This meta-analysis sought to identify the activation pattern of MNS during observation of stimuli with/without social-emotional components among individuals with ASD compared to age-matched typically developing controls. After a comprehensive literature search and the application of inclusion and exclusion criteria, 20 journal articles (with 24 experiments) were included in the meta-analysis. In summary, this coordinate-based fMRI meta-analysis indicated three main points. First, the MNS is impaired in ASD during action observation; Second, MNS dysfunction in ASD individuals is modulated by social-emotional components of the visual stimuli during action observation. Third, age seems to be an important factor in MNS function in ASD. The following discussion is divided into three parts, with each of the parts discussing the possible implications of the points listed above.
Abnormal activation within and beyond MNS in ASD
There has been a hot debate in the past decade on whether mirror neurons are “broken” in ASD, leading to the impairment in imitation in this group of individuals. The “broken mirror neuron theory” for autism asserts that the frontal and parietal brain regions with mirror neurons were found to be abnormally activated in individuals with ASD and given the property of mirror neurons (i.e., discharges during both activation observation and execution) that appears to support imitation, impaired imitation might be associated with the “broken” frontoparietal mirror neurons (35, 64). This meta-analysis indeed demonstrated abnormal MNS activations in individuals with ASD, evident by the hyperactivation of the right inferior frontal gyrus and left supplementary motor area, which supports the “broken mirror neuron theory”. However, at the same time, abnormal activations were also shown in brain regions beyond MNS, that is the hypoactivation of left precentral gyrus, amygdala and cerebellum. With the abnormalities shown in both MNS and other brain regions, one of the possible explanations is that the imitation deficits in ASD might not be solely attributable to the “broken mirror neurons”, but the interaction between MNS and other brain regions. One of the frameworks that studies the interplay between brain regions is neural connectivity (71). With regards to ASD, the disrupted connectivity theory posits that abnormal structural/functional connectivity in the brain might underlie the manifestation of behavioral phenotypes in ASD (72). Numerous empirical studies, including those using electrophysiological (73, 74) and neuroimaging (75) techniques, have pointed to disrupted neural connectivity in individuals with ASD. In particular, abnormal connectivity was found within the MNS. Regarding the neural connectivity between brain regions with mirror neurons, Rudie, Shehzad (76) recorded hypoconnectivity between the left inferior frontal gyrus pars opercularis and left parietal cortex in individuals with ASD relative to healthy controls. Regarding the connectivity between mirror neuron regions and the supporting MNS regions, it was revealed that the amygdala was hypoconnected with the frontal regions where the MNS is situated (77). This evidence collectively implies that atypical connectivity within the MNS contributes to dysfunction, which might be associated with the abnormal MNS activation patterns found in this meta-analysis.
MNS impairment during the observation of stimuli with/without social-emotional components
Two findings in the comparison between ASD and TD individuals, regarding the differences in activation patterns when they observe stimuli with and without social-emotional components, confirm our hypothesis that nature of the stimuli (with/without social-emotional components) might be one of the modulators of neural activation of the MNS. First, regarding the observation of stimuli without emotions, the MNS in the left hemisphere (i.e. left inferior parietal lobule and left supplementary motor area) were found hyperactivated, while the right middle occipital gyrus and the left postcentral gyrus were found hypoactivated in ASD when compared to TD individuals. Notably, the left-lateralized hyperactivation of MNS shown in our studies might be an important finding that enhance our understanding of the MNS deficits in ASD. Given an increase in neural activation measured by fMRI has been found to indicate an increase in mental effort during tasks to maintain behavioral performance (78), and the left-lateralized abnormalities in structural connectivity found in ASD in a previous study (79), it might be possible that individuals with ASD require extra mental effort for observation of stimuli without emotions, which could be attributable to the possible MNS deficits specifically in the left hemisphere. In terms of the MNS regions showing significant differences between ASD and TD, our results were largely consistent with that reported by Yang and Hofmann (28). However, we also noted some diverging results when compared to their study. Specifically, the hypoactivation in the left cerebellum Crus I and right postcentral gyrus were not found in their study, while our results reported highly significant hyperactivation in the left, but not the right, inferior parietal lobule. Given action observation and imitation was shown to engage the MNS as other brain regions in a differential manner (34), the diverging results could be due to the exclusion of the imitation studies in our meta-analysis. Furthermore, different methods of analysis and threshold levels might as well contribute to the inconsistency of results (80).
Second, regarding the observation of stimuli with emotions, our meta-analytic results indicated that the hyperactivation of the right inferior frontal gyrus in ASD individuals was statistically significant when compared to the TD individuals. Importantly, the hyperactivation of this brain region remained highly significant when mean age, mean IQ, gender and difference in body parts being observed as covariates. Given that the right inferior frontal gyrus was considered one of the core MNS regions (11, 14), our results might imply that the impaired right frontal MNS could be possibly associated with the dysfunctional social-emotional processing, one of the key characteristics of ASD (81). Indeed, among healthy individuals, it has been shown that the right inferior frontal gyrus is one of the possible neural correlates of social-emotional perception (82-85), in which its connectivity with the limbic system predicts the individual differences in successful emotional regulation (86). The brain-behavior relationship between the right inferior frontal gyrus and social-emotional processing in ASD could be further investigated to confirm our speculation.
