scholarly journals Intrathecal gene therapy rescues a model of demyelinating peripheral neuropathy

2016 ◽  
Vol 113 (17) ◽  
pp. E2421-E2429 ◽  
Author(s):  
Alexia Kagiava ◽  
Irene Sargiannidou ◽  
George Theophilidis ◽  
Christos Karaiskos ◽  
Jan Richter ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Peripheral Neuropathy ◽  
Schwann Cells ◽  
Peripheral Nerves ◽  
Lentiviral Vector ◽  
Intrathecal Injection ◽  
Specific Expression ◽  
Inherited Neuropathy ◽  
Spinal Roots ◽  
Demyelinating Peripheral Neuropathy

Inherited demyelinating peripheral neuropathies are progressive incurable diseases without effective treatment. To develop a gene therapy approach targeting myelinating Schwann cells that can be translatable, we delivered a lentiviral vector using a single lumbar intrathecal injection and a myelin-specific promoter. The human gene of interest, GJB1, which is mutated in X-linked Charcot–Marie–Tooth Disease (CMT1X), was delivered intrathecally into adult Gjb1-null mice, a genetically authentic model of CMT1X that develops a demyelinating peripheral neuropathy. We obtained widespread, stable, and cell-specific expression of connexin32 in up to 50% of Schwann cells in multiple lumbar spinal roots and peripheral nerves. Behavioral and electrophysiological analysis revealed significantly improved motor performance, quadriceps muscle contractility, and sciatic nerve conduction velocities. Furthermore, treated mice exhibited reduced numbers of demyelinated and remyelinated fibers and fewer inflammatory cells in lumbar motor roots, as well as in the femoral motor and sciatic nerves. This study demonstrates that a single intrathecal lentiviral gene delivery can lead to Schwann cell-specific expression in spinal roots extending to multiple peripheral nerves. This clinically relevant approach improves the phenotype of an inherited neuropathy mouse model and provides proof of principle for treating inherited demyelinating neuropathies.

scholarly journals Prohibitin 1 is essential to preserve mitochondria and myelin integrity in Schwann cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Gustavo Della-Flora Nunes ◽  
Emma R. Wilson ◽  
Leandro N. Marziali ◽  
Edward Hurley ◽  
Nicholas Silvestri ◽  
...  
Keyword(s):  
Peripheral Neuropathy ◽  
Schwann Cell ◽  
Schwann Cells ◽  
Peripheral Nerves ◽  
Mitochondrial Protein ◽  
Mitochondrial Damage ◽  
Homologous Protein ◽  
Demyelinating Peripheral Neuropathy ◽  
Prohibitin 1 ◽  
Cell Interface

AbstractIn peripheral nerves, Schwann cells form myelin and provide trophic support to axons. We previously showed that the mitochondrial protein prohibitin 2 can localize to the axon-Schwann-cell interface and is required for developmental myelination. Whether the homologous protein prohibitin 1 has a similar role, and whether prohibitins also play important roles in Schwann cell mitochondria is unknown. Here, we show that deletion of prohibitin 1 in Schwann cells minimally perturbs development, but later triggers a severe demyelinating peripheral neuropathy. Moreover, mitochondria are heavily affected by ablation of prohibitin 1 and demyelination occurs preferentially in cells with apparent mitochondrial loss. Furthermore, in response to mitochondrial damage, Schwann cells trigger the integrated stress response, but, contrary to what was previously suggested, this response is not detrimental in this context. These results identify a role for prohibitin 1 in myelin integrity and advance our understanding about the Schwann cell response to mitochondrial damage.

scholarly journals Blood–Nerve Barrier (BNB) Pathology in Diabetic Peripheral Neuropathy and In Vitro Human BNB Model

2020 ◽  
Vol 22 (1) ◽  
pp. 62
Author(s):  
Yukio Takeshita ◽  
Ryota Sato ◽  
Takashi Kanda
Keyword(s):  
Endothelial Cells ◽  
Peripheral Neuropathy ◽  
Schwann Cells ◽  
Cell Lines ◽  
Diabetic Peripheral Neuropathy ◽  
Peripheral Nerves ◽  
Vascular Endothelial Cells ◽  
Vascular Endothelial ◽  
Immortalized Cell

