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Upper urinary tract urolithiasis is an emerging disease in cats, with 98% of kidney stones composed of calcium oxalate. In humans, disturbances in the intestinal and urinary microbiota are suspected to contribute to the formation of calcium oxalate stones. We hypothesized that similar mechanisms may be at play in cats. This study examines the intestinal and urinary microbiota of nine cats with kidney stones compared to nine healthy cats before, during, and after treatment with the antibiotic cefovecin, a cephalosporin. Initially, cats with kidney stones displayed a less diverse intestinal microbiota. Antibiotic treatment reduced microbiota diversity in both groups. The absence of specific intestinal bacteria could lead to a loss of the functions these bacteria perform, such as oxalate degradation, which may contribute to the formation of calcium oxalate stones. This study confirms the presence of a distinct urobiome in cats with kidney stones, characterized by greater richness and diversity compared to healthy cats. These findings highlight the potential of microbiota modulation as a strategy to prevent renal lithiasis in cats.
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An 18-month-old neutered male domestic shorthair cat was presented for an emergency consultation after falling from the second floor. The cat sustained minor traumatic injuries but did not exhibit dyspnea. Routine radiographic examination raised suspicion of a diaphragmatic hernia, but the circumscribed nature of the soft tissues visible in the thorax was atypical for a classic traumatic diaphragmatic hernia. A positive contrast peritoneography highlighted the likely presence of a hernial sac, which strongly suggested a "true diaphragmatic hernia", also known as "pleuroperitoneal hernia". This diagnosis was confirmed during laparotomy, which allowed for the visualization of a 3 cm radial diaphragmatic defect in the right ventral quadrant of the pars sternalis. The diaphragm's edges were rounded. A portion of the falciform ligament and a part of the omentum were protruding through the defect and were contained within a hernial sac. Herniorrhaphy was performed. The cat recovered without complications. Given its presentation and location, ventrally and to the right, this anomaly is analogous to what is described in humans as "Morgagni hernia". Six other cases of Morgagni hernias have probably been reported in cats but were not identified as such. This case underscores the utility of peritoneography, a straightforward technique useful for diagnosing diaphragmatic hernias, which enables differentiation between acquired traumatic forms and congenital forms, particularly peritoneopericardial hernias and pleuroperitoneal hernias. True diaphragmatic hernias are almost always serendipitous discoveries.
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A 5-year-old recently castrated male Doberman dog presented for prolonged erection of one week's duration with associated pain and dysuria. This was the fourth episode within a year. Each episode was associated with an unusual event, which was stressful for the dog. Castration performed two months prior to the final episode did not prevent recurrence. Due to tissue necrosis, penile amputation and urethrostomy had to be performed. The dog recovered fully. Prolonged erection that persists beyond or that is unrelated to sexual stimulation is called "priapism". This term refers to the Greek god Priapus, a god of fertility, memorialized in sculptures for his giant phallus. In humans, depending on the mechanism involved, priapism is classified as nonischemic or ischemic. Because prognosis and treatment are different, priapism must be determined to be nonischemic or ischemic. Nonischemic priapism is a rare condition observed when an increase in penile arterial blood flow overwhelms the capacity of venous drainage; it is often associated with penile trauma, and does not require medical intervention. Ischemic priapism is associated with decreased venous return. In humans, ischemic priapism accounts for 95% of cases, the majority of which are idiopathic. Ischemic priapism is a urological emergency; simple conservative measures such as aspiration of blood from the corpora cavernosa and intracavernosal injection of an adrenergic agent are often successful. Stuttering priapism, also called recurrent or intermittent priapism, is a particular form of ischemic priapism reported in humans that is characterized by repetitive episodes of prolonged erections. Management consists of treating each new episode as an episode of acute ischemic priapism, and preventing recurrence with oral medications such as dutasteride and/or baclofen, gabapentin, or tadalafil. To the authors' knowledge, this case is the first report of stuttering priapism in a dog.
