PubMed Health⌕ Search

Biomedical subjects

A Blesch

Publications and source records attributed to A Blesch.

At least 19 recordsLinked to original sources
PubMed2005

Nerve replacement strategies for cavernous nerves.

OBJECTIVE: This article reviews novel restorative therapies for cavernous nerves that may be used to replace resected cavernous nerves at the time of pelvic surgery. METHODS: A literature-based presentation (Medline search) on current nerve replacement strategies was conducted with emphasis on neurobiological factors contributing to the restoration of erectile function after cavernous nerve injuries. RESULTS: A promising alternative to autologous nerve grafts for extending the length of successful nerve regeneration are artificial nerve guides. The addition of neurotrophic factors, extracellular matrix components and Schwann cells has been shown to promote cavernous nerve regeneration. Neurotrophic factors can be incorporated in the scaffold or can be supplied by cells seeded into the stroma. The regenerative capacity of these cells can be further enhanced by genetic modification with neurotrophic factor encoding genes. CONCLUSIONS: Artificial nerve guides, especially biodegradable ones containing growth-promoting factors or cells, are a promising option for the repair of cavernous nerve lesions.

Erectile Dysfunction↗
PubMed2004

[Tissue engineering of erectile nerves].

Dissection of the cavernous nerves eliminates spontaneous erections and may lead to irreversible erectile dysfunction due to degeneration of cavernous tissue. Novel procedures to reconstruct penile innervation include cavernous nerve interposition grafting and neurotrophic treatments to revitalize penile neural input, evaluated thus far in various preclinical models of cavernous nerve injury. Schwann cells crucially contribute to successful axonal regeneration by mechanical and paracrine mechanisms in the injured nerve, and Schwann cells seeded into guidance channels have been successfully employed to support regeneration in animal models of cavernous nerve injury. Gene therapy, tissue engineering, and reconstructive techniques have been combined to deliver neurotrophic factors and recover erectile function.

Absorbable Implants↗
PubMed2001

GDNF gene delivery to injured adult CNS motor neurons promotes axonal growth, expression of the trophic neuropeptide CGRP, and cellular protection.

Glial-cell-line--derived neurotrophic factor (GDNF) has been identified as a potent survival and differentiation factor for several neuronal populations in the central nervous system (CNS), but to date, distinct effects of GDNF on motor axon growth and regeneration in the adult have not been demonstrated. In the present study, ex vivo gene delivery was used to directly examine whether GDNF can influence axonal growth, expression of neuronal regeneration-related genes, and sustain the motor neuronal phenotype after adult CNS injury. Adult Fischer 344 rats underwent unilateral transections of the hypoglossal nerve, followed by intramedullary grafts of fibroblasts genetically modified to secrete GDNF. Control animals received lesions and grafts of cells expressing a reporter gene. Two weeks later, GDNF gene delivery (1) robustly promoted the growth of lesioned hypoglossal motor axons, (2) altered the expression and intracellular trafficking of the growth-related protein calcitonin gene-related peptide (CGRP), and (3) significantly sustained the cholinergic phenotype in 84 +/- 6% of hypoglossal neurons compared with 39 +/- 6% in control animals (P < 0.001). This is the first neurotrophic factor identified to increase the in vivo expression of the trophic peptide CGRP and the first report that GDNF promotes motor axonal growth in vivo in the adult CNS. Taken together with previous in vitro studies, these findings serve as the foundation for a model wherein GDNF and CGRP interact in a paracrine manner to regulate neuromuscular development and regeneration.

Animals↗
PubMed2001

Neurotrophism without neurotropism: BDNF promotes survival but not growth of lesioned corticospinal neurons.

Neurotrophic factors exert many effects on the intact and lesioned adult central nervous system (CNS). Among these effects are prevention of neuronal death (neurotrophism) and promotion of axonal growth (neurotropism) after injury. To date, however, it has not been established whether survival and axonal growth functions of neurotrophins can be independently modulated in injured adult neurons in vivo. To address this question, the ability of brain-derived neurotrophic factor (BDNF) to influence corticospinal motor neuronal survival and axonal growth was examined in two injury paradigms. In the first paradigm, a survival assay, adult Fischer 344 rats underwent subcortical lesions followed by grafts to the lesion cavity of syngenic fibroblasts genetically modified to secrete high amounts BDNF or, in control subjects, the reporter gene green fluorescent protein. In control subjects, only 36.2 +/- 7.0% of the retrogradely labeled corticospinal neurons survived the lesion, whereas 89.8 +/- 5.9% (P < 0.001) of the corticospinal neurons survived in animals that received BDNF-secreting grafts. However, in an axonal growth assay, BDNF-secreting cell grafts that were placed into either subcortical lesion sites or sites of thoracic spinal cord injury failed to elicit corticospinal axonal growth. Despite this lack of a neurotropic effect on lesioned corticospinal axons, BDNF-secreting cell grafts placed in the injured spinal cord significantly augmented the growth of other types of axons, including local motor, sensory, and coerulospinal axons. Immunolabeling for tyrosine kinase B (trkB) demonstrated that BDNF receptors were present on corticospinal neuronal somata and apical dendrites but were not detected on their projecting axons. Thus, single classes of neurons in the adult CNS appear to exhibit disparate survival and growth sensitivity to neurotrophic factors, potentially attributable at least in part to differential trafficking of neurotrophin receptors. The possibility of tropic/trophic divergence must be considered when designing strategies to promote CNS recovery from injury.