In diabetic peripheral neuropathy (DPN), metabolic disorder by hyperglycemia progresses in peripheral nerves. In addition to the direct damage to peripheral neural axons, the homeostatic mechanism of peripheral nerves is disrupted by dysfunction of the blood–nerve barrier (BNB) and Schwann cells. The disruption of the BNB, which is a crucial factor in DPN development and exacerbation, causes axonal degeneration via various pathways. Although many reports revealed that hyperglycemia and other important factors, such as dyslipidemia-induced dysfunction of Schwann cells, contributed to DPN, the molecular mechanisms underlying BNB disruption have not been sufficiently elucidated, mainly because of the lack of in vitro studies owing to difficulties in establishing human cell lines from vascular endothelial cells and pericytes that form the BNB. We have developed, for the first time, temperature-sensitive immortalized cell lines of vascular endothelial cells and pericytes originating from the BNB of human sciatic nerves, and we have elucidated the disruption to the BNB mainly in response to advanced glycation end products in DPN. Recently, we succeeded in developing an in vitro BNB model to reflect the anatomical characteristics of the BNB using cell sheet engineering, and we established immortalized cell lines originating from the human BNB. In this article, we review the pathologic evidence of the pathology of DPN in terms of BNB disruption, and we introduce the current in vitro BNB models.

scholarly journals Gene replacement therapy after neuropathy onset provides therapeutic benefit in a model of CMT1X

2019 ◽  
Vol 28 (21) ◽  
pp. 3528-3542 ◽  
Author(s):  
A Kagiava ◽  
J Richter ◽  
C Tryfonos ◽  
C Karaiskos ◽  
A J Heslegrave ◽  
...  
Keyword(s):  
Peripheral Neuropathy ◽  
Replacement Therapy ◽  
Lentiviral Vector ◽  
Intrathecal Injection ◽  
Gene Replacement ◽  
Therapeutic Benefit ◽  
Loss Of Function ◽  
Neurofilament Light ◽  
Junction Protein ◽  
Gene Replacement Therapy

Abstract X-linked Charcot-Marie-Tooth disease (CMT1X), one of the commonest forms of inherited demyelinating neuropathy, results from GJB1 gene mutations causing loss of function of the gap junction protein connexin32 (Cx32). The aim of this study was to examine whether delayed gene replacement therapy after the onset of peripheral neuropathy can provide a therapeutic benefit in the Gjb1-null/Cx32 knockout model of CMT1X. After delivery of the LV-Mpz.GJB1 lentiviral vector by a single lumbar intrathecal injection into 6-month-old Gjb1-null mice, we confirmed expression of Cx32 in lumbar roots and sciatic nerves correctly localized at the paranodal myelin areas. Gjb1-null mice treated with LV-Mpz.GJB1 compared with LV-Mpz.Egfp (mock) vector at the age of 6 months showed improved motor performance at 8 and 10 months. Furthermore, treated mice showed increased sciatic nerve conduction velocities, improvement of myelination and reduced inflammation in lumbar roots and peripheral nerves at 10 months of age, along with enhanced quadriceps muscle innervation. Plasma neurofilament light (NEFL) levels, a clinically relevant biomarker, were also ameliorated in fully treated mice. Intrathecal gene delivery after the onset of peripheral neuropathy offers a significant therapeutic benefit in this disease model, providing a proof of principle for treating patients with CMT1X at different ages.

scholarly journals Prohibitin 1 is essential to preserve mitochondria and myelin integrity in Schwann cells.

2020 ◽  
Author(s):  
Gustavo Della-Flora Nunes ◽  
Emma Wilson ◽  
Leandro Marziali ◽  
Edward Hurley ◽  
Nicholas Silvestri ◽  
...  
Keyword(s):  
Nervous System ◽  
Peripheral Neuropathy ◽  
Schwann Cells ◽  
Mitochondrial Protein ◽  
Mitochondrial Damage ◽  
System Function ◽  
Homologous Protein ◽  
Nervous System Function ◽  
Demyelinating Peripheral Neuropathy ◽  
Prohibitin 1

Abstract Myelin is required for nervous system function. In the peripheral nervous system, Schwann cells (SCs) form myelin and trophically support the axons they ensheath. We previously showed that the mitochondrial protein prohibitin 2 (PHB2) can localize to the axon-SC interface and is required for developmental myelination. Whether the homologous protein PHB1 has a similar role, and whether prohibitins also play important roles in SC mitochondria is unknown. Here we show that deletion of Phb1 in SCs only minimally perturbs development, but later triggers a severe demyelinating peripheral neuropathy. Moreover, mitochondria are heavily affected by ablation of Phb1 and demyelination occurs preferentially in cells with apparent mitochondrial loss. Furthermore, in response to mitochondrial damage, SCs trigger the integrated stress response (ISR), but, contrary to what was previously suggested, the ISR is not detrimental and may be beneficial in this context. These results identify a new role for PHB1 in myelin integrity and advance our understanding of how SCs respond to mitochondrial damage.

