Neural Tissue Research Articles

Efficient fabrication of 3D bioprinted functional sensory neurons using an inducible Neurogenin-2 human pluripotent stem cell line

Three-dimensional (3D) tissue models have gained recognition for their improved ability to mimic the native cell microenvironment compared to traditional two-dimensional models. This progress has been driven by advances in tissue-engineering technologies such as 3D bioprinting, a promising method for fabricating biomimetic living tissues. While bioprinting has succeeded in generating various tissues to date, creating neural tissue models remains challenging. In this context, we present an accelerated approach to fabricate 3D sensory neuron (SN) structures using a transgenic human pluripotent stem cell (hPSC)-line that contains an inducible Neurogenin-2 (NGN2) expression cassette. The NGN2 hPSC line was first differentiated to neural crest cell (NCC) progenitors, then incorporated into a cytocompatible gelatin methacryloyl-based bioink for 3D bioprinting. Upregulated NGN2 expression in the bioprinted NCCs resulted in induced SN (iSN) populations that exhibited specific cell markers, with 3D analysis revealing widespread neurite outgrowth through the scaffold volume. Calcium imaging demonstrated functional activity of iSNs, including membrane excitability properties and voltage-gated sodium channel (NaV) activity. This efficient approach to generate 3D bioprinted iSN structures streamlines the development of neural tissue models, useful for the study of neurodevelopment and disease states and offering translational potential.

Maximum-correntropy-based sequential method for fast neural population activity reconstruction in the cortex from incomplete abnormally-disturbed noisy measurements

This paper continues to explore the membrane potential reconstruction and pattern recognition problem in a neural tissue modeled by Stochastic Dynamic Neural Field (SDNF) equation. Although recent research has suggested an efficient solution based on the state-space approach through nonlinear Bayesian filtering framework, it is becoming extremely difficult to ignore the existence of non-Gaussian uncertainties in the SDNFs as well as the stability problem of neuronal population dynamics to outliers. Motivated by recent events in signal processing and mathematical neuroscience, this paper explores the SDNFs in a presence of non-Gaussian uncertainties, which is the shot noise case, where the corrupted data might appear due to broken sensors. We derive the “distributionally robust” state estimator for the membrane potential reconstruction process that is the Maximum Correntropy Criterion Extended Kalman Filter (MCC-EKF) as well as its fast and numerically robust (to roundoff) implementation method by using the sequential principle of processing the measurement vectors. The numerical experiments are provided to illustrate the performance of the novel estimation methods.

Glaucoma, More than Meets the Eye: Patterns of Demyelination Revealed in Human Postmortem Glaucomatous Optic Nerve.

Glaucoma is a neurodegenerative disease affecting millions worldwide, characterised by retinal ganglion cell (RGC) degeneration which leads to blindness in more advanced cases. Although the pathogenesis and underlying mechanisms of glaucoma are not fully understood, there are theories that hint at demyelination playing a role in the disease process. Demyelination, or the degeneration of the myelin sheath surrounding axons, has been found in previous studies using animal models of glaucoma and clinical assessments of glaucoma patients. However, this has not been fully realised or quantified in glaucoma patients. Utilising postmortem optic nerve samples from glaucoma and healthy subjects, various immunohistochemical and morphological assessments were performed to determine the extent, if any, of demyelination in glaucomatous optic nerves. Our findings revealed that alongside nerve shrinkage and degeneration of nerve tissue fascicles, there were significantly less myelin proteins, specifically myelin basic protein (MBP), in glaucoma optic nerves. Additionally, the loss of MBP was correlated with decreased oligodendrocyte (OLG) precursors and increasing glial activity. This further supports previous evidence that demyelination may be a secondary degenerative process associated with glaucoma disease progression. Not only do these results provide evidence for potential disease mechanisms, but this is also the first study to quantify optic nerve demyelination in glaucoma postmortem tissue.

Sequencing Sialic Acid Positioning in Gangliosides by High-Resolution Cyclic Ion Mobility Separations Coupled with Multiple Collision-Induced Dissociation-Based Tandem Mass Spectrometry Strategies.