Animals↗
PubMed2001

Modulation of neuronal survival and axonal growth in vivo by tetracycline-regulated neurotrophin expression.

Vector systems for the regulated and reversible expression of therapeutic genes are likely to improve the safety and efficacy of gene therapy for medical disease. In the present study, we investigated whether the expression of genes transferred into the central nervous system by ex vivo gene therapy can be regulated in vivo leading to controlled neuronal survival and axonal growth. Primary rat fibroblasts were transfected with a retrovirus containing a tetracycline responsive promoter for the expression of the neurotrophin nerve growth factor (NGF) or green fluorescent protein as a control (GFP). After lesions of basal forebrain cholinergic neurons, NGF-mediated neuronal rescue and axonal growth could be completely controlled over a 2-week period by the addition or removal of the tetracycline modulator doxycycline in the animals' drinking water. Further, continued expression of the reporter gene GFP could be reliably and repeatedly turned on and off in the injured CNS for at least 3 months post-grafting, the longest time point investigated. These data constitute the first report of regulated neuronal rescue and axonal growth by controlled neurotrophin gene delivery and long-term, regulated expression using ex vivo CNS gene therapy.

Analysis of Variance↗
PubMed2000

Neurite outgrowth can be modulated in vitro using a tetracycline-repressible gene therapy vector expressing human nerve growth factor.

The delivery of neurotrophic factors to the adult nervous system has potential applications for the treatment of neurodegenerative diseases and trauma. In vivo and ex vivo gene therapy offer a means of delivering growth factors and other therapeutic substances to the central nervous system (CNS) in an intraparenchymal, accurately targeted, and regionally restricted manner. Ideally, gene therapy delivery systems should also be regulatable, allowing exogenous control of amount of gene product delivery. In the present experiment, a tetracycline-regulatable gene expression system was generated to determine whether controllable release of nerve growth factor (NGF) and green fluorescent protein (GFP) from primary rat fibroblasts could modulate biological responses (neurite outgrowth) in vitro. Using a tetracycline-repressible construct, it was found that NGF mRNA, NGF protein, and NGF-induced neurite outgrowth could be tightly regulated within a 24 hour period, and in a dose-dependent fashion, by exposure to the tetracycline analog doxycycline. Similarly, levels of green fluorescence could be regulated in GFP-transfected cells. These findings in a neurobiological system lay the framework for future studies using regulated neurotrophin delivery in in vivo models of neurodegenerative diseases and CNS injury.

Animals↗
PubMed2000

Delivery of neurotrophic factors to neuronal targets: toward gene therapy in the CNS.

Neurotrophic factors are a large class of molecules that can influence neuronal survival, modulate neuronal function and induce axonal growth. They act on specific receptors in specific subsets of neurons and glia. Since their discovery, neurotrophic factors have received considerable interest as possible means for the treatment of neurodegenerative diseases and central nervous system (CNS) trauma. In vitro experiments, as well as studies in animal models of CNS disease, have supported the concept of neuroprotection as a new treatment strategy in degenerative neurologic disease, using neurotrophic factor delivery to CNS targets. Despite the encouraging results from animal studies, progress in adaptation of this treatment strategy to human disease has been slow and sometimes disappointing. One of the major keys for the successful use of neurotrophic factors in treating neurologic disease is the mode of delivery. Neurotrophic factors need to be specifically targeted and regionally restricted to the area of interest to achieve significant results without widespread, unwanted adverse effects. This article summarizes results from recent studies of neurotrophic factors in models of neurodegenerative diseases and CNS trauma and outlines current knowledge and future perspectives of trophic factor gene therapy in the CNS.

Journal Article↗
PubMed1999

Nerve growth factor-hypersecreting Schwann cell grafts augment and guide spinal cord axonal growth and remyelinate central nervous system axons in a phenotypically appropriate manner that correlates with expression of L1.

