RESUMO
BACKGROUND: Mitochondrial neurogastrointestinal encephalopathy is a rare multisystem autosomal recessive disease caused by mutations in the TYMP gene, that encodes for thymidine phosphorylase. Mitochondrial neurogastrointestinal encephalopathy is a progressive degenerative disease characterized by a distinctive tetrad of gastrointestinal dysmotility, peripheral neuropathy, ophthalmoplegia with ptosis, and asymptomatic leukoencephalopathy. It provides a diagnostic dilemma to physicians in regions like Pakistan because of a lack of genetic study availability and associated financial constraints of the population. However, with careful examination and a few basic investigations, mitochondrial neurogastrointestinal encephalopathy can be diagnosed by ruling out most of the close differentials. CASE PRESENTATION: We report the case of a 23-year-old Asian female whose chief complaints were epigastric pain, bilious emesis, weight loss for 3 months, and bilateral lower limb weakness for 20 days. All clinical signs and symptoms along with relevant investigations including nerve conduction studies, electromyography, and magnetic resonance imaging of the brain were highly suggestive of mitochondrial neurogastrointestinal encephalopathy syndrome. Because of financial constraints, genetic studies could not be performed. The patient was managed with a multidisciplinary approach involving gastroenterology, physiotherapy, and nutrition departments. Currently, therapeutic options for the disease include allogeneic hematopoietic stem cell transplant and carrier erythrocyte entrapped thymidine phosphorylase; however, these could not be provided to the patient owing to certain limitations. CONCLUSIONS: As misdiagnosis and delayed diagnosis are quite common in this disease, the prime objective of this case report is to increase the basic understanding of this disease, especially its signs and symptoms, and address the limitations regarding the diagnostic investigations and management of patients with mitochondrial neurogastrointestinal encephalopathy.
Assuntos
Pseudo-Obstrução Intestinal , Encefalomiopatias Mitocondriais , Distrofia Muscular Oculofaríngea , Adulto , Feminino , Humanos , Pseudo-Obstrução Intestinal/complicações , Pseudo-Obstrução Intestinal/diagnóstico , Pseudo-Obstrução Intestinal/terapia , Encefalomiopatias Mitocondriais/diagnóstico , Encefalomiopatias Mitocondriais/genética , Encefalomiopatias Mitocondriais/terapia , Distrofia Muscular Oculofaríngea/complicações , Distrofia Muscular Oculofaríngea/diagnóstico , Oftalmoplegia/congênito , Paquistão , Timidina Fosforilase/genética , Adulto JovemRESUMO
mitochondrial neuro-gastrointestinal encephalomyopathy (MNGIE) is a rare genetic disorder characterized by thymidine phosphorylase (TP) enzyme defect. The absence of TP activity induces the imbalance of mitochondrial nucleotide pool, leading to impaired mitochondrial DNA (mtDNA) replication and depletion. Since mtDNA is required to ensure oxidative phosphorylation, metabolically active tissues may not achieve sufficient energy production. The only effective life-saving approach in MNGIE has been the permanent replacement of TP via allogeneic hematopoietic stem cell or liver transplantation. However, the follow-up of transplanted patients showed that gut tissue changes do not revert and fatal complications, such as massive gastrointestinal bleeding, can occur. The purpose of this study was to clarify whether the reintroduction of TP after transplant can recover mtDNA copy number in a normal range. Using laser capture microdissection and droplet-digital-PCR, we assessed the mtDNA copy number in each layer of full-thickness ileal samples of a naive MNGIE cohort vs. controls and in a patient pre- and post-TP replacement. The treatment led to a significant recovery of gut tissue mtDNA amount, thus showing its efficacy. Our results indicate that a timely TP replacement is needed to maximize therapeutic success before irreversible degenerative tissue changes occur in MNGIE.
Assuntos
Transplante de Fígado , Erros Inatos do Metabolismo , Encefalomiopatias Mitocondriais , DNA Mitocondrial/genética , Humanos , Íleo , Microdissecção e Captura a Laser , Lasers , Encefalomiopatias Mitocondriais/genética , Encefalomiopatias Mitocondriais/terapiaRESUMO
Mitochondrial encephalomyopathies are disorders caused by mitochondrial and nuclear DNA mutations which affect the nervous and muscular systems. Current therapies for mitochondrial encephalomyopathies are inadequate and mostly palliative. However, stem cell-derived mitochondria transplantation has been demonstrated to play an key part in metabolic rescue, which offers great promise for mitochondrial encephalomyopathies. Here, we summarize the present status of stem cell therapy for mitochondrial encephalomyopathy and discuss mitochondrial transfer routes and the protection mechanisms of stem cells. We also identify and summarize future perspectives and challenges for the treatment of these intractable disorders based on the concept of mitochondrial transfer from stem cells.
