = 83) from owned and contracted clinics were surveyed. Each was asked about prescribing philosophies, behaviors, and barriers to providing counseling regarding HRT. RESULTS: Respondents reported HRT's greatest benefit to be in the prevention of osteoporosis (99%) and cardiovascular conditions (96%). Gynecologists were more likely to report the benefits of HRT for Alzheimer's than were clinicians in internal medicine or family practice (p < 0.05), and women providers were more likely than men to report this (p < 0.01). There was no statistical difference based on years in practice. Providers did not vary significantly by specialty or sex in their concerns of risk for breast cancer of endometrial cancer. However, those in family practice and internal medicine were significantly more likely to report concern about thromboembolism (p < 0.01). Only 42% of physicians claimed to initiate discussion with their patients more than 75% of the time. The two factors most often mentioned as barriers to counseling were time and lack of adequate knowledge. CONCLUSIONS: Providers want to be an integral part of their patient's education regarding HRT; however, time constrains and a need for adequate information make this difficult. Now health systems must examine models of education for both providers and patients to ensure that women have access to current information with which to make informed decisions.

Chronic haloperidol treatment does not affect structure of attention in schizophrenia.

Results of a number of investigations indicate attention is a multifactorial construct composed of four distinct cognitive factors including focus-execute, sustain, encode and shift abilities. While investigators have partially or fully replicated this attentional structure in a number of clinical and nonclinical populations, no study has adequately examined the structure of attention in patients with schizophrenia who are not treated with antipsychotics. In this study, we examined the four-factor theory of attention in patients with schizophrenia while they were stabilized on haloperidol (with no adjunctive antiparkinsonian/anticholinergic medications) and again when they were approximately 3 weeks drug free. Standard neuropsychological measures were used to assess attentional functions. Principal components analyses (varimax rotation) of neuropsychological test scores in medicated and drug-free conditions indicated that four factors accounted for 84.2 and 91.8 of total variance in medicated and unmedicated conditions, respectively. Based on these results, it appears that: (1) haloperidol does not appreciably affect structure of the attentional system in patients with schizophrenia; (2) unmedicated patients with schizophrenia exhibit a similar structure of attention as both medicated patients and controls, suggesting that attentional structure is 'normal' in schizophrenia; and (3) the four-factor attention theory is a useful and valid paradigm for evaluating attention in patients with schizophrenia, regardless of medication status.

Utility of WAIS-R short forms in schizophrenia.

Recently, short forms of the Wechsler Adult Intelligence Scale-Revised (Wechsler, 1981) have received increasing attention because of their ability to provide estimated IQ scores with substantial time savings (in some cases 85-90% savings). These short forms may have particular utility for individuals with schizophrenia because they require less time to administer and, as a result, are less taxing for these patients who often exhibit impaired attention and deficient motivation. In this study, we examine the psychometric properties of nine popular WAIS-R short forms in a group of 143 patients diagnosed with schizophrenia. Our results indicated that Kaufman's four subtest short form was the best overall estimator of Full Scale IQ (FSIQ) when a combination of administration time and psychometric properties were considered. However, Ward's seven subtest short form provided the closest estimation of FSIQ and had the lowest misclassification rate, while also providing estimates of Verbal and Performance IQs and yielding 46.5-49.7% time savings. All short forms had substantial misclassification rates, indicating that caution is warranted when using these forms to classify individuals according to standard levels of intellectual functioning (e.g., Average, Low Average, High Average). Clearly, the main consideration in selecting a short form is whether time savings or accuracy have priority.

Inter-rater reliability of the neurological examination in schizophrenia.

Neurological Examination Abnormalities (NEA) are prevalent in schizophrenia, but the significance of this is obscured by methodological problems. The Neurological Evaluation Scale (NES), the most widely used structured neurological examination in schizophrenia research, has had limited study of its inter-rater reliability (IRR). An augmented version of the NES was jointly administered (one examiner-rater and one observer-rater) by three pairs of psychiatrists to two populations of patients with idiopathic psychotic disorders. In addition to the ordinal and categorical data yielded by the original NES, continuous data were recorded in one of the series. Reliability analyses of our populations and a previously published study, reveal consistently adequate IRR in 12 of the 26 items assessed, and inconsistently adequate IRR in an additional 11. Consistent with studies using other NEA schedules, IRR was unacceptably low for some items that rely on subjective severity ratings. Certain rare abnormalities, which posed difficulties for the estimation of IRR, are probably not generally useful in the study of schizophrenia. Reliability estimates of continuous, ordinal and dichotomous data were comparable in most cases. We recommend that certain items from the NES be deleted, and that other studies of NEA in psychiatry follow similar procedures before undertaking further analyses.

Confirmation of a two-factor model of premorbid adjustment in males with schizophrenia.

Because schizophrenia is considered to be a neurodevelopmental disorder, premorbid adjustment is of particular interest. Premorbid adjustment is probably not a unitary construct but rather is expressed across a number of developmental domains. The current investigation examined the validity of a two-factor model that differentiated premorbid adjustment across social and academic domains and evaluated relationships between these premorbid adjustment domains and other variables of interest. Participants with schizophrenia (n = 141) underwent evaluation of premorbid adjustment (using the Premorbid Adjustment Scale), intellectual functioning, and psychiatric symptoms. Using confirmatory factor analysis, a two-factor model of premorbid adjustment was identified that included an academic domain and a social domain. The social domain was associated with symptom variables, while the academic domain was associated with measures of intelligence. Results provide evidence for at least two domains of premorbid adjustment in schizophrenia. Distinguishing between these two premorbid domains may be theoretically important because of potential differences in incidence rates and deterioration courses; some individuals with schizophrenia may exhibit adequate academic adjustment but poor social adjustment, while others may exhibit the opposite pattern.