RESUMEN
BACKGROUND: X-linked agammaglobulinemia (XLA) is an inborn error of immunity that renders boys susceptible to life-threatening infections due to loss of mature B cells and circulating immunoglobulins. It is caused by defects in the gene encoding the Bruton tyrosine kinase (BTK) that mediates the maturation of B cells in the bone marrow and their activation in the periphery. This paper reports on a gene editing protocol to achieve "knock-in" of a therapeutic BTK cassette in hematopoietic stem and progenitor cells (HSPCs) as a treatment for XLA. METHODS: To rescue BTK expression, this study employed a clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 system that creates a DNA double-strand break in an early exon of the BTK locus and an adeno-associated virus 6 virus that carries the donor template for homology-directed repair. The investigators evaluated the efficacy of the gene editing approach in HSPCs from patients with XLA that were cultured in vitro under B-cell differentiation conditions or that were transplanted in immunodeficient mice to study B-cell output in vivo. RESULTS: A (feeder-free) B-cell differentiation protocol was successfully applied to blood-mobilized HSPCs to reproduce in vitro the defects in B-cell maturation observed in patients with XLA. Using this system, the investigators could show the rescue of B-cell maturation by gene editing. Transplantation of edited XLA HSPCs into immunodeficient mice led to restoration of the human B-cell lineage compartment in the bone marrow and immunoglobulin production in the periphery. CONCLUSIONS: Gene editing efficiencies above 30% could be consistently achieved in human HSPCs. Given the potential selective advantage of corrected cells, as suggested by skewed X-linked inactivation in carrier females and by competitive repopulating experiments in mouse models, this work demonstrates the potential of this strategy as a future definitive therapy for XLA.
Asunto(s)
Agammaglobulinemia Tirosina Quinasa , Agammaglobulinemia , Linfocitos B , Edición Génica , Enfermedades Genéticas Ligadas al Cromosoma X , Células Madre Hematopoyéticas , Agammaglobulinemia/genética , Agammaglobulinemia/terapia , Agammaglobulinemia/inmunología , Animales , Agammaglobulinemia Tirosina Quinasa/genética , Enfermedades Genéticas Ligadas al Cromosoma X/genética , Enfermedades Genéticas Ligadas al Cromosoma X/terapia , Enfermedades Genéticas Ligadas al Cromosoma X/inmunología , Humanos , Linfocitos B/inmunología , Ratones , Masculino , Trasplante de Células Madre Hematopoyéticas , Diferenciación Celular/genética , Sistemas CRISPR-CasRESUMEN
Replacing a faulty gene with a correct copy has become a viable therapeutic option as a result of recent progress in gene editing protocols. Targeted integration of therapeutic genes in hematopoietic stem cells has been achieved for multiple genes using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system and Adeno-Associated Virus (AAV) to carry a donor template. Although this is a promising strategy to correct genetic blood disorders, it is associated with toxicity and loss of function in CD34+ hematopoietic stem and progenitor cells, which has hampered clinical application. Balancing the maximum achievable correction against deleterious effects on the cells is critical. However, multiple factors are known to contribute, and the optimization process is laborious and not always clearly defined. We have developed a flexible multidimensional Response Surface Methodology approach for optimization of gene correction. Using this approach, we could rapidly investigate and select editing conditions for CD34+ cells with the best possible balance between correction and cell/colony-forming unit (CFU) loss in a parsimonious one-shot experiment. This method revealed that using relatively low doses of AAV2/6 and CRISPR/Cas9 ribonucleoprotein complex, we can preserve the fitness of CD34+ cells and, at the same time, achieve high levels of targeted gene insertion. We then used these optimized editing conditions for the correction of p67phox-deficient chronic granulomatous disease (CGD), an autosomal recessive disorder of blood phagocytic cells resulting in severe recurrent bacterial and fungal infections and achieved rescue of p67phox expression and functional correction of CD34+-derived neutrophils from a CGD patient.
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Enfermedad Granulomatosa Crónica , Humanos , Enfermedad Granulomatosa Crónica/genética , Enfermedad Granulomatosa Crónica/terapia , Edición Génica , Terapia Genética/métodos , Antígenos CD34/genética , Células Madre Hematopoyéticas/metabolismo , Sistemas CRISPR-CasAsunto(s)
Síndromes de Inmunodeficiencia , Enfermedades de Inmunodeficiencia Primaria , Adulto , Humanos , Enfermedades de Inmunodeficiencia Primaria/diagnóstico , Síndromes de Inmunodeficiencia/complicaciones , Síndromes de Inmunodeficiencia/diagnóstico , Síndromes de Inmunodeficiencia/genética , Quinasas Asociadas a Receptores de Interleucina-1/genéticaRESUMEN
In recent years, gene-editing technologies have revolutionised precision medicine, and human trials of this technology have been reported in cell-based cancer therapies and other genetic disorders. The same techniques have the potential to reverse mutations in monogenic primary immunodeficiencies (PIDs), and transplantation of edited haematopoietic stem cells may provide a functional cure for these diseases. In this review, we discuss the methods of gene editing being explored and describe progress made so far with several PIDs. We also detail the remaining challenges, how to confidently detect off-target effects and chromosomal abnormalities in a timely manner, how to obtain long-term benefits, and how to achieve physiological levels of expression of the therapeutic gene. With advances in gene editing, we envisage a robust clinical translation of this technology in the coming decade.