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A 6-year-old mixed-breed male Papillon dog, castrated at the age of 7 months, presented for work-up of a difficulty walking associated with constipation and urinary incontinence. Ultrasonography and radiography were consistent with a tumor of the prostate and lymph node metastases. An irregular osteoproliferation of the ventral edges of L5-L6-L7 suggested tumor invasion. Periosteal proliferative lesions of the pelvis, the femur, the humerus, the tibia and the calcaneus were consistent with hypertrophic osteopathy. Necropsy and histological examination confirmed the diagnosis of prostatic adenocarcinoma with lymph node, pulmonary, liver and bone metastases, associated with hypertrophic osteopathy.
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Despite the ability to determine feline blood types, the transfusion of canine blood to cats is still practiced in some countries. Xenotransfusion is effective-even if its effects only last for a few days-and is not associated with serious adverse effects. It avoids the need for blood typing, and most importantly, it avoids the transmission of intraspecific infectious agents, notably the feline leukemia virus (FeLV). Transfusion with canine blood is easier, quicker and less costly than transfusion with feline blood; it is less disagreeable for the donor. In the light of these arguments, when feline blood collected according to current guidelines is not available, in particular when the donor is not confirmed to be negative for the FeLV provirus, the authors consider it to be judicious to use canine blood for feline transfusion in emergency situations; this practice is preferable to inaction and to the inoculation of an infectious agent. Allotransfusion remains preferable in non-emergency situations as a treatment of chronic compensated anaemiae or if an appropriate donor (negative for FeLV provirus) is available. However, 2-4 days after a xenotransfusion, if a clinical alteration and a significant decrease in haematocrit are observed, a transfusion with cat's blood confirmed to be negative for FeLV provirus should be performed. Xenotransfusion should never be used twice.
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Adeno-associated virus (AAV) vectors are considered efficient vectors for gene transfer, as illustrated by recent successful clinical trials targeting retinal or neurodegenerative disorders. However, limitations as host immune responses to AAV capsid or transduction of limited regions must still be overcome. Here, we focused on locoregional (LR) intravenous perfusion vector delivery that allows transduction of large muscular areas and is considered to be less immunogenic than intramuscular (IM) injection. To confirm this hypothesis, we injected 6 cynomolgus monkeys with an AAV serotype 8 (AAV8) vector encoding for the highly immunogenic GFP driven by either a muscle-specific promoter (n = 3) or a cytomegalovirus (CMV) promoter (n = 3). We report that LR delivery allows long-term GFP expression in the perfused limb (up to 1 year) despite the initiation of a peripheral transgene-specific immune response. The analysis of the immune status of the perfused limb shows that LR delivery induces persisting inflammation. However, this inflammation is not sufficient to result in transgene clearance and is balanced by resident regulatory T cells. Overall, our results suggest that LR delivery promotes persisting transgene expression by induction of Treg cells in situ and might be a safe alternative to IM route to target large muscle territories for the expression of secreted therapeutic factors.
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The identification of the most efficient method for whole central nervous system targeting that is translatable to humans and the safest route of adeno-associated virus (AAV) administration is a major concern for future applications in clinics. Additionally, as many AAV serotypes were identified for gene introduction into the brain and the spinal cord, another key to human gene-therapy success is to determine the most efficient serotype. In this study, we compared lumbar intrathecal administration through catheter implantation and intracerebroventricular administration in the cynomolgus macaque. We also evaluated and compared two AAV serotypes that are currently used in clinical trials: AAV9 and AAVrh10. We demonstrated that AAV9 lumbar intrathecal delivery using a catheter achieved consistent transgene expression in the motor neurons of the spinal cord and in the neurons/glial cells of several brain regions, whereas AAV9 intracerebroventricular delivery led to a consistent transgene expression in the brain. In contrast, AAVrh10 lumbar intrathecal delivery led to rare motor neuron targeting. Finally, we found that AAV9 efficiently targets respiratory and skeletal muscles after injection into the cerebrospinal fluid (CSF), which represents an outstanding new property that can be useful for the treatment of diseases affecting both the central nervous system and muscle.
