tacrolimus and Parkinson-Disease

GPI-1485 is a neuroimmunophilin ligand that binds to FK-506-binding proteins and is under development by Guilford for the potential treatment of erectile dysfunction following nerve injury during prostate resection and Parkinson's disease. In August 2002, phase II clinical trials investigating GPI-1485 for Parkinson's disease commenced, and in November 2003, further phase II trials commenced for post-prostatectomy erectile dysfunction.

Topics: Administration, Oral; Animals; Antiparkinson Agents; Clinical Trials, Phase II as Topic; Erectile Dysfunction; Humans; Male; Parkinson Disease; Patents as Topic; Prostatectomy; Structure-Activity Relationship; Tacrolimus

Optimizing assessments of rate of progression in Parkinson disease (PD) is important in designing clinical trials, especially of potential disease-modifying agents.. To examine the value of measures of impairment, disability, and quality of life in assessing progression in early PD.. Inception cohort analysis of data from 413 patients with early, untreated PD who were enrolled in 2 multicenter, randomized, double-blind clinical trials.. Participants were randomly assigned to 1 of 5 treatments (67 received creatine, 66 received minocycline, 71 received coenzyme Q10, 71 received GPI-1485, and 138 received placebo). We assessed the association between the rates of change in measures of impairment, disability, and quality of life and time to initiation of symptomatic treatment.. Time between baseline assessment and need for the initiation of symptomatic pharmaceutical treatment for PD was the primary indicator of disease progression.. After adjusting for baseline confounding variables with regard to the Unified Parkinson's Disease Rating Scale (UPDRS) Part II score, the UPDRS Part III score, the modified Rankin Scale score, level of education, and treatment group, we assessed the rate of change for the following measurements: the UPDRS Part II score; the UPDRS Part III score; the Schwab and England Independence Scale score (which measures activities of daily living); the Total Functional Capacity scale; the 39-item Parkinson's Disease Questionnaire, summary index, and activities of daily living subscale; and version 2 of the 12-item Short Form Health Survey Physical Summary and Mental Summary. Variables reaching the statistical threshold in univariate analysis were entered into a multivariable Cox proportional hazards model using time to symptomatic treatment as the dependent variable. More rapid change (ie, worsening) in the UPDRS Part II score (hazard ratio, 1.15 [95% CI, 1.08-1.22] for 1 scale unit change per 6 months), the UPDRS Part III score (hazard ratio, 1.09 [95% CI, 1.06-1.13] for 1 scale unit change per 6 months), and the Schwab and England Independence Scale score (hazard ratio, 1.29 [95% CI, 1.12-1.48] for 5 percentage point change per 6 months) was associated with earlier need for symptomatic therapy.. AND RELEVANCE In early PD, the UPDRS Part II score and Part III score and the Schwab and England Independence Scale score can be used to measure disease progression, whereas the 39-item Parkinson's Disease Questionnaire and summary index, Total Functional Capacity scale, and the 12-item Short Form Health Survey Physical Summary and Mental Summary are not sensitive to change.. clinicaltrials.gov Identifiers: NCT00063193 and NCT00076492.

Topics: Adult; Aged; Aged, 80 and over; Disease Progression; Double-Blind Method; Female; Humans; Male; Middle Aged; National Institutes of Health (U.S.); Parkinson Disease; Quality of Life; Severity of Illness Index; Surveys and Questionnaires; Tacrolimus; Treatment Outcome; United States

To assess the predictive value of baseline measures of impairment, disability, and quality of life for the timing of initiation of symptomatic treatment in early Parkinson disease (PD).. Inception cohort analysis.. Ambulatory population from multiple sites in the United States and Canada.. Four hundred thirteen patients with early, untreated PD who participated in 2 double-blind trials that assessed the potential of experimental drugs to serve as disease-modifying agents in PD. Intervention Participants were randomized into treatment groups: creatine (n = 67), minocycline (n = 66), coenzyme Q10 (n = 71), GPI-1485 (n = 71), and placebo (n = 138). Main Outcome Measure Time between baseline assessment and need for the initiation of symptomatic treatment for PD. The following baseline variables were assessed for their relation to the main outcome measure, while adjusting for possible treatment effect: sex; age; level of education; race/ethnicity; disease duration; occupational status; and Unified Parkinson Disease Rating Scale (UPDRS), Medical Outcomes Study Short Form Survey, Modified Rankin Scale, Schwab and England Activities of Daily Living Scale, Total Functional Capacity Scale, 39-item Parkinson Disease Questionnaire, and Geriatric Depression Scale scores. Variables reaching statistical threshold in univariate analyses (alpha = .15) were entered into a multivariable Cox proportional hazards regression model using time to symptomatic treatment as the dependent variable.. Approximately half (48.5%) of the participants reached end point within 12 months. Higher baseline impairment and disability, as determined by UPDRS III (motor section), UPDRS II (activities of daily living section, participant rating), and Modified Rankin Scale scores and level of education were independently associated with an earlier need for symptomatic treatment.. In early PD, greater impairment and disability and higher level of education are independently associated with an earlier need for symptomatic treatment.