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Edición Génica , Células Madre Hematopoyéticas , Humanos , Edición Génica/métodos , Terapia Genética/métodosRESUMEN
In March 2020, the United Kingdom Primary Immunodeficiency Network (UKPIN) established a registry of cases to collate the outcomes of individuals with PID and SID following SARS-CoV-2 infection and treatment. A total of 310 cases of SARS-CoV-2 infection in individuals with PID or SID have now been reported in the UK. The overall mortality within the cohort was 17.7% (n = 55/310). Individuals with CVID demonstrated an infection fatality rate (IFR) of 18.3% (n = 17/93), individuals with PID receiving IgRT had an IFR of 16.3% (n = 26/159) and individuals with SID, an IFR of 27.2% (n = 25/92). Individuals with PID and SID had higher inpatient mortality and died at a younger age than the general population. Increasing age, low pre-SARS-CoV-2 infection lymphocyte count and the presence of common co-morbidities increased the risk of mortality in PID. Access to specific COVID-19 treatments in this cohort was limited: only 22.9% (n = 33/144) of patients admitted to the hospital received dexamethasone, remdesivir, an anti-SARS-CoV-2 antibody-based therapeutic (e.g. REGN-COV2 or convalescent plasma) or tocilizumab as a monotherapy or in combination. Dexamethasone, remdesivir, and anti-SARS-CoV-2 antibody-based therapeutics appeared efficacious in PID and SID. Compared to the general population, individuals with PID or SID are at high risk of mortality following SARS-CoV-2 infection. Increasing age, low baseline lymphocyte count, and the presence of co-morbidities are additional risk factors for poor outcome in this cohort.
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Tratamiento Farmacológico de COVID-19 , COVID-19 , Síndromes de Inmunodeficiencia , Humanos , Anticuerpos Monoclonales Humanizados , Anticuerpos Neutralizantes , Anticuerpos Antivirales , COVID-19/terapia , Sueroterapia para COVID-19 , Dexametasona , Combinación de Medicamentos , Inmunización Pasiva , SARS-CoV-2 , Reino Unido/epidemiologíaRESUMEN
Allergic reactions frequently involve the production of immunoglobulin E (IgE) antibodies to proteins. However, reactions directed against carbohydrate moieties are increasingly being recognised. Tick bites can contribute to the development of immunoglobulin E to the galactose-1,3-galactose (alpha-gal) moiety on tick salivary proteins. These IgE molecules can cross-react with alpha-gal found in red meats, causing Type I IgE-mediated hypersensitivity reactions to these foods. We present three cases of delayed reactions to beef, pork and lamb in patients with prior tick bites and in the presence of a positive-specific IgE to alpha-gal. Patients were advised to avoid red meat consumption and to carry emergency treatment in the form of anti-histamines with or without adrenaline autoinjector devices. This is the first published report of red meat allergy caused by tick bites suffered in the UK.
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Infecciones por Burkholderia/diagnóstico , Infecciones por Burkholderia/etiología , Enfermedad Granulomatosa Crónica/complicaciones , Prostatitis/diagnóstico , Prostatitis/etiología , Adulto , Antibacterianos/uso terapéutico , Biomarcadores , Infecciones por Burkholderia/tratamiento farmacológico , Manejo de la Enfermedad , Susceptibilidad a Enfermedades , Humanos , Imagen por Resonancia Magnética , Masculino , Prostatitis/tratamiento farmacológico , Evaluación de SíntomasAsunto(s)
Anestesia General/efectos adversos , Anestésicos Generales/efectos adversos , Hipersensibilidad a las Drogas/diagnóstico , Pruebas Inmunológicas , Anciano , Anestésicos Generales/inmunología , Hipersensibilidad a las Drogas/inmunología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Estudios Retrospectivos , Factores de RiesgoRESUMEN
We present the case of a 19-year-old female with a mild form of Autosomal Dominant Hyper IgE syndrome (HIES) associated with a loss-of-function mutation in STAT3. Within the first years of life she developed multiple, Staphylococcus aureus associated abscesses in the neck and face requiring frequent incision and drainage. Respiratory tract infections were not a feature of the clinical phenotype and a high resolution thoracic CT scan was unremarkable. Retained dentition was noted but fungal nail disease and recurrent thrush were absent. The total IgE was 970 IU/L, Lymphocyte counts and immunoglobulin levels were normal (IgG borderline 18.5 gr/L). There was suboptimal response to test immunisation with Pneumovax II vaccine. Th17 cell phenotyping revealed low levels of IL-17 expressing cells (0.3% of total CD4 T Cells numbers). Genetic analysis identified a missense mutation, N567D, in a conserved region of the linker domain of STAT3. Functional studies in HEK293 cells reveal that this mutation potently inhibits STAT3 activity when compared to the wildtype protein. This is consistent with other reported mutations in STAT3 associated with HIES. However, surprisingly, the magnitude of inhibition was similar to another STAT3 mutation (V637M) which causes a much more severe form of the disease.