Gangliosides, a diverse class of glycosphingolipids, are highly abundant in neural tissue and have been implicated in numerous aging-related diseases. Their characterization with methods such as liquid chromatography-tandem mass spectrometry is often precluded by their structural complexity, isomeric heterogeneity, and lack of commercially available authentic standards. In this work, we coupled high-resolution cyclic ion mobility spectrometry with multiple collision-induced dissociation-based tandem mass spectrometry strategies to sequence the sialic acid positions in various ganglioside isomers. Initially, as a proof-of-concept demonstration, we were able to characterize the sialic acid positions in several GD1 and GT1 species. From there, we extended our approach to identify the location of N-glycolylneuraminic acid (NeuGc) residues in previously uncharacterized GD1 and GQ1 isomers. Our results highlight the potential of this presented methodology for the de novo characterization of gangliosides within complex biological matrices without the need for authentic standards.

A Genome-Wide Association Study Suggests New Susceptibility Loci for Primary Antiphospholipid Syndrome.

Primary antiphospholipid syndrome (PAPS) is a rare autoimmune disease characterized by the presence of antiphospholipid antibodies and the occurrence of thrombotic events and pregnancy complications. Our study aimed to identify novel genetic susceptibility loci associated with PAPS. We performed a genome-wide association study comprising 5,485 individuals (482 affected individuals) of European ancestry. Significant and suggestive independent variants from a meta-analysis of approximately 7 million variants were evaluated for functional and biological process enrichment. The genetic risk variability for PAPS in different populations was also assessed. Hierarchical clustering, Mahalanobis distance, and Dirichlet Process Mixtures with uncertainty clustering methods were used to assess genetic similarities between PAPS and other immune-mediated diseases. We revealed genetic associations with PAPS in a regulatory locus within the HLA class II region near HLA-DRA and in STAT1-STAT4 with a genome-wide level of significance; 34 additional suggestive genetic susceptibility loci for PAPS were also identified. The disease risk allele near HLA-DRA is associated with overexpression of HLA-DRB6, HLA-DRB9, HLA-DQA2, and HLA-DQB2 in immune cells, vascular tissue, and nervous tissue. This association is independent of the association between PAPS and HLA-DRB1*1302. Functional analyses highlighted immune-related pathways in PAPS-associated loci. The comparison with other immune-mediated diseases revealed a close genetic relatedness to neuromyelitis optica, systemic sclerosis, and Sjögren syndrome, suggesting co-localized causal variations close to STAT1-STAT4, TNPO3, and BLK. This study represents a comprehensive large-scale genetic analysis for PAPS and provides new insights into the genetic basis and pathophysiology of this rare disease.

Malformations of the Spinal Cord: From Genetics to Diagnosis and Rehabilitation

AbstractSpinal cord malformations, known as “spinal dysraphisms” encompass a diverse range of spinal abnormalities characterized by incomplete median closure of mesenchymal, bone, and nervous tissues. They are classified as “open,” involving both the spinal cord and overlying tissues, or “occult,” affecting only nervous system structures. Neurulation abnormalities along the neural tube, from the rostral to the caudal portions, primarily cause these malformations. Clinical presentations vary, including cutaneous manifestations like hemangiomas, dimples, hair tufts, and scoliosis. “Tethered cord syndrome,” often associated with these malformations, manifests as a clinical syndrome rather than a primary anomaly. Newborns are typically asymptomatic, with malformations often identified by associated skin abnormalities. Older children may experience pain, sensory/motor disturbances, urinary/anal sphincter abnormalities, and muscle weakness affecting mobility. Neuroimaging, crucial for diagnosis and treatment planning, includes ultrasound, CT, and MRI. Surgical intervention, tailored to specific malformation subtypes, may involve the repair of myelomeningocele soon after birth or conservative management for asymptomatic occult dysraphism. Rehabilitation encompasses physical, occupational, recreational, and speech therapies. Prevention is paramount, emphasizing the role of health care professionals in prenatal care and education. This review aims to provide a systematic classification of spinal cord malformations to aid clinicians in diagnosis and management.

Deposition of Gadolinium in the Central and Peripheral Nervous Systems and Its Effects on Sensory, Cognitive, and Athletic Implications after Multiple Injections of Gadolinium-Based Contrast Agents in Rats.