Assuntos
Mitocôndrias/transplante , Encefalomiopatias Mitocondriais/terapia , Transplante de Células-Tronco/métodos , Animais , DNA Mitocondrial/imunologia , DNA Mitocondrial/metabolismo , Vesículas Extracelulares/imunologia , Vesículas Extracelulares/metabolismo , Humanos , Mitocôndrias/imunologia , Mitocôndrias/metabolismo , Encefalomiopatias Mitocondriais/imunologia , Encefalomiopatias Mitocondriais/metabolismo , Nanotubos , Células-Tronco/imunologia , Células-Tronco/metabolismoRESUMO
We report the longest follow-up of clinical and biochemical features of two previously reported adult mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) patients treated with liver transplantation (LT), adding information on a third, recently transplanted, patient. All three patients overcame the early post-operative period and tolerated immunosuppressive therapy. Plasma nucleoside levels dramatically decreased, with evidence of clinical improvement of ambulation and neuropathy. Conversely, other features of MNGIE, as gastrointestinal dysmotility, low weight, ophthalmoparesis, and leukoencephalopathy were essentially unchanged. A similar picture characterized two patients treated with allogenic hematopoietic stem cell transplantation (AHSCT). In conclusion, LT promptly and stably normalizes nucleoside imbalance in MNGIE, stabilizing or improving some clinical parameters with marginal periprocedural mortality rate as compared to AHSCT. Nevertheless, restoring thymidine phosphorylase (TP) activity, achieved by both LT and AHSCT, does not allow a full clinical recovery, probably due to consolidated cellular damage and/or incomplete enzymatic tissue replacement.
Assuntos
Transplante de Fígado , Encefalomiopatias Mitocondriais , Oftalmoplegia , Adulto , Seguimentos , Humanos , Encefalomiopatias Mitocondriais/terapia , Timidina FosforilaseRESUMO
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a fatal disorder characterized by progressive gastrointestinal dysmotility, peripheral neuropathy, leukoencephalopathy, skeletal myopathy, ophthalmoparesis, and ptosis. MNGIE stems from deficient thymidine phosphorylase activity (TP) leading to toxic elevations of plasma thymidine. Hematopoietic stem cell transplant (HSCT) restores TP activity and halts disease progression but has high transplant-related morbidity and mortality. Liver transplant (LT) was reported to restore TP activity in two adult MNGIE patients. We report successful LT in four additional MNGIE patients, including a pediatric patient. Our patients were diagnosed between ages 14 months and 36 years with elevated thymidine levels and biallelic pathogenic variants in TYMP. Two patients presented with progressive gastrointestinal dysmotility, and three demonstrated progressive peripheral neuropathy with two suffering limitations in ambulation. Two patients, including the child, had liver dysfunction and cirrhosis. Following LT, thymidine levels nearly normalized in all four patients and remained low for the duration of follow-up. Disease symptoms stabilized in all patients, with some manifesting improvements, including intestinal function. No patient died, and LT appeared to have a more favorable safety profile than HSCT, especially when liver disease is present. Follow-up studies will need to document the long-term impact of this new approach on disease outcome. Take Home Message: Liver transplantation is effective in stabilizing symptoms and nearly normalizing thymidine levels in patients with mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) and may have an improved safety profile over hematopoietic stem cell transplant.