Topics: Activities of Daily Living; Adult; Aged; Antiparkinson Agents; Creatine; Disability Evaluation; Disease Management; Drug Administration Schedule; Endpoint Determination; Female; Humans; Male; Middle Aged; Minocycline; Mobility Limitation; Neuropsychological Tests; Parkinson Disease; Predictive Value of Tests; Severity of Illness Index; Surveys and Questionnaires; Tacrolimus; Time Factors; Ubiquinone

To determine if future studies of coenzyme Q(10) and GPI-1485 in Parkinson disease (PD) may be warranted.. We conducted a randomized, double-blind, calibrated futility clinical trial of coenzyme Q10 and GPI-1485 in early untreated PD using placebo data from the DATATOP study to establish the futility threshold.. The primary outcome measure (change in total Unified Parkinson's Disease Rating Scale scores over 1 year) did not meet the prespecified criteria for futility for either agent. Secondary analyses using calibration controls and other more recent placebo data question the appropriateness of the predetermined definition of futility, and suggest that a more restrictive threshold may be needed.. Coenzyme Q(10) and GPI-1485 may warrant further study in Parkinson disease, although the data are inconsistent. Additional factors (cost, availability of other agents, more recent data on placebo outcomes, other ongoing trials) should also be considered in the selection of agents for Phase III studies.

Topics: Aged; Coenzymes; Double-Blind Method; Female; Headache; Humans; Male; Middle Aged; Nausea; Parkinson Disease; Tacrolimus; Ubiquinone

The relevance of the immunosuppressive drug tacrolimus in the prevention of rejection and graft-versus-host disease in transplanted patients is beyond all doubt. However, tacrolimus often has neurotoxic effects, including severe conditions such as posterior reversible leukoencephalopathy syndrome.. A 75-year-old male who had undergone a kidney transplantation five years earlier, for which he was receiving treatment with tacrolimus and mycophenolate. He also had advanced Parkinson's disease, treated with several dopamine agonists. The patient visited the emergency department after a week-long history of visual hallucinations, delirium, expansive mood, confusion and headache. The focal psychogeriatric examination revealed psychosis secondary to dopaminergic agonists as the first diagnostic option, without excluding other possible iatrogenic causes despite the tacrolimus being within the therapeutic range (8.3 ng/mL). Both cranial computed tomography, which did not show any significant findings, and a magnetic resonance scan, in which a bilateral parietooccipital oedema was observed, were performed, this latter finding being compatible with posterior reversible leukoencephalopathy syndrome. While the patient was in hospital, tacrolimus was replaced by everolimus, and the dopaminergic medication was adjusted, resulting in a swift and full remission of the clinical signs and symptoms.. The diagnosis of posterior reversible leukoencephalopathy syndrome should be considered in patients with a history of organ transplantation treated with immunosuppressive drugs who have an acute onset condition with neurological or psychiatric symptoms.. Sindrome de leucoencefalopatia posterior reversible en un paciente con enfermedad de Parkinson y sintomatologia inicial psiquiatrica: una presentacion clinica compleja.. Introduccion. La relevancia del farmaco inmunosupresor tacrolimus en la prevencion del rechazo y la enfermedad de injerto contra huesped en pacientes trasplantados es indiscutible. Sin embargo, con frecuencia, el tacrolimus presenta efectos neurotoxicos, incluyendo cuadros graves, como el sindrome de leucoencefalopatia posterior reversible. Caso clinico. Varon de 75 años, con antecedentes de trasplante renal hace cinco años, en tratamiento con tacrolimus y micofenolato, y de enfermedad de Parkinson avanzada, en tratamiento con varios agonistas dopaminergicos. Acudio a urgencias por un cuadro de una semana de evolucion consistente en alucinaciones visuales, delirios, animo expansivo, confusion y cefalea. La exploracion psicogeriatrica por focos mostro como primera opcion diagnostica una psicosis secundaria a agonistas dopaminergicos, sin excluir otras causas yatrogenas a pesar de encontrarse el tacrolimus en el rango terapeutico (8,3 ng/mL). Se realizaron una tomografia computarizada craneal, que no mostro hallazgos significativos, y una resonancia magnetica, en la que se visualizo un edema parietooccipital bilateral, hallazgo compatible con un sindrome de leucoencefalopatia posterior reversible. Durante el ingreso se sustituyo el tacrolimus por everolimus y se ajusto la medicacion dopaminergica, con lo que se produjo de forma rapida una remision completa del cuadro. Conclusiones. El diagnostico de sindrome de leucoencefalopatia posterior reversible debe considerarse en los pacientes con antecedentes de trasplante de organo en tratamiento con farmacos inmunosupresores que presentan un cuadro de instauracion aguda con sintomas neurologicos o psiquiatricos.

