Table 1 Summary of studies using in vitro model to study ASD
From: Human in vitro models for understanding mechanisms of autism spectrum disorder
Mutation type | Study | Gene/Syndrome | Number of individuals (ASD/Control) | Isogenic? | Proliferation | Cell death | Neuronal differentiation | Electrophysiological properties |
|---|---|---|---|---|---|---|---|---|
Idiopathic | Mariani et al. [59] | Idiopathic with macrocephaly | 4/8 | No | Decreased cell-cycle length in NPCs and early stages of neuronal differentiation | NA | Increased MAP2 in neurons More synaptic puncta Increased proportion of GABAergic neurons | Reduced peak sodium current |
Liu et al. [115] | Idiopathic (no seizures or ID) | 3/3 | No | NA | NA | No change in cell proportions No difference in the number of primary neurites | Reduced sEPSC frequency and half width while amplitude and rise time were not changed Decreased Na+ and fast K+ voltage-gated currents | |
Marchetto et al. [60] | Idiopathic with macrocephaly | 8/5 | No | NPCs proliferated faster | NA | Reduced proportion of BRN2+ and NGN2+ cells Increased proportion of GABAergic cells Fewer excitatory glutamatergic synapses Reduced maturation | No difference was observed in the frequency of spontaneous action potentials Reduced number of synchronized bursts No increase in the number of spikes with maturation | |
Russo et al. [116] | Idiopathic (without seizures) | 3/3 | No | No differences in proliferation | NA | No change in cell proportions Reduced synaptogenesis as an interaction between astrocytes and neurons | Decreased spontaneous spike rate | |
DeRosa et al. [112] | Idiopathic | 5/5 | No | NA | NA | NA | Fewer spontaneous spikes Fewer spontaneous calcium transients | |
Griesi-Oliveira et al. [113] | Idiopathic | 13/8 | No | NA | NA | NA | NA | |
Schafer et al. [114] | Idiopathic | 8/5 | No | NA | NA | Acceleration differentiation of neurons More complex neurite branching patterns | NA | |
Lewis et al. [143] | Idiopathic | 1 multiplex family - 1 affected individual/1 intermediate phenotype relative/1 unaffected relative and 1 unaffected control | No | No changes in cell cycle | Increased apoptosis of both excitatory and inhibitory neurons | DEG were enriched for GO terms related to neuron differentiation | NA | |
Moore et al. [118] | Idiopathic | 3/3 | No | Increased proliferation | NA | Decrease in the proportion of neurons Shorter neurites | NA | |
Adhya et al. [117] preprint | Idiopathic/NRXN1/3p deletion syndrome | 6/2/1/3 controls | No | Upregulation of genes associated with cell proliferation | Upregulation of genes associated with regulation of apoptosis | Delayed neuronal maturation Fewer excitatory and inhibitory NPCs but more GABAergic neurons | NA | |
Wang et al. [119] | Idiopathic with macrocephaly (subset of [60]) | 3/3 | No | Increased proliferation leading to an increase in double stranded breaks | No | Decreased cell migration Defects in polarity and adherence junctions | NA | |
Griesi-Oliveira et al. [120] | Idiopathic | 6/6 | No | Upregulation of genes associated with cell proliferation in NPCs | NA | Upregulation of genes associated with synapse and neurotransmitter release Shorter neurites with fewer ramifications | NA | |
| Â | Deshpande et al. [125] | 16p11.2 deletion and duplication | 3 deletion/3 duplication/4 control | No | No changes in proliferation | NA | 16p11.2 deletion neurons have increased total dendrite length and more extensive dendritic arbors compared with controls The 16p11 duplication neurons exhibit the opposite phenotype with significantly reduced total dendrite length Lower density of synaptic puncta, in both 16p11.2 deletion and 16p11.2 duplication | Reduced excitability in 16p11.2 deletion No difference in the kinetics or frequency of the mEPSCs in 16p11.2 deletion and 16p11 duplication neurons |
CNV | Lin et al. [144] | 22q11.2 deletion (DiGeorge syndrome) | 8/7 | No | Downregulated DEGs were enriched for cell cycle GO terms Decreased proliferation | Upregulated DEGs were enriched for apoptosis | NA | NA |
Toyoshima et al. [124] | 22q11.2 deletion (DiGeorge syndrome) | 2/3 | No | Smaller neurospheres | NA | The fraction of neurons was reduced while the fraction of astrocytes was increased Shorter neurites | Â | |
Fink et al. [122] preprint | Dup15q syndrome (15q11-q13 duplication) Angelman syndrome (15q11-q13/UBE3A maternal deletion) 15q11-q13 paternal duplication | 4 Dup15q/3 Angelman syndrome/1 paternal duplication/6 controls | No | NA | No change in cell death in Dup15q | No differences in dendritic complexity in Dup15q Decrease dendritic complexity in Angelman syndrome | Delayed maturation of action potential Increased frequency and amplitude of synaptic events Increased frequency of spontaneous firing of action potentials | |
Fink et al. [121] | Angelman syndrome (15q11-q13/UBE3A maternal deletion) | 3/4 | Yes | NA | No change in cell death | No changes in cell proportions | Impaired maturation of resting membrane potential Decreased spontaneous excitatory synaptic activity | |
