Meningeal Lymphatics Research Articles

A functional role of meningeal lymphatics in sex difference of stress susceptibility in mice

Major depressive disorder is one of the most common mental health conditions. Meningeal lymphatics are essential for drainage of molecules in the cerebrospinal fluid to the peripheral immune system. Their potential role in depression-like behaviour has not been investigated. Here, we show in mice, sub-chronic variable stress as a model of depression-like behaviour impairs meningeal lymphatics in females but not in males. Manipulations of meningeal lymphatics regulate the sex difference in the susceptibility to stress-induced depression- and anxiety-like behaviors in mice, as well as alterations of the medial prefrontal cortex and the ventral tegmental area, brain regions critical for emotional regulation. Together, our findings suggest meningeal lymphatic impairment contributes to susceptibility to stress in mice, and that restoration of the meningeal lymphatics might have potential for modulation of depression-like behaviour.

Update on the current knowledge of lymphatic drainage system and its emerging roles in glioma management.

The draining of brain interstitial fluid (ISF) to cerebrospinal fluid (CSF) and the subsequent draining of CSF to meningeal lymphatics is well-known. Nonetheless, its role in the development of glioma is a remarkable finding that has to be extensively understood. The glymphatic system (GS) collects CSF from the subarachnoid space and brain ISF through aquaporin-4 (AQP4) water channels. The glial limiting membrane and the perivascular astrocyte-end-feet membrane both have elevated levels of AQP4. CSF is thought to drain through the nerve sheaths of the olfactory and other cranial nerves as well as spinal meningeal lymphatics via dorsal or basal lymphatic vessels. Meningeal lymphatic vessels (MLVs) exist below the skull in the dorsal and basal regions. In this view, MLVs offer a pathway to drain macromolecules and traffic immunological cells from the CNS into cervical lymph nodes (CLNs), and thus can be used as a candidate curing strategy against glioma and other associated complications, such as neuro-inflammation. Taken together, the lymphatic drainage system could provide a route or approach for drug targeting of glioma and other neurological conditions. Nevertheless, its pathophysiological role in glioma remains elusive, which needs extensive research. The current review aims to explore the lymphatic drainage system, its role in glioma progression, and possible therapeutic techniques that target MLVs in the CNS.

Glymphatic System: Emerging Therapeutic Target for Neurological Diseases.

The newly discovered glymphatic system acts as pseudolymphatic vessels subserving brain waste clearance and is functionally dependent on astrocytic aquaporin-4 channels. The glymphatic system primarily functions during sleep as an interchange between cerebrospinal fluid and interstitial fluid, with cerebrospinal fluid flowing into the parenchyma via the perivascular spaces and then exchanging with interstitial fluid. The discovery of meningeal lymphatics helps refine the conceptual framework of glymphatic pathway, as certain waste products collected alongside perivascular spaces ultimately drain into the cervical lymph nodes via meningeal lymphatics, whose function regulates the functioning of the glymphatic system. The glymphatic and meningeal lymphatic systems are critical for the homeostasis of central nervous system, and their malfunctions complicate cerebral dysfunction and diseases. The present review will shed light on the structure, regulation, functions, and interrelationships of the glymphatic system and meningeal lymphatics. We will also expound on their impairments and corresponding targeted intervention in neurodegenerative diseases, traumatic brain injury, stroke, and infectious/autoimmune diseases, offering valuable references for future research.

Meningeal lymphatics mediate drainage of viruses from the CNS.

Meningeal lymphatics mediate drainage of viruses from the CNS.

Drainage of senescent astrocytes from brain via meningeal lymphatic routes

Recent progress on the central lymphatic system has greatly increased our understanding of how the brain maintains its own waste homeostasis. Here, we showed that perivascular spaces and meningeal lymphatic vessels form a functional route for clearance of senescent astrocytes from the aging brain. Blocking meningeal lymphatic drainage by ligation of the deep cervical lymph nodes impaired clearance of senescent astrocytes from brain parenchyma, subsequently increasing neuroinflammation in aged mice. By contrast, enhancing meningeal lymphatic vessel diameter by a recombinant adeno-associated virus encoding mouse vascular endothelial growth factor-C (VEGF-C) improved clearance of senescent astrocytes and mitigated neuroinflammation. Mechanistically, VEGF-C was highly expressed in senescent astrocytes, contributing themselves to migrate across lymphatic vessels along C–C motif chemokine ligand 21 (CCL21) gradient by interacting with VEGF receptor 3. Moreover, intra-cisternal injection of antibody against CCL21 hampered senescent astrocytes into the lymphatic vessels and exacerbated short memory defects of aged mice. Together, these findings reveal a new perspective for the meningeal lymphatics in the removal of senescent astrocytes, thus offering a valuable target for therapeutic intervention.