Topics: Aged; Humans; Immunosuppressive Agents; Male; Mental Disorders; Parkinson Disease; Posterior Leukoencephalopathy Syndrome; Tacrolimus

Mitochondrial and lysosomal dysfunction have been implicated in substantia nigra dopaminergic neurodegeneration in Parkinson's disease (PD), but how these pathways are linked in human neurons remains unclear. Here we studied dopaminergic neurons derived from patients with idiopathic and familial PD. We identified a time-dependent pathological cascade beginning with mitochondrial oxidant stress leading to oxidized dopamine accumulation and ultimately resulting in reduced glucocerebrosidase enzymatic activity, lysosomal dysfunction, and α-synuclein accumulation. This toxic cascade was observed in human, but not in mouse, PD neurons at least in part because of species-specific differences in dopamine metabolism. Increasing dopamine synthesis or α-synuclein amounts in mouse midbrain neurons recapitulated pathological phenotypes observed in human neurons. Thus, dopamine oxidation represents an important link between mitochondrial and lysosomal dysfunction in PD pathogenesis.

Topics: alpha-Synuclein; Animals; Antioxidants; Calcineurin Inhibitors; Cell Line; Disease Models, Animal; Dopamine; Dopaminergic Neurons; Glucosylceramidase; Humans; Lysosomes; Melanins; Mesencephalon; Mice; Mice, Knockout; Mitochondria; Oxidation-Reduction; Oxidative Stress; Parkinson Disease; Protein Deglycase DJ-1; Substantia Nigra; Tacrolimus

Topics: alpha-Synuclein; Animals; Calcineurin; Disease Models, Animal; Parkinson Disease; Phosphoproteins; Proteome; Rats; Rats, Sprague-Dawley; Tacrolimus; Tacrolimus Binding Protein 1A

Reactive microglia have been associated with the histological changes that occur in Parkinson's disease brains and mouse models of the disease. Multiple studies from autopsy brains have verified the presence of microgliosis in several brain regions including substantia nigra, striatum, hippocampus and various cortical areas. MPTP injections in rodents have also shown striato-nigral microgliosis correlating with the loss of dopaminergic neurons. However, consistent data with respect to cytokine and immune cell changes during Parkinson's disease have not been fully defined.. In order to improve understanding of the role of neuroinflammation in Parkinson's disease, we employed the MPTP injection model using humanized CD34+ mice along with age-matched C57BL/6 mice. NSG mice engrafted with hu-CD34+ hematopoietic stem cells were injected with MPTP to quantify cytokine changes, neuron loss, gliosis, and behavioral dysfunction. The mice were also treated with or without the calcineurin/NFAT inhibitor, FK506, to determine whether modulating the immune response could attenuate disease. MPTP injections produced impairment of motor performance, increased microgliosis, elevated brain cytokine levels, and reduced tyrosine hydroxylase immunoreactivity in the substantia nigra and striatum of both humanized CD34+ mice and C57BL/6 mice with a strikingly different profile of human versus mouse cytokine elevations observed in each. Interestingly, FK506 injections significantly attenuated the MPTP-induced effects in the humanized CD34+ mice compared the C57BL/6 mice. In addition, analyses of human plasma from Parkinson's disease donors compared to age-matched, healthy controls demonstrated an increase in a number of pro-inflammatory cytokines in female patients similar to that observed in MPTP-injected female CD34+ mice.. This study demonstrates for the first time, induction of Parkinson's disease-like symptoms in female humanized CD34+ mice using MPTP. The profile of cytokine changes in the serum and brains of the humanized CD34+ mice following MPTP injection differed significantly from that occurring in the more commonly used C57BL/6 strain of mice. Moreover, several cytokine elevations observed in the MPTP injected humanized CD34+ mice were similarly increased in plasma of PD patients suggesting that these mice offer the more relevant model for the inflammatory aspects of human disease. Consistent with this, the effects of MPTP on loss of tyrosine hydroxylase immunoreactivity, loss of motor strength, and increase in proinflammatory cytokines were attenuated using an immunosuppressant drug, FK506, in the humanized CD34+ but not the C57BL/6 mice. Collectively, these findings suggest that MPTP injected, humanized CD34+ mice represent a more accurate model for assessing inflammatory changes in PD.