Schwann cells contribute to efficient axonal regeneration after peripheral nerve injury and, when grafted to the central nervous system (CNS), also support a modest degree of central axonal regeneration. This study examined (1) whether Schwann cells grafted to the CNS exhibit normal patterns of differentiation and association with spinal axons and what signals putatively modulate these interactions, and (2) whether Schwann cells overexpressing neurotrophic factors enhance axonal regeneration. Thus, primary Schwann cells were transduced to hypersecrete human nerve growth factor (NGF) and were grafted to spinal cord injury sites in adult rats. Comparisons were made to nontransfected Schwann cells. From 3 days to 6 months later, grafted Schwann cells exhibited a phenotypic and temporal course of differentiation that matched patterns normally observed after peripheral nerve injury. Schwann cells spontaneously aligned into regular spatial arrays within the cord, appropriately remyelinated coerulospinal axons that regenerated into grafts, and appropriately ensheathed but did not myelinate sensory axons extending into grafts. Coordinate expression of the cell adhesion molecule L1 on Schwann cells and axons correlated with establishment of appropriate patterns of axon-Schwann cell ensheathment. Transduction of Schwann cells to overexpress NGF robustly increased axonal growth but did not otherwise alter the nature of interactions with growing axons. These findings suggest that signals expressed on Schwann cells that modulate peripheral axonal regeneration and myelination are also recognized in the CNS and that the modification of Schwann cells to overexpress growth factors significantly augments their capacity to support extensive axonal growth in models of CNS injury.

Animals↗
PubMed1999

Leukemia inhibitory factor augments neurotrophin expression and corticospinal axon growth after adult CNS injury.

The cytokine leukemia inhibitory factor (LIF) modulates glial and neuronal function in development and after peripheral nerve injury, but little is known regarding its role in the injured adult CNS. To further understand the biological role of LIF and its potential mechanisms of action after CNS injury, effects of cellularly delivered LIF on axonal growth, glial activation, and expression of trophic factors were examined after adult mammalian spinal cord injury. Fibroblasts genetically modified to produce high amounts of LIF were grafted to the injured spinal cords of adult Fischer 344 rats. Two weeks after injury, animals with LIF-secreting cells showed a specific and significant increase in corticospinal axon growth compared with control animals. Furthermore, expression of neurotrophin-3, but not nerve growth factor, brain-derived neurotrophic factor, glia cell line-derived neurotrophic factor, or ciliary neurotrophic factor, was increased at the lesion site in LIF-grafted but not in control subjects. No differences in astroglial and microglial/macrophage activation were observed. Thus, LIF can directly or indirectly modulate molecular and cellular responses of the adult CNS to injury. These findings also demonstrate that neurotrophic molecules can augment expression of other trophic factors in vivo after traumatic injury in the adult CNS.

Animals↗
PubMed1997

Cellular delivery of neurotrophin-3 promotes corticospinal axonal growth and partial functional recovery after spinal cord injury.

The injured adult mammalian spinal cord shows little spontaneous recovery after injury. In the present study, the contribution of projections in the dorsal half of the spinal cord to functional loss after adult spinal cord injury was examined, together with the effects of transgenic cellular delivery of neurotrophin-3 (NT-3) on morphological and functional disturbances. Adult rats underwent bilateral dorsal column spinal cord lesions that remove the dorsal corticospinal projections or underwent more extensive resections of the entire dorsal spinal cord bilaterally that remove corticospinal, rubrospinal, and cerulospinal projections. Long-lasting functional deficits were observed on a motor grid task requiring detailed integration of sensorimotor skills, but only in animals with dorsal hemisection lesions as opposed to dorsal column lesions. Syngenic primary rat fibroblasts genetically modified to produce NT-3 were then grafted to acute spinal cord dorsal hemisection lesion cavities. Up to 3 months later, significant partial functional recovery occurred in NT-3-grafted animals together with a significant increase in corticospinal axon growth at and distal to the injury site. These findings indicate that (1) several spinal pathways contribute to loss of motor function after spinal cord injury, (2) NT-3 is a neurotrophic factor for the injured corticospinal projection, and (3) functional deficits are partially ameliorated by local cellular delivery of NT-3. Lesions of the corticospinal projection may be necessary, but insufficient in isolation, to cause sensorimotor dysfunction after spinal cord injury in the rat.

Animals↗
PubMed1997

Robust growth of chronically injured spinal cord axons induced by grafts of genetically modified NGF-secreting cells.

Little spontaneous regeneration of axons occurs after acute and chronic injury to the CNS. Previously we have shown that the continuous local delivery of neurotrophic factors to the acutely injured spinal cord induces robust growth of spinal and supraspinal axons. In the present study we examined whether chronically injured axons also demonstrate significant neurotrophin responsiveness. Adult rats underwent bilateral dorsal hemisection lesions that axotomize descending supraspinal pathways, including the corticospinal, rubrospinal, and cerulospinal tracts, and ascending dorsal spinal sensory projections. One to three months later, injured rats received grafts of syngenic fibroblasts genetically modified to produce nerve growth factor (NGF). Control subjects received unmodified cell grafts or cells transduced to express the reporter gene beta-galactosidase. Three to five months after grafting, animals that received NGF-secreting grafts showed dense growth of putative cerulospinal axons and primary sensory axons of the dorsolateral fasciculus into the grafted lesion site. Growth from corticospinal, raphaespinal, and local motor axons was not detected. Thus, robust growth of defined populations of supraspinal and spinal axons can be elicited in chronic stages after spinal cord injury by localized, continuous transgenic delivery of neurotrophic factors.