Assuntos
Transplante de Fígado/métodos , Mitocôndrias/metabolismo , Encefalomiopatias Mitocondriais/terapia , Timidina Fosforilase/genética , Adolescente , Adulto , Transtornos da Motilidade Esofágica/genética , Feminino , Transplante de Células-Tronco Hematopoéticas/mortalidade , Humanos , Lactente , Transplante de Fígado/mortalidade , Imageamento por Ressonância Magnética , Masculino , Mitocôndrias/enzimologia , Mitocôndrias/patologia , Encefalomiopatias Mitocondriais/diagnóstico por imagem , Encefalomiopatias Mitocondriais/genética , Encefalomiopatias Mitocondriais/fisiopatologia , Doenças do Sistema Nervoso Periférico/genética , Timidina/sangue , Sequenciamento do ExomaRESUMO
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a devastating disease caused by mutations in TYMP, which encodes thymidine phosphorylase (TP). In MNGIE patients, TP dysfunction results in systemic thymidine and deoxyuridine overload, which interferes with mitochondrial DNA replication. Preclinical studies have shown that gene therapy using a lentiviral vector targeted to hematopoietic stem cells or an adeno-associated virus (AAV) vector transcriptionally targeted to liver are feasible approaches to treat MNGIE. Here, we studied the effect of various promoters (thyroxine-binding globulin [TBG], phosphoglycerate kinase [PGK], hybrid liver-specific promoter [HLP], and alpha-1-antitrypsin [AAT]) and DNA configuration (single stranded or self complementary) on expression of the TYMP transgene in the AAV8 serotype in a murine model of MNGIE. All vectors restored liver TP activity and normalized nucleoside homeostasis in mice. However, the liver-specific promoters TBG, HLP, and AAT were more effective than the constitutive PGK promoter, and the self-complementary DNA configuration did not provide any therapeutic advantage over the single-stranded configuration. Among all constructs, only AAV-AAT was effective in all mice treated at the lowest dose (5 × 1010 vector genomes/kg). As use of the AAT promoter will likely minimize the dose needed to achieve clinical efficacy as compared to the other promoters tested, we propose using the AAT promoter in the vector eventually designed for clinical use.
Assuntos
Dependovirus/genética , Expressão Gênica , Terapia Genética , Vetores Genéticos/genética , Encefalomiopatias Mitocondriais/genética , Encefalomiopatias Mitocondriais/terapia , Regiões Promotoras Genéticas , alfa 1-Antitripsina/genética , Animais , Modelos Animais de Doenças , Ordem dos Genes , Terapia Genética/métodos , Vetores Genéticos/administração & dosagem , Homeostase , Fígado/metabolismo , Fígado/patologia , Camundongos , Camundongos Knockout , Mitocôndrias Hepáticas/genética , Mitocôndrias Hepáticas/metabolismo , Encefalomiopatias Mitocondriais/metabolismo , Timidina Fosforilase/genética , Transdução Genética , alfa 1-Antitripsina/metabolismoRESUMO
BACKGROUND: Gastrointestinal complications are the main cause of death in patients with mitochondrial neurogastrointestinal encephalomyopathy (MNGIE). Available treatments often restore biochemical homeostasis, but fail to cure gastrointestinal symptoms. METHODS: We evaluated the small intestine neuromuscular pathology of an untreated MNGIE patient and two recipients of hematopoietic stem cells, focusing on enteric neurons and glia. Additionally, we evaluated the intestinal neuromuscular pathology in a mouse model of MNGIE treated with hematopoietic stem cell gene therapy. Quantification of muscle wall thickness and ganglion cell density was performed blind to the genotype with ImageJ. Significance of differences between groups was determined by two-tailed Mann-Whitney U test (P < 0.05). RESULTS: Our data confirm that MNGIE presents with muscle atrophy and loss of Cajal cells and CD117/c-kit immunoreactivity in the small intestine. We also show that hematopoietic stem cell transplantation does not benefit human intestinal pathology at least on short-term. CONCLUSIONS: We suggest that hematopoietic stem cell transplantation may be insufficient to restore intestinal neuropathology, especially at later stages of MNGIE. As interstitial Cajal cells and their networks play a key role in development of gastrointestinal dysmotility, alternative therapeutic approaches taking absence of these cells into account could be required.