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Neutralizing antibodies directed against adeno-associated virus (AAV) are commonly found in humans. In seropositive subjects, vector administration is not feasible as antibodies neutralize AAV vectors even at low titers. Consequently, a relatively large proportion of humans is excluded from enrollment in clinical trials and, similarly, vector redosing is not feasible because of development of high-titer antibodies following AAV vector administration. Plasmapheresis has been proposed as strategy to remove anti-AAV antibodies from the bloodstream. Although safe and relatively effective, the technology has some limitations mainly related to the nonspecific removal of all circulating IgG. Here we developed an AAV-specific plasmapheresis column which was shown to efficiently and selectively deplete anti-AAV antibodies without depleting the total immunoglobulin pool from plasma. We showed the nearly complete removal of anti-AAV antibodies from high titer purified human IgG pools and plasma samples, decreasing titers to levels that allow AAV vector administration in mice. These results provide proof-of-concept of a method for the AAV-specific depletion of neutralizing antibodies in the setting of in vivo gene transfer.
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Anticuerpos Neutralizantes/aislamiento & purificación , Anticuerpos Antivirales/aislamiento & purificación , Cápside , Dependovirus/inmunología , Vectores Genéticos/inmunología , Inmunoglobulina G/aislamiento & purificación , Plasmaféresis/métodos , Animales , Técnicas de Transferencia de Gen , Humanos , RatonesRESUMEN
Limb-girdle muscular dystrophy type 2A (LGMD2A or LGMDR1) is a neuromuscular disorder caused by mutations in the calpain 3 gene (CAPN3). Previous experiments using adeno-associated viral (AAV) vector-mediated calpain 3 gene transfer in mice indicated cardiac toxicity associated with the ectopic expression of the calpain 3 transgene. Here, we performed a preliminary dose study in a severe double-knockout mouse model deficient in calpain 3 and dysferlin. We evaluated safety and biodistribution of AAV9-desmin-hCAPN3 vector administration to nonhuman primates (NHPs) with a dose of 3 × 1013 viral genomes/kg. Vector administration did not lead to observable adverse effects or to detectable toxicity in NHP. Of note, the transgene expression did not produce any abnormal changes in cardiac morphology or function of injected animals while reaching therapeutic expression in skeletal muscle. Additional investigation on the underlying causes of cardiac toxicity observed after gene transfer in mice and the role of titin in this phenomenon suggest species-specific titin splicing. Mice have a reduced capacity for buffering calpain 3 activity compared to NHPs and humans. Our studies highlight a complex interplay between calpain 3 and titin binding sites and demonstrate an effective and safe profile for systemic calpain 3 vector delivery in NHP, providing critical support for the clinical potential of calpain 3 gene therapy in humans.
