Autism Spectrum Disorder (ASD) is a pervasive neurodevelopmental condition affecting approximately 1 in 36 children globally, with a rising prevalence due to improved diagnostic tools and increased awareness [1,2]. Males are diagnosed with ASD more frequently than females, with a ratio of approximately 4:1. However, evidence suggests that subtler symptom presentations or masking behaviors may lead to underdiagnosing in females [3,4]. ASD is characterized by clinically considerable heterogeneity. Core symptoms include difficulties with social communication, restricted interests, and repetitive behavior [5]. These core features often present alongside a spectrum of associated comorbidities, such as global developmental delay, intellectual disabilities, various forms of epilepsy, gastrointestinal issues, anxiety, and sleep disturbances [6,7]. The severity of symptoms can range from mild, allowing for independent living and functional communication, to profound, requiring lifelong support [8].

ASD’s clinical manifestations typically emerge by the age of three years, although subtle signs may be observed as early as infancy. Early indicators include delayed speech and language development, reduced eye contact, atypical social interactions, and repetitive motor behaviors such as hand-flapping, rocking, or spinning [9]. Sensory sensitivities, including aversion to certain sounds, textures, or lights, are also common [10,11]. The clinical trajectory varies widely, with some children showing marked improvement with early intervention, while others experience persistent or worsening symptoms [12].

The diagnosis of ASD is clinical and based on criteria specified in the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) [13]. Key diagnostic criteria include persistent deficits in social communication and interaction across multiple contexts, as well as restricted, repetitive patterns of behavior, interest, or activity [14]. Diagnostic tools commonly used include structured interviews like the Autism Diagnostic Interview-Revised (ADI-R) and standardized assessments like the Autism Diagnostic Observation Schedule (ADOS) [12,15]. A multidisciplinary team consisting of pediatricians, neurologists, psychologists, and speech therapists often conducts developmental, behavioral, and cognitive assessments as part of their evaluations [14]. Neuroimaging, laboratory, and genetic testing are not diagnostic but can provide supportive evidence [5,16]. MRI scans of the brain often show structural and functional problems in children with ASD. These problems include different connections in the default mode network and less gray matter in the amygdala and prefrontal cortex [17,18].

Various forms of genetic testing are part of the diagnostic algorithm for ASD. Testing typically follows established clinical guidelines that aim to detect syndromic forms of autism. These syndromes are characterized by well-documented genetic mutations that contribute to ASD symptoms and other associated features [19].

For suspected monogenic syndromes with the clinical presentation of ASD, such as fragile X syndrome, targeted tests like FMR1 gene analysis are often the first choice [20]. The testing algorithm usually proceeds with chromosomal microarray analysis (CMA), which is widely recommended as a first-tier test due to its ability to identify submicroscopic chromosomal deletions or duplications that are frequently associated with ASD [21]. Additionally, next-generation sequencing (NGS), including whole-exome sequencing (WES) or panel testing of autism-related genes, is increasingly used to identify mutations in single genes linked to ASD [22]. While these tests have significantly advanced our understanding and diagnostic capabilities, their overall detection rate for pathogenic variants in children with isolated, non-syndromic forms of ASD remains approximately 10–20% [16,23].

Chromosomal abnormalities, single-gene mutations, and copy number variations (CNV) have been identified as a potential key contributor to ASD, with many of these genetic changes impacting synaptic function, neurogenesis, and neuronal migration [16,19]. A detailed overview of the major genes involved, their functions, and their impact on neurodevelopment is provided in Table 1.

Environmental factors, including advanced parental age, prenatal infections, and exposure to toxins like valproic acid, have been linked to an elevated risk of ASD [24,25,26]. Epigenetic modifications, such as DNA methylation and histone acetylation, provide a valuable link between genetic predisposition and environmental triggers [25,26,27]. These modifications regulate gene expression without altering the underlying DNA sequence, allowing dynamic responses to environmental influences. For instance, changes in DNA methylation patterns can silence or activate key genes involved in neurodevelopmental processes, while histone acetylation alters chromatin structure, influencing the accessibility of transcriptional machinery [27,28,29]. Epigenetic mechanisms like these are a major part of how prenatal environmental exposures affect glial function and neurodevelopment [30]. Emerging evidence suggests that maternal immune activation during pregnancy may alter fetal brain development via cytokine-driven neuroinflammatory mechanisms. These immune-related disruptions are particularly concerning during critical developmental windows, as they may interfere with processes such as synaptic pruning and myelination, potentially leading to long-term neurodevelopmental consequences [31].