Assuntos
Gastroenteropatias/patologia , Gastroenteropatias/terapia , Terapia Genética , Transplante de Células-Tronco Hematopoéticas , Intestino Delgado/patologia , Encefalomiopatias Mitocondriais/patologia , Encefalomiopatias Mitocondriais/terapia , Adolescente , Animais , Criança , Modelos Animais de Doenças , Humanos , Células Intersticiais de Cajal/patologia , Camundongos , Atrofia Muscular/patologia , Adulto JovemRESUMO
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a rare, autosomal-recessive mitochondrial disorder caused by TYMP mutations presenting with a multisystemic, often lethal syndrome of progressive leukoencephalopathy, ophthalmoparesis, demyelinating neuropathy, cachexia and gastrointestinal dysmotility. Hemodialysis (HMD) has been suggested as a treatment to reduce accumulation of thymidine and deoxyuridine. However, all studies so far have failed to measure the toxic metabolites in cerebrospinal fluid (CSF), which is the crucial compartment for CNS damage.Our study is the first prospective, longitudinal investigation, exploiting detailed serial testing of predefined clinical and molecular outcome parameters (including serial CSF assessments) in a 29-year-old MNGIE patient undergoing 1 year of extensive HMD. We demonstrate that HMD only transiently restores increased serum and urine levels of thymidine and deoxyuridine, but fails to reduce CSF levels of the toxic metabolites and is ineffective to influence neurological function. These findings have direct important implications for clinical practice: They prevent a burdensome, long-term invasive, but ultimately probably ineffective procedure in future MNGIE patients.
Assuntos
Doenças do Sistema Nervoso Central/etiologia , Desoxiuridina/sangue , Pseudo-Obstrução Intestinal/terapia , Encefalomiopatias Mitocondriais/terapia , Diálise Renal , Timidina/sangue , Adulto , Doenças do Sistema Nervoso Central/patologia , Doenças do Sistema Nervoso Central/terapia , Desoxiuridina/urina , Humanos , Pseudo-Obstrução Intestinal/genética , Pseudo-Obstrução Intestinal/patologia , Masculino , Encefalomiopatias Mitocondriais/genética , Encefalomiopatias Mitocondriais/patologia , Distrofia Muscular Oculofaríngea , Mutação , Oftalmoplegia/congênito , Doenças Raras , Timidina/urina , Timidina FosforilaseRESUMO
Mitochondrial neurogastrointestinal encephalopathy (MNGIE), usually an autosomal-recessive inherited condition, causes gastrointestinal dysmotility, ophthalmoplegia, ptosis, leukoencephalopathy and neuropathy. The chromosome 22 disorder, due to mutations in the nuclear gene TYMP encoding thymidine phosphorylase (TP), leads to the accumulation of thymidine and deoxyuridine, with mitochondrial dysfunction.This report describes a patient with an MNGIE-like syndrome with a heterozygous TYMP mutation who showed marked, but transient improvement postallogeneic haematopoietic stem cell transplantation (HSCT).The patient, showing ptosis and ophthalmoplegia, was initially managed for myasthenia gravis. She developed gastrointestinal symptoms, dysarthria, dysphagia and weakness, and MNGIE was considered due to its low TP levels and improvement after platelet transfusions. She underwent HSCT, with dramatic improvement, but regressed 18 months later despite normal TP levels, platelet counts and full chimerism.MNGIE may encompass a spectrum of disorders. TP deficiency alone is unlikely to explain all clinical signs, and other factors, including the possible development of anti-TP antibodies, which may play a role in the pathophysiology.
Assuntos
Transplante de Células-Tronco Hematopoéticas/métodos , Pseudo-Obstrução Intestinal/terapia , Encefalomiopatias Mitocondriais/terapia , Timidina Fosforilase/genética , Adulto , Feminino , Humanos , Pseudo-Obstrução Intestinal/genética , Encefalomiopatias Mitocondriais/genética , Distrofia Muscular Oculofaríngea , Mutação , Oftalmoplegia/congênito , Transplante Homólogo , Resultado do TratamentoAssuntos
Alocação de Recursos para a Atenção à Saúde/ética , Encefalomiopatias Mitocondriais/terapia , Transferência de Pacientes/legislação & jurisprudência , Medicina Estatal/legislação & jurisprudência , Terapias em Estudo/economia , Medicina Baseada em Evidências , Alocação de Recursos para a Atenção à Saúde/economia , Alocação de Recursos para a Atenção à Saúde/legislação & jurisprudência , Humanos , Lactente , Masculino , Encefalomiopatias Mitocondriais/economia , Cuidados Paliativos , Transferência de Pacientes/economia , Medicina Estatal/economia , Medicina Estatal/ética , Reino UnidoRESUMO
Recent clinical trials have shown that in vivo and ex vivo gene therapy strategies can be an option for the treatment of several neurological disorders. Both strategies require efficient and safe vectors to 1) deliver the therapeutic gene directly into the CNS or 2) to genetically modify stem cells that will be used as Trojan horses for the systemic delivery of the therapeutic protein. A group of target diseases for these therapeutic strategies are mitochondrial encephalopathies due to mutations in nuclear DNA genes. In this study, we have developed a lentiviral vector (CCoq9WP) able to overexpress Coq9 mRNA and COQ9 protein in mouse embryonic fibroblasts (MEFs) and hematopoietic progenitor cells (HPCs) from Coq9R239X mice, an animal model of mitochondrial encephalopathy due to primary Coenzyme Q (CoQ) deficiency. Ectopic over-expression of Coq9 in both cell types restored the CoQ biosynthetic pathway and mitochondrial function, improving the fitness of the transduced cells. These results show the potential of the CCoq9WP lentiviral vector as a tool for gene therapy to treat mitochondrial encephalopathies.