Topics: Animals; Antigens, CD34; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunohistochemistry; Immunosuppressive Agents; Inflammation; Mice; Mice, Inbred C57BL; Mice, Inbred NOD; Mice, SCID; Parkinson Disease; Parkinsonian Disorders; Tacrolimus

Alpha-synuclein (α-synuclein) is considered a key player in Parkinson's disease (PD), but the exact relationship between α-synuclein aggregation and dopaminergic neurodegeneration remains unresolved. There is increasing evidence that neuroinflammatory processes are closely linked to dopaminergic cell death, but whether the inflammatory process is causally involved in PD or rather reflects secondary consequences of nigrostriatal pathway injury is still under debate. We evaluated the therapeutic effect of the immunophilin ligand FK506 in a rAAV2/7 α-synuclein overexpression rat model. Treatment with FK506 significantly increased the survival of dopaminergic neurons in a dose-dependent manner. No reduction in α-synuclein aggregation was apparent in this time window, but FK506 significantly lowered the infiltration of both T helper and cytotoxic T cells and the number and subtype of microglia and macrophages. These data suggest that the anti-inflammatory properties of FK506 decrease neurodegeneration in this α-synuclein-based PD model, pointing to a causal role of neuroinflammation in the pathogenesis of PD.

Topics: alpha-Synuclein; Animals; Anti-Inflammatory Agents; Cell Death; Disease Models, Animal; Dopamine; Dopaminergic Neurons; Dose-Response Relationship, Drug; Female; Gene Expression; Immunosuppressive Agents; Inflammation; Male; Microglia; Parkinson Disease; Protein Aggregates; Rats, Wistar; Tacrolimus

Calcineurin (CN) is a highly conserved Ca(2+)-calmodulin (CaM)-dependent phosphatase that senses Ca(2+) concentrations and transduces that information into cellular responses. Ca(2+) homeostasis is disrupted by α-synuclein (α-syn), a small lipid binding protein whose misfolding and accumulation is a pathological hallmark of several neurodegenerative diseases. We report that α-syn, from yeast to neurons, leads to sustained highly elevated levels of cytoplasmic Ca(2+), thereby activating a CaM-CN cascade that engages substrates that result in toxicity. Surprisingly, complete inhibition of CN also results in toxicity. Limiting the availability of CaM shifts CN's spectrum of substrates toward protective pathways. Modulating CN or CN's substrates with highly selective genetic and pharmacological tools (FK506) does the same. FK506 crosses the blood brain barrier, is well tolerated in humans, and is active in neurons and glia. Thus, a tunable response to CN, which has been conserved for a billion years, can be targeted to rebalance the phosphatase's activities from toxic toward beneficial substrates. These findings have immediate therapeutic implications for synucleinopathies.

Topics: alpha-Synuclein; Animals; Calcineurin; Calcineurin Inhibitors; Calcium Signaling; Calmodulin; Cells, Cultured; Gene Knockdown Techniques; Humans; Lewy Body Disease; Mice; Mice, Transgenic; Models, Neurological; Neurons; NFATC Transcription Factors; Parkinson Disease; Phosphoric Monoester Hydrolases; Rats; Recombinant Proteins; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Tacrolimus

Aggregation of alpha-synuclein (alpha-SYN) plays a key role in Parkinson's disease. We have previously shown that aggregation of alpha-SYN in vitro is accelerated by addition of FK506 binding proteins (FKBP) and that this effect can be counteracted by FK506, a specific inhibitor of these enzymes. In this paper, we investigated in detail the effect of FKBP12 on early aggregation and on fibril formation of wild-type, A53T and A30P alpha-SYN. FKBP12 has a much smaller effect on the fibril formation of these two clinical mutants alpha-SYN. Using an inactive enzyme, we were able to discriminate between catalytic and non-catalytic effects that differentially influence the two processes. A model explaining non-linear concentration dependencies is proposed.