Neuroinflammation creates an immune regulatory niche at the meningeal lymphatic vasculature near the cribriform plate.

Meningeal lymphatics near the cribriform plate undergo lymphangiogenesis during neuroinflammation to drain excess fluid. Here, we hypothesized that lymphangiogenic vessels may acquire an altered phenotype to regulate immunity. Using single-cell RNA sequencing of meningeal lymphatics near the cribriform plate from healthy and experimental autoimmune encephalomyelitis in the C57BL/6 model, we report that neuroinflammation induces the upregulation of genes involved in antigen presentation such as major histocompatibility complex class II, adhesion molecules including vascular cell adhesion protein 1 and immunoregulatory molecules such as programmed cell death 1 ligand 1, where many of these changes are mediated by interferon-γ. The inflamed lymphatics retain CD11c+ cells and CD4 T cells where they capture and present antigen, creating an immunoregulatory niche that represents an underappreciated interface in the regulation of neuroinflammation. We also found discontinuity of the arachnoid membrane near the cribriform plate, which provides unrestricted access to the cerebrospinal fluid. These findings highlight a previously unknown function of local meningeal lymphatics in regulating immunity that has only previously been characterized in draining lymph nodes.

The meningeal lymphatic vasculature in neuroinflammation.

The lymphatic vasculature is a unidirectional network of lymphatic endothelial cells, whose main role is to maintain fluid homeostasis along with the absorption of dietary fat in the gastrointestinal organs and management and coordination of immune cell trafficking into lymph nodes during homeostasis and under inflammatory conditions. In homeostatic conditions, immune cells, such as dendritic cells, macrophages, or T cells can enter into the lymphatic vasculature and move easily through the lymph reaching secondary lymph nodes where immune cell activation or peripheral tolerance can be modulated. However, under inflammatory conditions such as pathogen infection, increased permeabilization of lymphatic vessels allows faster immune cell migration into inflamed tissues following a chemokine gradient, facilitating pathogen clearance and the resolution of inflammation. Interestingly, since the re-discovery of lymphatic vasculature in the central nervous system, known as the meningeal lymphatic vasculature, the role of these lymphatics as a key player in several neurological disorders has been described, with emphasis on the neurodegenerative process. Alternatively, less has been discussed about meningeal lymphatics and its role in neuroinflammation. In this review, we discuss current knowledge about the anatomy and function of the meningeal lymphatic vasculature and specifically analyze its contribution to different neuroinflammatory processes, highlighting the potential therapeutic target of meningeal lymphatic vasculature in these pathological conditions.

Early life adversity drives sex-specific anhedonia and meningeal immune gene expression through mast cell activation

Exposure to early life adversity (ELA) in the form of physical and/or psychological abuse or neglect increases the risk of developing psychiatric and inflammatory disorders later in life. It has been hypothesized that exposure to ELA results in persistent, low grade inflammation that leads to increased disease susceptibility by amplifying the crosstalk between stress-processing brain networks and the immune system, but the mechanisms remain largely unexplored. The meninges, a layer of three overlapping membranes that surround the central nervous system (CNS)- dura mater, arachnoid, and piamater – possess unique features that allow them to play a key role in coordinating immune trafficking between the brain and the peripheral immune system. These include a network of lymphatic vessels that carry cerebrospinal fluid from the brain to the deep cervical lymph nodes, fenestrated blood vessels that allow the passage of molecules from blood to the CNS, and a rich population of resident mast cells, master regulators of the immune system. Using a mouse model of ELA consisting of neonatal maternal separation plus early weaning (NMSEW), we sought to explore the effects of ELA on sucrose preference behavior, dura mater expression of inflammatory markers and mast cell histology in adult male and female C57Bl/6 mice. We found that NMSEW alone does not affect sucrose preference behavior in males or females, but it increases the dura mater expression of the genes coding for mast cell protease CMA1 (cma1) and the inflammatory cytokine TNF alpha (tnf alpha) in females. When NMSEW is combined with an adult mild stress (that does not affect behavior or gene expression in NH animals) females show reduced sucrose preference and even greater increases in meningeal cma1 levels. Interestingly, systemic administration of the mast cell stabilizer Ketotifen before exposure to adult stress prevents both, reduction in sucrose preference an increases in cma1 expression in NMSEW females, but facilitates stress-induced sucrose anhedonia in NMSEW males and NH females. Finally, histological analyses showed that, compared to males, females have increased baseline activation levels of mast cells located in the transverse sinus of the dura mater, where the meningeal lymphatics run along, and that, in males and females exposed to adult stress, NMSEW increases the number of mast cells in the interparietal region of the dura mater and the levels of mast cell activation in the sagittal sinus regions of the dura mater. Together, our results indicate that ELA induces long-term meningeal immune gene changes and heightened sensitivity to adult stress-induced behavioral and meningeal immune responses and that these effects could mediated via mast cells.

