Modelling urea cycle disorders using iPSCs

The urea cycle is a liver-based pathway enabling disposal of nitrogen waste. Urea cycle disorders (UCDs) are inherited metabolic diseases caused by deficiency of enzymes or transporters involved in the urea cycle and have a prevalence of 1:35,000 live births. Patients present recurrent acute hyperammonaemia, which causes high rate of death and neurological sequelae. Long-term therapy relies on a protein-restricted diet and ammonia scavenger drugs. Currently, liver transplantation is the only cure. Hence, high unmet needs require the identification of effective methods to model these diseases to generate innovative therapeutics. Advances in both induced pluripotent stem cells (iPSCs) and genome editing technologies have provided an invaluable opportunity to model patient-specific phenotypes in vitro by creating patients' avatar models, to investigate the pathophysiology, uncover novel therapeutic targets and provide a platform for drug discovery. This review summarises the progress made thus far in generating 2- and 3-dimensional iPSCs models for UCDs, the challenges encountered and how iPSCs offer future avenues for innovation in developing the next-generation of therapies for UCDs

Inherited and acquired vitamin B12 deficiencies: Which administration route to choose for supplementation?

Vitamin B12 or cobalamin deficiency is a commonly encountered clinical scenario and most clinicians will have familiarity prescribing Vitamin B12 to treat their patients. Despite the high prevalence of this condition, there is widespread heterogeneity regarding routes, schedules and dosages of vitamin B12 administration. In this review, we summarise the complex metabolic pathway of Vitamin B12, the inherited and acquired causes of Vitamin B12 deficiency and subsequently highlight the disparate international practice of prescribing Vitamin B12 replacement therapy. We describe the evidence base underpinning the novel sublingual, intranasal and subcutaneous modes of B12 replacement in comparison to intramuscular and oral routes, with their respective benefits for patient compliance and cost-saving

Gene therapy for Urea Cycle Defects: an update from historical perspectives to future prospects

Urea cycle defects are severe inherited metabolic diseases with high unmet needs, which present a permanent risk of hyperammonaemic decompensation and subsequent acute death or neurological sequelae, when treated with conventional dietetic and medical therapies. Liver transplantation is currently the only curative option, but has the potential to be supplanted by highly effective gene therapy interventions without the attendant need for life-long immunosuppression or limitations imposed by donor liver supply. Over the last three decades, pioneering genetic technologies have been explored to circumvent the consequences of urea cycle defects, improve quality of life and long-term outcome: adenoviral vectors, adeno-associated viral (AAV) vectors, gene editing, genome integration and non-viral technology with messenger RNA (mRNA). In this review, we present a summarised view of this historical path, which include some seminal milestones of the gene therapy's epic. We provide an update about the state of the art of gene therapy technologies for urea cycle defects and the current advantages and pitfalls driving future directions for research and development. This article is protected by copyright. All rights reserved

Prolonged respiratory failure responds to conventional therapy in isolated homocysteine remethylation defects

Isolated remethylation defects are rare inherited diseases caused by a defective remethylation of homocysteine to methionine, preventing various essential methylation reactions to occur. Patients present with a systemic phenotype, which can especially affect the central and peripheral nervous systems leading to epileptic encephalopathy, developmental delay and peripheral neuropathy. Respiratory failure has been described in some cases, caused by both central and peripheral neurological involvement. In published cases, the genetic diagnosis and initiation of appropriate therapy were rapidly performed following respiratory failure and led to a rapid recovery of respiratory insufficiency within days. Here, we present two infantile-onset cases of isolated remethylation defects, cobalamine (Cbl)G and methylenetetrahydrofolate reductase (MTHFR) deficiencies, which were diagnosed after several months of respiratory failure. Disease modifying therapy based on hydroxocobalamin and betaine was initiated and shows a progressive improvement and enabled weaning off respiratory support after 21 and 17 months in CblG and MTHFR patients respectively. We show that prolonged respiratory failure responds to conventional therapy in isolated remethylation defects, but can require a sustained period of time before observing a full response to therapy

Migration of a moonlet in a ring of solid particles : Theory and application to Saturn's propellers

