RESUMEN
[Figure: see text].
Asunto(s)
Aorta/efectos de los fármacos , Enfermedades de la Aorta/prevención & control , Aterosclerosis/prevención & control , Colesterol/metabolismo , Proteínas de Choque Térmico/administración & dosificación , Chaperonas Moleculares/administración & dosificación , Proproteína Convertasa 9/metabolismo , Receptores de LDL/metabolismo , Vacunas Sintéticas/administración & dosificación , Anciano , Anciano de 80 o más Años , Animales , Anticuerpos/sangre , Aorta/enzimología , Aorta/inmunología , Aorta/patología , Enfermedades de la Aorta/enzimología , Enfermedades de la Aorta/inmunología , Enfermedades de la Aorta/patología , Aterosclerosis/enzimología , Aterosclerosis/inmunología , Aterosclerosis/patología , Estudios de Casos y Controles , Modelos Animales de Enfermedad , Femenino , Proteínas de Choque Térmico/inmunología , Proteínas de Choque Térmico/metabolismo , Células Hep G2 , Humanos , Inmunogenicidad Vacunal , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados para ApoE , Persona de Mediana Edad , Chaperonas Moleculares/inmunología , Chaperonas Moleculares/metabolismo , Placa Aterosclerótica , Receptores de LDL/genética , Vacunación , Vacunas Sintéticas/inmunologíaRESUMEN
AIMS: The estrogen-inducible protein Heat Shock Protein 27 (HSP27) as well as anti-HSP27 antibodies are elevated in healthy subjects compared to cardiovascular disease patients. Vaccination of ApoE-/- mice with recombinant HSP25 (rHSP25, the murine ortholog), boosts anti- HSP25 levels and attenuates atherogenesis. As estrogens promote HSP27 synthesis, cellular release and blood levels, we hypothesize that menopause will result in loss of HSP27 atheroprotection. Hence, the rationale for this study is to compare the efficacy of rHSP25 vaccination vs. estradiol (E2) therapy for the prevention of post-menopausal atherogenesis. METHODS AND RESULTS: ApoE-/- mice subjected to ovariectomy (OVX) showed a 65 % increase atherosclerotic burden compared to sham mice after 5 weeks of a high fat diet. Relative to vaccination with rC1, a truncated HSP27 control peptide, atherogenesis was reduced by 5-weekly rHSP25 vaccinations (-43 %), a subcutaneous E2 slow release pellet (-52 %) or a combination thereof (-82 %). Plasma cholesterol levels declined in parallel with the reductions in atherogenesis, but relative to rC1/OVX mice plasma PCSK9 levels were 52 % higher in E2/OVX and 41 % lower in rHSP25/OVX mice (p < 0.0001 for both). Hepatic LDLR mRNA levels did not change with E2 treatment but increased markedly with rHSP25 vaccination. Conversely, hepatic PCSK9 mRNA increased 148 % with E2 treatment vs. rC1/OVX but did not change with rHSP25 vaccination. In human HepG2 hepatocytes E2 increased PCSK9 promoter activity 303 %, while the combination of [rHSP27 + PAb] decreased PCSK9 promoter activity by 64 %. CONCLUSION: The reduction in post-OVX atherogenesis and cholesterol levels with rHSP25 vaccination is associated with increased LDLR but not PCSK9 expression. Surprisingly, E2 therapy attenuates atherogenesis and cholesterol levels post-OVX without altering LDLR but increases PCSK9 expression and promoter activity. This is the first documentation of increased PCSK9 expression with E2 therapy and raises questions about balancing physiological estrogenic / PCSK9 homeostasis and targeting PCSK9 in women - are there effects beyond cholesterol?
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Aterosclerosis/prevención & control , Colesterol/sangre , Estradiol/administración & dosificación , Terapia de Reemplazo de Estrógeno , Proteínas de Choque Térmico HSP27/metabolismo , Proteínas de Choque Térmico/administración & dosificación , Hígado/efectos de los fármacos , Chaperonas Moleculares/administración & dosificación , Proproteína Convertasa 9/metabolismo , Receptores de LDL/metabolismo , Vacunas/administración & dosificación , Animales , Aterosclerosis/sangre , Aterosclerosis/enzimología , Aterosclerosis/inmunología , Biomarcadores/sangre , Modelos Animales de Enfermedad , Regulación hacia Abajo , Implantes de Medicamentos , Femenino , Proteínas de Choque Térmico/inmunología , Células Hep G2 , Humanos , Hígado/enzimología , Menopausia , Ratones Endogámicos C57BL , Ratones Noqueados para ApoE , Chaperonas Moleculares/inmunología , Ovariectomía , Proproteína Convertasa 9/genética , Receptores de LDL/genética , VacunaciónRESUMEN
Fabry disease is characterized by deficient activity of α-galactosidase A, which results in accumulation of glycolipids, such as globotriaosylceremide, in various tissue. Clinical symptoms are varied. In childhood, pain in extremities, hypohidrosis, and angiokeratoma are main symptoms, In adulthood, renal, cardiac and cerebrovascular symptoms are occurred In past, only symptomatic treatments were available. In early 2000th, enzyme replacement therapy was developed after positive results of clinical trials. Ten years after approval, the data of long term safety and efficacy of enzyme replacement.