Astrocytes, microglia, and oligodendrocytes are vital components of the central nervous system, contributing to both the structural and functional maturation of the brain. These neuroglial cells regulate fundamental processes, including synaptogenesis, immune homeostasis, and neuronal support, all of which are essential for establishing and maintaining neural architecture [32]. Given their diverse roles, disruptions in neuroglial function can significantly impact brain connectivity and plasticity [33,34]. A detailed overview of the key functions of astrocytes, microglia, and oligodendrocytes, along with specific alterations observed in ASD pathophysiology, is provided in Table 2.

Astrocytes play a critical role in metabolic exchange by linking the vasculature and neurons through the delivery of glucose and lactate, essential substrates for neuronal function. They play a crucial role in neurovascular coupling by modulating blood flow in the brain in response to neuronal activity [35,36].

Microglia serve as the primary immune effector cells of the central nervous system, playing a vital role in maintaining immune homeostasis. They are integral to synaptic pruning, a developmental process that refines neural networks by eliminating redundant or unnecessary synapses [33,37]. Oligodendrocytes, on the other hand, are essential for preserving axonal functionality and structural integrity. They achieve this by synthesizing myelin sheaths, which facilitate rapid propagation of action potentials and provide metabolic support to axons [38].

The coordinated actions of these neuroglial cells provide the dynamic processes that shape neural architecture, which form the basis for higher cognitive and behavioral functions [32,39]. Neuroglia, once considered merely supportive elements for neurons, are currently considered major players in shaping neural connectivity, regulating synaptic dynamics, and maintaining homeostatic balance within the central nervous system [40].

Recent progress in imaging and molecular biology has provided a new way to look at glial abnormalities in ASD, shedding light on their roles in dysregulated synapses, neuroinflammation, and changed neural circuitry. These findings not only deepen our understanding of ASD pathophysiology but also point to neuroglia as promising targets for novel therapeutic strategies [18,33,41].

Dysregulation of neuroglial function in ASD profoundly affects brain development and activity. Impaired glial cell function contributes to core features of ASD, including deficits in social communication, repetitive behaviors, and cognitive impairments [39]. Recent research highlights reactive astrogliosis, hyperactive microglial pruning, and impaired myelination as critical glial abnormalities contributing to ASD pathophysiology [62]. These glial changes alter neuronal signaling and network dynamics, leading to impaired cognitive and behavioral outcomes [39,62]. A summary of key molecular changes observed in astrocytes, microglia, and oligodendrocytes in ASD is presented in Table 3.

Astrocytes are essential for preserving synaptic homeostasis, providing neuronal support, and maintaining general cerebral equilibrium [36,42]. In ASD, astrocytic dysfunction disrupts these processes, contributing to pathophysiological changes observed in the condition [36,62].

Astrocytes express Glial Fibrillary Acidic Protein (GFAP), a cytoskeletal protein indicative of their reactive state [63]. Post-mortem analyses of ASD brains reveal elevated GFAP levels, consistent with chronic astrogliosis and associated neuroinflammation [64]. Reactive astrocytes demonstrate altered calcium signaling, disrupting their ability to respond appropriately to synaptic activity. This dysregulation impairs critical functions, including neurotransmitter recycling (e.g., glutamate–glutamine cycling) and ion homeostasis, leading to excitatory–inhibitory imbalances in neural circuits [65].

Emerging evidence suggests that distinct reactive astrocyte subtypes, such as A1 (neurotoxic) and A2 (neuroprotective), may differentially contribute to ASD pathophysiology. Such changes further exacerbate the disruption of synaptic plasticity, neuronal connectivity, and overall network stability [65].

