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Psychiatric Times. Vol. 29 No. 3 February 28, 2012 Psychiatric Times. Vol. 29 No. 3VANGUARD ISSUES IN PSYCHIATRY Are Animal Models Relevant in Modern
Psychiatry?
By Andre Menache, BSc(Hons), BVSc, MRCVS | February 28, 2012Dr Menache is Director of Antidote Europe and is a Member of the Belgian Deontological Committeeon Animal Research.
The use of animals in medical research became firmly established in 1865, withthe publication of An Introduction to the Study of Experimental Medicine byClaude Bernard. This scientific discourse laid the groundwork for the study of comparative physiology between animals and humans. Half a century later, theanimal model was introduced into the behavioral sciences by early theoristsPavlov (classical conditioning), Watson (behaviorism), and Skinner (operantconditioning). Later, the animal model was used to investigate conditionsranging from maternal deprivation to depression and learned helplessness.
Most early psychiatric drugs were discovered through serendipity rather thanthrough the use of animal models. Isoniazid(Drug information on isoniazid),originally used to treat tuberculosis, was found to possess mood-alteringproperties and was marketed as the first antidepressant in 1957.2,3 MAO inhibitors originated from an effort to develop antituberculosis medications; they were superseded bytricyclic antidepressants (TCAs), which were discovered by clinical observation.
The potential psychotropic effect of chlorpromazine(Drug information on chlorpromazine), originallyused as an anesthetic adjunct in a Paris hospital in 1952, was discovered by a military surgeon later inthe same year.4,5 Thus, the phenothiazines came from a search for better pre-anesthetic agents.
The Australian physician John Cade reported the calming effect of lithium) in humans in 1949. The first benzodiazepine, chlordiazepoxide(Drug information onchlordiazepoxide) (Librium), was discovered accidentally in 1955. The first studies of benzodiazepines were unsuccessful attempts to treat patients with schizophrenia.8 In contrast to discoveries made through chance observation, newer psychotropic drugs, such as SSRIs,were discovered through the process of rational drug design. Five SSRIs (citalopram, fluvoxamine(Druginformation on fluvoxamine), fluoxetine(Drug information on fluoxetine), paroxetine(Drug informationon paroxetine), sertraline(Drug information on sertraline)) were produced independently by 5 differentcompanies. Rational drug design remains the main driving force behind the development of modern http://www.psychiatrictimes.com/display/article/10168/2039315 Psychiatric Times. Vol. 29 No. 3 February 28, 2012 Animals as model systems in psychiatry
Since the mid-20th century, researchers have designed animal models of stress, anxiety, depression, andobsessive-compulsive conditions in the laboratory to develop, test, and validate drugs to treat humandisorders.
Rats and mice are most commonly used in specific behavioral tests, such as the despair test, tail suspension test, and open field test.
Current animal models of human psychiatric conditions face the same methodological limitations asthey did 30 years ago. According to Beach : The validity of interspecific generalization can never exceed the reliability of intraspecificanalysis; and the latter is an indispensable antecedent of the former. . . . Significant comparison ofa particular type of behavior in two different species is impossible unless and until the behaviorhas been adequately analyzed in each species by itself.
This hypothesis is conditional on the existence (or availability) of animal models to accurately mimichuman psychiatric conditions. In reality, the overwhelming majority of mental disorders recognized byDSM, the International Statistical Classification of Diseases and Related Health Problems (published bythe World Health Organization), and the American Psychiatric Association do not have a counterpart inlaboratory animals. For those human conditions that are considered to have animal homologues, thereoften exist critical causal mechanisms that differ between humans and animals, which raisemethodological questions about the soundness and relevance of these animal models.15 Because of the multifactorial nature of conditions such as depression and anxiety and the ambiguitiesinherent in psychiatric diagnosis and treatment, the use of animal models in psychiatry presents uniquechallenges—unlike those found in other medical disciplines. In most cases, animal models represent acompromise because the cause and mechanism of the human condition under investigation may not befully understood. In addition, researchers are using a relatively simple system (receptor activation orinactivation) to represent a more complex and less readily studied system (human mental disorders).