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1-Desoxinojirimicina/análogos & derivados , Terapia de Reemplazo Enzimático/tendencias , Enfermedad de Fabry/tratamiento farmacológico , Isoenzimas/administración & dosificación , Chaperonas Moleculares/administración & dosificación , Proteínas Recombinantes/administración & dosificación , alfa-Galactosidasa/administración & dosificación , 1-Desoxinojirimicina/administración & dosificación , Animales , Cromosomas Humanos X/enzimología , Aprobación de Drogas , Terapia de Reemplazo Enzimático/métodos , Enfermedad de Fabry/enzimología , Enfermedad de Fabry/genética , Enfermedad de Fabry/metabolismo , Femenino , Globósidos/metabolismo , Humanos , Masculino , Ratones , Mutación , Ensayos Clínicos Controlados Aleatorios como Asunto , Resultado del Tratamiento , alfa-Galactosidasa/genéticaRESUMEN
BACKGROUND: Patients with Fabry disease (FD) and amenable mutations can be treated with the chaperone migalastat to restore endogenous α-galactosidase A (AGAL) activity. However, certain amenable mutations do not respond biochemically in vivo as expected. Here, we aimed to establish a patient-specific and mutation-specific cell model to evaluate the amenability to chaperone therapy in FD. METHODS: Since current tests to determine amenability are limited to heterologous mutation expression in HEK293T cells with endogenous AGAL activity, we generated CRISPR/Cas9-mediated AGAL-deficient HEK293T cells as a basis for mutant overexpression. Furthermore, primary urinary cells from patients were isolated and immortalised as a patient-specific cell model system to evaluate the amenability to chaperone therapy. RESULTS: Under treatment (>13 months), carriers of p.N215S (n=6) showed a significant reduction of plasma lyso-Gb3 (p<0.05). Lyso-Gb3 levels in carriers of p.L294S increased (p<0.05) and two patients developed severe albuminuria. Both missense mutations were amenable in wild-type HEK293T cells (p<0.05), but presented different responses in CRISPR/Cas9-mediated AGAL knockouts and immortalised urinary cells. Chaperone incubation resulted in increased AGAL activity (p<0.0001) and intracellular globotriaosylceramide (Gb3) reduction (p<0.05) in immortalised p.N215S cells but not in p.L294S and IVS2+1 G>A cells. CONCLUSION: We conclude that repeated AGAL activity measurements in patients' white blood cells are mandatory to assess the in vivo amenability to migalastat. Plasma lyso-Gb3 might be an appropriate tool to measure the biochemical response to migalastat. Patients with low AGAL activities and increasing lyso-Gb3 levels despite in vitro amenability might not benefit sufficiently from chaperone treatment.
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Enfermedad de Fabry/genética , alfa-Galactosidasa/genética , 1-Desoxinojirimicina/administración & dosificación , 1-Desoxinojirimicina/análogos & derivados , Tratamiento Basado en Trasplante de Células y Tejidos/métodos , Terapia de Reemplazo Enzimático/métodos , Enfermedad de Fabry/metabolismo , Enfermedad de Fabry/terapia , Edición Génica , Células HEK293 , Humanos , Chaperonas Moleculares/administración & dosificación , Medicina de Precisión/métodos , Trihexosilceramidas/metabolismo , alfa-Galactosidasa/metabolismoRESUMEN
Heat shock proteins (HSPs) participate in the regulation of different cell activities in response to stimuli. By applying different strategies, the modulation of heat shock proteins is at the center of attention. Conventional delivery approaches are not fully encouraged due to cytotoxicity and immunogenicity issues. Exosomes are touted as bio-shuttles for delivery of distinct biomolecules inside the cells. Here, we aimed to HSP27 small interfering RNA (siRNA)-tagged exosomes for the inhibition of Hsp27 in human neuroblastoma cell line SH-SY5Y and explored differentiation into neuron-like cells. Exosomes were isolated, characterized by scanning electron microscope (SEM) and CD63 then enriched with siRNA against Hsp27. Neuroblastoma cells were incubated with exosomes carrying siRNA for 48 hr. Exosome uptake was monitored by immunofluorescence assay. The cell viability and proliferation were analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and bromodeoxyuridine/5-bromo-2'-deoxyuridine incorporation assays. The ability of cells to form colonies was evaluated by clonogenic assay. The cell potential to express NeuN, a mature neuron factor, was studied by flow cytometry analysis. SEM showed the nano-sized particles and a high level of CD63 after enrichment. Immunofluorescence imaging revealed an appropriate transfection rate in cell exposed to Hsp27 siRNA tagged exosomes. The cell viability and proliferation were reduced compared to cells received nude exosomes ( p < 0.05). Clonogenic activity of cells was diminished by the inhibition of Hsp27. Flow cytometry analysis revealed that the inhibition of Hsp27 prohibited NeuN content, showing the maturation of SH-SY5Y cells to mature cells compared to control. These data confirmed that exosomes could be used as appropriate bio-shuttles for the inhibition of Hsp27-aborted cell differentiation toward mature neuron.