Astrocytes play a crucial role in the glutamate–glutamine cycle by converting synaptic glutamate into glutamine, thereby preventing excitotoxicity and maintaining synaptic homeostasis [66]. In ASD, disruptions in glutamate uptake have been associated with reduced expression of excitatory amino acid transporter 2 (EAAT2), the primary astrocytic glutamate transporter. This reduction leads to elevated extracellular glutamate levels, resulting in excitotoxic neuronal injury and altered neural circuit functionality. Such dysregulation of glutamate signaling has been implicated in the emergence of core ASD features, including repetitive behaviors and sensory processing abnormalities [67,68].

Neurovascular coupling is successfully mediated by the coordinated activity of astrocytes. They mediate communication between neurons and blood vessels to regulate cerebral blood flow and meet metabolic demands [42,44]. In ASD, astrocytic dysfunction reduces cerebral blood flow, particularly in regions critical for higher cognitive and social functions [69,70]. This impaired neurovascular coupling compromises metabolic support to neurons, further exacerbating neural circuit dysregulation [60]. Neuroimaging studies consistently report hypoperfusion in key areas such as the prefrontal cortex and anterior cingulate cortex, regions associated with executive function and social cognition [71].

Beyond astrocytic dysfunction, microglia also exhibit profound abnormalities in ASD [33]. These cells play a critical role in synaptic pruning, immune surveillance, and inflammatory regulation, yet in ASD, they often display chronic activation and excessive complement-mediated synapse elimination [33,45]. The growing body of research suggests that microglial hyperactivity contributes to altered neuronal circuit formation, excessive synaptic connectivity, and persistent neuroinflammation, all of which are hallmarks of ASD neuropathology [33].

Microglial pruning of synaptic connections is tightly regulated by complement proteins (e.g., C1q and C3). Dysregulation of this pathway in ASD leads to excessive or insufficient pruning, resulting in either hyperconnectivity or synaptic deficits [47]. The latest study investigated the role of complement in ASD development using multiple reaction monitoring (MRM). The study has successfully identified 16 out of 33 proteins within these pathways as differentially expressed in the plasma of children with ASD compared to controls. Notably, CFHR3, C4BPB, C4BPA, CFH, C9, SERPIND1, C8A, F9, and F11 were reported as altered in ASD plasma for the first time. Among these, SERPIND1 expression showed a positive correlation with ASD severity, as measured by the Childhood Autism Rating Scale (CARS). Furthermore, machine learning analysis identified a panel of 12 differentially expressed proteins with potential diagnostic utility for ASD [72].

Animal models demonstrate that imbalances in microglial activity contribute to behavioral rigidity and sensory hypersensitivity, trades that are commonly observed in ASD patients [73]. Interestingly, human postmortem and imaging studies have also reported alterations in microglial activation patterns in ASD, further bridging preclinical and clinical findings [74].

In addition to astrocytes and microglia, oligodendrocytes—the myelinating cells of the central nervous system—also exhibit significant dysfunction in ASD [75]. Proper oligodendrocyte maturation and myelination are essential for efficient neuronal communication, yet studies indicate that ASD is associated with delayed oligodendrocyte precursor maturation, impaired myelin production, and reduced white matter integrity [38,49,52]. Diffusion tensor imaging (DTI) studies have consistently demonstrated structural abnormalities in major white matter tracts, further supporting the role of oligodendrocyte dysfunction in the pathophysiology of ASD [76].

Hypomyelination is a prominent feature observed in ASD brains [50,62]. Studies indicate delayed or reduced oligodendrocyte differentiation and myelination in key brain regions, such as the corpus callosum and prefrontal cortex [77]. Longitudinal neuroimaging studies show that impaired white matter integrity often appears early in ASD and persists throughout development [78]. The convergence of neuroimaging and genetic studies highlights a critical role for disrupted myelination in ASD, further expanding our understanding of how neuroglial dysfunction contributes to altered brain connectivity. These deficits correlate with impairments in executive function, social behavior, and sensory processing, linking structural abnormalities to core behavioral features of ASD [77,79].