Animals↗
PubMed1996

Transforming growth factor-beta-mediated autocrine growth regulation of gliomas as detected with phosphorothioate antisense oligonucleotides.

Transforming growth factors-beta 1 and -beta 2 (TGF-beta 1 and -beta 2) are important growth-regulatory proteins for astroglial neoplasms. We analyzed their role in tumor-cell proliferation in 12 glioma cell lines, employing phosphorothioate antisense oligodeoxynucleotides (S-ODNs, 14 mer), specifically targeted against the coding sequences of TGF-beta 1-mRNA and TGF-beta 2-mRNA. TGF-beta 1-S-ODNs inhibited cell proliferation in 5 of 12 gliomas, whereas TGF-beta 2-S-ODNs reduced the cell proliferation in all glioma cell lines, compared to nonsense-S-ODN-treated and S-ODN-untreated cells as controls. The efficacy and specificity of antisense effects was validated by Northern-blot analysis and determination of protein concentrations in culture supernatants (ELISA). Exogenous hrTGF-beta 1 either stimulated or inhibited the cell lines, whereas pnTGF-beta 2 stimulated the proliferation of most glioma cells. Blocking the extracellular pathway of TGF-beta by neutralizing antibodies only slightly inhibited those cell lines, which were markedly stimulated by TGF-betas. As the effects of TGF-beta 2-S-ODNs were much stronger than those of TGF-beta neutralizing antibodies, we postulate that the endogenously produced TGF-beta 2 control glioma-cell proliferation, in part by an intracellular loop.

Base Sequence↗
PubMed1995

Transforming growth factor-beta-mediated regulation of human peripheral blood mononuclear cell proliferation as detected with phosphorothioate antisense oligodeoxynucleotides.

Transforming growth factor-beta (TGF-beta) is a potent cytokine that has influence upon immunosuppressive as well as proinflammatory processes. In this context we have investigated the role of endogenous TGF-beta in human peripheral blood mononuclear cells (PBMCs) by TGF-beta 1 phosphorothioate antisense oligodeoxynucleotides (TGF-beta 1-S-ODNs). In short-term cultures (up to 3 days), TGF-beta 1-S-ODNs-treated, interleukin 2-activated PBMCs displayed a growth advantage (up to 148%) compared to nonsense or untreated controls as measured by cell counting and [3H]thymidine incorporation. Long-term antagonization of TGF-beta production (up to 12 days) showed no significant differences between both antisense- and nonsense-treated PBMCs. The efficacy and specificity of TGF-beta 1-S-ODNs effects was validated by gene expression studies (Northern blot and ELISA) and analysis of cellular uptake of BrdU-labeled S-ODN (Immunocytochemistry). TGF-beta 1-S-ODNs at the final concentration of 1 microM reduced TGF-beta 1 protein concentration up to 65% in PBMCs cultures without significant changes in TGF-beta 1 mRNA expression. These experiments demonstrate that TGF-beta 1-S-ODNs specifically antagonize TGF-beta 1-mediated autocrine suppression of IL-2-dependent PBMC activation. TGF-beta 1-specific antisense oligonucleotides may represent a promising immunoresponse modifying agent that could be used for T-cell directed immunostimulation.

Base Sequence↗
PubMed1994

Cloning of a novel malignant melanoma-derived growth-regulatory protein, MIA.

Growth and progression of malignant melanoma cells is influenced by a complex network of growth-stimulating and -inhibiting factors produced by both the tumor cells and the local environment. Here we report the purification and molecular cloning of a novel growth regulating protein, designated melanoma inhibitory activity (MIA) and provide a preliminary functional characterization. MIA is translated as a 131-amino acid precursor and processed into a mature 107-amino acid protein after cleavage of a putative secretion signal. A murine complementary DNA was isolated that encoded a MIA-protein with 88% amino acid identity. MIA is secreted into the culture supernatant by several malignant melanoma cell lines as an M(r) 11,000 autocrine growth factor and acts as a potent tumor cell growth inhibitor for malignant melanoma cells and some other neuroectodermal tumors, including gliomas. MIA has no homology to any other known protein and, therefore, represents a novel type of growth-regulatory factor. Furthermore, we describe a molecular approach to express functionally active MIA in Escherichia coli, which might be attractive as a future antitumor therapeutical substance.

Amino Acid Sequence↗