Assuntos
Fibroblastos/metabolismo , Terapia Genética , Células-Tronco Hematopoéticas/metabolismo , Encefalomiopatias Mitocondriais/genética , Proteínas Mitocondriais/genética , Animais , Transplante de Medula Óssea , Modelos Animais de Doenças , Expressão Gênica , Terapia Genética/métodos , Vetores Genéticos/genética , Lentivirus/genética , Camundongos , Camundongos Knockout , Mitocôndrias/genética , Mitocôndrias/metabolismo , Encefalomiopatias Mitocondriais/metabolismo , Encefalomiopatias Mitocondriais/terapia , Proteínas Mitocondriais/metabolismo , Aptidão Física , Transdução Genética , Ubiquinona/biossínteseRESUMO
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a metabolic disorder caused by mutations in TYMP, encoding thymidine phosphorylase (TP). In MNGIE patients, TP dysfunction produces systemic thymidine and deoxyuridine accumulation, which ultimately impairs mitochondrial DNA replication and results in mitochondrial dysfunction. To date, only allogeneic hematopoietic stem cell transplantation has demonstrated long-term clinical efficacy, but high morbidity and mortality associated with this procedure necessitate the search for safer alternatives. In a previous study, we demonstrated that hematopoietic stem cell gene therapy using a lentiviral vector containing the coding sequence of TYMP restored the biochemical homeostasis in an animal model of MNGIE. In the present follow-up study, we show that ectopic expression of TP in the hematopoietic system restores normal nucleoside levels in plasma, as well as in tissues affected in MNGIE such as small intestine, skeletal muscle, brain, and liver. Mitochondrial dNTP pool imbalances observed in liver of the animal model were also corrected by the treatment. The biochemical effects were maintained at least 20 months even with low levels of chimerism. No alterations in the blood cell counts or other toxic effects were observed in association with the lentiviral transduction or TP overexpression. These results further support the notion that gene therapy is a feasible treatment option for MNGIE.
Assuntos
Terapia Genética , Vetores Genéticos/administração & dosagem , Transplante de Células-Tronco Hematopoéticas , Pseudo-Obstrução Intestinal/terapia , Lentivirus/genética , Encefalomiopatias Mitocondriais/terapia , Nucleosídeos/metabolismo , Timidina Fosforilase/genética , Animais , Terapia Combinada , Modelos Animais de Doenças , Feminino , Homeostase , Pseudo-Obstrução Intestinal/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Encefalomiopatias Mitocondriais/genética , Distrofia Muscular Oculofaríngea , Oftalmoplegia/congênitoRESUMO
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a rare autosomal recessive disorder. The mutation in the ECGF1 gene causes severe deficiency of thymidine phosphorylase (TP), which in turn increases thymidine and deoxyuridine in the blood, serum, and tissue. The toxic levels of these products cause malfunction of the mitochondrial respiratory chain and mitochondrial DNA. Commonly, patients become symptomatic between 15 and 20 years of age (range 5 months to 35 years). The most commonly affected systems are gastrointestinal, followed by ocular, and nervous system. The disease is often fatal; high mortality rate is reported between 20 and 40 years of age. Treatment modalities that can increase thymidine phosphorylase activity and decrease thymidine and deoxy-uridine have shown symptomatic improvements in patients with MNGIE. Platelet transfusion, hemodialysis, peritoneal dialysis or allogeneic hematopoietic stem cell transplantation (HSCT) have been tried. The survival and long-term benefits of these measures are still not clear. Engrafted patients after stem cell transplantation have showed improvements in serum thymidine and deoxyuridine. We are reporting a case of MNGIE from Saudi Arabia, who underwent allogeneic hematopoietic stem cell transplantation. No MNGIE case has been previously reported from Saudi Arabia or the Gulf Arab countries. From the available literature, so far only 11 patients with MNGIE have undergone stem cell transplantation.