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Calpaína/genética , Calpaína/uso terapéutico , Cardiotoxicidad/etiología , Conectina/genética , Terapia Genética/efectos adversos , Proteínas Musculares/genética , Proteínas Musculares/uso terapéutico , Distrofia Muscular de Cinturas/genética , Distrofia Muscular de Cinturas/terapia , Empalme del ARN/genética , Animales , Sitios de Unión , Biomarcadores/sangre , Cardiotoxicidad/sangre , Conectina/química , Dependovirus/genética , Disferlina/deficiencia , Disferlina/metabolismo , Estabilidad de Enzimas , Regulación de la Expresión Génica , Ratones Noqueados , MicroARNs/genética , MicroARNs/metabolismo , Músculo Esquelético/metabolismo , Distrofia Muscular de Cinturas/sangre , Distrofia Muscular de Cinturas/patología , Miocardio/metabolismo , Miocardio/patología , Primates , Dominios Proteicos , Proteolisis , Especificidad de la Especie , Distribución Tisular , TransgenesRESUMEN
Growing demonstrations of regenerative potential for some stem cells led recently to promising therapeutic proposals for neuromuscular diseases. We have shown that allogeneic MuStem cell transplantation into Golden Retriever muscular dystrophy (GRMD) dogs under continuous immunosuppression (IS) leads to persistent clinical stabilization and muscle repair. However, long-term IS in medical practice is associated with adverse effects raising safety concerns. Here, we investigate whether the IS removal or its restriction to the transplantation period could be considered. Dogs aged 4-5 months old received vascular infusions of allogeneic MuStem cells without IS (GRMDMU/no-IS) or under transient IS (GRMDMU/tr-IS). At 5 months post-infusion, persisting clinical status improvement of the GRMDMU/tr-IS dogs was observed while GRMDMU/no-IS dogs exhibited no benefit. Histologically, only 9-month-old GRMDMU/tr-IS dogs showed an increased muscle regenerative activity. A mixed cell reaction with the host peripheral blood mononucleated cells (PBMCs) and corresponding donor cells revealed undetectable to weak lymphocyte proliferation in GRMDMU/tr-IS dogs compared with a significant proliferation in GRMDMU/no-IS dogs. Importantly, any dog group showed neither cellular nor humoral anti-dystrophin responses. Our results show that transient IS is necessary and sufficient to sustain allogeneic MuStem cell transplantation benefits and prevent host immunity. These findings provide useful critical insight to designing therapeutic strategies.
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Enfermedades de los Perros/terapia , Terapia de Inmunosupresión/métodos , Distrofia Muscular Animal/terapia , Trasplante de Células Madre/métodos , Células Alogénicas/inmunología , Animales , Perros , Distrofina/inmunología , Masculino , Distrofia Muscular Animal/inmunología , Células Madre/citología , Células Madre/inmunología , Trasplante Homólogo/métodosRESUMEN
Duchenne muscular dystrophy (DMD) is an incurable X-linked muscle-wasting disease caused by mutations in the dystrophin gene. Gene therapy using highly functional microdystrophin genes and recombinant adeno-associated virus (rAAV) vectors is an attractive strategy to treat DMD. Here we show that locoregional and systemic delivery of a rAAV2/8 vector expressing a canine microdystrophin (cMD1) is effective in restoring dystrophin expression and stabilizing clinical symptoms in studies performed on a total of 12 treated golden retriever muscular dystrophy (GRMD) dogs. Locoregional delivery induces high levels of microdystrophin expression in limb musculature and significant amelioration of histological and functional parameters. Systemic intravenous administration without immunosuppression results in significant and sustained levels of microdystrophin in skeletal muscles and reduces dystrophic symptoms for over 2 years. No toxicity or adverse immune consequences of vector administration are observed. These studies indicate safety and efficacy of systemic rAAV-cMD1 delivery in a large animal model of DMD, and pave the way towards clinical trials of rAAV-microdystrophin gene therapy in DMD patients.
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Distrofina/genética , Técnicas de Transferencia de Gen , Músculo Esquelético/metabolismo , Distrofia Muscular Animal/terapia , Distrofia Muscular de Duchenne/genética , Administración Intravenosa , Animales , Dependovirus , Modelos Animales de Enfermedad , Perros , Terapia Genética , Vectores Genéticos , Masculino , Músculo Esquelético/fisiopatología , Distrofia Muscular Animal/genética , Distrofia Muscular Animal/metabolismo , Distrofia Muscular Animal/fisiopatología , Distrofia Muscular de Duchenne/metabolismo , Distrofia Muscular de Duchenne/fisiopatología , TransgenesRESUMEN
Recombinant adeno-associated virus (AAV) has emerged as a promising vector for retinal gene delivery to restore visual function in certain forms of inherited retinal dystrophies. Several studies in rodent models have shown that intravitreal injection of the AAV2/2 vector is the optimal route for efficient retinal ganglion cell (RGC) transduction. However, translation of these findings to larger species, including humans, is complicated by anatomical differences in the eye, a key difference being the comparatively smaller volume of the vitreous chamber in rodents. Here, we address the role of the vitreous body as a potential barrier to AAV2/2 diffusion and transduction in the RGCs of dogs and macaques, two of the most relevant preclinical models. We intravitreally administered the AAV2/2 vector carrying the CMV-eGFP reporter cassette in dog and macaque eyes, either directly into the vitreous chamber or after complete vitrectomy, a surgical procedure that removes the vitreous body. Our findings suggest that the vitreous body appears to trap the injected vector, thus impairing the diffusion and transduction of AAV2/2 to inner retinal neurons. We show that vitrectomy before intravitreal vector injection is an effective means of overcoming this physical barrier, improving the transduction of RGCs in dog and macaque retinas. These findings support the use of vitrectomy in clinical trials of intravitreal gene transfer techniques targeting inner retinal neurons.