MNS activation in different age groups
Another set of analyses we performed was to investigate the activation patterns of MNS and other brain regions in the adolescent and adult subgroups. In addition, we conducted meta-regression with mean age as the regressor within the ASD group to preliminary observe the MNS developmental trajectory in this group of individuals. In the adolescent subgroup, bilateral frontoparietal MNS were shown to be hyperactivated in the individuals with ASD; the extent of this hyperactivation was shown to remain only in the frontal MNS regions (i.e., left supplementary motor area and right inferior frontal gyrus pars orbitalis) in the adult subgroup. One explanation for the difference in the activation patterns of MNS between the adolescent and adult subgroups could be due to the greater variation in the adult subgroup, which in turn may reduce the statistical significance of the observed effects. This limitation notwithstanding, our results provided preliminary evidence in favor of the age effect on MNS functioning in ASD. Further research is necessary to substantiate the present findings and examine the role that age plays on the MNS functioning in ASD.
In addition, using the linear meta-regression method, we identified a significant linear decrease in the activation of the left inferior temporal gyrus as age increased in the ASD group. The meta-analytic results also reflected that hyperactivation in the left temporal gyrus was only shown in the adolescent subgroup but not in the adult subgroup. This echoed the results in a previous study, in which the inferior temporal gyrus gray matter volume changed from leftward asymmetry (i.e. thicker gray matter on the left side) toward symmetry in individuals with autism from childhood through adulthood (87). Conversely, the right cerebellum appeared to exhibit a linear age-related increase in activation. In the adolescent subgroup, the right cerebellum exhibited hyperactivation, and the degree of hyperactivation was further increased in the adult subgroup. The increase in the intensity of hyperactivation may be a consequence of increased lateralization with age (98). Although the two brain areas are not considered to be core MNS regions, our findings seem to suggest that the age-related changes in other brain regions beyond MNS could also influence the activation of MNS. More studies are needed to delineate the reciprocal influences and interplay between the inferior temporal gyrus, cerebellum and the MNS over time.
Regarding literature search, we have attempted to obtain a comprehensive set of relevant fMRI data for meta-analysis by extending our scope of search. Using multiple search engines and manual searches from the reference lists of multiple previous review papers, we found 20 suitable studies that provided us whole brain analytic data for meta-analysis. In addition to these 20 studies, we found an addition of 12 studies that could have been included if whole brain analysis data were available. However, these papers only included region of interest analyses in their papers, or the whole brain analysis data were not obtainable from the corresponding authors. If the whole-brain results were available, the power of this meta-analysis would have been increased, and a more comprehensive picture regarding MNS functioning in ASD could have been presented. Regarding study inclusion, our meta-analysis only included action observation studies, which might limit the generalizability of the results to the imitation deficits in ASD. However, as a number of previous studies provided support that action observation, in contrast to action execution, is one of the key neural processes supporting imitation (11, 34). Moreover, evidence yielded from single neuron recording in humans also showed that the MNS contained different subtypes of mirror neurons, including those discharging both during action observation and execution, but also those discharging during action observation/execution only (15). Showing MNS impairment in ASD could be regarded as one of the important steps in understanding the neural mechanism underlying imitation deficits in ASD, hence potentially guiding treatment development. Future meta-analysis may investigate how imitation and action observation might differ when more imitation studies with more homogeneous study design is available.
In addition, the age of the participants in the included studies ranged from 11.3–37 for ASD individuals and 11.5–37 for TD individuals. Thus, the age effect on MNS can be examined within this age range only; the difference between the ASD and TD individuals in MNS activation remains unknown for populations beyond this age range. Future fMRI studies related to the MNS that target these age ranges are recommended so that the age effect on the MNS can be examined more holistically. Moreover, the meta-regression we performed using the ES-SDM software was a linear regression, which assumed the variable of interest (i.e., mean age) has a linear relationship with brain activation. However, a previous study has shown that the developmental trajectory of the ASD brain appeared to be u-shaped (31), suggesting that linear meta-regression method may not be ideal for articulating the association between age and MNS activation; together with the fact that the statistical significances found in the reported brain regions were driven by limited number of studies, results have to be treated with caution.
Heterogeneity of IQ and gender in our study samples is another limitation as some of the MNS abnormal activations disappeared when IQ and age were included as covariates. Future studies with a more homogeneous sample should be warranted to confirm the observation in our meta-analysis. In addition, the heterogeneity of ASD symptomatology and the overrepresentation of adult age-group may limit the generalization of the findings to the entire patient population with ASD. Future studies with a broader age range, and including different levels of functioning would help to delineate the intricate relationship between IQ, age, and levels of symptom severity in wider patient population with the disorder. Furthermore, it should be noted that our included studies generally had a small sample with a range of 5 to 21 participants per group. Given that studies with small sample sizes tend to overestimate the study effect (50), it might be possible that the MNS abnormality in ASD might have been overestimated. Thus, the generalization of the findings to individuals with ASD in general may be limited by the relatively small sample-size in our meta-analysis.