Assuntos
Encéfalo/patologia , Encefalomiopatias Mitocondriais/patologia , Encefalomiopatias Mitocondriais/terapia , Músculos/patologia , Transplante de Células-Tronco , Adulto , Humanos , Masculino , Encefalomiopatias Mitocondriais/sangue , Encefalomiopatias Mitocondriais/diagnóstico , Transplante de Células-Tronco/métodosRESUMO
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a rare disease caused by thymidine phosphorylase deficiency which leads to toxic accumulations of thymidine (dThd) and deoxyuridine (dUrd). It lacks an established treatment and the prognosis is traditionally poor. We report a case of a young female patient with normal renal function and MNGIE treated by peritoneal dialysis (PD) and allogeneic bone marrow transplantation (BMT). PD was effective in reducing dThd and dUrd plasma levels and in improving clinical symptoms. To our knowledge, this is the first report on the beneficial effects of PD regarding MNGIE neurological symptoms. PD, therefore, should be considered especially in medically compromised patients as a supportive treatment to improve clinical conditions before BMT.
Assuntos
Transplante de Medula Óssea , Pseudo-Obstrução Intestinal/terapia , Encefalomiopatias Mitocondriais/terapia , Diálise Peritoneal , Evolução Fatal , Feminino , Humanos , Pseudo-Obstrução Intestinal/diagnóstico , Encefalomiopatias Mitocondriais/diagnóstico , Distrofia Muscular Oculofaríngea , Oftalmoplegia/congênito , Adulto JovemRESUMO
INTRODUCTION: Mitochondrial neuro-gastrointestinal encephalomyopathy syndrome (MNGIE syndrome) is a rare genetic disorder that is defined clinically by severe gastrointestinal dysmotility, cachexia, peripheral neuropathy, ptosis and/or ophthalmoparesis, and leukoencephalopathy. CASE REPORT: We report a case of a 26-year-old man with MNGIE syndrome with a unique clinical picture consisting of recurrent episodes of spontaneous bowel perforation and multiple intra-abdominal abscesses. The patient was admitted to our hospital several times in the past few years and underwent urgent laparotomies due to perforations in the small bowel. CONCLUSION: Case reports regarding bowel perforations in MNGIE syndrome are scarce and mostly relate to a single perforation and attributed to chronic pseudo-obstruction of the small bowel. To the best of our knowledge, there are no case reports regarding recurrent perforations and abscesses. Surgical management of these cases is challenging as there is no curative option for this genetic disorder. Primary care physicians and treating gastroenterologists should be aware of the potential surgical emergencies associated with this disorder.
Assuntos
Abscesso Abdominal/diagnóstico , Abscesso Abdominal/cirurgia , Gastroenteropatias/diagnóstico , Perfuração Intestinal/diagnóstico , Perfuração Intestinal/cirurgia , Encefalomiopatias Mitocondriais/diagnóstico , Abdome Agudo/diagnóstico , Abdome Agudo/etiologia , Adulto , Diagnóstico Diferencial , Serviço Hospitalar de Emergência , Tratamento de Emergência , Gastroenteropatias/terapia , Humanos , Laparotomia/métodos , Masculino , Encefalomiopatias Mitocondriais/terapia , Doenças Raras , Recidiva , Medição de Risco , Índice de Gravidade de Doença , Síndrome , Tomografia Computadorizada por Raios X/métodosRESUMO
Suicide gene therapy (SGT) is a promising strategy for treating cancer. In this work, we show that thymidine phosphorylase (TP) deficiency, the underlying genetic defect in mitochondrial neurogastrointestinal encephalomyopathy (MNGIE), presents an opportunity to apply SGT using capecitabine, a commonly used prodrug that is converted into 5-fluorouracil by TP. Using an immortalised B-lymphoblastoid cell line from a patient with MNGIE, the tumourigenic EL-4 cell line, lentiviral vectors encoding TP and a double knockout (Tymp(-/-)Upp1(-/-)) murine model, we found that EL-4 cell-derived TP(+) tumours were exquisitely sensitive to capecitabine and generated a significant local bystander effect. In addition, we detected a spontaneous cytolytic immune response in a significant fraction of the animals surviving more than 20 days after termination of the therapy. These data indicate that, in individuals lacking TP expression, TP is a highly specific suicide gene, which can be used to treat tumours that could hypothetically arise in MNGIE patients undergoing gene therapy, as these tumours will likely originate from the gene-modified cells and will be selectively targeted by capecitabine. These observations have important implications for gene therapy for MNGIE.