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Terapia Genética , Vectores Genéticos/uso terapéutico , Células Ganglionares de la Retina , Animales , Dependovirus/genética , Perros , Técnicas de Transferencia de Gen , Proteínas Fluorescentes Verdes , Humanos , Inyecciones Intravítreas , Macaca , Retina/patología , Retina/trasplante , Transducción Genética , VitrectomíaRESUMEN
We previously reported that subretinal injection of AAV2/5 RK.cpde6ß allowed long-term preservation of photoreceptor function and vision in the rod-cone dysplasia type 1 (rcd1) dog, a large animal model of naturally occurring PDE6ß deficiency. The present study builds on these earlier findings to provide a detailed assessment of the long-term effects of gene therapy on the spatiotemporal pattern of retinal degeneration in rcd1 dogs treated at 20 days of age. We analyzed the density distribution of the retinal layers and of particular photoreceptor cells in 3.5-year-old treated and untreated rcd1 dogs. Whereas no rods were observed outside the bleb or in untreated eyes, gene transfer halted rod degeneration in all vector-exposed regions. Moreover, while gene therapy resulted in the preservation of cones, glial cells and both the inner nuclear and ganglion cell layers, no cells remained in vector-unexposed retinas, except in the visual streak. Finally, the retinal structure of treated 3.5-year-old rcd1 dogs was identical to that of unaffected 4-month-old rcd1 dogs, indicating near complete preservation. Our findings indicate that gene therapy arrests the degenerative process even if intervention is initiated after the onset of photoreceptor degeneration, and point to significant potential of this therapeutic approach in future clinical trials.
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Fosfodiesterasas de Nucleótidos Cíclicos Tipo 6/genética , Terapia Genética/métodos , Degeneración Retiniana/terapia , Células Fotorreceptoras Retinianas Bastones/patología , Animales , Fosfodiesterasas de Nucleótidos Cíclicos Tipo 6/deficiencia , Dependovirus/genética , Modelos Animales de Enfermedad , Perros , Vectores Genéticos/administración & dosificación , Humanos , Retina/fisiopatología , Degeneración Retiniana/genética , Degeneración Retiniana/patologíaRESUMEN
A 20 kg German shepherd dog was presented to a French veterinary teaching hospital for seizures and hyperthermia. The dog had returned 1 month previously from a six-month stay in Senegal and sub-Saharan Africa. Biochemistry and haematology showed severe hypoglycaemia (0.12 g/L), anaemia and thrombocytopenia. Despite administration of large amounts of glucose (30 mL of 30% glucose IV and 10 mL of 70% sucrose by gavage tube hourly), 26 consecutive blood glucose measurements were below 0.25 g/L (except one). Routine cytological examination of blood smears revealed numerous free extracytoplasmic protozoa consistent with Trypanosoma congolense. PCR confirmed a Trypanosoma congolense forest-type infection. Treatment consisted of six injections of pentamidine at 48-hour intervals. Trypanosomes had disappeared from the blood smears four days following the first injection. Clinical improvement was correlated with the normalization of laboratory values. The infection relapsed twice and the dog was treated again; clinical signs and parasites disappeared and the dog was considered cured; however, 6 years after this incident, serological examination by ELISA T. congolense was positive. The status of this dog (infected or non-infected) remains unclear. Hypoglycaemia was the most notable clinical feature in this case. It was spectacular in its severity and in its refractory nature; glucose administration seemed only to feed the trypanosomes, indicating that treatment of hypoglycaemia may in fact have been detrimental.