After repeat administration of gadolinium-based contrast agents (GBCAs), the association between gadolinium retention in the central and peripheral nervous systems and the main manifestations of myelopathy and progressive neurologic symptoms remains unclear. We investigated the effects of the repeat administration of GBCAs on gadolinium retention in the central and peripheral nervous systems and the sensory, cognitive, and athletic implications. Forty-eight male Wistar rats (6 weeks of age) were randomly divided into 4 experimental groups (12 rats in each group): the gadodiamide group (linear and nonionic GBCAs), the gadopentetate dimeglumine group (linear and ionic GBCAs), the gadoterate meglumine group (macrocyclic and ionic GBCAs), and the control group (0.9% saline solution). The brains of the rats were scanned using 9.4T MRI. Sensory behavioral tests were performed to assess the effect of GBCAs on pain sensitivity function. Gadolinium deposition in the brain, spinal cord, and peripheral nerves was determined by inductively coupled plasma mass-spectrometry. Transmission electron microscopy was used to observe the microscopic distribution of gadolinium after deposition in the spinal cord. The histopathologic features in the spinal cord were analyzed by H&E staining, Nissl staining, glial fibrillary acidic protein staining, and neuron-specific enolase staining after administration of GBCAs. All GBCAs resulted in gadolinium deposition in the central and peripheral nerve tissues, with the highest deposition in the sciatic nerve tissue (mean, 62.86 [SD, 12.56] nmol/g). Decreased muscle power, impairment of spatial cognitive function power, and pain hypersensitivity to thermal and mechanical stimuli were observed after exposure to gadodiamide. At the spinal cord, transmission electron microscopy found that the region of gadolinium depositions had a spheric structure similar to "sea urchins" and was mainly located near the vascular basement membrane. Multiple injections of GBCAs caused gadolinium deposition in the brain, spinal cord, and peripheral nerves, especially in the spinal cords of the gadodiamide group. Gadodiamide led to pain hypersensitivity and decreased muscle power and cognitive ability. For the patients who are hypersensitive to pain and need multiple MRI examinations, we recommend using macrocyclic GBCAs and the lowest dose possible.

Evaluation of the Efficiency of Neural Enolase (NSA) in the Diagnosis of Viral Encephalitis: A Medical Review

Encephalitis is an inflammation that occurs in the brain when a virus in some cases, bacteria attack it directly, and can also be triggered by other factors. It is an inflammatory process of the meninges, the membranes that cover the central nervous system. Enolase isoenzyme found in neuronal and neuroendocrine tissues. Its levels in tissues other than erythrocytes are negligible. Its measurement is useful for monitoring tumors of neuroendocrine origin, such as neuroblastoma, pheochromocytoma, medullary thyroid carcinoma, small cell lung carcinoma, melanoma, and some pancreatic tumors. NSE concentrations in cerebrospinal fluid (CSF) are often elevated in diseases that result in relatively rapid neuronal destruction. This review aims to evaluate the efficiency of neural enolase (NSA) in diagnosing viral encephalitis. The methodology used an integrative literature review and a synthesis process to develop the study, expand the understanding of knowledge, and achieve the expected results. The integrative literature review is a method that aims to synthesize results obtained in research on a topic or issue, in a systematic, ordered, and comprehensive way. It is called integrative because it provides broader information on a subject/problem, thus constituting a body of knowledge. To carry out the study, a search for scientific articles was carried out through the Virtual Health Library, in the SCIELO, LILACS, and PUBMED databases, using the terminologies registered in the Health Sciences descriptors, encephalitis, enolase, virus, and diagnosis.