Meningeal lymphatics regulate radiotherapy efficacy through modulating anti-tumor immunity

As a first-line treatment, radiotherapy (RT) is known to modulate the immune microenvironment of glioma, but it is unknown whether the meningeal lymphatic vessel (MLV)-cervical lymph node (CLN) network regulates the process or influences RT efficacy. Here, we show that the MLV-CLN network contributes to RT efficacy in brain tumors and mediates the RT-modulated anti-tumor immunity that is enhanced by vascular endothelial growth factor C (VEGF-C). Meningeal lymphatic dysfunction impaired tumor-derived dendritic cell (DC) trafficking and CD8+ T cell activation after RT, whereas tumors overexpressing VEGF-C with meningeal lymphatic expansion were highly sensitive to RT. Mechanistically, VEGF-C-driven modulation of RT-triggered anti-tumor immunity was attributed to C-C Motif Chemokine Ligand 21 (CCL21)-dependent DC trafficking and CD8+ T cell activation. Notably, delivery of VEGF-C mRNA significantly enhanced RT efficacy and anti-tumor immunity in brain tumors. These findings suggest an essential role of the MLV-CLN network in RT-triggered anti-tumor immunity, and highlight the potential of VEGF-C mRNA for brain tumor therapy.

The cellular and molecular mechanisms of cerebrovascular regeneration and repair

<p indent=0mm>Stroke is a severe acute cerebrovascular disease that can cause a number of neuronal deaths and brain edema. The treatment of ischemic stroke mainly includes hyperacute vascular recanalization and later pro-angiogenesis therapy. At present, the promotion of angiogenesis is mainly through the combined application of vascular growth factors and guidance molecules, and the transplantation of endothelial progenitor cells to promote vascular regeneration and repair. Pericytes can promote angiogenesis and rebuild brain-blood-barrier. Activated platelets shed microparticles, which contain a variety of growth factors central to angiogenesis and neurogenesis during brain vascular regeneration. The newly discovered meningeal lymphatic vessels can invade the injured parenchyma to resolve edema and act as “growing tracks” to guide the growth of nascent blood vessels. These functions of pericytes, platelets, and the meningeal lymphatics suggest their potential as novel pro-angiogenic targets for ischemic stroke. In addition to ischemic stroke, microbleeds resulting from ruptures in microvessels are also a very extensive chronic cerebrovascular disease. After cerebral microbleeds, microglia can mediate the repair of brain vascular rupture through direct physical adhesion and mechanical traction. Here, we review the cellular and molecular mechanisms in cerebral vascular regeneration and repair. In addition, we provide our perspectives for future mechanistic studies and treatment strategies for neurovascular diseases.

Not Just Blood: Brain Fluid Systems and Their Relevance to Cerebrovascular Diseases.

Not Just Blood: Brain Fluid Systems and Their Relevance to Cerebrovascular Diseases.

Specificity of Adaptive Immune Responses in Central Nervous System Health, Aging and Diseases.