Hundred meter sized objects have been identified by the Cassini spacecraft in Saturn's A ring through the so-called "propeller" features they create in the ring. These moonlets should migrate, due to their gravitational interaction with the ring ; in fact, some orbital variation have been detected. The standard theory of type I migration of planets in protoplanetary disks can't be applied to the ring system, as it is pressureless. Thus, we compute the differential torque felt by a moonlet embedded in a two-dimensional disk of solid particles, with flat surface density profile, both analytically and numerically. We find that the corresponding migration rate is too small to explain the observed variations of the propeller's orbit in Saturn's A-ring. However, local density fluctuations (due to gravity wakes in the marginally gravitationally stable A-ring) may exert a stochastic torque on a moonlet. Our simulations show that this torque can be large enough to account for the observations, depending on the parameters of the rings. We find that on time scales of several years the migration of propellers is likely to be dominated by stochastic effects (while the former, non-stochastic migration dominates after ~ 10^{4-5} years). In that case, the migration rates provided by observations so far suggests that the surface density of the A ring should be of the order of 700 kg/m^2. The age of the propellers shouldn't exceed 1 to 100 million years, depending on the dominant migration regime.Comment: 17 pages, 5 figures, submitted to Astronomical Journal on february, the 23

Natural history of epilepsy in argininosuccinic aciduria provides new insights into pathophysiology: A retrospective international study

OBJECTIVE: Argininosuccinate lyase (ASL) is integral to the urea cycle, which enables nitrogen wasting and biosynthesis of arginine, a precursor of nitric oxide. Inherited ASL deficiency causes argininosuccinic aciduria, the second most common urea cycle defect and an inherited model of systemic nitric oxide deficiency. Patients present with developmental delay, epilepsy, and movement disorder. Here we aim to characterize epilepsy, a common and neurodebilitating comorbidity in argininosuccinic aciduria. METHODS: We conducted a retrospective study in seven tertiary metabolic centers in the UK, Italy, and Canada from 2020 to 2022, to assess the phenotype of epilepsy in argininosuccinic aciduria and correlate it with clinical, biochemical, radiological, and electroencephalographic data. RESULTS: Thirty-seven patients, 1-31 years of age, were included. Twenty-two patients (60%) presented with epilepsy. The median age at epilepsy onset was 24 months. Generalized tonic-clonic and focal seizures were most common in early-onset patients, whereas atypical absences were predominant in late-onset patients. Seventeen patients (77%) required antiseizure medications and six (27%) had pharmacoresistant epilepsy. Patients with epilepsy presented with a severe neurodebilitating disease with higher rates of speech delay (p = .04) and autism spectrum disorders (p = .01) and more frequent arginine supplementation (p = .01) compared to patients without epilepsy. Neonatal seizures were not associated with a higher risk of developing epilepsy. Biomarkers of ureagenesis did not differ between epileptic and non-epileptic patients. Epilepsy onset in early infancy (p = .05) and electroencephalographic background asymmetry (p = .0007) were significant predictors of partially controlled or refractory epilepsy. SIGNIFICANCE: Epilepsy in argininosuccinic aciduria is frequent, polymorphic, and associated with more frequent neurodevelopmental comorbidities. We identified prognostic factors for pharmacoresistance in epilepsy. This study does not support defective ureagenesis as prominent in the pathophysiology of epilepsy but suggests a role of central dopamine deficiency. A role of arginine in epileptogenesis was not supported and warrants further studies to assess the potential arginine neurotoxicity in argininosuccinic aciduria

Liver transplantation in ornithine transcarbamylase deficiency: A retrospective multicentre cohort study