Topics: alpha-Synuclein; Amino Acid Substitution; Brain; Catalytic Domain; Cell Line, Tumor; Humans; Immunosuppressive Agents; Mutation; Nerve Degeneration; Neurofibrillary Tangles; Neurons; Nonlinear Dynamics; Parkinson Disease; Tacrolimus; Tacrolimus Binding Protein 1A; Time Factors

A progressive model of Parkinson's disease has been recently developed in the rat where a unilateral excitotoxic injection into the globus pallidus leads to a gradual loss of dopaminergic neurons in the ipsilateral substantia nigra over a period of at least 6 weeks. In this model microglial activation is observed in the ipsilateral substantia nigra 3 weeks after the lesion and could contribute to neuronal death at this time. The immunosuppressant drug tacrolimus (FK506) reduces dopamine cell death at 3 weeks following a globus pallidus lesion, but not thereafter. Tacrolimus-mediated neuroprotection could result from suppression of microglial activation but the microglial activation at three weeks post-lesion was not much reduced. Microglial activation was observed even in the apparent absence of neuronal death, prompting the suggestion that tacrolimus may prevent, or at least delay, the release of toxic cytokines from activated microglia. By 6 weeks after the GP lesion, even this mechanism fails to protect the dopamine cells from damage.

Topics: Animals; Cell Death; Disease Models, Animal; Disease Progression; Dopamine; Gliosis; Globus Pallidus; Microglia; Neurons; Neuroprotective Agents; Parkinson Disease; Rats; Substantia Nigra; Tacrolimus; Time Factors

The mechanism of action of the neurotoxin 6-hydroxydopamine (6-OHDA) is thought to involve the generation of free radicals and subsequent apoptotic processes. We have demonstrated in vitro that the neuroimmunophilin, FK506 (10-100 nM), dose dependently and significantly restored the ROS production to the control level, increased the Bcl-2 protein level, partly inhibited the cytochrome C release from mitochondria and reduced the caspase-3 activation in SH-SY5Y cells. On the other hand, there was no significant restoration of the ATP level by FK506 and the toxin activated proteins, p53 and Bax, were not normalized by FK506. In support of these latter results, daily administration of FK506 for 7 days to rats (0.5, 1 and 3 mg/kg i.p.) did not significantly prevent the apomorphine-induced contralateral circling, measured 2 weeks after unilateral nigral lesioning. Moreover, FK506 pretreatment did not significantly lower the toxin elevated lipid peroxidation levels, indicating that oxidative stress was present even after the FK506 treatment in the lesioned striatum. Taken together, our results with FK506 are inconsistent. We confirm the antioxidant nature of FK506, that is, it blocks ROS production in SH-SY5Y cells. However, there were no significant protective effects in any apoptotic analyses in SH-SY5Y cells and in animal studies, a 7-day FK506 pre-treatment was not able to reverse the toxic effect of 6-OHDA in a rat model of Parkinson's disease.

Topics: Animals; Apomorphine; Behavior, Animal; Caspase 3; Caspases; Cell Death; Cell Line, Tumor; Disease Models, Animal; Dopamine; Dopamine Agonists; Dose-Response Relationship, Drug; Immunophilins; Male; Motor Activity; Nerve Degeneration; Neuroblastoma; Neurons; Neuroprotective Agents; Oxidopamine; Parkinson Disease; Random Allocation; Rats; Rats, Wistar; Tacrolimus