Furthermore, neuroimaging studies of individuals with ASD consistently report reduced fractional anisotropy (FA), a key measure of white matter integrity that reflects microstructural disruptions in neural connectivity. These alterations are particularly pronounced in major white matter tracts, including the corpus callosum, which is crucial for interhemispheric communication, and the superior longitudinal fasciculus, which facilitates long-range connectivity between frontal, parietal, and temporal regions. Reduced FA in these pathways has been linked to deficits in social communication, executive function, and sensory processing, further underscoring the role of white matter abnormalities in ASD pathophysiology. Such findings align with the broader hypothesis that disrupted neural connectivity is a core feature of ASD, contributing to atypical information processing and cognitive function [80,81].

Dysregulated signaling pathways, chronic neuroinflammation, and genetic and epigenetic factors intricately interact to shape neuroglial dysfunction in ASD. These interconnected mechanisms disrupt synaptic homeostasis, neuronal–glial communication, and overall brain connectivity, contributing to the hallmark features of ASD [27,62,65].

Among these pathways, the mammalian target of the rapamycin (mTOR) signaling pathway and the wingless-related integration site/β-catenin (Wnt/β-catenin) signaling pathway signaling cascades emerge as crucial regulators of neurodevelopmental processes that are often altered in ASD [82,83]. The mammalian target of the rapamycin (mTOR) pathway, a central controller of cellular metabolism, homeostasis, integrating signals from nutrients, growth factors, energy availability, and synaptic plasticity, is highly sensitive to environmental and genetic cues that shape early brain development. It functions through two distinct complexes, the mammalian target of rapamycin complex 1 and 2 (mTORC1 and mTORC2), each with specific physiological roles. mTORC1 controls protein synthesis, lipid metabolism, and autophagy by phosphorylating downstream effectors such as S6 kinase (S6K) and 4E-binding protein 1 (4E-BP1), promoting anabolic processes crucial for neuronal and glial function. It is highly active in synaptic plasticity, regulating dendritic spine formation and the remodeling of synaptic connections. mTORC2, in contrast, governs cytoskeletal organization and cell survival through the activation of Akt and protein kinase C (PKC), playing a role in neuronal differentiation and migration. In neuroglial physiology, mTOR signaling modulates astrocytic metabolic support, microglial immune responses, and oligodendrocyte-driven myelination, ensuring proper neuronal connectivity and brain network stability [84].

Persistent hyperactivation of mTOR in ASD has been associated with excessive synaptogenesis, disrupted neuronal connectivity, and impaired synaptic pruning. This dysregulation affects both neurons and glial cells, leading to an imbalance in excitatory and inhibitory neurotransmission [85,86]. Microglial dysfunction, driven by unchecked mTOR signaling, alters synaptic remodeling by either failing to eliminate redundant synapses or excessively pruning functional connections, both of which contribute to the atypical network architecture characteristic of ASD. Additionally, astrocytes, which play a vital role in maintaining synaptic homeostasis and metabolic support, exhibit abnormal glutamate clearance, altered calcium signaling, and disrupted neurovascular coupling due to mTOR dysregulation. The net effect is a hyperconnected but functionally inefficient neural circuitry, manifesting as the core behavioral and cognitive symptoms of ASD [87,88].

In parallel, the Wnt/β-catenin pathway plays an essential role in orchestrating neurodevelopmental processes, including synaptic assembly, glial differentiation, and myelination [89,90]. The pathway is initiated when Wnt proteins, a family of secreted glycoproteins, bind to Frizzled receptors and low-density lipoprotein receptor-related proteins (LRP5/6) on the cell membrane. This interaction inhibits the activity of the β-catenin destruction complex, which includes adenomatous polyposis coli (APC), axin, and glycogen synthase kinase-3β (GSK-3β). As a result, β-catenin accumulates in the cytoplasm and translocates into the nucleus, where it interacts with T-cell factor/lymphoid enhancer-binding factor (TCF/LEF) transcription factors to regulate the expression of genes involved in cell proliferation, differentiation, and survival [91,92].