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Diferenciación Celular/fisiología , Proteínas de Choque Térmico/antagonistas & inhibidores , Chaperonas Moleculares/antagonistas & inhibidores , Células-Madre Neurales/citología , Neurogénesis/fisiología , Neuronas/citología , Línea Celular Tumoral , Proliferación Celular/fisiología , Supervivencia Celular/fisiología , Exosomas , Vectores Genéticos , Proteínas de Choque Térmico/administración & dosificación , Humanos , Chaperonas Moleculares/administración & dosificación , Neuroblastoma , Neuronas/metabolismo , ARN Interferente Pequeño/administración & dosificación , TransfecciónRESUMEN
Oculocutaneous albinism (OCA) is an autosomal recessive disease characterized by the reduction or complete lack of melanin pigment in the skin, hair, and eyes. No effective treatment for OCA is available at present. OCA type 1 is caused by mutations that disrupt the function of tyrosinase (TYR), the rate-limiting enzyme of melanin synthesis. Recently, it was shown that tyrosinase in some patients with OCA type 1 mutation is retained in the endoplasmic reticulum and that its catalytic activity is lost, a phenomenon known as endoplasmic reticulum retention. However, to our knowledge, the intracellular localization of tyrosinase in Japanese patients with OCA type 1 missense mutations has not been reported. In this study, we first investigated the intracellular localization of Japanese OCA type 1A missense mutant tyrosinases using Western blotting and immunohistochemical staining. R77Q, R239W, D383N, and P431L mutant tyrosinases were retained in the endoplasmic reticulum, and H211Y mutant tyrosinase was partially transported to the Golgi apparatus. Second, we explored the possibility of chemical chaperone therapy for Japanese patients with OCA type 1A missense mutations and found that HeLa cells expressing P431L mutant tyrosinase have restored tyrosinase activity after treatment with a low-dose tyrosinase inhibitor, as a chemical chaperone, in a dose-dependent manner. These results provide the basis for a possible chemical chaperone therapy to recover tyrosinase activities in patients with OCA type 1A patients.
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Albinismo Oculocutáneo/genética , Melaninas/metabolismo , Chaperonas Moleculares/administración & dosificación , Terapia Molecular Dirigida/métodos , Mutación Missense/genética , Albinismo Oculocutáneo/terapia , Células Cultivadas , Predisposición Genética a la Enfermedad , Células HeLa , Humanos , Japón , Monofenol Monooxigenasa/genética , Enfermedades Raras , Resultado del TratamientoRESUMEN
Acute kidney injury (AKI) is a medical condition characterized by kidney damage with a rapid decline of renal function, which is associated with high mortality and morbidity. Recent research has further established an intimate relationship between AKI and chronic kidney disease. Perturbations of kidney cells in AKI result in the accumulation of unfolded and misfolded proteins in the endoplasmic reticulum (ER), leading to unfolded protein response (UPR) or ER stress. In this review, we analyze the role and regulation of ER stress in AKI triggered by renal ischemia-reperfusion and cisplatin nephrotoxicity. The balance between the two major components of UPR, the adaptive pathway and the apoptotic pathway, plays a critical role in determining the cell fate in ER stress. The adaptive pathway is evoked to attenuate translation, induce chaperones, maintain protein homeostasis and promote cell survival. Prolonged ER stress activates the apoptotic pathway, resulting in the elimination of dysfunctional cells. Therefore, regulating ER stress in kidney cells may provide a therapeutic target in AKI. KEY MESSAGES Perturbations of kidney cells in acute kidney injury result in the accumulation of unfolded and misfolded proteins in ER, leading to unfolded protein response (UPR) or ER stress. The balance between the adaptive pathway and the apoptotic pathway of UPR plays a critical role in determining the cell fate in ER stress. Modulation of ER stress in kidney cells may provide a therapeutic strategy for acute kidney injury.