Hybrid construction of tissue-engineered nerve graft using skin derived precursors induced neurons and Schwann cells to enhance peripheral neuroregeneration

Peripheral nerve injury is a major challenge in clinical treatment due to the limited intrinsic capacity for nerve regeneration. Tissue engineering approaches offer promising solutions by providing biomimetic scaffolds and cell sources to promote nerve regeneration. In the present work, we investigated the potential role of skin-derived progenitors (SKPs), which are induced into neurons and Schwann cells (SCs), and their extracellular matrix in tissue-engineered nerve grafts (TENGs) to enhance peripheral neuroregeneration. SKPs were induced to differentiate into neurons and SCs in vitro and incorporated into nerve grafts composed of a biocompatible scaffold including chitosan neural conduit and silk fibroin filaments. In vivo experiments using a rat model of peripheral nerve injury showed that TENGs significantly enhanced nerve regeneration compared to the scaffold control group, catching up with the autograft group. Histological analysis showed improved axonal regrowth, myelination and functional recovery in animals treated with these TENGs. In addition, immunohistochemical staining confirmed the presence of induced neurons and SCs within the regenerated nerve tissue. Our results suggest that SKP-induced neurons and SCs in tissue-engineered nerve grafts have great potential for promoting peripheral nerve regeneration and represent a promising approach for clinical translation in the treatment of peripheral nerve injury. Further optimization and characterization of these engineered constructs is warranted to improve their clinical applicability and efficacy.

The impact of nutrition on people's cognitive development

Background: Adequate nutrition of the brain maintains the structural and functional integrity of the brain and nervous tissue, which improves cognitive ability, improving the relationships we have every day and, above all, preventing cognitive deterioration associated with aging.

Applications of 2D Nanomaterials in Neural Interface.

Neural interfaces are crucial conduits between neural tissues and external devices, enabling the recording and modulation of neural activity. However, with increasing demand, simple neural interfaces are no longer adequate to meet the requirements for precision, functionality, and safety. There are three main challenges in fabricating advanced neural interfaces: sensitivity, heat management, and biocompatibility. The electrical, chemical, and optical properties of 2D nanomaterials enhance the sensitivity of various types of neural interfaces, while the newly developed interfaces do not exhibit adverse reactions in terms of heat management and biocompatibility. Additionally, 2D nanomaterials can further improve the functionality of these interfaces, including magnetic resonance imaging (MRI) compatibility, stretchability, and drug delivery. In this review, we examine the recent applications of 2D nanomaterials in neural interfaces, focusing on their contributions to enhancing performance and functionality. Finally, we summarize the advantages and disadvantages of these nanomaterials, analyze the importance of biocompatibility testing for 2D nanomaterials, and propose that improving and developing composite material structures to enhance interface performance will continue to lead the forefront of this field.

Topical application of daphnetin hydrogel for traumatic brain injury.

Traumatic brain injury (TBI) causes neuronal cell damage and dysfunction. According to previous studies, daphnetin (Dap) has a protective effect in neurological injury. However, the in vivo bioavailability of daphnetin is not high. The purpose of this study was to determine whether administering daphnetin directly into the site of injury via a hydrogel drug carrier could improve its therapeutic impact. Tripolycerol monostearates / daphnetin (TM/Dap) hydrogels were prepared and characterised using water bath heating, scanning electron microscopy (SEM) and small animal in vivo imaging techniques. The TBI model was established using the Feeney free fall impact method. Using the Morris water maze test, the mNSS neurological deficit rating scale, haematoxylin-eosin staining, and liver and kidney function tests, the therapeutic benefit of TM/Dap and its toxic side effects were assessed. The therapeutic effects of TM/Dap were further investigated using wet and dry gravimetric methods, Evans blue staining, protein immunoblotting, immunofluorescence staining techniques and ELISA. The efficacy of the TM/Dap hydrogel in gradually releasing daphnetin in the context of traumatic brain damage was shown by both in vitro and in vivo tests. Behavioral experiments showed that the learning and spatial memory abilities of TM/Dap hydrogel treated mice were significantly improved in the water maze experiment. And TM/Dap hydrogel has high biosafety for organisms. The results of the therapeutic mechanism of action showed that TM/Dap hydrogel showed more significant efficacy in reducing the neuroinflammatory response caused by TNF-α, IL-6 and other factors, as well as promoting the recovery of post-traumatic neurological function. The use of hydrogel as a drug carrier for daphnetin showed more significant efficacy in reducing neuroinflammatory response, protecting nerve tissue and promoting post-traumatic neurological recovery compared with traditional drug delivery methods.