scholarly journals AAV9-mediated Schwann cell-targeted gene therapy rescues a model of demyelinating neuropathy

Gene Therapy ◽  
2021 ◽  
Author(s):  
Alexia Kagiava ◽  
Christos Karaiskos ◽  
Jan Richter ◽  
Christina Tryfonos ◽  
Matthew J. Jennings ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Schwann Cells ◽  
Intrathecal Injection ◽  
Demyelinating Neuropathy ◽  
Gene Encoding ◽  
Therapy Approach ◽  
Gene Therapy Approach ◽  
Charcot Marie Tooth Disease ◽  
Targeted Expression ◽  
Protein Zero

AbstractMutations in the GJB1 gene, encoding the gap junction (GJ) protein connexin32 (Cx32), cause X-linked Charcot-Marie-Tooth disease (CMT1X), an inherited demyelinating neuropathy. We developed a gene therapy approach for CMT1X using an AAV9 vector to deliver the GJB1/Cx32 gene under the myelin protein zero (Mpz) promoter for targeted expression in Schwann cells. Lumbar intrathecal injection of the AAV9-Mpz.GJB1 resulted in widespread biodistribution in the peripheral nervous system including lumbar roots, sciatic and femoral nerves, as well as in Cx32 expression in the paranodal non-compact myelin areas of myelinated fibers. A pre-, as well as post-onset treatment trial in Gjb1-null mice, demonstrated improved motor performance and sciatic nerve conduction velocities along with improved myelination and reduced inflammation in peripheral nerve tissues. Blood biomarker levels were also significantly ameliorated in treated mice. This study provides evidence that a clinically translatable AAV9-mediated gene therapy approach targeting Schwann cells could potentially treat CMT1X.

scholarly journals mTORC1 and JUN are activated after deletion of Prohibitin 1 in Schwann cells and may link mitochondrial dysfunction to demyelination

2020 ◽  
Author(s):  
Gustavo Della-Flora Nunes ◽  
Emma R. Wilson ◽  
Edward Hurley ◽  
Bin He ◽  
Bert W. O’Malley ◽  
...  
Keyword(s):  
Nervous System ◽  
Peripheral Neuropathy ◽  
Schwann Cell ◽  
Mitochondrial Dysfunction ◽  
Schwann Cells ◽  
Critical Role ◽  
Mitochondrial Protein ◽  
Important Regulator ◽  
Demyelinating Peripheral Neuropathy ◽  
Prohibitin 1

AbstractSchwann cell (SC) mitochondria are quickly emerging as an important regulator of myelin maintenance in the peripheral nervous system (PNS). However, the mechanisms underlying demyelination in the context of mitochondrial dysfunction in the PNS are incompletely understood. We recently showed that conditional ablation of the mitochondrial protein Prohibitin 1 (Phb1) in SCs causes a severe and fast progressing demyelinating peripheral neuropathy, but the mechanism that causes failure of myelin maintenance remained unknown. Here, we report that mTORC1 and JUN are continuously activated in the absence of Phb1, likely due to mitochondrial damage. Moreover, we demonstrate that these pathways are involved in the demyelination process, and that inhibition of mTORC1 using rapamycin partially rescues the demyelinating pathology. Therefore, we propose that mTORC1 and JUN may play a critical role as executioners of demyelination in the context of perturbations to SC mitochondria.

scholarly journals Activation of mTORC1 and c-Jun by Prohibitin1 loss in Schwann cells may link mitochondrial dysfunction to demyelination

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Gustavo Della Flora Nunes ◽  
Emma R Wilson ◽  
Edward Hurley ◽  
Bin He ◽  
Bert W O'Malley ◽  
...  
Keyword(s):  
Nervous System ◽  
Peripheral Neuropathy ◽  
Schwann Cell ◽  
Mitochondrial Dysfunction ◽  
Schwann Cells ◽  
Critical Role ◽  
Mitochondrial Protein ◽  
Important Regulator ◽  
Demyelinating Peripheral Neuropathy ◽  
Prohibitin 1

