Maria Grazia Natali Sora

Lentiviral haemopoietic stem-cell gene therapy in early-onset metachromatic leukodystrophy: an ad-hoc analysis of a non-randomised, open-label, phase 1/2 trial

BACKGROUND Metachromatic leukodystrophy (a deficiency of arylsulfatase A [ARSA]) is a fatal demyelinating lysosomal disease with no approved treatment. We aimed to assess the long-term outcomes in a cohort of patients with early-onset metachromatic leukodystrophy who underwent haemopoietic stem-cell gene therapy (HSC-GT). METHODS This is an ad-hoc analysis of data from an ongoing, non-randomised, open-label, single-arm phase 1/2 trial, in which we enrolled patients with a molecular and biochemical diagnosis of metachromatic leukodystrophy (presymptomatic late-infantile or early-juvenile disease or early-symptomatic early-juvenile disease) at the Paediatric Clinical Research Unit, Ospedale San Raffaele, in Milan. Trial participants received HSC-GT, which consisted of the infusion of autologous HSCs transduced with a lentiviral vector encoding ARSA cDNA, after exposure-targeted busulfan conditioning. The primary endpoints of the trial are safety (toxicity, absence of engraftment failure or delayed haematological reconstitution, and safety of lentiviral vector-tranduced cell infusion) and efficacy (improvement in Gross Motor Function Measure [GMFM] score relative to untreated historical controls, and ARSA activity, 24 months post-treatment) of HSC-GT. For this ad-hoc analysis, we assessed safety and efficacy outcomes in all patients who had received treatment and been followed up for at least 18 months post-treatment on June 1, 2015. This trial is registered with ClinicalTrials.gov, number NCT01560182. FINDINGS Between April, 2010, and February, 2013, we had enrolled nine children with a diagnosis of early-onset disease (six had late-infantile disease, two had early-juvenile disease, and one had early-onset disease that could not be definitively classified). At the time of analysis all children had survived, with a median follow-up of 36 months (range 18-54). The most commonly reported adverse events were cytopenia (reported in all patients) and mucositis of different grades of severity (in five of nine patients [grade 3 in four of five patients]). No serious adverse events related to the medicinal product were reported. Stable, sustained engraftment of gene-corrected HSCs was observed (a median of 60·4% [range 14·0-95·6] lentiviral vector-positive colony-forming cells across follow-up) and the engraftment level was stable during follow-up; engraftment determinants included the duration of absolute neutropenia and the vector copy number of the medicinal product. A progressive reconstitution of ARSA activity in circulating haemopoietic cells and in the cerebrospinal fluid was documented in all patients in association with a reduction of the storage material in peripheral nerve samples in six of seven patients. Eight patients, seven of whom received treatment when presymptomatic, had prevention of disease onset or halted disease progression as per clinical and instrumental assessment, compared with historical untreated control patients with early-onset disease. GMFM scores for six patients up to the last follow-up showed that gross motor performance was similar to that of normally developing children. The extent of benefit appeared to be influenced by the interval between HSC-GT and the expected time of disease onset. Treatment resulted in protection from CNS demyelination in eight patients and, in at least three patients, amelioration of peripheral nervous system abnormalities, with signs of remyelination at both sites. INTERPRETATION Our ad-hoc findings provide preliminary evidence of safety and therapeutic benefit of HSC-GT in patients with early-onset metachromatic leukodystrophy who received treatment in the presymptomatic or very early-symptomatic stage. The results of this trial will be reported when all 20 patients have achieved 3 years of follow-up. FUNDING Italian Telethon Foundation and GlaxoSmithKline.