Clinical Analysis and Treatment Strategies About Cartilage Scaffold Exposure After Auricular Reconstruction With the Expanded Two-Flap Method.

Objectives: Soft tissue expansion is one of the main methods for autologous cartilage auricular reconstruction. The aim of this study was to analyze the risk factors for cartilage exposure after this method and to describe a surgical method for this complication. Methods: From January 2018 to December 2020, 853 patients (908 sides) underwent auricular reconstruction with an expanded two-flap method at our center. Thirty-two patients experienced cartilage exposure postoperatively. These patients were set as the case group, and 1:1 matched sampling was performed among patients who did not have cartilage exposure. The matched sample of 32 cases was set as the control group. All 64 patients were evaluated according to the Orbit, Mandible, Ear, Nerve, and Soft tissue (OMENS) classification system to analyze the correlation between cartilage exposure and hemifacial microsomia (HFM) and OMENS subtypes. The complication was repaired with superficial temporal fascial flap combined with skin graft. Results: HFM might be a risk factor for scaffold cartilage exposure, and there was a significant correlation between cartilage exposure and orbital malformation, facial nerve dysplasia, and soft tissue developmental malformation. The use of a superficial temporal fascial flap combined with a split-thickness skin graft to repair the complication achieved satisfactory outcomes. Conclusions: There is a correlation between cartilage scaffold exposure and the severity of HFM. Temporoparietal fascial flap transfer combined with skin grafting proved to be an effective method for cartilage exposure.

Recent advances and applications of human brain models.

Recent advances in human pluripotent stem cell (hPSC) technologies have prompted the emergence of new research fields and applications for human neurons and brain organoids. Brain organoids have gained attention as an in vitro model system that recapitulates the higher structure, cellular diversity and function of the brain to explore brain development, disease modeling, drug screening, and regenerative medicine. This progress has been accelerated by abundant interactions of brain organoid technology with various research fields. A cross-disciplinary approach with human brain organoid technology offers a higher-ordered advance for more accurately understanding the human brain. In this review, we summarize the status of neural induction in two- and three-dimensional culture systems from hPSCs and the modeling of neurodegenerative diseases using brain organoids. We also highlight the latest bioengineered technologies for the assembly of spatially higher-ordered neural tissues and prospects of brain organoid technology toward the understanding of the potential and abilities of the human brain.

Mesoporous manganese-doped nano-antioxidant suppresses pyroptosis for effective ischemic stroke therapy

Pyroptosis mediates neuroinflammation following ischemic stroke and contributes to its development, indicating that targeting pyroptosis-related pathways could be a potential therapeutic strategy for stroke. In this study, a mesoporous manganese-doped cerium oxide (MMC) nano-antioxidant encapsulated with caspase-1 inhibitor VX765 is rationally designed and engineered to relieve ischemic stroke by integrating the suppression of pyroptosis activity with reactive oxygen species (ROS) scavenging capability. Through the surface modification of Angiopoietin-2 (Ang2), the nano-antioxidant can specifically bind to the low-density lipoprotein receptor-related protein, cross the blood–brain barrier, and ultimately aggregate in ischemic neuronal tissues. In mice subjected to middle cerebral artery occlusion/reperfusion, the nano-antioxidant alleviates oxidative stress, inhibits cell pyroptosis, reduces inflammatory cytokines, and regulates microglial polarization toward M2, ultimately mitigating neuroinflammation and reperfusion injury in brain tissue. Mechanistic studies reveal that the pyroptosis-associated NOD-like receptor signaling pathway, the inflammation-associated TNF signaling pathway, the neutrophil chemotaxis-associated Toll-like receptor signaling pathway, and the oxidative stress-associated NF-kappa B signaling pathway are involved in the antioxidant-mediated stroke therapy. Endowed with pyroptosis-reversing ability and nano-antioxidant activities, this engineered antioxidant presents a novel therapeutic paradigm for minimizing reperfusion injury and enhancing stroke treatment efficacy.

Conductive collagen/polypyrrole-b-polycaprolactone hydrogel for bioprinting of neural tissue constructs.