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Lesión Renal Aguda/patología , Estrés del Retículo Endoplásmico/efectos de los fármacos , Riñón/patología , Insuficiencia Renal Crónica/patología , Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/tratamiento farmacológico , Animales , Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Cisplatino/efectos adversos , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/metabolismo , Retículo Endoplásmico/patología , Humanos , Precondicionamiento Isquémico/métodos , Riñón/irrigación sanguínea , Riñón/efectos de los fármacos , Chaperonas Moleculares/administración & dosificación , Pliegue de Proteína/efectos de los fármacos , Insuficiencia Renal Crónica/prevención & control , Daño por Reperfusión/complicaciones , Tapsigargina/administración & dosificación , Resultado del Tratamiento , Tunicamicina/administración & dosificación , Respuesta de Proteína Desplegada/efectos de los fármacosRESUMEN
Type 2 diabetes is a chronic inflammatory disease. Autophagy, the dynamic process of lysosomal degradation of damaged organelles and proteins, may protect ßcells from destruction by inflammation in type 2 diabetes. The present study investigated the role of autophagy, inflammation and endoplasmic reticulum (ER) stress in type 2 diabetes. INS1 cells were incubated with lipopolysaccharide. The chemical chaperone 4phenylbutyric acid was used to inhibit ER stress, and 3methyadenine (3MA) was used to inhibit autophagy. Apoptosis was detected by flow cytometry and cell proliferation using Cell Counting kit8 solution. Light chain3B, interleukin (IL) 1ß, caspase1 and C/EBP homologous protein production were assessed by western blotting, and rat activating transcription factor 4 and rat binding immunoglobulin heavy chain protein gene expression were determined by realtime reverse transcriptionpolymerase chain reaction. The results showed that inhibiting autophagy with 3MA unexpectedly contributed to cell death in ßcells. This response was associated with an increase in inflammatory cytokines, including IL1ß and caspase1. Inhibiting ER stress with 4phenylbutyric acid led to a decrease in cell apoptosis. These results showed that autophagy may have a protective effect by reducing inflammatory cytokines in ßcells. In addition, the inositolrequiring enzyme 1 pathway mediated the ER stress associated with autophagy and inflammatory cytokines (IL1ß and caspase1). Therefore, inflammatory cytokines may be critical signalling nodes, which are associated with ER stressmediated ßcell death.
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Diabetes Mellitus Tipo 2/genética , Estrés del Retículo Endoplásmico/genética , Inflamación/genética , Factor de Transcripción Activador 4/genética , Adenina/administración & dosificación , Adenina/análogos & derivados , Animales , Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Autofagia/genética , Caspasa 1/genética , Recuento de Células , Proliferación Celular/efectos de los fármacos , Diabetes Mellitus Tipo 2/complicaciones , Diabetes Mellitus Tipo 2/patología , Estrés del Retículo Endoplásmico/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Proteínas de Choque Térmico/genética , Humanos , Inflamación/complicaciones , Inflamación/patología , Células Secretoras de Insulina/efectos de los fármacos , Células Secretoras de Insulina/metabolismo , Interleucina-1beta/genética , Lipopolisacáridos/toxicidad , Chaperonas Moleculares/administración & dosificación , Fenilbutiratos/administración & dosificación , Ratas , Factor de Transcripción CHOP/genéticaRESUMEN
Regorafenib (RG) was an oral multi-kinase inhibitor with poor water solubility. In order to enhance the drug's solubility, dissolution and bioavailability, the binary molecular chaperone drug between RG and ß-cyclodetrin (ß-CD) had prepared by co-crystallization. The structure of RG-ß-CD was characterized by thermal analysis, powder X-ray diffraction, infrared spectroscopy and nuclear magnetic resonance. Phase-solubility study revealed the higher solubility and complexing ability of ß-CDwith RG.The solubility and dissolution of RG-ß-CD was significantly enhanced in pH 1.2 medium, pH 6.8 PBS buffer solution and distilled water compared to RG. In vivo distribution and antitumor studies revealed that the bioavailability of RG-ß-CD was increased when ß-CD mated with RG. Therefore, these findings could provide a suitable pharmaceutical dosage to enhanced therapeutic activity.