In the central nervous system, Wnt/β-catenin signaling is essential for oligodendrocyte differentiation and myelination, as it modulates the balance between precursor cell proliferation and maturation. It influences axonal guidance and neuronal migration during early development, ensuring the proper establishment of cortical layers and functional neuronal networks. Additionally, Wnt/β-catenin signaling plays a crucial role in dendritic arborization, regulating synaptic stability and adaptability in response to environmental stimuli. Astrocytic function is also modulated by this pathway, as Wnt/β-catenin signaling controls astrocyte-mediated glutamate uptake, ion homeostasis, and trophic support for neurons. Furthermore, Wnt/β-catenin signaling regulates microglial activity, particularly in synaptic pruning and neuroinflammatory responses, helping maintain synaptic efficiency and circuit refinement [85,93].

Beyond its developmental functions, Wnt/β-catenin signaling remains active in the adult brain, contributing to synaptic maintenance, plasticity, and repair mechanisms in response to injury. Proper modulation of this pathway is critical for maintaining white matter integrity and overall brain connectivity, underscoring its significance in both neurodevelopment and ongoing neuronal function [94].

Altered Wnt/β-catenin activity in ASD has been associated with impaired white matter integrity, as evidenced by diffusion tensor imaging (DTI) studies, which reveal disrupted connectivity in major associative tracts such as the corpus callosum and superior longitudinal fasciculus [80]. Dysfunctional Wnt/β-catenin signaling also impacts astrocytic function, diminishing their ability to regulate synapse formation and plasticity while concurrently influencing microglial activation states [95]. Additionally, disruptions in Wnt/β-catenin signaling have been linked to the dysregulation of neural progenitor cell fate determination, further exacerbating the structural and functional abnormalities characteristic of ASD [96].

The interplay between these signaling pathways underscores the complexity of neuroglial dysfunction in ASD. Emerging evidence suggests that these pathways interact at multiple levels, with mTOR influencing Wnt/β-catenin activity and vice versa [86,95]. Dysregulation within one pathway may cascade into alterations in the other, compounding neurodevelopmental impairments. Understanding these intricate molecular interactions will be essential for advancing knowledge of ASD pathophysiology and identifying potential therapeutic targets. Given the central role of these pathways in regulating neuroglial homeostasis, targeting their dysregulation has become a promising avenue for therapeutic intervention [97]. Experimental studies demonstrate that pharmacological modulation of these pathways can restore glial function and improve behavioral outcomes in ASD models [98].

Besides dysregulation of the key signaling pathways, chronic neuroinflammation is another complex pathophysiological element of ASD. The intricate crosstalk between immune signaling and neuroglial function further complicates the pathophysiology of the disorder [99,100]. A growing body of evidence suggests that persistent activation of microglia and astrocytes contributes to aberrant synaptic pruning, impaired neuronal plasticity, and disruptions in excitatory/inhibitory balance, which are hallmark features of ASD [101]. Elevated levels of pro-inflammatory cytokines such as IL-6, TNF-α, and IL-1β not only disrupt neuroglial communication but also exacerbate synaptic dysfunction by altering glutamatergic and GABAergic signaling, leading to cognitive and behavioral impairments [65,102].

Another form of the neuroinflammatory process connected with the pathogenesis of ASD is prenatal exposure to maternal immune activation (MIA) [31]. This exposure has been shown to induce long-term neuroglial abnormalities and ASD-like behavior in offspring, with experimental models demonstrating that maternal infection, autoimmunity, or other immune insults can lead to persistent alterations in microglial morphology and function. These immune-mediated changes are accompanied by an increased susceptibility to environmental stressors, further amplifying neurodevelopmental vulnerabilities [103,104].

However, neuroinflammation does not act in isolation but rather intersects with genetic and epigenetic mechanisms that govern neuroglial function [101]. Inflammatory signaling can influence gene expression patterns by modifying epigenetic landscapes, while genetic mutations in key ASD-related genes further exacerbate glial dysregulation [27,33].