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Enfermedades de los Perros/parasitología , Hipoglucemia/veterinaria , Parasitemia/veterinaria , Trypanosoma congolense/aislamiento & purificación , Tripanosomiasis Africana/veterinaria , África del Sur del Sahara , Animales , Antiprotozoarios/uso terapéutico , Enfermedades de los Perros/sangre , Perros , Femenino , Fiebre/etiología , Fiebre/veterinaria , Glucosa/efectos adversos , Glucosa/uso terapéutico , Hipoglucemia/tratamiento farmacológico , Hipoglucemia/etiología , Parasitemia/sangre , Parasitemia/diagnóstico , Parasitemia/tratamiento farmacológico , Pentamidina/uso terapéutico , Recurrencia , Convulsiones/etiología , Convulsiones/veterinaria , Senegal , Viaje , Tripanosomiasis Africana/sangre , Tripanosomiasis Africana/diagnóstico , Tripanosomiasis Africana/tratamiento farmacológicoRESUMEN
BACKGROUND: Several adult stem cell populations exhibit myogenic regenerative potential, thus representing attractive candidates for therapeutic approaches of neuromuscular diseases such as Duchenne Muscular Dystrophy (DMD). We have recently shown that systemic delivery of MuStem cells, skeletal muscle-resident stem cells isolated in healthy dog, generates the remodelling of muscle tissue and gives rise to striking clinical benefits in Golden Retriever Muscular Dystrophy (GRMD) dog. This global effect, which is observed in the clinically relevant DMD animal model, leads us to question here the molecular pathways that are impacted by MuStem cell transplantation. To address this issue, we compare the global gene expression profile between healthy, GRMD and MuStem cell treated GRMD dog muscle, four months after allogenic MuStem cell transplantation. RESULTS: In the dystrophic context of the GRMD dog, disease-related deregulation is observed in the case of 282 genes related to various processes such as inflammatory response, regeneration, calcium ion binding, extracellular matrix organization, metabolism and apoptosis regulation. Importantly, we reveal the impact of MuStem cell transplantation on several molecular and cellular pathways based on a selection of 31 genes displaying signals specifically modulated by the treatment. Concomitant with a diffuse dystrophin expression, a histological remodelling and a stabilization of GRMD dog clinical status, we show that cell delivery is associated with an up-regulation of genes reflecting a sustained enhancement of muscle regeneration. We also identify a decreased mRNA expression of a set of genes having metabolic functions associated with lipid homeostasis and energy. Interestingly, ubiquitin-mediated protein degradation is highly enhanced in GRMD dog muscle after systemic delivery of MuStem cells. CONCLUSIONS: Overall, our results provide the first high-throughput characterization of GRMD dog muscle and throw new light on the complex molecular/cellular effects associated with muscle repair and the clinical efficacy of MuStem cell-based therapy.