While examples can be found to demonstrate common and conserved modes of action ofneurotransmitter chemicals throughout phylogenetically remote organisms, this approach has its limitswhen studying complex systems, such as the human CNS. According to molecular biologist Marc van The reductionist method of dissecting biological systems into their constituent parts has beeneffective in explaining the chemical basis of numerous living processes. However, manybiologists now realize that this approach has reached its limit. Biological systems are extremelycomplex and have emergent properties that cannot be explained, or even predicted, by studyingtheir individual parts. The reductionist approach—although successful in the early days ofmolecular biology—underestimates this complexity and therefore has an increasingly detrimentalinfluence on many areas of biomedical research, including drug discovery and vaccinedevelopment.
Animal models, in general, have not been subjected to the rigors of evidence-based medicine. Fewsystematic reviews or meta-analyses have been conducted to compare treatment outcomes in laboratoryanimals with outcomes in clinical trials. Overall, the animal model has performed poorly as a predictivemodality of human outcome in these reviews.18-22 Examples of animal models in psychiatry
http://www.psychiatrictimes.com/display/article/10168/2039315 Psychiatric Times. Vol. 29 No. 3 February 28, 2012 Behavioral despair. The behavioral despair model is commonly used to screen candidateantidepressants. Like other TCAs, imipramine(Drug information on imipramine) was screened using thePorsolt forced swim test. In this test, the animal (a rat or a mouse) is placed in a container with coldwater and is forced to swim until exhausted; it is then briefly taken out of the water. This is repeateduntil the animal reaches a state of helplessness and stops swimming. Although immobility time isreduced by antidepressant agents such as imipramine, significant strain differences have been reported.
23 The Porsolt forced swim test has been criticized on the grounds that the state of helplessness is more astrategy of survival than a sense of “despair.” The increased immobility simply demonstrates a positivebehavioral adaptation, in which the animal has learned that it cannot escape and is conserving energyuntil it is removed from the water. In addition, while the test is reported to distinguish antidepressantsand neuroleptics from anxiolytics, false positives have been reported for a number of other compounds,including stimulants, convulsants, anticholinergics, pentobarbital, and opiates.24,25 Canine acral lick dermatitis. This is considered by some researchers to be a suitable animal model forthe study of obsessive-compulsive disorder (OCD) in humans. However, most cases of acral lick dermatitis in animals have an underlying allergic cause; if the allergen is eliminated, the conditionresolves. In humans, OCD is an anxiety disorder characterized by intrusive thoughts, unrelated toallergies. Although SSRI treatment may be effective for both acral lick dermatitis in animals and OCDin humans because of a shared neurotransmitter response, this does not confer homology on the animalmodel. According to Nonneman and Woodruff, “If every aspect is fully isomorphic between the animal model and the human condition, including cause and mechanism, the model is homologous.”Clearly, the OCD dog model does not fulfill these criteria. In addition, the dog model ignores theexistence of a genetic component and the presence of comorbidities, which are thought to play animportant role in human OCD.28,29 Transgenic mice and the study of anxiety-like endophenotypes. Humans and mice share approximately97% of their working DNA and approximately 24,000 genes per body cell. In light of these facts, scientists have turned their attention to the study of differential gene expression patterns in geneticallyaltered mice to investigate anxiety through the corticotropin-releasing hormone system, the serotoninsystem, and the GABA system. However, because genes work in networks and animals are examples of complex systems, small changes at the gene level can have major consequences for the individual.
Thus, it is irrelevant to point to observed similarities in genetic makeup (including transgenes) betweenspecies, since the details of the differences are in the interactions between conserved genes, not in thegenes themselves.32 As Hirst and colleagues have shown in the example of the serotonin system, it is unlikely that serotonin type 6 knockout mice will be useful for validating the serotonin type 6 receptor as atherapeutic target because of unexpected species differences in both receptor regional distribution andpharmacological profiles. It is now evident that slight variations in rodent and human amino acidsequences can lead to unexpectedly large differences in the pharmacology of the receptors, withpotentially disastrous effects for drug development. What has not been clearly documented until thestudy by Hirst and colleagues is that mouse receptors could be significantly different from rat receptors.