The immunosuppressive drugs tacrolimus (TAC) and rapamycin (RAPA) have both been found to have neuroprotective effects on dopaminergic neurons. The purpose of the present study was to investigate whether liposomal formulations of these drugs administered directly into the brain improve cell survival and fiber outgrowth. Rats with unilateral 6-hydroxydopamine lesions were transplanted with 800,000 fetal rat ventral mesencephalic cells and randomly divided to one of four groups. Group 1 received a transplant containing cells only; group 2 received a cell suspension containing 0.68 microM liposomal RAPA (LRAPA); group 3 received a cell suspension containing 2.0 microM liposomal TAC (LTAC); and group 4 received a cell suspension containing a liposomal formulation of both 0.68 microM RAPA and 2.0 microM TAC (LRAPATAC). Rats were sacrificed after 6 weeks, and cell survival and fiber outgrowth were assessed using tyrosine hydroxylase (TH) immunohistochemistry. The animals receiving a cell suspension containing either LTAC or LRAPATAC were found to have significantly more surviving TH-immunoreactive (TH-ir) cells than the control group receiving cells only. The group receiving LTAC had significantly longer fibers, the group receiving LRAPA had significantly more fibers close to the graft, and the group receiving LRAPATAC had significantly more fibers at all distances. This study shows the feasibility of using liposomal formulations of neuroimmunophilins directly in the brain at the time of implantation to improve graft survival and fiber outgrowth. Furthermore, we have shown that the combination of LTAC and LRAPA has a synergistic effect. These compounds may play an important role in optimizing graft survival and host reinnervation in cell-mediated brain repair strategies for the treatment of neurological conditions.

Topics: Adrenergic Agents; Animals; Brain; Cell Growth Processes; Cell Survival; Cell Transplantation; Dopamine; Female; Immunohistochemistry; Immunosuppressive Agents; Liposomes; Mesencephalon; Neurons; Neuroprotective Agents; Oxidopamine; Parkinson Disease; Rats; Rats, Wistar; Sirolimus; Tacrolimus; Tyrosine 3-Monooxygenase; Water

We investigated both the antioxidant activities of GPI1046, a non-immunosuppressive derivative of FK506, and the in vivo neuroprotective properties against toxicity of intracerebroventricular 6-hydroxydopamine (6-OHDA) in mice. The 6-OHDA-induced reduction in dopamine and its metabolites in the striatum was significantly normalized by daily administration of GPI1046. Moreover, GPI1046 significantly reduced lipid peroxidation in vivo. Further, GPI1046 significantly increased striatal glutathione (GSH) levels by activating GSH synthesis, although the striatal catalase and superoxide dismutase activities did not change. We conclude that GPI1046 may have neuroprotective effects both in cell cultures and in vivo.

Topics: Animals; Antioxidants; Catalase; Dopamine; Dopamine Agents; Glutamate-Cysteine Ligase; Glutathione; Glutathione Peroxidase; Glutathione Transferase; Immunophilins; Immunosuppressive Agents; Male; Mice; Mice, Inbred ICR; Neostriatum; Neurons; Neuroprotective Agents; Oxidative Stress; Oxidopamine; Parkinson Disease; Pyrrolidines; Superoxide Dismutase; Tacrolimus

alpha-Synuclein is a presynaptic protein of unknown function that has been implicated in the pathogenesis of Parkinson's disease. To gain insight into the function of alpha-synuclein, the present study examined the association between alpha-synuclein and the following Bcl-2 family proteins: Bcl-2; Bcl-XL; Bcl-associated death promoter (BAD); and Bcl-2-associated X-protein. The results of a binding assay using gluthathione S-transferase (GST) fusion alpha-synuclein protein and an immunoprecipitation assay revealed that wild-type or mutant (A30P and A53T) alpha-synuclein (approximately 16 kDa) does not bind to any of these members of the Bcl-2 family. Furthermore, no binding was observed between alpha-synuclein and BAD, regardless of the phosphorylation state of the serine residue in BAD. In contrast, alpha-synuclein was observed to bind to synphilin-1. Although alpha-synuclein has been reported to bind to BAD, modification of alpha-synuclein might be required for such binding to occur.

