Jul 2026· Neurobiology of Disease· pp.
107544
· 0 citations· 63 references
Medicine
TL;DR
RNA-sequencing on the patient derived neuroepithelial stem cells (NESCs) found that the PHOX2B-PARM has a profound impact on the transcriptional profile of the cells, highlighting the use of a suitable model of CCHS/HS and providing a clear path for future experimental validation.
Abstract
Mutation in paired-like homeobox 2B (PHOX2B) is used as the diagnostic marker of Congenital Central Hypoventilation Syndrome (CCHS). The mutant gene/protein affects neural crest cells embryonic development which leads to congenital central hypoventilation syndrome (CCHS). When individuals also have Hirschsprung's disease (HSCR) with CCHS it is known as Haddad syndrome (HS). Previous studies on CCHS/HS have mainly focused on the conformational dynamics of the mutant protein and have remained controversial. Here we performed RNA-sequencing on the patient derived neuroepithelial stem cells (NESCs), pertinent to the neurodevelopmental phenotype in CCHS/HS, and found that the PHOX2B-PARM has a profound impact on the transcriptional profile of the cells. The single copy of PHOX2B-PARM in heterozygote cells led to >8 fold differentially expressed genes. This include genes e.g., STMN2, L1CAM, ONECUT2 and NFASC, that are reported to have role in neurodevelopment. In the patient cells there was a significant enrichment of genes related to neuronal development and synapse organization mainly driven by L1CAM interactions and synaptogenesis signaling pathway. Our results not only highlight the use of a suitable model of CCHS/HS but also provide a clear path for future experimental validation.
This work differentiated a genome-wide heterozygous loss-of-function CRISPR library into neural progenitor cells (NPCs) and defined about 250 genes essential for neural differentiation in a haploinsufficient manner, and offers new insights into the embryonic molecular basis of ASD and other NDDs driven by gene dosage imbalance.
Roni Sarel-Gallily, Assa Sherman, Daniel Pollak et al.· Cell Death and Disease· 1 citation
It is demonstrated that SYNGAP1 haploinsufficiency disrupts early human brain development and accelerates intrinsic neuronal maturation, with pathogenic mechanisms emerging before synaptogenesis and extending beyond SYNGAP1’s established synaptic role.
Montanna Waters, Lucas Teasdale, Sean Byars et al.· bioRxiv· 0 citations
Plasma proteomic analysis of patients with CILD40 identified significant enrichment of pathways related to platelet activation, complement and coagulation cascades and differential signatures in CILD40, highlighting the potential of plasma proteomics for understanding distinct pathogenic mechanisms across PCD subtypes.
Siming Kong, Mingshuo Wang, Xuedong Wang et al.· Phenomics· 0 citations
OBJECTIVE
SCN2A pathogenic mutations, such as the recurrent heterozygous Nav1.2-L1342P, are monogenic causes of epilepsy. In this human-induced pluripotent stem cell-derived model system, we aim to investigate the molecular and cellular mechanisms underlying SCN2A-L1342P-associated pathology.
METHODS
Using a human male induced pluripotent stem cell (iPSC) reference line (KOLF) carrying the Nav1.2-L1342P mutation, we generated three-dimensional (3D) cortical organoids for functional studies. Patch-clamp, multi-electrode array (MEA) recordings, immunocytochemistry, and RNA sequencing were used to characterize the disease phenotypes.
RESULTS
Nav1.2-L1342P organoid neurons displayed increased intrinsic excitability and amplified excitatory post-synaptic currents, which are consistent with an increase in excitatory synapse formation revealed by SYN1/PSD95 immunostaining. Moreover, elevated network firing activity, as demonstrated by MEA, indicates a pronounced network hyperexcitability. Transcriptomic profiling of organoids carrying the Nav1.2-L1342P mutation further revealed significant alterations in synaptic, glutamatergic, developmental, and senescence/apoptotic pathways.