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Perfilación de la Expresión Génica , Músculo Esquelético/patología , Distrofia Muscular Animal/genética , Distrofia Muscular Animal/terapia , Trasplante de Células Madre , Animales , Modelos Animales de Enfermedad , Perros , Estudios de Seguimiento , Humanos , Músculo Esquelético/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , Control de Calidad , Reacción en Cadena en Tiempo Real de la Polimerasa , Reproducibilidad de los ResultadosRESUMEN
No treatment is available for early-onset forms of metachromatic leukodystrophy (MLD), a lysosomal storage disease caused by autosomal recessive defect in arylsulfatase A (ARSA) gene causing severe demyelination in central and peripheral nervous systems. We have developed a gene therapy approach, based on intracerebral administration of AAVrh.10-hARSA vector, coding for human ARSA enzyme. We have previously demonstrated potency of this approach in MLD mice lacking ARSA expression. We describe herein the preclinical efficacy, safety, and biodistribution profile of intracerebral administration of AAVrh.10-hARSA to nonhuman primates (NHPs). NHPs received either the dose planned for patients adjusted to the brain volume ratio between child and NHP (1×dose, 1.1×10(11) vg/hemisphere, unilateral or bilateral injection) or 5-fold this dose (5×dose, 5.5×10(11) vg/hemisphere, bilateral injection). NHPs were subjected to clinical, biological, and brain imaging observations and were euthanized 7 or 90 days after injection. There was no toxicity based on clinical and biological parameters, nor treatment-related histological findings in peripheral organs. A neuroinflammatory process correlating with brain MRI T2 hypersignals was observed in the brain 90 days after administration of the 5×dose, but was absent or minimal after administration of the 1×dose. Antibody response to AAVrh.10 and hARSA was detected, without correlation with brain lesions. After injection of the 1×dose, AAVrh.10-hARSA vector was detected in a large part of the injected hemisphere, while ARSA activity exceeded the normal endogenous activity level by 14-31%. Consistently with other reports, vector genome was detected in off-target organs such as liver, spleen, lymph nodes, or blood, but not in gonads. Importantly, AAVrh.10-hARSA vector was no longer detectable in urine at day 7. Our data demonstrate requisite safe and effective profile for intracerebral AAVrh.10-hARSA delivery in NHPs, supporting its clinical use in children affected with MLD.
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Cerebrósido Sulfatasa/genética , Dependovirus/genética , Leucodistrofia Metacromática/terapia , Animales , Encéfalo/metabolismo , Cerebrósido Sulfatasa/metabolismo , Niño , Terapia Genética , Vectores Genéticos , Humanos , Macaca fascicularis , MasculinoRESUMEN
This study investigated the long-term effectiveness and safety of a variant of the transobturator vaginal tape inside-out technique for acquired urinary incontinence. Twelve spayed female dogs were operated over a 2 yr period. No intraoperative complications were encountered. Transient dysuria was the most common postoperative complication (7 out of 12 dogs). On the 12th day postoperatively, incontinence was completely cured in 11 out of 12 dogs (92%). At the time of the second evaluation (median follow-up time was 21 mo), patients classified as "cured," "greatly improved," or "improved" were 25, 50, and 25% of the total, respectively. At the time of either the fourth evaluation or at the time of death (median follow-up time was 52 mo), 50% of the bitches (6 out of 12) had the same results as previously but the other 50% had leakage that reappeared sporadically. A fistula appeared on the path of the tape in two bitches at 28 and 32 mo postsurgically. The technique presented is effective and more cost effective than the standard technique and could constitute an attractive alternative; however, it could be associated with an immediate postoperative dysuria, delayed fistula formation, and a partial recurrence of clinical signs.