Studies have shown that mutations in genes like PTEN, MECP2, and CNTNAP2 disrupt critical processes such as synaptic pruning, inflammation regulation, and myelination [105]. These genetic predispositions do not fully account for the complexity of ASD pathology [106]. Increasing evidence suggests that epigenetic mechanisms serve as a critical interface between genetic predispositions and environmental factors, fine-tuning neuroglial gene expression [107]. Modifications such as DNA methylation and histone acetylation dynamically regulate transcription in response to developmental cues, potentially mediating the long-term impact of prenatal exposures on neuroglial integrity [108].

Epigenetic mechanisms such as DNA methylation and histone acetylation modulate gene expression in response to environmental factors, linking prenatal exposures to long-term neuroglial deficits [29]. DNA methylation, primarily occurring at CpG islands, is a key epigenetic modification that typically leads to transcriptional silencing by altering chromatin structure and preventing transcription factor binding. In contrast, histone acetylation relaxes chromatin and facilitates gene transcription by neutralizing the positive charge of histone proteins, thereby allowing greater accessibility of regulatory elements [109].

These modifications are highly dynamic and responsive to external stimuli, including maternal stress, dietary factors, infections, inflammations and exposure to environmental toxins during critical periods of neurodevelopment. Disruptions in these epigenetic processes can lead to persistent changes in the gene expression involved in synaptic plasticity, glial differentiation, and neuroinflammation, thereby contributing to long-lasting alterations in brain structure and function. Understanding this delicate interplay between epigenetic regulation and neuroglial integrity provides valuable insight into how early-life environmental influences shape neurodevelopmental trajectories [27,29].

Advances in genome-editing tools have enabled precise investigations into these genetic contributions, allowing researchers to dissect the functional impact of specific epigenetic modifications and their role in neurodevelopmental processes. Techniques such as Clustered Regularly Interspaced Short Palindromic Repeats- Cas9 (CRISPR-Cas9) and base editing provide unprecedented control over gene regulation, facilitating the development of targeted therapeutic approaches. By refining our ability to manipulate epigenetic landscapes, these tools hold promise for correcting aberrant gene expression patterns associated with neuroglial dysfunction, paving the way for precision medicine strategies in neurological disorders [110].

Despite significant advances in understanding the neuroglial contributions to ASD, several limitations hinder the direct translation of these findings into clinical practice. One of the most pressing challenges is the substantial heterogeneity of ASD, both in terms of clinical presentation and underlying pathophysiological mechanisms. This variability complicates the development of universal therapeutic strategies, as individuals with ASD exhibit diverse genetic, epigenetic, and neurobiological profiles that influence their response to treatment. Current interventions, while promising, often yield highly variable outcomes, underscoring the need for biomarker-driven precision medicine [144].

A major obstacle in achieving targeted ASD therapies is the lack of reliable biomarkers that can effectively stratify patients into clinically meaningful subgroups. Biomarkers derived from genomic, epigenetic, proteomic, or neuroimaging data hold the potential for improving diagnostic specificity, monitoring treatment efficacy, and minimizing adverse effects. However, many candidate biomarkers fail to demonstrate consistent predictive value across different studies and patient cohorts, limiting their clinical applicability. Future research should prioritize large-scale, multicenter validation studies to establish reproducible and clinically relevant biomarkers [145].

Another emerging approach with transformative potential is the application of artificial intelligence (AI) and machine learning (ML) in ASD research. These advanced computational tools can analyze high-dimensional datasets to identify novel correlations between genetic mutations, neuroglial dysfunction, and clinical phenotypes. AI-driven models may refine individualized treatment predictions, optimize therapeutic strategies, and uncover previously unrecognized molecular targets for intervention. However, translating these methods into clinical practice requires robust validation, standardized methodologies, and interdisciplinary collaboration between data scientists, neuroscientists, and clinicians [146,147].

Beyond scientific and technical challenges, logistical, ethical, and financial constraints present significant barriers to implementing emerging ASD therapies. Glial-targeted treatments, such as stem cell-based interventions and gene-editing approaches, require rigorous safety and efficacy evaluations, as well as ethical frameworks that balance innovation with patient safety. Additionally, the high cost of personalized medicine approaches raises concerns about equitable access, necessitating policy-driven initiatives to ensure that these therapies benefit a broad spectrum of individuals rather than a privileged few [148].