The field of neuroimmunology endorses the involvement of the adaptive immune system in central nervous system (CNS) health, disease, and aging. While immune cell trafficking into the CNS is highly regulated, small numbers of antigen-experienced lymphocytes can still enter the cerebrospinal fluid (CSF)-filled compartments for regular immune surveillance under homeostatic conditions. Meningeal lymphatics facilitate drainage of brain-derived antigens from the CSF to deep cervical lymph nodes to prime potential adaptive immune responses. During aging and CNS disorders, brain barriers and meningeal lymphatic functions are impaired, and immune cell trafficking and antigen efflux are altered. In this context, alterations in the immune cell repertoire of blood and CSF and T and B cells primed against CNS-derived autoantigens have been observed in various CNS disorders. However, for many diseases, a causal relationship between observed immune responses and neuropathological findings is lacking. Here, we review recent discoveries about the association between the adaptive immune system and CNS disorders such as autoimmune neuroinflammatory and neurodegenerative diseases. We focus on the current challenges in identifying specific T cell epitopes in CNS diseases and discuss the potential implications for future diagnostic and treatment options.

The Brain-Nose Interface: A Potential Cerebrospinal Fluid Clearance Site in Humans.

The human brain functions at the center of a network of systems aimed at providing a structural and immunological layer of protection. The cerebrospinal fluid (CSF) maintains a physiological homeostasis that is of paramount importance to proper neurological activity. CSF is largely produced in the choroid plexus where it is continuous with the brain extracellular fluid and circulates through the ventricles. CSF movement through the central nervous system has been extensively explored. Across numerous animal species, the involvement of various drainage pathways in CSF, including arachnoid granulations, cranial nerves, perivascular pathways, and meningeal lymphatics, has been studied. Among these, there is a proposed CSF clearance route spanning the olfactory nerve and exiting the brain at the cribriform plate and entering lymphatics. While this pathway has been demonstrated in multiple animal species, evidence of a similar CSF egress mechanism involving the nasal cavity in humans remains poorly consolidated. This review will synthesize contemporary evidence surrounding CSF clearance at the nose-brain interface, examining across species this anatomical pathway, and its possible significance to human neurodegenerative disease. Our discussion of a bidirectional nasal pathway includes examination of the immune surveillance in the olfactory region protecting the brain. Overall, we expect that an expanded discussion of the brain-nose pathway and interactions with the environment will contribute to an improved understanding of neurodegenerative and infectious diseases, and potentially to novel prevention and treatment considerations.

The growth and expansion of meningeal lymphatic networks are affected in craniosynostosis.

Skull malformations are associated with vascular anomalies that can impair fluid balance in the central nervous system. We previously reported that humans with craniosynostosis and mutations in TWIST1 have dural venous sinus malformations. It is still unknown whether meningeal lymphatic networks, which are patterned alongside the venous sinuses, are also affected. We now show that the growth and expansion of meningeal lymphatics are perturbed in Twist1 craniosynostosis models. Changes to the local meningeal environment, including hypoplastic dura and venous malformations, affect the ability of lymphatic networks to sprout and remodel. Dorsal networks along the transverse sinus are hypoplastic with reduced branching. By contrast, basal networks closer to the skull base are more variably affected, showing exuberant growth in some animals, suggesting they are compensating for vessel loss in dorsal networks. Injecting a molecular tracer into cerebrospinal fluid reveals significantly less drainage to the deep cervical lymph nodes, which is indicative of impaired lymphatic function. Collectively, our results show that meningeal lymphatic networks are affected in craniosynostosis, suggesting that the clearance of β-amyloid and waste from the central nervous system may be impeded.

Influence of sleep disruption on protein accumulation in neurodegenerative diseases

Abnormal accumulation of disease proteins in the central nervous system is a neuropathological feature in neurodegenerative disorders. Recently, a growing body of evidence has supported a role of disruption of the sleep-wake cycle in disease development, pathological changes and abnormal protein accumulation in neurodegenerative diseases, especially in Alzheimer’s disease and Parkinson’s disease. Sleep deprivation promotes abnormal accumulation of disease proteins. Interestingly, amyloid-β (Aβ) has daily oscillations in human cerebral spinal fluid (CSF) and is cleared more in sleep. Both circadian genes and circadian hormones are associated with disease protein deposition. Recently, the glymphatic pathway and meningeal lymphatics have been shown to play a critical role in Aβ clearance, which is mediated by the aquaporin (AQP-4) water channel on astrocytes. The rate of the clearance of Aβ by the glymphatic pathway is different during the sleep/wake cycle. Most importantly, circadian rhythms facilitate glymphatic clearance of solutes and Aβ in the CSF and interstitial fluid in an AQP-4-dependent manner, which further provides evidence for the involvement of circadian rhythms in disease protein clearance.