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Enfermedades de los Perros/cirugía , Cabestrillo Suburetral/veterinaria , Incontinencia Urinaria/veterinaria , Animales , Perros , Femenino , Histerectomía/efectos adversos , Histerectomía/veterinaria , Ovariectomía/efectos adversos , Ovariectomía/veterinaria , Complicaciones Posoperatorias/veterinaria , Resultado del Tratamiento , Incontinencia Urinaria/cirugíaRESUMEN
CASE SERIES SUMMARY: In October 2011, an abnormally large morbidity and mortality event was noted in the intensive care unit (ICU) of a veterinary school hospital in Nantes, France. Cats, and cats only, transferred from the emergency room presented with fever, ulcers on the tongue and cutaneous lesions around venepuncture or surgical incision sites, leading to suspicion of a feline calicivirus-associated virulent systemic disease confirmed with reverse transcriptase-polymerase chain reaction. A total of 14 cats were suspected. The clinical features and the origin of the contamination were described for each cat. The median length of incubation was 4.5 days. Fifty-seven percent of the cats were euthanased (8/14) and 21% died (3/14), with a combined mortality of 79% (11/14) - the highest ever reported. Median survival was 12 days. The recovery rate was 21% (3/14). RELEVANCE AND NOVEL INFORMATION: Eight outbreaks have been reported, in veterinary clinics or in group-housed cats. The main unusual aspects of the present outbreak were: (1) the extreme flare-up of lesions at sites of skin breach, precluding any puncture/incision; (2) the suggested better survival rate at home than in hospital; and (3) the immediate control of the outbreak after recognition of the disease. Other striking but less unusual features of this outbreak were: (4) the increasing of the virulence of the calicivirus with the passage of time; and (5) the primary role that the caregivers' hands played in the spread of the outbreak.
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Duchenne muscular dystrophy (DMD) is a severe muscle-wasting disorder caused by mutations in the dystrophin gene, without curative treatment yet available. Our study provides, for the first time, the overall safety profile and therapeutic dose of a recombinant adeno-associated virus vector, serotype 8 (rAAV8) carrying a modified U7snRNA sequence promoting exon skipping to restore a functional in-frame dystrophin transcript, and injected by locoregional transvenous perfusion of the forelimb. Eighteen Golden Retriever Muscular Dystrophy (GRMD) dogs were exposed to increasing doses of GMP-manufactured vector. Treatment was well tolerated in all, and no acute nor delayed adverse effect, including systemic and immune toxicity was detected. There was a dose relationship for the amount of exon skipping with up to 80% of myofibers expressing dystrophin at the highest dose. Similarly, histological, nuclear magnetic resonance pathological indices and strength improvement responded in a dose-dependent manner. The systematic comparison of effects using different independent methods, allowed to define a minimum threshold of dystrophin expressing fibers (>33% for structural measures and >40% for strength) under which there was no clear-cut therapeutic effect. Altogether, these results support the concept of a phase 1/2 trial of locoregional delivery into upper limbs of nonambulatory DMD patients.
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Dependovirus/genética , Distrofina/genética , Miembro Anterior/fisiopatología , Distrofia Muscular de Duchenne/terapia , ARN Nuclear Pequeño/genética , Animales , Estudios de Cohortes , Modelos Animales de Enfermedad , Perros , Relación Dosis-Respuesta a Droga , Exones , Terapia Genética , Vectores Genéticos/administración & dosificación , Humanos , Infusiones Intravenosas , Distrofia Muscular de Duchenne/genética , Distrofia Muscular de Duchenne/fisiopatología , ARN Nuclear Pequeño/metabolismoRESUMEN
Preventing untoward immune responses against a specific antigen is a major challenge in different clinical settings such as gene therapy, transplantation, or autoimmunity. Following intramuscular delivery of recombinant adeno-associated virus (rAAV)-derived vectors, transgene rejection can be a roadblock to successful clinical translation. Specific immunomodulation strategies potentially leading to sustained transgene expression while minimizing pharmacological immunosuppression are desirable. Tolerogenic dendritic cells (TolDC) are potential candidates but have not yet been evaluated in the context of gene therapy, to our knowledge. Following intramuscular delivery of rAAV-derived vectors expressing an immunogenic protein in the nonhuman primate model, we assessed the immunomodulating potential of autologous bone marrow-derived TolDC generated in the presence of IL10 and pulsed with the transgene product. TolDC administered either intradermally or intravenously were safe and well tolerated. While the intravenous route showed a modest ability to modulate host immunity against the transgene product, intradermally delivery resulted in a robust vaccination of the macaques when associated to intramuscular rAAV-derived vectors-based gene transfer. These findings demonstrate the critical role of TolDC mode of injection in modulating host immunity. This study also provides the first evidence of the potential of TolDC-based immunomodulation in gene therapy.