Role of non-human primates in psychiatric and neurological research. The use of non-human primatesby Harlow and associates to demonstrate the effects of maternal deprivation has been well documented. In experiments conducted between 1957 and 1963, they removed baby rhesus monkeys http://www.psychiatrictimes.com/display/article/10168/2039315 Psychiatric Times. Vol. 29 No. 3 February 28, 2012 from their mothers and observed the effects of partial and total social isolation. Some of the monkeyswere kept in solitary confinement for up to15 years. These studies have been criticized on both ethicaland methodological grounds.15 Diagnosing “depression” in a monkey is at odds with the successful ongoing process of clarifyingpsychiatric diagnoses by using DSM criteria. It is not possible to ascertain feelings of worthlessness andexcessive guilt, indecisiveness, and thoughts of death from observations of monkeys. Harlow’s student Stephen Suomi became more circumspect in 1995, when he carefully referred to the condition inmonkeys as “something equivalent to depression” rather than “depression.” The non-human primate is considered by some researchers to be the mostappropriate model for the study of brain function. Among non-humanprimates, the rhesus macaque is the animal of choice for cognitivestudies. While there may be similarities between the brains of humansand non-human primates, the monkey brain is not a scaled down versionof the human brain. Rather, each primate brain is the unique result of evolutionary biology, molded over millions of years in response to
environmental, social, and genetic influences (Figure). With the human
brain, the effects of cultural evolution are also considered.
There are numerous differences in the anatomy and physiology of theCNS in monkeys and humans, including differences in locations ofspecialized areas in the brain. The visual 1 area accounts for 10% of thetotal cortex in the monkey but only 3% in humans, and anatomically corresponding visual areas inmonkeys and humans can perform very different functions. The human brain’s architecture and physiology is far more complex than that of the monkey brain. One indication of this is the length oftime it takes for the brain to develop in its major phase: 136 days for monkeys and 470 days for humans.
Other significant differences include the number of synapses a human neuron makes (between 7000 and 10,000) compared with the number a rhesus monkey neuron makes (between 2000 and 6000) andthe expression of at least 91 genes involved in a variety of neural mechanisms that differ betweenmonkeys and humans.37,39 According to Kreiman and associates : Even though the hippocampus appears to be one of the most conserved areas of the brain (mostsimilar among mammals), there are still considerable differences. Neurotransmitter receptordistribution varies widely between species. For example, there is an additional small layer ofhigh-density kainate receptors in the deepest part of the hippocampal molecular layer in themonkey, but not in humans. The inhibitory GABA receptors are located with high density in the human CA 1 hippocampal region, but not in the same region in monkeys. These resultsdemonstrate considerable changes of the regional and laminar distribution of important signalingmolecules in an otherwise evolutionary conservative brain region.
The validity of the animal model in regulatory toxicology
From a regulatory perspective, psychiatric drugs are required to undergo animal testing to demonstratesafety and efficacy. Safety relates to toxicity testing, which requires a rodent species (usually a rat) and a non-rodent species (usually a dog) to undergo single-dose (acute), repeated-dose, and long-termexposure to a candidate compound. The acute toxic dose (LD50) is the median lethal dose of acompound required to kill 50% of a group of test animals. Repeated-dose exposures usually require 14- http://www.psychiatrictimes.com/display/article/10168/2039315 Psychiatric Times. Vol. 29 No. 3 February 28, 2012 to 28-day studies, while long-term exposure requires up to 90 days in the rat and up to 12 months in thedog. Significant variations in the toxicity profile of a test compound may occur because of speciesdifferences in absorption, distribution, metabolism, and excretion (ADME).
Human lethal overdose figures are compiled by national poisons
information centers. These are empirical data obtained from human
accidental or deliberate poisoning and overdose (A. Vale, personal
communication, 2009). Human lethal overdose cannot be predicted from
animal LD50 values (Table). Animal studies are in fact poor predictors
of human outcome with respect to drug toxicity.41-44The use of
retrospective correlation is often inadvertently confused with prediction.