Bioprinting is increasingly being used for fabrication of engineered tissues for regenerative medicine, drug testing, and other biomedical applications. The success of this technology lies with the development of suitable bioinks and hydrogels that are specific to the intended tissue application. For applications such as neural tissue engineering, conductivity plays an important role in determining the neural differentiation and neural tissue regeneration. Although several conductive hydrogels based on metal nanoparticles (NPs) such as gold and silver, carbon-based materials such as graphene and carbon nanotubes and conducting polymers such as polypyrrole (PPy) and polyaniline were used, they possess several disadvantages. The long-term cytotoxicity of metal nanoparticles (NPs) and carbon-based materials restricts their use in regenerative medicine. The conductive polymers, on the other hand, are non-biodegradable and possess weak mechanical properties limiting their printability into three-dimensional constructs. The aim of this study is to develop a biodegradable, conductive, and printable hydrogel based on collagen and a block copolymer of PPy and polycaprolactone (PCL) (PPy-block-poly(caprolactone) [PPy-b-PCL]) for bioprinting of neural tissue constructs. The printability, including the influence of the printing speed and material flow rate on the printed fiber width; rheological properties; and cytotoxicity of these hydrogels were studied. The results prove that the collagen/PPy-b-PCL hydrogels possessed better printability and biocompatibility. Thus, the collagen/PPy-b-PCL hydrogels reported this study has the potential to be used in the bioprinting of neural tissue constructs for the repair of damaged neural tissues and drug testing or precision medicine applications.

One‐dimensional nanomaterials for nerve tissue engineering to repair spinal cord injury

AbstractIn recent years, tissue engineering has emerged as a cutting‐edge approach for the treatment of spinal cord injury (SCI) owing to its remarkable capabilities. It can create living tissues with robust vitality, achieve maximal tissue repair with minimal cell usage, and facilitate seamless reconstruction with unmatched plasticity, all while addressing immune rejection issues. Among these advancements, one‐dimensional (1D) materials have garnered significant attention. Their morphology closely resembles the extracellular matrix environment, thereby fostering the elongation of dendrites and axons on neurons and greatly enhancing the prospects for SCI repair. With a keen focus on the latest advancements in the application of 1D nanomaterials in nerve tissue engineering for spinal nerve repair, this review delves into several key aspects. Firstly, it explores the “bottom‐up” approach to synthesizing 1D nanomaterials. Secondly, it examines the mechanisms by which these nanomaterials influence neural tissue engineering. Thirdly, it presents various cutting‐edge strategies aimed at optimizing the morphology and performance of 1D materials, thereby enhancing the efficiency of nerve tissue injury repair. Lastly, it discusses the current challenges and future prospects facing this fascinating field. We aspire that this comprehensive review will provide a profound understanding of the development of 1D materials in neural tissue engineering and inspire a wider audience with its potential.

Biomimetic design and integrated biofabrication of an in-vitro three-dimensional multi-scale multilayer cortical model

The lack of accurate and reliable in vitro brain models hinders the development of brain science and research on brain diseases. Owing to the complex structure of the brain tissue and its highly nonlinear characteristics, the construction of brain-like in vitro tissue models remains one of the most challenging research fields in the construction of living tissues. This study proposes a multi-scale design of a brain-like model with a biomimetic cortical structure, which includes the macroscopic structural features of six layers of different cellular components, as well as micrometer-scale continuous fiber structures running through all layers vertically. To achieve integrated biomanufacturing of such a complex multi-scale brain-like model, a multi-material composite printing/culturing integrated bioprinting platform was developed in-house by integrating cell-laden hydrogel ink direct writing printing and electrohydrodynamic fiber 3D printing technologies. Through integrated bioprinting, multi-scale models with different cellular components and fiber structural parameters were prepared to study the effects of macroscopic and microscopic structural features on the directionality of neural cells, as well as the interaction between glial cells and neurons within the tissue model in a three-dimensional manner. The results revealed that the manufactured in vitro biomimetic cortical model achieved morphological connections between the layers of neurons, reflecting the structure and cellular morphology of the natural cortex. Micrometer-scale (10 μm) cross-layer fibers effectively guided and controlled the extension length and direction of the neurites of surrounding neural cells but had no significant effect on the migration of neurons. In contrast, glial cells significantly promoted the migration of surrounding PC12 cells towards the glial layer but did not contribute to the extension of neurites. This study provides a basis for the design and manufacture of accurate brain-like models for the functionalization of neuronal tissues.