Assuntos
Genes Transgênicos Suicidas , Terapia Genética/métodos , Pseudo-Obstrução Intestinal/genética , Pseudo-Obstrução Intestinal/terapia , Lentivirus/genética , Encefalomiopatias Mitocondriais/genética , Encefalomiopatias Mitocondriais/terapia , Timidina Fosforilase/metabolismo , Animais , Capecitabina , Linhagem Celular Tumoral , Terapia Combinada , Desoxicitidina/análogos & derivados , Desoxicitidina/metabolismo , Desoxicitidina/uso terapêutico , Modelos Animais de Doenças , Fluoruracila/análogos & derivados , Fluoruracila/metabolismo , Fluoruracila/uso terapêutico , Técnicas de Inativação de Genes , Vetores Genéticos/administração & dosagem , Humanos , Pseudo-Obstrução Intestinal/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Encefalomiopatias Mitocondriais/patologia , Distrofia Muscular Oculofaríngea , Oftalmoplegia/congênito , Timidina Fosforilase/genéticaRESUMO
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is an autosomal recessive disorder caused by mutations in TYMP, enconding thymidine phosphorylase (TP). TP deficiency results in systemic accumulation of thymidine and deoxyuridine, which interferes with mitochondrial DNA (mtDNA) replication and leads to mitochondrial dysfunction. To date, the only treatment available for MNGIE patients is allogeneic hematopoietic stem cell transplantation, which is associated with high morbidity and mortality. Here, we report that AAV2/8-mediated transfer of the human TYMP coding sequence (hcTYMP) under the control of a liver-specific promoter prevents the biochemical imbalances in a murine model of MNGIE. hcTYMP expression was restricted to liver, and a dose as low as 2 × 10(11) genome copies/kg led to a permanent reduction in systemic nucleoside levels to normal values in about 50% of treated mice. Higher doses resulted in reductions to normal or slightly below normal levels in virtually all mice treated. The nucleoside reduction achieved by this treatment prevented deoxycytidine triphosphate (dCTP) depletion, which is the limiting factor affecting mtDNA replication in this disease. These results demonstrate that the use of AAV to direct TYMP expression in liver is feasible as a potentially safe gene therapy strategy for MNGIE.
Assuntos
Terapia Genética , Pseudo-Obstrução Intestinal/genética , Pseudo-Obstrução Intestinal/terapia , Encefalomiopatias Mitocondriais/genética , Encefalomiopatias Mitocondriais/terapia , Timidina Fosforilase/genética , Animais , DNA Mitocondrial/genética , Dependovirus/genética , Modelos Animais de Doenças , Vetores Genéticos , Homeostase/genética , Humanos , Pseudo-Obstrução Intestinal/patologia , Fígado/metabolismo , Camundongos , Encefalomiopatias Mitocondriais/patologia , Distrofia Muscular Oculofaríngea , Mutação , Oftalmoplegia/congênito , Timidina/metabolismo , Timidina Fosforilase/biossínteseRESUMO
Mitochondrial DNA (mtDNA) depletion syndromes (MDS) are a genetically and clinically heterogeneous group of autosomal recessive disorders that are characterized by a severe reduction in mtDNA content leading to impaired energy production in affected tissues and organs. MDS are due to defects in mtDNA maintenance caused by mutations in nuclear genes that function in either mitochondrial nucleotide synthesis (TK2, SUCLA2, SUCLG1, RRM2B, DGUOK, and TYMP) or mtDNA replication (POLG and C10orf2). MDS are phenotypically heterogeneous and usually classified as myopathic, encephalomyopathic, hepatocerebral or neurogastrointestinal. Myopathic MDS, caused by mutations in TK2, usually present before the age of 2 years with hypotonia and muscle weakness. Encephalomyopathic MDS, caused by mutations in SUCLA2, SUCLG1, or RRM2B, typically present during infancy with hypotonia and pronounced neurological features. Hepatocerebral MDS, caused by mutations in DGUOK, MPV17, POLG, or C10orf2, commonly have an early-onset liver dysfunction and neurological involvement. Finally, TYMP mutations have been associated with mitochondrial neurogastrointestinal encephalopathy (MNGIE) disease that typically presents before the age of 20 years with progressive gastrointestinal dysmotility and peripheral neuropathy. Overall, MDS are severe disorders with poor prognosis in the majority of affected individuals. No efficacious therapy is available for any of these disorders. Affected individuals should have a comprehensive evaluation to assess the degree of involvement of different systems. Treatment is directed mainly toward providing symptomatic management. Nutritional modulation and cofactor supplementation may be beneficial. Liver transplantation remains controversial. Finally, stem cell transplantation in MNGIE disease shows promising results.