Mini review: Prospective therapeutic targets of Alzheimer's disease.

Alzheimer's disease is a neurological condition that causes the disruption of neuronal connections in the human brain. It is progressive and targets about 10% of the United States population over the age of 65.3 to date, there is no cure to the disease. Physicians can treat symptoms but lack the ability to stop the progression of the disease. However, promising research has come to the surface in recent years. A collection of these therapeutic targets, which have yielded positive results in mice models, are presented in this article. They include targets such as meningeal lymphatics, mitochondrial homeostasis, genomic instability, calcium homeostasis, and cold-shock proteins such as RNA-binding motif protein 3 and reticulon-3, high-density lipoprotein, and antibodies.

Glymphatic System Dysfunction in Mild Traumatic Brain Injury

To assess the clinical role of the glymphatic system in mild traumatic brain injury (mTBI) and post-concussive syndrome (PCS). Clinical manifestations of mTBI, or concussion, involve a wide array of cognitive, behavioral, and mechanical impairments that commonly spontaneously resolve within weeks. When these symptoms persist, it defines a class of mTBI known as post-concussive syndrome. A multifaceted approach for diagnosing concussion and PCS, heavily reliant on a neurocognitive screening, has become the standard in suspected cases. Conventional imaging protocols are occasionally implemented for exclusion of structural injury, rarely revealing substantial evidence in otherwise uncomplicated mTBI. A CNS-specific lymphatic network, termed glymphatic, has shown to play a critical role in immune surveillance and drainage of cellular debris. Moreover, recent evidence points to glymphatic dysfunction in TBI, including mild cases, as its anatomical layout becomes better understood. Here, we review the current literature on glymphatic function and imaging modalities, with an emphasis on implications in mTBI. Literature was compiled primarily using various keyword searches (glymphatic + imaging, meningeal lymphatics, glymphatic + concussion, etc.) via Pubmed and the NIH/NLM archive. Inclusion criteria involved limiting to studies on human patients or tissue. The glymphatic system displays critical function in healthy patients and in disease, with activity that suggests a diurnal sleep-cycle. Advanced imaging methodologies, most notably, the use of various MRI techniques, have identified impairments in meningeal lymphatic dysfunction in TBI, however, the clinical application of glymphatic imaging has yet to be well-studied and shows challenges in providing definitive data. Nonetheless, the potential for glymphatic imaging to expand our understanding of mTBI and PCS warrants further investigation. Anatomical and functional properties of the glymphatic network make an appealing target for concussion diagnosis, observing recovery, and exposing impact-related microstructural injuries, however the implementation of imaging in a clinical setting has yet to be well-characterized.

COVID-19 and Alzheimer's disease: Meninges-mediated neuropathology.

BackgroundSARS‐CoV‐2 the causative agent of COVID‐19 displays a broad range of pathophysiology. Cytokine storms associated with COVID‐19 damage the blood‐brain barrier (BBB) and allow pro‐inflammatory factors to invade the brain, further promoting neurodegeneration. While SARS‐CoV‐2 viral RNA and proteins have been detected in brain tissues, the mechanisms of neuroinvasion remain unknown. COVID‐19 has had a disproportionate impact on those suffering from neurodegenerative disorders such as Alzheimer’s disease (AD). Understanding the mechanisms of SARS‐CoV‐2 neuroinvasion is crucial to study the long‐term neurocognitive effects of COVID‐19 on AD pathology. Viruses can infiltrate the brain through the meninges via infected immune cells. The meninges regulate the immune surveillance of the brain and play a key role in the efflux of pathogens from the brain. Meningeal dysfunction has been demonstrated to exacerbate amyloid‐beta pathogenesis. In this study, we explore the neuroinvasion pathway of SARS‐CoV‐2 through the meninges and its effect on AD pathology.Method5x FAD x hACE2 mice were inoculated intranasally with a sublethal dose of SARS‐CoV‐2. The mice were maintained for 2 weeks. Mouse brains and meninges were harvested. The tissue was stained and immunofluorescence imaging was conducted to study viral proliferation and immune responses. Histo‐cytometry was conducted for quantitative imaging analysis. Gene expression studies were done using Nanostring assays. All experiments involving the SARS‐Cov‐2 virus were carried out in a BSL3 facility.ResultThis ongoing study demonstrates the proliferation of the SARS‐CoV‐2 virus in the brain via meningeal lymphatics. SARS‐CoV‐2 infection resulted in increased neuroinflammation. Additionally, inflammatory responses induced meningeal dysfunction resulting in increased amyloid‐beta pathology and cerebrospinal fluid drainage.ConclusionGiven the increasing evidence for a viral hypothesis of Alzheimer’s Disease it is extremely important to study the neurodegenerative effects of COVID‐19 which has affected millions worldwide. We demonstrate that SARS‐CoV‐2 infiltrates the brain via the meninges promoting neuroinflammation. Furthermore, amyloid‐beta pathologies are exacerbated by COVID‐19 in animal models providing preclinical evidence of the long‐term neurodegenerative effects of COVID‐19.