Ornithine transcarbamylase deficiency (OTCD) is an X-linked defect of ureagenesis and the most common urea cycle disorder. Patients present with hyperammonemia causing neurological symptoms, which can lead to coma and death. Liver transplantation (LT) is the only curative therapy, but has several limitations including organ shortage, significant morbidity and requirement of lifelong immunosuppression. This study aims to identify the characteristics and outcomes of patients who underwent LT for OTCD. // We conducted a retrospective study for OTCD patients from 5 UK centres receiving LT in 3 transplantation centres between 2010 and 2022. Patients' demographics, family history, initial presentation, age at LT, graft type and pre- and post-LT clinical, metabolic, and neurocognitive profile were collected from medical records.// A total of 20 OTCD patients (11 males, 9 females) were enrolled in this study. 6/20 had neonatal and 14/20 late-onset presentation. 2/20 patients had positive family history for OTCD and one of them was diagnosed antenatally and received prospective treatment. All patients were managed with standard of care based on protein-restricted diet, ammonia scavengers and supplementation with arginine and/or citrulline before LT. 15/20 patients had neurodevelopmental problems before LT. The indication for LT was presence (or family history) of recurrent metabolic decompensations occurring despite standard medical therapy leading to neurodisability and quality of life impairment. Median age at LT was 10.5 months (6–24) and 66 months (35–156) in neonatal and late onset patients, respectively. 15/20 patients had deceased donor LT (DDLT) and 5/20 had living related donor LT (LDLT). Overall survival was 95% with one patient dying 6 h after LT. 13/20 had complications after LT and 2/20 patients required re-transplantation. All patients discontinued dietary restriction and ammonia scavengers after LT and remained metabolically stable. Patients who had neurodevelopmental problems before LT persisted to have difficulties after LT. 1/5 patients who was reported to have normal neurodevelopment before LT developed behavioural problems after LT, while the remaining 4 maintained their abilities without any reported issues. // LT was found to be effective in correcting the metabolic defect, eliminates the risk of hyperammonemia and prolongs patients' survival

The beta Pictoris system: Setting constraints on the planet and the disk structures at mid-IR wavelengths with NEAR

[abridged] We analyzed mid-infrared high-contrast coronagraphic images of the beta Pictoris system, taking advantage of the NEAR experiment using the VLT/VISIR instrument. The goal of our analysis is to investigate both the detection of the planet beta Pictoris b and of the disk features at mid-IR wavelengths. In addition, by combining several epochs of observation, we expect to constrain the position of the known clumps and improve our knowledge on the dynamics of the disk. To evaluate the planet b flux contribution, we extracted the photometry and compared it to the flux published in the literature. In addition, we used previous data from T-ReCS and VISIR, to study the evolution of the position of the southwest clump that was initially observed in the planetary disk back in 2003. While we did not detect the planet b, we were able to put constraints on the presence of circumplanetary material, ruling out the equivalent of a Saturn-like planetary ring around the planet. The disk presents several noticeable structures, including the known southwest clump. Using a 16-year baseline, sampled with five epochs of observations, we were able to examine the evolution of the clump: the clump orbits in a Keplerian motion with an sma of 56.1+-0.4 au. In addition to the known clump, the images clearly show the presence of a second clump on the northeast side of the disk and fainter and closer structures that are yet to be confirmed. We found correlations between the CO clumps detected with ALMA and the mid-IR images. If the circumplanetary material were located at the Roche radius, the maximum amount of dust determined from the flux upper limit around beta Pictoris b would correspond to the mass of an asteroid of 5 km in diameter. Finally, the Keplerian motion of the southwestern clump is possibly indicative of a yet-to-be-detected planet or signals the presence of a vortex.Comment: Accepted in Astronomy and Astrophysic

Assessment of pre-clinical liver models based on their ability to predict the liver-tropism of AAV vectors

The liver is a prime target for in vivo gene therapies using recombinant adeno-associated viral vectors (rAAV). Multiple clinical trials have been undertaken for this target in the past 15 years, however we are still to see market approval of the first liver-targeted AAV-based gene therapy. Inefficient expression of the therapeutic transgene, vector-induced liver toxicity and capsid, and/or transgene-mediated immune responses reported at high vector doses are the main challenges to date. One of the contributing factors to the insufficient clinical outcomes, despite highly encouraging preclinical data, is the lack of robust, biologically- and clinically-predictive preclinical models. To this end, this study reports findings of a functional evaluation of six AAV vectors in twelve preclinical models of the human liver, with the aim to uncover which combination of models is the most relevant for the identification of AAV capsid variant for safe and efficient transgene delivery to primary human hepatocytes. The results, generated by studies in models ranging from immortalized cells, iPSC-derived and primary hepatocytes, and primary human hepatic organoids to in vivo models, increased our understanding of the strengths and weaknesses of each system. This should allow the development of novel gene therapies targeting the human liver