Intra-arterial transplantation of HLA-matched donor mesoangioblasts in Duchenne muscular dystrophy

Intra‐arterial transplantation of mesoangioblasts proved safe and partially efficacious in preclinical models of muscular dystrophy. We now report the first‐in‐human, exploratory, non‐randomized open‐label phase I–IIa clinical trial of intra‐arterial HLA‐matched donor cell transplantation in 5 Duchenne patients. We administered escalating doses of donor‐derived mesoangioblasts in limb arteries under immunosuppressive therapy (tacrolimus). Four consecutive infusions were performed at 2‐month intervals, preceded and followed by clinical, laboratory, and muscular MRI analyses. Two months after the last infusion, a muscle biopsy was performed. Safety was the primary endpoint. The study was relatively safe: One patient developed a thalamic stroke with no clinical consequences and whose correlation with mesoangioblast infusion remained unclear. MRI documented the progression of the disease in 4/5 patients. Functional measures were transiently stabilized in 2/3 ambulant patients, but no functional improvements were observed. Low level of donor DNA was detected in muscle biopsies of 4/5 patients and donor‐derived dystrophin in 1. Intra‐arterial transplantation of donor mesoangioblasts in human proved to be feasible and relatively safe. Future implementation of the protocol, together with a younger age of patients, will be needed to approach efficacy.

Analyzing Histopathological Features of Rare Charcot-Marie-Tooth Neuropathies to Unravel Their Pathogenesis

BACKGROUND Charcot-Marie-Tooth (CMT) neuropathies are very heterogeneous disorders from both a clinical and genetic point of view. The CMT genes identified so far encode different proteins that are variably involved in regulating Schwann cells and/or axonal functions. However, the function of most of these proteins still remains to be elucidated. OBJECTIVE To characterize a large cohort of patients with demyelinating, axonal, and intermediate forms of CMT neuropathy. DESIGN A cohort of 131 unrelated patients were screened for mutations in 12 genes responsible for CMT neuropathies. Demyelinating, axonal, and intermediate forms of CMT neuropathy were initially distinguished as usual on the basis of electrophysiological criteria and clinical evaluation. A sural nerve biopsy was also performed for selected cases. Accordingly, patients underwent first-level analysis of the genes most frequently mutated in each clinical form of CMT neuropathy. RESULTS Although our cohort had a particularly high percentage of cases of rare axonal and intermediate CMT neuropathies, we found mutations in 40% of patients. Among identified changes, 7 represented new mutations occurring in the MPZ, GJB1, EGR2, MFN2, NEFL, and HSBP1/HSP27 genes. Histopathological analysis performed in selected cases revealed morphological features, which correlated with the molecular diagnosis and provided evidence of the underlying pathogenetic mechanism. CONCLUSION Clinical and pathological analysis of patients with CMT neuropathies contributes to our understanding of the molecular mechanisms of CMT neuropathies.

Paths of cognitive and language development in healthy preterm infants

OBJECTIVE Despite the presence of many studies on difficulties related to premature birth, findings on developmental outcomes are heterogeneous. This could be explained from a biological and environmental point of view, but also from a methodological one. The aims of this study were as follows: assess cognitive and linguistic performance using the BSID-III in a population of healthy preterm infants at 24 and 36 months (corrected age); analyze whether the correction for prematurity should be applied, decide when to stop using corrected age and evaluate possible improvements between 24 and 36 months. METHODS Developmental outcome was assessed at 24 and 36 months (corrected age) with the BSID-III in 75 healthy preterm (GA=32.5±1.97; BW=1631.55±453.92) and 69 term-born children (GA=39.77±1.00; BW=3298.95±457.27). RESULTS Preterm infants had significantly lower scores than those of term infants in Cognitive (COG) and Language (LANG REC, LANG EC) scales of the BSID-III at both 24 and 36 months, considering both corrected (CA) and chronological (UCA) age. At 24 months, significant differences between corrected and chronological scores were found for each BSID-III scale, while at 36 months, significant differences between corrected and chronological scores were found for LANG scales. Only the scores in the COG scale were statistically different between 24 and 36 months (F=4.894, P=0.009, η(2)=0.075). Considering only the preterm sample at 24 months, the differences between CA and UCA scores in the COG scale were significantly correlated to GA (p=0.000) and days in hospital (p=0.002;), while differences between CA and UCA scores in the LANG ESP scale were significantly correlated to GA (p=0.010), days in hospital (p=0.001), and birth weight (p=0.007). At 36 months, no significant correlations were found. CONCLUSIONS Preterm birth is followed by poorer cognitive and language outcomes during infancy than full-term birth. Age correction of prematurity is useful if the child is under 2 years of age; however, our findings raise concerns about the need for age correction, considering that at later ages, healthy preterm children have a higher rate of developmental delay compared with term infants. With regard to cognitive development, preterm children seem to recover from their initial disadvantage; however, with regard to linguistic development, data confirm that preterm infants are at risk for language difficulties.

