Atypical neurogenesis in induced pluripotent stem cell (iPSC) from autistic individuals

Abstract

30Autism is a heterogenous collection of disorders with a complex molecular underpinning. 31Evidence from post mortem studies using adult brains have identified atypical co-expression 32 of genes which may occur during the prenatal period. Recent studies using induced pluripotent 33 stem cells (iPSCs) generated from autistic individuals have suggested that prenatal 34 development is a critical period for the emergence of pathophysiology associated with this 35 condition. However, how early during development these differences emerge and whether such 36 alterations can be seen across the development of multiple brain regions is unclear. In this study 37 we investigated whether early prenatal stages of neurodevelopment differ between iPSCs 38 generated from typically developing and autistic individuals. We specifically selected autistic 39 individuals unrelated to each other with a heterogeneous genetic background and no known 40 comorbidities, to probe for common molecular phenotypes. Differentiation of iPSCs towards a 41 cortical lineage revealed abnormal cell fate acquisition from an early stage of development. 42Interestingly, abnormal differentiation occurred in the absence of alteration in cell proliferation 43 during cortical differentiation, differing from previous studies. Moreover, these effects 44 appeared specific for the acquisition of a cortical fate, as differentiation of iPSCs towards a 45 midbrain lineage was not accompanied by differences in neurogenesis between typically 46 developing and autism iPSC lines. RNA-sequencing on a subset of our cohort further revealed 47 autism-specific signatures during cortical differentiation similar to that observed in post 48 mortem studies, indicating a potential common biological mechanism. Together, these data 49 suggest unique developmental differences associated with autism may establish at an early 50 prenatal stage. 51 52 53 4 54 Recent methodological advances in the field of induced pluripotent stem cell (iPSC) 75technology [15][16][17][18][19][20] has made it possible to study prenatal cellular behaviour in autism in detail, 76 something that was not possible using post mortem brains. IPSCs have similar abilities to 77 5 embryonic stem cells to generate any tissue of the body. These can then be differentiated into 78 neurons of various lineages 15-17, 19, 20 . As the neurons contain the same genetic information as 79 the individual from whom it was derived, typical or autistic, its cellular behaviours is 80 influenced by its genetic background. Using these methods, studies have shown significant 81 anomalies in cellular/molecular behaviour during prenatal-equivalent periods of development 82 in autistic individuals [21][22][23] . One such study generated iPSCs from autistic individuals who were 83 comorbid for macrocephaly, and demonstrated: (1) atypical cortical differentiation and 84 increased cell proliferation of cortical neural precursor cells (NPCs) from iPSCs, and (2) an 85 imbalance in excitatory (glutamate-producing) and inhibitory (GABA-produ...

GenTree, an integrated resource for analyzing the evolution and function of primate-specific coding genes

“…ZNF430 in the European population (Cheng et al 2017). Lastly, the association between ZNF430 and autism is reproduced recently in a transcriptome study based on iPSC neurons of autism patients (Adhya et al 2018).…”

Interferon-γ signaling in human iPSC–derived neurons recapitulates neurodevelopmental disorder phenotypes

“…Two ASD lines were generated using CytoTune Sendai reprogramming kits (Thermo Fisher Scientific, A16517). HiPSC reprogramming was validated as previously described ( 24 , 27 , 28 , 56 ). Briefly, genome-wide expression profiling using Illumina BeadChip v4 and the bioinformatics tool “Pluritest” was used to confirm hiPSC identity.…”

“…HiPSCs were differentiated as previously reported ( 24 ). HiPSCs were grown until 95 to 100% confluent and then neuralized using a modified dual SMAD inhibition protocol ( 24 , 27 , 28 , 56 ). For neuralization, StemFlex was replaced with a 1:1 mixture of N2- and B27-supplemented medium, made following the manufacturer’s guidelines (Thermo Fisher Scientific, 17502048 and 17504044) to which SMAD and Wnt inhibitors (10 μM SB431542, 1 μM dorsomorphin, and 2 μM XAV939) were added (hereafter known as N2:B27+++).…”

Shifting Developmental Trajectories During Critical Periods of Brain Formation

“…In order to fill the gap between human disease and model organisms, the development of stem cell technologies—both embryonic and induced pluripotent stem cells (iPSCs)—have given access to functional readouts typical of early stages of human brain development (Dolmetsch and Geschwind, 2011 ). iPSCs have provided insights into the cellular alterations underlying neuropsychiatric disorders such as autism and schizophrenia (Marchetto et al, 2017 ; Adhya et al, 2020 ; reviewed in Ben-Reuven and Reiner, 2016 ; Vitrac and Cloëz-Tayarani, 2018 ). Although concerns have been raised regarding the clinical applications of iPSCs as a valuable source for cell transplantation therapy and the significant reprogramming variability of human-derived iPSCs, the simplicity of two-dimensional cultures is suitable for mechanistic studies, large-scale screening, or high-throughput drug testing.…”