A classic study by Olson and colleagues looked at the concordance between adverse events seen in humans and data obtained frompreclinical animal toxicity studies. While this study measured sensitivity,it ignored specificity; therefore, its conclusions are largely irrelevant to the great prediction debate.32Although this study has been quoted in support of animal use as a predictive modality, Olson stated that “this study did not attempt to assess the predictability of preclinical experimental data to humans.” In addition to the challenges presented by differences in ADME between species, other variables caninfluence the outcome of animal studies. The laboratory environment is inherently stressful for theanimals. In addition to caged housing, which provides little or no environmental enrichment, evenroutine procedures can lead to significant changes in physiological parameters correlated with stress (eg,serum or plasma concentrations of corticosterone or glucose, heart rate, and blood pressure). While these changes may not be considered to be of much significance with respect to routine drug testing, thesame cannot be said of drugs specifically designed to treat stress and similar conditions. According toZinberg and Robertson, the difference between an animal’s natural setting and an artificial laboratory environment cannot be underestimated; it may produce major pharmacological effects. Variables such asgroup housing versus single housing, type of bedding, light-dark cycle, and handling by humans canaffect the outcome of a study.48-50 Evidence-based psychiatry
Evidence-based medicine aims to apply the best available evidence gained from the scientific method toclinical decision making. Clinical research is the solid ground of medicine, whereas biological theory isa necessary but changing superstructure. In an era of the human genome and advanced noninvasive screening techniques, it is perhaps timely to examine the relevance of the animal model in modernpsychiatry. This becomes all the more interesting when we consider the notion that psychiatry is theonly medical discipline that attempts to treat a conceptual abstraction—namely the human mind—inaddition to organs and physiological processes. Although molecular psychiatry provides some insight into the mechanism of mental illness, external psychosocial factors that influence human behavior fallwell outside of its scope.52 Some researchers continue to justify the need for invasive cognitive studies in animals (particularly thenon-human primate) on grounds that single neuron studies are not feasible in humans.53,54 That is nolonger the case, since single neuron activity can now be studied in patients with Parkinson disease whoundergo deep brain stimulation and in patients with epilepsy in whom electrode probes are used toidentify epileptic foci. We need to reconcile ourselves with the fact that the level of detail that has beenachieved in the study of the macaque brain will likely never be achieved in studies of the human brain.55 http://www.psychiatrictimes.com/display/article/10168/2039315 Psychiatric Times. Vol. 29 No. 3 February 28, 2012 Current trends indicate that diagnosis and treatment of psychiatric disorders in the 21st century will relymore on an integrative approach, including the best of genomic advances (eg, human braintranscriptome; pharmacogenomics in drug development geared toward personalized medicine);noninvasive screening techniques combined with ethical pharmacological studies, such aspharmaco-magnetoencephalography56,57; and clinical observation and other relevant human-basedmethodologies.
Empathy in psychiatry
There is an urgent need to acknowledge the role of empathy in science. The inclusion of empathy as afundamental tenet is especially important in psychiatry and psychology, as an antidote to themechanistic view of disease and the undervaluing or even ignoring of psychological and spiritualcomponents. Animal researchers have traditionally been circumspect about including any notion of empathy in their work, for fear of being stigmatized as anthropomorphic.
In contrast, some researchers have openly expressed the moral conflict they face when using non-humanprimates in experiments. These researchers occasionally acknowledge the very negative effects of family separation, isolation, and boredom, in addition to the pain and suffering inflicted on the animalsduring the experiments.
Several studies have indicated that mammalian species, especially the chimpanzee, have a sense of selfand self-awareness.59,60 This has led some scientists, particularly behaviorists, to suggest that theseanimals should be afforded special protection from pain, suffering, and incarceration. In addition, studies of other mammalian species, including mice and rats, suggest that they also possess awareness ofself and even more subtle “human” qualities associated with empathy and social joy. All of this empirical evidence points to a need to embrace a paradigm of “ultimate concern in science.”63 References
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