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Antineoplásicos/farmacología , Chaperonas Moleculares/química , Neoplasias Experimentales/tratamiento farmacológico , Compuestos de Fenilurea/química , Compuestos de Fenilurea/farmacología , Piridinas/química , Piridinas/farmacología , beta-Ciclodextrinas/química , beta-Ciclodextrinas/farmacología , Administración Oral , Animales , Antineoplásicos/administración & dosificación , Antineoplásicos/química , Antineoplásicos/farmacocinética , Disponibilidad Biológica , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Cristalización , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Chaperonas Moleculares/administración & dosificación , Estructura Molecular , Neoplasias Experimentales/patología , Compuestos de Fenilurea/administración & dosificación , Compuestos de Fenilurea/farmacocinética , Piridinas/administración & dosificación , Piridinas/farmacocinética , Solubilidad , Relación Estructura-Actividad , beta-Ciclodextrinas/farmacocinéticaRESUMEN
BACKGROUND: Fabry disease is an X-linked lysosomal storage disorder caused by GLA mutations, resulting in α-galactosidase (α-Gal) deficiency and accumulation of lysosomal substrates. Migalastat, an oral pharmacological chaperone being developed as an alternative to intravenous enzyme replacement therapy (ERT), stabilises specific mutant (amenable) forms of α-Gal to facilitate normal lysosomal trafficking. METHODS: The main objective of the 18-month, randomised, active-controlled ATTRACT study was to assess the effects of migalastat on renal function in patients with Fabry disease previously treated with ERT. Effects on heart, disease substrate, patient-reported outcomes (PROs) and safety were also assessed. RESULTS: Fifty-seven adults (56% female) receiving ERT (88% had multiorgan disease) were randomised (1.5:1), based on a preliminary cell-based assay of responsiveness to migalastat, to receive 18â months open-label migalastat or remain on ERT. Four patients had non-amenable mutant forms of α-Gal based on the validated cell-based assay conducted after treatment initiation and were excluded from primary efficacy analyses only. Migalastat and ERT had similar effects on renal function. Left ventricular mass index decreased significantly with migalastat treatment (-6.6â g/m2 (-11.0 to -2.2)); there was no significant change with ERT. Predefined renal, cardiac or cerebrovascular events occurred in 29% and 44% of patients in the migalastat and ERT groups, respectively. Plasma globotriaosylsphingosine remained low and stable following the switch from ERT to migalastat. PROs were comparable between groups. Migalastat was generally safe and well tolerated. CONCLUSIONS: Migalastat offers promise as a first-in-class oral monotherapy alternative treatment to intravenous ERT for patients with Fabry disease and amenable mutations. TRIAL REGISTRATION NUMBER: NCT00925301; Pre-results.
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1-Desoxinojirimicina/análogos & derivados , Enfermedad de Fabry/tratamiento farmacológico , Chaperonas Moleculares/administración & dosificación , alfa-Galactosidasa/genética , 1-Desoxinojirimicina/administración & dosificación , 1-Desoxinojirimicina/efectos adversos , Administración Oral , Adolescente , Adulto , Anciano , Terapia de Reemplazo Enzimático/efectos adversos , Enfermedad de Fabry/metabolismo , Enfermedad de Fabry/fisiopatología , Femenino , Humanos , Lisosomas/genética , Lisosomas/patología , Masculino , Persona de Mediana Edad , Chaperonas Moleculares/efectos adversos , Resultado del TratamientoRESUMEN
GBA gene mutations are the greatest cause of Parkinson disease (PD). GBA encodes the lysosomal enzyme glucocerebrosidase (GCase) but the mechanisms by which loss of GCase contributes to PD remain unclear. Inhibition of autophagy and the generation of endoplasmic reticulum (ER) stress are both implicated. Mutant GCase can unfold in the ER and be degraded via the unfolded protein response, activating ER stress and reducing lysosomal GCase. Small molecule chaperones that cross the blood brain barrier help mutant GCase refold and traffic correctly to lysosomes are putative treatments for PD. We treated fibroblast cells from PD patients with heterozygous GBA mutations and Drosophila expressing human wild-type, N370S and L444P GBA with the molecular chaperones ambroxol and isofagomine. Both chaperones increased GCase levels and activity, but also GBA mRNA, in control and mutant GBA fibroblasts. Expression of mutated GBA in Drosophila resulted in dopaminergic neuronal loss, a progressive locomotor defect, abnormal aggregates in the ER and increased levels of the ER stress reporter Xbp1-EGFP. Treatment with both chaperones lowered ER stress and prevented the loss of motor function, providing proof of principle that small molecule chaperones can reverse mutant GBA-mediated ER stress in vivo and might prove effective for treating PD.