Abstract 1122‐000183: Assessing Meningeal Lymphatic MRI Biomarkers in Acute Large Vessel Occlusion Stroke Treated with Thrombectomy

Introduction : Until recently, there was very little known about the brain’s waste removal system. Unlike the peripheral organ systems, the CNS lacked a clearly defined lymphatic vasculature and waste clearance mechanism. However, in 2015, the meningeal lymphatic system (MLS) was first visualized in the dura of humans (1). Since then, a number of studies have investigated the MLS’s potential impact on various neurological diseases, including Alzheimer’s, Parkinson’s, Multiple Sclerosis, hydrocephalus, and epilepsy (2‐6). Other studies have also demonstrated the MLS’s profound influence on stroke (7‐10). However, there is little research that characterizes MLS imaging biomarkers in relation to stroke. Our study evaluates currently proposed MLS MRI biomarkers in large vessel occlusion stroke treated with thrombectomy. Methods : Participants: We recruited twenty patients who had an acute ischemic stroke treated with mechanical thrombectomy (MT) in a large intracranial artery (middle cerebral artery, internal carotid artery, basilar and/or vertebral). The mean age of our population was 69.9 ± 15.5 years. 12 (60%) of our patients had been treated with a combined approach stentriever + aspiration catheter, 4 (20%) with stentriever only, and 4 (20%) with contact aspiration only. Study Design: Basic demographic data, clinical comorbidities, information related to the stroke, technical details about the MT and outcome were collected from electronic medical records. We implemented High Resolution MR Vessel Wall Imaging (HR‐VWI) with a black blood T1‐weighted sequence to characterize changes in the enhancing vessels around the superior sagittal sinus that have been described as potential markers for meningeal lymphatic vessels (1). Within 24 hours after the MT procedure, patients underwent a 45‐minute 3T‐ brain MRI with contrast using a 20‐channel head coil (MAGNETOM Skyra, Siemens) with a sagittal isotropic black blood sequence included with the standard protocol. Our field of view included the superior sagittal sinus. Coronal reconstructed pre‐and post‐contrast back blood T1 sequences were qualitatively evaluated by a board certified Neuroradiologist with 8 years of academic experience evaluating the cerebrovasculature for the presence of purported meningeal lymphatic prominence or asymmetry. Results : There was no significant prominence or asymmetric difference seen in the purported meningeal lymphatics based on 3D high‐resolution T1 black blood MRI sequences in acute stroke large vessel occlusion post‐MT subjects. Conclusions : While in our study no observable differences were seen in the purported meningeal lymphatics based on current MR techniques, it marks an important initial step in development of a useful biomarker for the CNS lymphatic system. Future studies are needed to evaluate alternative meningeal lymphatic quantitative biomarkers. Our study also highlights the need for improved standardization of biomarkers for the meningeal lymphatics. By continuing to evaluate imaging biomarkers, we can further understand the MLS’s potentially profound role in complex neurological diseases.

The meningeal lymphatics: regulators of Aβ immunotherapy?

A new study by Da Mesquita et al. reports on how meningeal lymphatic modulation may influence amyloid-beta immunotherapy and microglial function in mouse models of Alzheimer's disease (AD). This research has broad implications for unraveling the role meningeal lymphatics may play in regulating immunity in the brain during AD pathology and treatment.