Longitudinal MRI quantification of muscle degeneration in Duchenne muscular dystrophy

The aim of this study was to evaluate the usefulness of magnetic resonance imaging (MRI) in detecting the progression of Duchenne muscular dystrophy (DMD) by quantification of fat infiltration (FI) and muscle volume index (MVI, a residual‐to‐total muscle volume ratio).

Use of Defibrotide to help prevent post-transplant endothelial injury in a genetically predisposed infant with metachromatic leukodystrophy undergoing hematopoietic stem cell gene therapy

Valeria Calbi ● Francesca Fumagalli ● Giulia Consiglieri ● Rachele Penati ● Serena Acquati ● Daniela Redaelli ● Vanessa Attanasio ● Facchini Marcella ● Maria Pia Cicalese ● Maddalena Migliavacca ● Federica Barzaghi ● Francesca Ferrua ● Andrea Assanelli ● Paolo Silvani ● Matteo Zoccolillo ● Giovanna Chidini ● Robert Chiesa ● Ruchi Arora ● Francesca Ciotti ● Marina Sarzana ● Gigliola Antonioli ● Cristina Baldoli ● Francesco Morena ● Sabata Martino ● Gian Luigi Ardissino ● Maria Grazia Natali Sora ● Luigi Naldini ● Fabio Ciceri ● Alessandro Aiuti ● Maria Ester Bernardo

Quantitative analysis of heart rate variability signal in diabetic subjects

An automatic procedure is presented for processing heart rate variability (HRV, taken from the ECG) and respiration of diabetic subjects with or without neuropathy. Spectral analysis is carried out using autospectra, cross-spectra, and coherence parametric methods (based on autoregressive modeling). Spectral parameters, and in particular the power associated with low-frequency and high-frequency bands, as well as the contribution of the HRV spectrum coherent with respiration, seem to discriminate satisfactorily between diabetic subjects with and without neuropathy. These results were obtained from the control population when resting/standing and when respiration was controlled. This fact reflects the impact of neural control systems on heart rate and respiration.<<ETX>>

Primary progressive multiple sclerosis presenting with severe predominant cognitive impairment and psychiatric symptoms: A challenging case

Severe cognitive dysfunction is a frequent feature of multiple sclerosis (MS), normally associated with later stages of the disease in adult population. Nevertheless, progressive cognitive and neuropsychiatric disturbances might rarely be the presenting and predominant symptom. In order to better characterize this peculiar phenotype of MS, we report on the case of a 38-year-old man who referred to our hospital with the suspect of hereditary leukodystrophy after 5 years of behavioral and mood abnormalities, global cognitive dysfunction, clumsiness, and very mild pyramidal and cerebellar signs. Brain and spinal magnetic resonance imaging (MRI) combined with cerebrospinal fluid (CSF) analysis prompted the diagnosis of MS.

Vocal cord paralysis in Charcot–Marie–Tooth type 4b1 disease associated with a novel mutation in the myotubularin-related protein 2 gene: A case report and review of the literature

Highlights • Vocal cord paralysis is a relevant symptom of Charcot–Marie–Tooth type 4B1.• Patients harboring MTMR2 mutations should be investigated for laryngeal function.• A new mutation in the MTMR2 gene is described.• The frequency of vocal cord paralysis in early-onset CMT subtypes is explored.