Asunto(s)
Glucosilceramidasa/genética , Chaperonas Moleculares/administración & dosificación , Mutación , Enfermedad de Parkinson/tratamiento farmacológico , Ambroxol/administración & dosificación , Ambroxol/farmacología , Animales , Modelos Animales de Enfermedad , Neuronas Dopaminérgicas/efectos de los fármacos , Neuronas Dopaminérgicas/patología , Drosophila melanogaster/genética , Retículo Endoplásmico/metabolismo , Estrés del Retículo Endoplásmico/efectos de los fármacos , Humanos , Iminopiranosas/administración & dosificación , Iminopiranosas/farmacología , Chaperonas Moleculares/farmacología , Enfermedad de Parkinson/genética , Enfermedad de Parkinson/patologíaRESUMEN
Salmonella infection is an increasingly important public health problem owing to the emergence of multidrug resistance and the lack of broadly efficient vaccines. Novel strategies of vaccination are required to induce protective immune responses at mucosal surfaces and in the circulation, to limit bacteria entry and dissemination. To this aim, intranasal anti-Salmonella vaccination with an innovative formulation composed of gas-filled microbubbles and the pathogen-derived protective protein serodominant secreted effector protein B (SseB-MB) was evaluated in a mouse infection model. Intranasal application of SseB-MB induced gut and systemic immunoglobulin A, T-helper type 17 cell (Th17), and Th1 responses, all of which are associated with natural immunity against Salmonella In vaccinated mice, a significant reduction in bacterial load was observed in intestinal tissues and the spleen after an otherwise lethal oral infection. Therefore, MB serve as an efficient carrier for nasal delivery of a Salmonella antigen that results in protection upon activation of the common mucosal immune system.
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Antígenos Bacterianos/inmunología , Proteínas Bacterianas/inmunología , Portadores de Fármacos/administración & dosificación , Tracto Gastrointestinal/inmunología , Chaperonas Moleculares/inmunología , Infecciones por Salmonella/prevención & control , Vacunas contra la Salmonella/inmunología , Salmonella/inmunología , Administración Intranasal , Animales , Anticuerpos Antibacterianos/análisis , Anticuerpos Antibacterianos/sangre , Antígenos Bacterianos/administración & dosificación , Carga Bacteriana , Proteínas Bacterianas/administración & dosificación , Modelos Animales de Enfermedad , Femenino , Tracto Gastrointestinal/microbiología , Inmunidad Mucosa , Inmunoglobulina A/análisis , Inmunoglobulina A/sangre , Ratones Endogámicos BALB C , Chaperonas Moleculares/administración & dosificación , Infecciones por Salmonella/inmunología , Vacunas contra la Salmonella/administración & dosificación , Células TH1/inmunología , Células Th17/inmunología , Resultado del TratamientoRESUMEN
PURPOSE: The purpose of this article was to review recent advances in the applications of nanotechnology in cataract treatment and prevention strategies. DESIGN: A literature review on the use of nanotechnology for the prevention and treatment of cataract was done. METHODS: Research articles about nanotechnology-based treatments and prevention technologies for cataract were searched on Web of Science, and the most recent advances were reported. RESULTS: Nonsteroid anti-inflammatory drugs, natural antioxidants, biologic and chemical chaperones, and chaperones such as molecules have found great application in preventing and treating cataracts. Current scientific research on new treatment strategies, which focuses on the biochemical basis of the disease, will likely result in new anticataract agents. However, none of the drug formulations will be approved for use unless efficient delivery is promised. Nanoparticle engineering together with biomimetic strategies enable the development of next-generation, more efficient, less complex, and personalized treatments. CONCLUSIONS: The only currently available treatment for cataracts, surgical replacement of the opacified lens, is not an easily accessible option in developing countries. New treatment strategies based on topical drugs would enable treatment to reach massive populations facing the threat of blindness and more effectively deal with the postsurgical complications. Nanotechnology plays a key role in improving drug delivery systems with enhanced controlled release, targeted delivery, and bioavailability to overcome diffusion limitations in the eye.