Schwann cell (SC) mitochondria are quickly emerging as an important regulator of myelin maintenance in the peripheral nervous system (PNS). However, the mechanisms underlying demyelination in the context of mitochondrial dysfunction in the PNS are incompletely understood. We recently showed that conditional ablation of the mitochondrial protein Prohibitin 1 (PHB1) in SCs causes a severe and fast progressing demyelinating peripheral neuropathy in mice, but the mechanism that causes failure of myelin maintenance remained unknown. Here, we report that mTORC1 and c-Jun are continuously activated in the absence of Phb1, likely as part of the SC response to mitochondrial damage. Moreover, we demonstrate that these pathways are involved in the demyelination process, and that inhibition of mTORC1 using rapamycin partially rescues the demyelinating pathology. Therefore, we propose that mTORC1 and c-Jun may play a critical role as executioners of demyelination in the context of perturbations to SC mitochondria.

Galc ablation in Schwann cells produces a demyelinating peripheral neuropathy characterized by psychosine formation but lacking globoid cells

2018 ◽  
Vol 123 (2) ◽  
pp. S147
Author(s):  
Nadav I. Weinstock ◽  
Daesung Shin ◽  
Ernesto R. Bongarzone ◽  
Duc Nguyen ◽  
Nicholas J. Silvestri ◽  
...  
Keyword(s):  
Peripheral Neuropathy ◽  
Schwann Cells ◽  
Demyelinating Peripheral Neuropathy ◽  
Globoid Cells

scholarly journals Development of a forward-oriented therapeutic lentiviral vector for hemoglobin disorders

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Naoya Uchida ◽  
Matthew M. Hsieh ◽  
Lydia Raines ◽  
Juan J. Haro-Mora ◽  
Selami Demirci ◽  
...  
Keyword(s):  
Gene Therapy ◽  
High Efficiency ◽  
Lentiviral Vector ◽  
Transduction Efficiency ◽  
Specific Expression ◽  
Hematopoietic Stem ◽  
Rev Response Element ◽  
High Level ◽  
Intron 2

Abstract Hematopoietic stem cell (HSC) gene therapy is being evaluated for hemoglobin disorders including sickle cell disease (SCD). Therapeutic globin vectors have demanding requirements including high-efficiency transduction at the HSC level and high-level, erythroid-specific expression with long-term persistence. The requirement of intron 2 for high-level β-globin expression dictates a reverse-oriented globin-expression cassette to prevent its loss from RNA splicing. Current reverse-oriented globin vectors can drive phenotypic correction, but they are limited by low vector titers and low transduction efficiencies. Here we report a clinically relevant forward-oriented β-globin-expressing vector, which has sixfold higher vector titers and four to tenfold higher transduction efficiency for long-term hematopoietic repopulating cells in humanized mice and rhesus macaques. Insertion of Rev response element (RRE) allows intron 2 to be retained, and β-globin production is observed in transplanted macaques and human SCD CD34+ cells. These findings bring us closer to a widely applicable gene therapy for hemoglobin disorders.

Human Neurofibromas in Co-Culture with Mouse Neurons

1982 ◽  
Vol 40 ◽  
pp. 194-195
Author(s):  
R.L. Martuza ◽  
T. Liszczak ◽  
A. Okun ◽  
T-Y Wang
Keyword(s):  
Schwann Cell ◽  
Schwann Cells ◽  
Genetic Disorder ◽  
Cranial Nerves ◽  
Sympathetic Nerves ◽  
Acoustic Neuromas ◽  
Spinal Roots ◽  
Neural Crest Origin ◽  
Multiple Abnormalities ◽  
Other Neoplasms

Neurofibromatosis (NF) is an autosomal dominant genetic disorder with a prevalence of 1/3,000 births. The NF mutation causes multiple abnormalities of various cells of neural crest origin. Schwann cell tumors (neurofibromas, acoustic neuromas) are the most common feature of neurofibromatosis although meningiomas, gliomas, and other neoplasms may be seen. The schwann cell tumors commonly develop from the schwann cells associated with sensory or sympathetic nerves or their ganglia. Schwann cell tumors on ventral spinal roots or motor cranial nerves are much less common. Since the sensory neuron membrane is known to contain a mitogenic factor for schwann cells, we have postulated that neurofibromatosis may be due to an abnormal interaction between the nerve and the schwann cell and that this interaction may be hormonally modulated. To test this possibility a system has been developed in which an enriched schwannoma cell culture can be obtained and co-cultured with pure neurons.

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