RESUMEN
Background: Chronic spontaneous urticaria (CSU) is a condition characterised by the presence of hives with/without angioedema, that affects individuals on more days than not for 6 weeks or more. The role of infection as a potential trigger for CSU is well described, but the current clinical guidelines do not recommend routine screening for underlying infections. Main observations: We report a case of severe prolonged chronic spontaneous urticaria in a 19-year-old, that went into rapid remission following the treatment of dental infection. Conclusions: Clinicians should recognise the potential role that infection can have in causing chronic urticaria. There should be a low threshold to treat infection in such circumstances.
Asunto(s)
Angioedema , Urticaria , Enfermedad Crónica , Antagonistas de los Receptores Histamínicos , Humanos , Enfermedades Maxilomandibulares , Masculino , Omalizumab , Adulto JovenRESUMEN
A 54-year-old man with end-stage renal failure attended for dialysis. Within seconds of applying 2% w/v chlorhexidine (ChloraPrep 3 mL Wand Applicator) to the skin surrounding the insertion point of his dialysis catheter (Tesio catheter), he developed pruritus, urticaria, shortness of breath, hypotension and reduced responsiveness. Treatment for anaphylaxis was initiated with rapid improvement of his symptoms, and he made a full recovery. Allergy to chlorhexidine was confirmed with skin testing, and the patient was warned against all future exposure to chlorhexidine. Subsequent dialysis without chlorhexidine was uneventful.
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Anafilaxia/inducido químicamente , Clorhexidina/efectos adversos , Desinfectantes/efectos adversos , Fallo Renal Crónico/terapia , Administración Cutánea , Clorhexidina/administración & dosificación , Desinfectantes/administración & dosificación , Humanos , Masculino , Persona de Mediana Edad , Diálisis RenalRESUMEN
BACKGROUND: Different gastrojejunal anastomotic (GJA) techniques have been described in laparoscopic Roux-en-Y gastric bypass (LRYGB). There is conflicting data on whether one technique is superior to the other. We aimed to compare hand-sewn (HSA), circular-stapled (CSA) and linear-stapled (LSA) anastomotic techniques in terms of stricture rates and their impact on subsequent weight loss. METHODS: A prospectively collected database was used to identify patients undergoing LRYGB surgery between March 2005 and May 2012. Anastomotic technique (HSA, CSA, LSA) was performed according to individual surgeon preference. The database recorded patient demographics, relevant comorbidities and the type of GJA performed. Serial weight measurements and percentage excess weight loss (%EWL) were available at defined follow-up intervals. RESULTS: Included in the data were 426 patients, divided between HSA (n = 174, 40.8%), CSA (n = 110, 25.8%) and LSA (n = 142, 33.3%). There was no significant difference in the stricture rates (HSA n = 17, 9.72%; CSA n = 9, 8.18%; LSA n = 8, 5.63%; p = 0.4006). Weight loss was similar between the three techniques (HSA, CSA and LSA) at 3 months (40.6% ± 16.2% vs 35.92% ± 21.42% vs 48.21 % ± 14.79%; p = 0.0821), 6 months (61.48% ± 23.94% vs 58.16 % ± 27.31% vs 60.18% ± 22.26%; p = 0.2296), 12 months (72.94% ± 19.93% vs 69.72 ± 21.42% vs 66.05% ± 17.75%; p = 0.0617) and 24 months (73.29% ± 22.31% vs 68.75 % ± 24.71% vs 69.40% ± 23.10%; p = 0.7242), respectively. The stricture group lost significantly greater weight (%EWL) within the first 3 months compared to the non-stricture group (45.39% ± 16.82 % vs 39.22 % ± 21.93%; p = 0.0340); however, this difference had resolved at 6 months (61.29% ± 18.50 % vs 59.79% ± 23.03%; p = 0.8802) and 12 months (71.59 % ± 18.67 % vs 68.69 % ± 22.19 %; p = 0.5970). CONCLUSIONS: There was no significant difference in the rate of strictures between the three techniques, although the linear technique appears to have the lowest requirement for post-operative dilatation. The re-intervention rate will, in part, be dictated by the threshold for endoscopy, which will vary between units. Weight loss was similar between the three anastomotic techniques. Surgeons should use techniques that they are most familiar with, as stricture and weight loss rates are not significantly different.