Finally, one of the greatest translational challenges is the difficulty of bridging the gap between preclinical models and human neuroglial function. Many ASD studies rely on rodent models, which fail to fully recapitulate the complexity of human neurodevelopment. The development of induced pluripotent stem cell (iPSC) models and organoid systems offers a more relevant human-specific platform for testing glial-targeted treatments before clinical translation. Future research should expand the use of these advanced models to improve the predictability of therapeutic responses in ASD patients [149].

To overcome these challenges and accelerate the translation of neuroglial-targeted therapies into clinical practice, future research must prioritize several key areas. First, the development and validation of reproducible biomarkers will be critical for predicting treatment response and stratifying ASD subtypes based on distinct neurobiological signatures. Advances in genomic, epigenetic, and neuroimaging-based biomarkers may enable more precise patient classification, ultimately guiding the selection of personalized therapeutic interventions [150].

Additionally, long-term clinical trials are necessary to assess the sustained neurodevelopmental impact of emerging neuroglial-targeted therapies. While several experimental treatments have shown promise in preclinical models, their efficacy and safety over extended periods remain largely unknown. Future studies should incorporate comprehensive longitudinal designs, evaluating not only immediate symptom relief but also long-term cognitive, behavioral, and functional outcomes in individuals with ASD [151].

Another crucial step is the refinement of human-based models that better replicate neuroglial interactions in ASD pathology. Induced pluripotent stem cell (iPSC)-derived neuroglia and brain organoid systems provide promising platforms for studying patient-specific glial dysfunction and screening potential therapeutic compounds. Expanding the use of these advanced models will improve translational validity, bridging the gap between basic research and clinically applicable treatments [152].

In parallel, multi-modal therapeutic strategies should be explored to address the complex and multifactorial nature of ASD. Future interventions should integrate anti-inflammatory, neurotrophic, and synaptic-modulating approaches, potentially combining pharmacological agents, immunotherapies, and neuromodulation techniques to enhance treatment efficacy. Combinatorial strategies may prove particularly beneficial in tailoring interventions to the specific glial dysfunction profiles present in different ASD subtypes [144].

Finally, as the field moves toward more advanced and individualized treatments, it is imperative to establish ethical and regulatory frameworks that guide the responsible implementation of stem cell-based and gene-editing therapies in ASD. Considerations surrounding genetic privacy, patient safety, long-term monitoring, and equitable access must be addressed to ensure that cutting-edge interventions benefit a broad spectrum of individuals rather than a select few. Close collaboration between scientists, clinicians, bioethicists, and policymakers will be essential in developing responsible, widely accessible therapeutic solutions that align with the highest standards of medical ethics and patient care [153].

The growing recognition of neuroglial dysfunction as a key component of ASD pathophysiology has opened new avenues for therapeutic intervention. The interplay between astrocytes, microglia, and oligodendrocytes in shaping neural connectivity, synaptic plasticity, and neuroimmune regulation underscores the complexity of ASD and the need for multifaceted treatment strategies. While current research has identified several promising molecular targets, including mTOR, Wnt/β-catenin, and NF-κB pathways, significant work remains in refining these approaches to ensure safety, specificity, and long-term efficacy [96,97].

The advent of precision medicine in ASD represents a paradigm shift in how we approach diagnosis and treatment. The integration of biomarkers, neuroimaging tools, and AI-driven analytics holds the potential to tailor interventions to individual patients, moving beyond the one-size-fits-all model. Advances in stem cell therapy, gene editing, and pharmacological modulation of neuroglial function offer hope for targeted interventions that address the underlying biological mechanisms of ASD rather than merely alleviating symptoms [134,139].

However, translating these innovations into widespread clinical use will require overcoming scientific, logistical, and ethical hurdles. Collaborative efforts among researchers, clinicians, industry leaders, and policymakers will be essential to bridge the gap between bench-side discoveries and real-world applications. Ensuring equitable access to emerging therapies, particularly in underserved populations, must remain a priority to avoid widening healthcare disparities [144,150].