Topics: alpha-Synuclein; Animals; Apoptosis; bcl-2-Associated X Protein; bcl-Associated Death Protein; bcl-X Protein; Binding Sites; Brain; Calcineurin; Calcineurin Inhibitors; Carrier Proteins; COS Cells; Immunosuppressive Agents; Nerve Tissue Proteins; Neurons; Parkinson Disease; Phosphorylation; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-bcl-2; Synucleins; Tacrolimus

Astrocytes secreting a large amount of 3,4-dihydroxyphenylalanine (dopa) were generated by adenoviral transduction of the human tyrosine hydroxylase (TH) gene. After characterizing in vitro, the effect of transplantation of these astrocytes to the striatum of hemiparkinsonian model rats was investigated. Subconfluent cortical astrocytes were infected by replication-defect adenovirus type 5 carrying the human TH-1 gene or the LacZ reporter gene under the promoter of the glial fibrillary acidic protein (AdexGFAP-HTH-1, AdexGFAP-NL-LacZ). Dopa secretion was not evident at 3 days after the transduction of the HTH-1 gene but it increased from 7 days up to at least 4 months. The secretion was substrate (tyrosine)-dependent, and was enhanced by loading tetrahydrobioputerin (BH4) concentration-dependently. One-third of the hemiparkinsonian model rats, that were transplanted the HTH-1 gene-transduced astrocytes or introduced the direct injection of the viral vector to the striatum, showed a reduction of methamphetamine-induced rotations for at least 6 weeks. Apomorphine-induced rotation was decreased to the 50% level of the control's, but the reduction was obtained equally by the transplantation of HTH-1 gene-transduced or LacZ reporter gene-transduced astrocytes, or by the introduction of HTH-1 or LacZ gene carrying adenovirus. Treatment with FK506 for 3 weeks improved the late-phase apomorphine-induced rotations following the introduction of the HTH-1 gene carrying adenovirus. Histological examination revealed that, in animals that showed a reduction of methamphetamine-rotation, the TH positive astrocytes-like cells were distributed widely in the host striatum for at least 4 weeks. The number of TH positive astrocytes-like cells and their immunoreactivity decreased after 6 weeks when OX-41 positive microglias/macrophages were infiltrated. Data indicate that the adenoviral transduction of the human TH gene to astrocytes and its introduction to the striatum is a promising approach for the treatment of Parkinson's disease. However, the further technical improvements are required to optimize the adenoviral gene delivery, such as the control of viral toxicity and the regulation of the immune response.

Topics: Adenoviridae; Animals; Animals, Newborn; Apomorphine; Astrocytes; beta-Galactosidase; Cells, Cultured; Corpus Striatum; Dihydroxyphenylalanine; Female; Genetic Vectors; Graft Rejection; Humans; Immunosuppressive Agents; Methamphetamine; Mice; Mice, Inbred ICR; Parkinson Disease; Postural Balance; Rats; Rats, Wistar; Sensation Disorders; Tacrolimus; Transfection; Tyrosine 3-Monooxygenase

Although immunosuppressant immunophilin ligands promote neurite outgrowth in vitro, their neurotrophic activities are clearly independent of their immunosuppressive activity. In the present report, a novel nonimmunosuppressive immunophilin ligand, GPI-1046 (3-(3-pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate+ ++) is described. In vitro, GPI-1046 bound to FK506 binding protein-12 and elicited neurite outgrowth from sensory neuronal cultures with picomolar potency with maximal effects comparable to nerve growth factor. In vivo, GPI-1046 stimulated the regeneration of lesioned sciatic nerve axons and myelin levels. In the central nervous system, GPI-1046 promoted protection and/or sprouting of serotonin-containing nerve fibers in somatosensory cortex following parachloroamphetamine treatment. GPI-1046 also induced regenerative sprouting from spared nigrostriatal dopaminergic neurons following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity in mice or 6-hydroxydopamine (6-OHDA) toxicity in rats. The rotational abnormality in 6-OHDA treated rats was alleviated by GPI-1046. These neurotrophic actions in multiple models suggest therapeutic utility for GPI-1046 in neurodegenerative diseases.

Topics: 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; Animals; Carrier Proteins; Cells, Cultured; Chickens; Disease Models, Animal; DNA-Binding Proteins; Dopamine Agents; Heat-Shock Proteins; Male; Mice; Nerve Crush; Nerve Regeneration; Nervous System Diseases; Neurites; Neurons; Oxidopamine; Parkinson Disease; Pyrrolidines; Rats; Rats, Sprague-Dawley; Sciatic Nerve; Serotonin; Tacrolimus; Tacrolimus Binding Proteins; Tyrosine 3-Monooxygenase

Topics: Animals; Carrier Proteins; Disease Models, Animal; DNA-Binding Proteins; Heat-Shock Proteins; Humans; Hydroxydopamines; Ligands; Neurodegenerative Diseases; Parkinson Disease; Peptidylprolyl Isomerase; Tacrolimus; Tacrolimus Binding Proteins