Multi-Dimensional Morphometric and Volumetric Analysis of the Posterior Cranial Fossa to Study Type I Chiari Malformation

Chiari malformation type I (CM-1) is a complex disorder in which tonsillar herniation through the foramen magnum manifests with a spectrum of clinical symptoms. This work analyzes morphometric and volumetric characteristics of CM-1 patients. With Institutional Review Board (IRB) approval, we retrospectively reviewed a total of 72 adult CM-1 patients and 26 healthy adult volunteers who underwent volumetric magnetic resonance brain imaging. Clinical data was retrospectively extracted from the electronic medical record. We analyzed multi-dimensional morphometric and volumetric features within the posterior cranial fossa and correlated these features with syrinx formation and the decision to undergo surgical decompression. In our study, CM-1 patients had decreased cerebellar, brainstem and 4th ventricular volumes but larger tonsillar volume with increased total tonsillar length. CM-1 patients who underwent surgery had significantly more neural tissue within the cross-sectional area of cisterna magna. Logistic regression demonstrated that combining neural tissue at foramen magnum with cerebellar & 4th ventricular volumes led to great degree of correlation with syrinx formation (AUC 0.911). Our findings suggest that the amount of tissue at the foramen magnum correlates with CM-1 patients who underwent decompressive surgery, more so than tonsillar length. Additionally, the combination of neural tissue at foramen magnum, cerebellar & 4th ventricular volumes led to great degree of correlation with syrinx formation. Together, these findings suggest that a global compressive phenomenon within the posterior fossa leads to CM-1 symptomatology and syrinx formation.

Spatial enrichment and genomic analyses reveal the link of NOMO1 with amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a severe motor neuron disease with uncertain genetic predisposition in most sporadic cases. The spatial architecture of cell types and gene expression are the basis of cell-cell interactions, biological function and disease pathology, but are not well investigated in the human motor cortex, a key ALS-relevant brain region. Recent studies indicated single nucleus transcriptomic features of motor neuron vulnerability in ALS motor cortex. However, the brain regional vulnerability of ALS-associated genes and the genetic link between region-specific genes and ALS risk remain largely unclear. Here, we developed an entropy-weighted differential gene expression matrix-based tool (SpatialE) to identify the spatial enrichment of gene sets in spatial transcriptomics. We benchmarked SpatialE against another enrichment tool (multimodal intersection analysis) using spatial transcriptomics data from both human and mouse brain tissues. To investigate regional vulnerability, we analysed three human motor cortex and two dorsolateral prefrontal cortex tissues for spatial enrichment of ALS-associated genes. We also used Cell2location to estimate the abundance of cell types in ALS-related cortex layers. To dissect the link of regionally expressed genes and ALS risk, we performed burden analyses of rare loss-of-function variants detected by whole-genome sequencing in ALS patients and controls, then analysed differential gene expression in the TargetALS RNA-sequencing dataset. SpatialE showed more accurate and specific spatial enrichment of regional cell type markers than multimodal intersection analysis in both mouse brain and human dorsolateral prefrontal cortex. Spatial transcriptomic analyses of human motor cortex showed heterogeneous cell types and spatial gene expression profiles. We found that 260 manually curated ALS-associated genes are significantly enriched in layer 5 of the motor cortex, with abundant expression of upper motor neurons and layer 5 excitatory neurons. Burden analyses of rare loss-of-function variants in Layer 5-associated genes nominated NOMO1 as a novel ALS-associated gene in a combined sample set of 6814 ALS patients and 3324 controls (P = 0.029). Gene expression analyses in CNS tissues revealed downregulation of NOMO1 in ALS, which is consistent with a loss-of-function disease mechanism. In conclusion, our integrated spatial transcriptomics and genomic analyses identified regional brain vulnerability in ALS and the association of a layer 5 gene (NOMO1) with ALS risk.