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Microglial tunneling nanotubes: an intercellular transfer facilitating mitochondrial dysfunction and
Abstract:
Cerebral malaria (CM), the most severe neurological manifestation of Plasmodium infection, is characterized by microglial activation that plays a pivotal role in initiating pathogenic neuroinflammatory cascades. Tunneling nanotubes (TNTs) are dynamic F-actin-based intercellular connections which transfer mitochondria and pathogenic factors. Although TNTs have been implicated in various neuropathological conditions, their precise involvement in CM pathogenesis, particularly in relation to microglial activation, remains undefined. In this study, single-cell RNA-sequencing (scRNA-seq) revealed significant dysregulation of TNT-associated genes and actin cytoskeleton pathway remodeling in microglia of ECM model. In vitro studies demonstrated that Plasmodium-infected red blood cells (pRBCs)-stimulated primary microglia formed extensive F-actin-rich tunneling nanotubes, which mediated the bidirectional transfer for mitochondria and facilitated intercellular trafficking of lysosomal contents and malarial pigment. These TNT-mediated intercellular communication amplified microglial activation, as evidenced by: (i) lipid peroxidation, (ii) mitochondrial dysfunction, and (iii) autophagosome (LC3+) accumulation. This process further amplifies neuroinflammation through TNFα/IL-6 secretion and expansion of CD45high microglial populations. Pharmacological TNT inhibition restores microglial homeostasis in ECM model. In conclusion, TNTs mediate neuroinflammation in the ECM model by transferring mitochondria and malarial pigment between microglia. Although mitochondrial transfer may transiently support cellular homeostasis, progressive malarial pigment accumulation triggers lipid metabolism dysregulation and amplified neuroinflammation. Inhibiting TNTs formation attenuates microglial hyperactivation, highlighting targeted regulation of TNT-mediated intercellular communication as a potential therapeutic approach for CM-associated neuropathology.
Insights
Tunneling nanotubes (TNTs) facilitate neuroinflammation in cerebral malaria (CM) by transferring mitochondria and malarial pigment between microglia. Inhibiting TNTs restores microglial balance, offering a potential therapeutic target for CM neuropathology.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Cerebral malaria (CM) involves microglial activation, a key driver of neuroinflammation.
- Tunneling nanotubes (TNTs) are actin-based structures involved in intercellular communication and material transfer, implicated in neuropathology.
- The role of TNTs in CM pathogenesis and their connection to microglial activation are not well understood.
Purpose of the Study:
- To investigate the involvement of TNTs in microglial activation and neuroinflammation during cerebral malaria.
- To elucidate the mechanisms of intercellular communication mediated by TNTs in CM.
- To evaluate the therapeutic potential of targeting TNTs in CM.
Main Methods:
- Single-cell RNA-sequencing (scRNA-seq) analysis of microglia in a mouse model of experimental cerebral malaria (ECM).
- In vitro studies using primary microglia stimulated with Plasmodium-infected red blood cells (pRBCs).
- Assessment of mitochondrial and lysosomal content transfer via TNTs.
- Measurement of microglial activation markers, neuroinflammation (TNFα, IL-6), and lipid peroxidation.
- Pharmacological inhibition of TNT formation in the ECM model.
Main Results:
- scRNA-seq revealed dysregulation of TNT-associated genes and actin cytoskeleton pathways in microglia from the ECM model.
- Primary microglia stimulated with pRBCs formed F-actin-rich TNTs, enabling bidirectional transfer of mitochondria and intercellular trafficking of lysosomal contents and malarial pigment.
- TNT-mediated communication amplified microglial activation, leading to lipid peroxidation, mitochondrial dysfunction, autophagosome accumulation, increased TNFα/IL-6 secretion, and expansion of CD45high microglia.
- Pharmacological inhibition of TNTs restored microglial homeostasis in the ECM model.
Conclusions:
- TNTs play a critical role in mediating neuroinflammation in experimental cerebral malaria by facilitating the transfer of mitochondria and malarial pigment between microglia.
- While mitochondrial transfer may offer transient support, progressive malarial pigment accumulation drives lipid metabolism dysregulation and exacerbates neuroinflammation.
- Targeting TNT formation and the associated intercellular communication presents a promising therapeutic strategy for mitigating CM-associated neuropathology.
