KIF5C deficiency causes abnormal cortical neuronal migration, dendritic branching, and spine morphology in mice.

BACKGROUND: Malformation of cortical development (MCD) includes a variety of developmental disorders that are common causes of neurodevelopmental delay and epilepsy. Most recently, clinical studies found that patients carrying KIF5C mutations present early-onset MCD; however, the underlying mechanisms remain elusive. METHODS: KIF5C expression level was examined in mouse primary cortical neurons and human ips-derived forebrain organoids. We studied the cortical neuronal migration, dendritic branching, and dendritic spine growth after knocking down the KIF5C gene by electroporation in vitro and in vivo. Then, we studied the transcriptome differences between the knockdown and control groups through RNA sequencing. RESULTS: We observed high KIF5C expression in neurons during the early developmental stage in mice and the human brain. Kif5c deficiency results in disturbed cortical neuronal migration, dendritic, and spine growth. Finally, we found that Kif5c knockdown affected several genes associated with cortical neuronal development in vitro. CONCLUSIONS: These results suggested a critical role for Kif5c in cortical development, providing insights into underlying pathogenic factors of kinesins in MCD. IMPACT: KIF5C mutation-related MCD might be caused by abnormal early cortical neuronal development. Kif5c deficiency led to abnormal cortical neuronal dendritic and spine growth and neuronal migration. Our findings explain how Kif5c deficiency is involved in the aberrant development of cortical neurons and provide a new perspective for the pathology of MCD.

Transcriptional networks identify synaptotagmin-like 3 as a regulator of cortical neuronal migration during early neurodevelopment.

Human brain development is a complex process involving neural proliferation, differentiation, and migration that are directed by many essential cellular factors and drivers. Here, using the NetBID2 algorithm and developing human brain RNA sequencing dataset, we identify synaptotagmin-like 3 (SYTL3) as one of the top drivers of early human brain development. Interestingly, SYTL3 exhibits high activity but low expression in both early developmental human cortex and human embryonic stem cell (hESC)-derived neurons. Knockout of SYTL3 (SYTL3-KO) in human neurons or knockdown of Sytl3 in embryonic mouse cortex markedly promotes neuronal migration. SYTL3-KO causes an abnormal distribution of deep-layer neurons in brain organoids and reduces presynaptic neurotransmitter release in hESC-derived neurons. We further demonstrate that SYTL3-KO-accelerated neuronal migration is modulated by high expression of matrix metalloproteinases. Together, based on bioinformatics and biological experiments, we identify SYTL3 as a regulator of cortical neuronal migration in human and mouse developing brains.

Mutations of CNTNAP1 led to defects in neuronal development.

Mutations of CNTNAP1 were associated with myelination disorders, suggesting the role of CNTNAP1 in myelination processes. Whether CNTNAP1 may have a role in early cortical neuronal development is largely unknown. In this study, we identified 4 compound heterozygous mutations of CNTNAP1 in 2 Chinese families. Using mouse models, we found that CNTNAP1 is highly expressed in neurons and is located predominantly in MAP2+ neurons during the early developmental stage. Importantly, Cntnap1 deficiency results in aberrant dendritic growth and spine development in vitro and in vivo, and it delayed migration of cortical neurons during early development. Finally, we found that the number of parvalbumin+ neurons in the cortex and hippocampus of Cntnap1-/- mice is strikingly increased by P15, suggesting that excitation/inhibition balance is impaired. Together, this evidence elucidates a critical function of CNTNAP1 in cortical development, providing insights underlying molecular and circuit mechanisms of CNTNAP1-related disease.

A novel deletion with two pathogenic variants of UGT1A1 causing Crigler-Najjar syndrome in two unrelated Chinese.