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Catarata/tratamiento farmacológico , Nanotecnología/métodos , Antiinflamatorios/administración & dosificación , Antioxidantes/administración & dosificación , Factores Biológicos/administración & dosificación , Catarata/prevención & control , Extracción de Catarata/métodos , Sistemas de Liberación de Medicamentos , Humanos , Chaperonas Moleculares/administración & dosificación , Complicaciones Posoperatorias/prevención & controlRESUMEN
Fabry disease is an X-linked lysosomal storage disorder caused by mutations in the gene that encodes α-galactosidase A and is characterized by pathological accumulation of globotriaosylceramide and globotriaosylsphingosine. Earlier, the authors demonstrated that oral coadministration of the pharmacological chaperone AT1001 (migalastat HCl; 1-deoxygalactonojirimycin HCl) prior to intravenous administration of enzyme replacement therapy improved the pharmacological properties of the enzyme. In this study, the authors investigated the effects of coformulating AT1001 with a proprietary recombinant human α-galactosidase A (ATB100) into a single intravenous formulation. AT1001 increased the physical stability and reduced aggregation of ATB100 at neutral pH in vitro, and increased the potency for ATB100-mediated globotriaosylceramide reduction in cultured Fabry fibroblasts. In Fabry mice, AT1001 coformulation increased the total exposure of active enzyme, and increased ATB100 levels in cardiomyocytes, cardiac vascular endothelial cells, renal distal tubular epithelial cells, and glomerular cells, cell types that do not show substantial uptake with enzyme replacement therapy alone. Notably, AT1001 coformulation also leads to greater tissue globotriaosylceramide reduction when compared with ATB100 alone, which was positively correlated with reductions in plasma globotriaosylsphingosine. Collectively, these data indicate that intravenous administration of ATB100 coformulated with AT1001 may provide an improved therapy for Fabry disease and thus warrants further investigation.
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Enfermedad de Fabry/tratamiento farmacológico , Chaperonas Moleculares/administración & dosificación , Oligopéptidos/administración & dosificación , alfa-Galactosidasa/administración & dosificación , Animales , Modelos Animales de Enfermedad , Combinación de Medicamentos , Terapia de Reemplazo Enzimático , Enfermedad de Fabry/patología , Fibroblastos/efectos de los fármacos , Humanos , Ratones , Mutación , Especificidad por SustratoRESUMEN
Key tissue targets in treating exfoliation syndrome (XFS) and the associated glaucoma include lens, iris, and ciliary body, which produce the exfoliative material, and the trabecular meshwork, which may be impaired by the exfoliative material. In addition to antiglaucoma drug therapy, strategies for treating the disease include approaches for preventing formation of exfoliative material as well as those aimed at digesting exfoliative material. A variety of drug molecules including small molecules, protein drugs, and nucleic acids are potential candidates for treating XFS. Potential drug classes include antioxidants, lysyl oxidase-like 1 enhancers, antifibrotics, anti-inflammatory agents, proteases, and chaperones. However, the delivery of these agents to the target tissues in the anterior segment is hindered by protective static and dynamic barriers of the eye. Thus, unique drug delivery approaches are needed for each drug type (small molecules, proteins, and nucleic acids). In addition, there is a need for sustaining drug therapy for treating XFS, which can potentially be addressed by using nanoparticles, microparticles, implants, and contact lens delivery systems. This article provides an overview of drug delivery challenges and opportunities in treating XFS with the focus being on nanomedicines.
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Antihipertensivos/administración & dosificación , Antioxidantes/administración & dosificación , Sistemas de Liberación de Medicamentos , Síndrome de Exfoliación/tratamiento farmacológico , Glaucoma de Ángulo Abierto/tratamiento farmacológico , Chaperonas Moleculares/administración & dosificación , HumanosAsunto(s)
Galactosa/metabolismo , Glucosa/metabolismo , Enfermedades por Almacenamiento Lisosomal/tratamiento farmacológico , Enfermedades por Almacenamiento Lisosomal/genética , Chaperonas Moleculares/uso terapéutico , Terapia Molecular Dirigida , Administración Oral , Animales , Enfermedad de Fabry , Galactosidasas , Gangliosidosis GM1 , Glucosidasas , Humanos , Leucodistrofia de Células Globoides , Enfermedades por Almacenamiento Lisosomal/diagnóstico , Enfermedades por Almacenamiento Lisosomal/metabolismo , Chaperonas Moleculares/administración & dosificación , Patología Molecular , Deficiencias en la ProteostasisRESUMEN
Gaucher disease (GD) is a rare monogenetic disorder leading to dysfunction of acid ß-glucosidase (ß-glucocerebrosidase; GCase) and accumulation of glucosylceramide in lysosomes, especially in macrophages (Gaucher cells). Many of the mutations at the origin of GD do not impair the catalytic activity of GCase, but cause misfolding and subsequent degradation by the quality control system at the endoplasmic reticulum. Pharmacological chaperones (PCs) capable of restoring the correct folding and trafficking of the endogenous mutant enzyme represent promising alternatives to the currently available enzyme replacement and substrate reduction therapies (ERT and SRT, respectively), but unfavorable biodistribution and potential side-effects remain important issues. We have now designed a strategy to enhance the controlled delivery of PCs to macrophages that exploit the formation of ternary complexes between the PC, a trivalent mannosylated ß-cyclodextrin (ßCD) conjugate and the macrophage mannose receptor (MMR). First, PC candidates with appropriate relative avidities towards the ßCD cavity and the GCase active site were selected to ensure efficient transfer of the PC cargo from the host to the GCase active site. Control experiments confirmed that the ßCD carrier was selectively recognized by mannose-specific lectins and that the corresponding PC:mannosylated ßCD supramolecular complex retained both the chaperoning activity, as confirmed in human GD fibroblasts, and the MMR binding ability. Finally, fluorescence microscopy techniques proved targeting and cellular uptake of the PC-loaded system in macrophages. Altogether, the results support that combined cyclodextrin encapsulation and glycotargeting may improve the efficacy of PCs for GD.