Assuntos
DNA Mitocondrial/genética , DNA Mitocondrial/fisiologia , Doenças Mitocondriais/genética , Doenças Mitocondriais/fisiopatologia , Doenças do Sistema Nervoso/genética , Doenças do Sistema Nervoso/fisiopatologia , Replicação do DNA/genética , Replicação do DNA/fisiologia , DNA Mitocondrial/metabolismo , Humanos , Transplante de Fígado , Doenças Mitocondriais/terapia , Encefalomiopatias Mitocondriais/genética , Encefalomiopatias Mitocondriais/patologia , Encefalomiopatias Mitocondriais/terapia , Miopatias Mitocondriais/genética , Miopatias Mitocondriais/patologia , Miopatias Mitocondriais/terapia , Mutação/genética , Mutação/fisiologia , Doenças do Sistema Nervoso/terapia , Nucleotídeos/metabolismo , Apoio Nutricional , Timidina/metabolismoRESUMO
Mitochondrial neurogastrointestinal encephalopathy (MNGIE) is caused by deficiency in thymidine phosphorylase (TP), that regulates thymidine (dThd) and deoxyuridine (dUrd). Toxic levels of dThd and dUrd can lead to mitochondrial dysfunction by impairing mitochondrial DNA replication, causing GI and neurologic deterioration. We studied the impact of bone marrow transplant (BMT) and platelets, as a source of TP on the clinical outcome of MNGIE. We report a case of MNGIE, who presented with severe vomiting. Over time, he was non-ambulatory and his GI symptoms got progressively worse with severe dysphagia, abdominal pain episodes, persistent vomiting and diarrhea. Being unfit for intense conditioning regimen, he received a mini BMT, with mild conditioning regimen. Bone marrow was obtained from his HLA fully matched brother. One month after transplantation, donor chimerism in peripheral blood was 33%. Excellent clinical responses were achieved 3 months after transplantation and circulating donor cell chimerism decreased to 24% with a significant increase in platelet TP activity. Ten months post transplant the patient's symptoms recurred and fresh single donor platelets were infused, with a significant increase in platelet TP activity. Mini BMT and platelet transfusion can transiently increase circulating TP activity and might prevent progress of this fatal disease.
Assuntos
Transplante de Medula Óssea , Pseudo-Obstrução Intestinal/terapia , Encefalomiopatias Mitocondriais/terapia , Transfusão de Plaquetas , Condicionamento Pré-Transplante , Adulto , Aloenxertos , Humanos , Pseudo-Obstrução Intestinal/sangue , Masculino , Encefalomiopatias Mitocondriais/sangue , Distrofia Muscular Oculofaríngea , Oftalmoplegia/congênitoRESUMO
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a rare multisystemic autosomal recessive disorder characterized by ptosis, gastrointestinal dysmotility, cachexia, peripheral neuropathy, and leukoencephalopathy. We aimed to raise awareness in radiologists regarding this difficult-to-diagnose syndrome, which occurs in the presence of coexistent gastrointestinal dysmotility, cachexia, and neurologic manifestations. We report imaging and clinical findings of three patients with MNGIE. Our findings indicate that early diagnosis of the disease, together with the timely treatment of acute intercurrent illnesses, may retard the progression of MNGIE.