Two Chinese female infants from two unrelated families were diagnosed with Crigler-Najjar syndromes-I (CNS-I) and CNS-II respectively. The CNS-I patient had Serum Total Bilirubin Concentration (STBC) peaked at 26.1 mg/dL. She was not responsive to Phenobarbital and received liver transplantation at 2-year-old. The CNS-II patient's STBC fluctuated between 10.2 mg/dL and 17.4 mg/dL and had a milder phenotype. Sequencing of Uridine Diphosphate Glucuronosyl Transferase 1A1 (UGT1A1) revealed the CNS-I patient carried a heterozygous pathogenic variant in c.392 T > C (p.Leu131Pro) and the CNS-II patient carried a heterozygous pathogenic variant in c.1456 T > G (p.Tyr486Asp). Furthermore, a novel deletion spanning exons 2-4 of UGT1A1 were detected in both patients. We studied two family members' genotyping results of UGT1A1 to clarify the inheritance of this microdeletion. To our knowledge, this is probably the first time showing 2 CNS cases both carrying compound heterozygous variations of a known pathogenic variant and a novel microdeletion.

De Novo and Inherited SETD1A Variants in Early-onset Epilepsy.

Early-onset epilepsy is a neurological abnormality in childhood, and it is especially common in the first 2 years after birth. Seizures in early life mostly result from structural or metabolic disorders in the brain, and the genetic causes of idiopathic seizures have been extensively investigated. In this study, we identified four missense mutations in the SETD1A gene (SET domain-containing 1A, histone lysine methyltransferase): three de novo mutations in three individuals and one inherited mutation in a four-generation family. Whole-exome sequencing indicated that all four of these mutations were responsible for the seizures. Mutations of SETD1A have been implicated in schizophrenia and developmental disorders, so we examined the role of the four mutations (R913C, Q269R, G1369R, and R1392H) in neural development. We found that their expression in mouse primary cortical neurons affected excitatory synapse development. Moreover, expression of the R913C mutation also affected the migration of cortical neurons in the mouse brain. We further identified two common genes (Neurl4 and Usp39) affected by mutations of SETD1A. These results suggested that the mutations of SETD1A play a fundamental role in abnormal synaptic function and the development of neurons, so they may be pathogenic factors for neurodevelopmental disorders.

Arsenic speciation in rice cereals for infants.

The aim of this study was to conduct a survey of arsenic (As) content in rice cereals for infants. The analysis was based on the FDA Elemental Analysis Manual (EAM 4.11). An inductively coupled plasma mass spectrometer (ICP-MS) was used to determine total As. Due to the different toxicities of the chemical forms of arsenic, the ICP-MS coupled to a high-performance liquid chromatograph (HPLC) was used to perform As speciation. The total and speciated arsenic was determined in 31 different infant rice cereals sold in U.S. supermarkets. The mass fraction of total inorganic As (iAs; sum of arsenite As(III) and arsenate As(V)) concentrations ranged between 55.5 ± 1.3 and 158.0 ± 6.0 µg/kg. The average total arsenic and iAs concentrations in infant rice cereal were 174.4 and 101.4 µg/kg, respectively. There was no substantial difference in iAs levels between organic and conventional rice cereals. The mixed-grain rice cereal contained the least total (105 µg/kg) and inorganic arsenic (63 µg/kg). The major detected organoarsenical species was dimethylarsinic acid (DMA). Monomethylarsonic acid (MMA) was not detected, or only trace levels were found. Spiked sample percent recoveries for iAs, DMA, and MMA ranged from a low of 97.3% for iAs to a high of 115.0% for DMA. Results for speciated and total As in the National Institute of Standards and Technology standard reference material rice flour (NIST SRM 1568) were in good agreement with certified values. In the NIST SRM 1568 sample (n = 5) repeatability (%RSD) was 2.8% for iAs, 1.7% for DMA and species sum, and 5.3% for the total arsenic by As total method. The average percent mass balance was 99.9 ± 6.3% for the NIST SRM 1568 sample. This study provides new and much needed information on arsenic levels in rice-based infant cereals.