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Portadores de Fármacos/administración & dosificación , Portadores de Fármacos/química , Sistemas de Liberación de Medicamentos , Enfermedad de Gaucher/tratamiento farmacológico , Macrófagos/metabolismo , Chaperonas Moleculares/administración & dosificación , beta-Ciclodextrinas/química , Conformación de Carbohidratos , Enfermedad de Gaucher/patología , Humanos , Macrófagos/efectos de los fármacos , Microscopía Fluorescente , Chaperonas Moleculares/farmacología , Chaperonas Moleculares/uso terapéutico , beta-Ciclodextrinas/administración & dosificaciónAsunto(s)
Antígenos de Neoplasias/inmunología , Vacunas contra el Cáncer/inmunología , Chaperonas Moleculares/inmunología , Neoplasias/terapia , Animales , Presentación de Antígeno , Antígenos de Neoplasias/administración & dosificación , Vacunas contra el Cáncer/administración & dosificación , Humanos , Inmunidad Innata , Chaperonas Moleculares/administración & dosificación , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/inmunologíaRESUMEN
Heat shock protein 90 (HSP90) is involved in protein folding and functions as a chaperone for numerous client proteins, many of which are important in non-small cell lung cancer (NSCLC) pathogenesis. We sought to define preclinical effects of the HSP90 inhibitor NVP-AUY922 and identify predictors of response. We assessed in vitro effects of NVP-AUY922 on proliferation and protein expression in NSCLC cell lines. We evaluated gene expression changes induced by NVP-AUY922 exposure. Xenograft models were evaluated for tumor control and biological effects. NVP-AUY922 potently inhibited in vitro growth in all 41 NSCLC cell lines evaluated with IC50 < 100 nmol/L. IC100 (complete inhibition of proliferation) < 40 nmol/L was seen in 36 of 41 lines. Consistent gene expression changes after NVP-AUY922 exposure involved a wide range of cellular functions, including consistently decreased dihydrofolate reductase after exposure. NVP-AUY922 slowed growth of A549 (KRAS-mutant) xenografts and achieved tumor stability and decreased EGF receptor (EGFR) protein expression in H1975 xenografts, a model harboring a sensitizing and a resistance mutation for EGFR-tyrosine kinase inhibitors in the EGFR gene. These data will help inform the evaluation of correlative data from a recently completed phase II NSCLC trial and a planned phase IB trial of NVP-AUY922 in combination with pemetrexed in NSCLCs.
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Apoptosis/efectos de los fármacos , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Isoxazoles/administración & dosificación , Resorcinoles/administración & dosificación , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/patología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Ensayos Clínicos como Asunto , Receptores ErbB/antagonistas & inhibidores , Receptores ErbB/metabolismo , Proteínas HSP90 de Choque Térmico/genética , Proteínas HSP90 de Choque Térmico/metabolismo , Humanos , Chaperonas Moleculares/administración & dosificación , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Peptide chaperon TD1 was discovered to facilitate several proteins' transdermal delivery via topical co-administration. To design a practical, safe system for advanced transdermal peptide, a novel method was carried out. Human epidermal growth factor (hEGF) was selected as the model biological agent and a fusion protein: TD1-hEGF was designed. Study showed that TD1-hEGF not only had the similar bioactivity with native hEGF, but also possessed considerable higher transdermal ability than hEGF and a co-administration of TD1 and hEGF. These results provided convincing evidence for the advantages of TD1-hEGF in cosmetic and medical applications. Moreover, the fusion pattern between the cargoes and TD1 offered a new approach to facilitate other hydrophilic drugs' transdermal delivery for therapeutic application.