Gillentine et al. [126] | CNRNA7(15q13.3 deletion) deletion and duplication | 3 duplication/3 deletion/3 control | No | NA | NA | NA | Decreased α7 nAChR-associated calcium flux in both deletions and duplications | |
Deneault et al. [145] | 16p11.2 deletion, Nrxn1, DLGAP, CNTN5, AGBL4, GLI, CAPRIN, VIP, ANOS1, EHMT2, THRA, SET | 53 lines from 26 individuals 15 ASD/11 control (1 individual from each, 2 Mz from SET) | Yes | NA | NA | NA | Increased neuronal activity in glutamatergic neurons with CNTN5 or EHMT2 mutations | |
Single gene | Pasca et al. [142] | CACNA1C (Timothy syndrome) | 2/3 | No | NA | NA | Decreased fraction of neurons expressing lower-layer markers and an increased fraction of neurons expressing upper-layer markers. More neurons expressed tyrosine hydroxylase (TH) which was not caused by shift in cell fate | No differences in the action potential threshold or amplitude, resting membrane potential, input resistance or capacitance Wider action potentials Increase in the sustained Ca2+ rise after depolarization |
Wang et al. [131] | CHD8 | 1 individual 2 control lines/4 heterozygous lines (Crispr) | Yes | NA | NA | DEG were enriched for neurogenesis, neuronal differentiation and forebrain development | NA | |
Sugathan et al. [132] | CHD8 (knockdown) | 1 individual | Yes | NA | NA | Downregulated DEG were enriched for synapse formation and neuron differentiation | NA | |
Frega et al. [139] | EHMT1 (Kleefstra syndrome) | 3/3 | Yes | NA | NA | No difference in dendritic morphology or synaptic density | No differences in AMPA-related mEPSCs Fewer, less regular, and longer network bursts which were caused by NMDAR activity | |
Marro et al. [138] | NLGN4 | 1 ES line/1 knockout line/1 mutant line | Yes | NA | NA | Increased number of excitatory synapses in mutant line | Increased frequency and amplitude of mEPSCs in mutant line | |
Zeng et al. [134] | NRXN1 | 1 ES line/1 iPSC line | Yes | NA | NA | Downregulation of astrocyte differentiation DEG were enriched for neuron differentiation-related GO terms | NA | |
Pak et al. [133] | NRXN1 | 2 induced mutations in 1 line | Yes | NA | NA | No changes in differentiation No changes in synaptic density No change in the number if readily releasable pool of vesicles | No change in intrinsic membrane properties Decreased frequency of spontaneous mEPSCs Impaired evoked neurotransmitter release | |
Lam et al. [135] | NRXN1 | 1/4 | No | Slower proliferation | NA | Decreased excitatory neuronal maturation Higher proportions of astroglia | Action potentials had lower amplitude and lower rise time Lower calcium concentration in response to depolarization | |
Avazzadeh et al. [136] | NRXN1 | 3/5 | No | NA | NA | No difference in cell proportions | Increased frequency, duration, and amplitude of calcium transients associated with action potentials | |
Flaherty et al. [137] | NRXN1 | 4/4 | Yes | NA | NA | Decreased proportion of mature neurons Decreased neurite number and length | Decrease in the number of spontaneous action potentials (using two maturation methods) | |
Ross et al. [140] | PTCHD1-AS | 2/2 | Yes | No changes in proliferation | NA | Increased number of synapses in one of the subjects and decreased dendrite complexity in the other | No changes in membrane potential Decreased frequency of mEPSCs Decreased NMDA-evoked current amplitude | |
Zaslavsky et al. [146] | SHANK2 | 2/4 | Yes | Downregulation of cell cycle genes | NA | Increased synapse numbers, dendrite length, and neuron complexity Increased number of functional excitatory connections | Increased sEPSC frequency | |
Yi et al. [127] | SHANK3 | 1 line | Yes | NA | NA | Decrease length and branching of neurites No change in the density or size of synapses | Increased input resistance with no change in capacitance Decreased evoked excitatory postsynaptic currents Decreased amplitude of spontaneous miniature EPSCs Hyperexcitability caused by impaired Ih currents | |
Kathuria et al. [128] | SHANK3 | 2/3 1 ES line | Yes | NA | NA | Smaller cell soma and more primary neurites in olfactory placodal neurons but not in cortical neurons Shorter neurites in cortical neurons | NA | |
Gouder et al. [129] | SHANK3 | 4/3 | No | NA | NA | Reduced dendritic spine densities and spine volume | NA | |
Huang et al. [130] | SHANK3 | 2 lines | Yes | NA | NA | Reduced neuronal soma size, neurite length, and neurite branch number | Reduced frequency of sEPSC | |
Sun et al. [141] | UBE3A (Angelman’s syndrome) | 1 ES cell line | Yes | NA | NA | No changes in neuron morphology | Increased firing frequency of action potentials Increased synchronization | |
Deneault et al. [147] | Many genes (isogenic) AFF2/FMR2, ANOS1, ASTN2, ATRX, CACNA1C, CHD8, DLGAP2, KCNQ2, SCN2A, TENM1 | 1 control individual/1 line | Yes | NA | NA | NA | Reduced sEPSCs and in 5 out of 10 mutations often (4 out of 5 mutations) accompanied by reduced burst frequency |