SIGNIFICANCE
Our findings demonstrate that the Nav1.2-L1342P mutation drives a multifaceted disease phenotype, including network hyperexcitability and disruption of pathways related to neuronal and synaptic functions. These results advance our understanding of SCN2A-related developmental and epileptic encephalopathy (DEE), laying a foundation for personalized interventions.
M. I. Olivero-Acosta, Morgan Robinson, Zhefu Que et al.· Epilepsia· 0 citations
The 574-kilobase pair 16p11.2 microdeletion raises a person’s odds for neurodevelopmental and energy balance conditions, particularly autism and obesity, with considerable clinical heterogeneity, and how much this reflects genetic versus environmental or stochastic factors is unclear. GABAergic forebrain interneurons originate from progenitors residing in the ventricular zones of the fetal ventral telencephalon, and their perturbation is implicated in 16p11.2 phenotypes, prompting investigation of how the 16p11.2 microdeletion impacts their development. Here we studied human-induced pluripotent stem cell (IPSC) derived ventral telencephalic interneuron progenitors in two-dimensional culture, comparing IPSCs isogenic except for a heterozygous 16p11.2 microdeletion to minimize confounding effects of genetic background. Single-cell RNA sequencing generated single-cell transcriptome populations for comparative bioinformatics, revealing hundreds of differentially expressed transcripts, many associated with cell signaling, chromatin biology, and neurodevelopmental conditions. Pertinently, we find that transcript level variation is significantly greater between 16p11.2 heterozygous progenitors than their isogenic wild type counterparts both for sets of genes comprising regulons, gene-sets functionally connected by transcription factor regulation, and for randomly selected gene sets. This indicates that the 16p11.2 locus itself has a genome-wide property in stabilizing transcription between cells. Regulons with the greatest increased variability in 16p11.2 heterozygous progenitors exhibit strong enrichment for cell cycle-related genes, and many are regulated by transcription factors themselves associated with autism and/or obesity, suggesting the hypothesis that enhanced transcriptional variation contributes to 16p11.2 microdeletion phenotypes.
Yifei Yang, Idoia Quintana Urzainqui, T. Pratt· Frontiers in Molecular Neuro...· 0 citations
Genome-wide association studies report single nucleotide polymorphisms (SNPs) in the PKHD1-TFAP2B genomic interval are associated with primary open-angle glaucoma (POAG) but do not distinguish the causal gene. While neural crest cell (NCC)-specific Tfap2b inactivation causes anterior segment dysgenesis (ASD) and congenital glaucoma (CG), PKHD1 mutations cause autosomal recessive polycystic kidney disease. We now show that Pkhd1del3-67/del3-67 mice also exhibit CG and ASD. Integrating genetic, epigenetic, bioinformatic and developmental analyses, we show that Pkhd1del3-67/del3-67 mice lack Tfap2b and AP-2β expression in a subset of periocular mesenchymal cells at E13.5 and its derivatives. Our data suggest the Pkhd1del3-67 deletion disrupts features of the Pkhd1-Tfap2b genomic architecture essential for Tfap2b cell-specific function. Consistent with this model, Pkhd1del3-67/+;Tfap2bko/+mice develop ASD and lack Tfap2b and AP-2β in relevant cell-types. Our results suggest POAG-associated SNPs at this complex locus may impact disease risk by altering TFAP2B regulatory landscapes, providing a mechanistic link and highlighting regulatory complexity of disease-associated regions. Genetic studies have linked the PKHD1–TFAP2B locus to glaucoma risk, but the underlying mechanism is unclear. Here, the authors show that deleting Pkhd1 in mice disrupts Tfap2b regulation in developing eye tissues, causing congenital glaucoma-like defects and implicating TFAP2B dysregulation in disease risk.
Y. Ishimoto, Luis F. Menezes, Naoki Nakaya et al.· Nature Communications· 0 citations
