Francesc Carmona

Microarray expression analysis in idiopathic and LRRK2-associated Parkinson's disease

LRRK2 mutations are the most common genetic cause of Parkinsons disease (PD). We performed a whole-genome RNA profiling of putamen tissue from idiopathic PD (IPD), LRRK2-associated PD (G2019S mutation), neurologically healthy controls and one asymptomatic LRRK2 mutation carrier, by using the Genechip Human Exon 1.0-ST Array. The differentially expressed genes found in IPD revealed an alteration of biological pathways related to long-term potentiation (LTP), GABA receptor signalling, and calcium signalling pathways, among others. These pathways are mainly related with cell signalling cascades and synaptic plasticity processes. They were also altered in the asymptomatic LRRK2 mutation carrier but not in the LRRK2-associated PD group. The expression changes seen in IPD might be attributed to an adaptive consequence of a dysfunction in the dopamine transmission. The lack of these altered molecular pathways in LRRK2-associated PD patients suggests that these cases could show a different molecular response to dopamine transmission impairment.

Cellular transplants in amyotrophic lateral sclerosis patients: an observational study.

BACKGROUND AIMS Cytotherapy is a promising option for neurodegenerative disease treatment. Because of the fatal prognosis and imperative need for effective treatment, amyotrophic lateral sclerosis (ALS) patients request this therapy before its effectiveness has been verified. The increase in clinics offering cytotherapies but providing little scientific information has prompted considerable medical tourism. We present an observational study of Spanish ALS patients receiving cytotherapy, analyzing the experiences arising from the treatment (TX) and considering two progression markers, FVC and ALSFRS-R. METHODS Twelve ALS patients with a mean age of 48.6 years (SD 12.8) received cytotherapy 26.9 months (SD 15.8) after clinical onset. ALSFRS-R and FVC at TX were 32.3 (SD 6.8) and 63.4% (SD 15.3), respectively. TX involved transplants of olfactory ensheathing cells in three patients, and autologous mesenchymal stromal cells in the remainder. RESULTS One patient died 33 months post-TX after surviving for 49 months. Five required mechanical non-invasive home ventilation 7.4 months post-TX. Two required invasive ventilation 13 months post-TX. Five patients needed gastrostomy feeding 23.3 months post-TX. Survival between clinical onset and the study end date was 50 months (SD 17.2). No significant adverse events or changes in the decline of FVC and ALSFRS-R compared with the diseases natural history were observed. CONCLUSIONS Our observations suggest that these therapies do not halt the course of the disease. Cytotherapy cannot yet be considered a curative treatment for ALS.

Altered Clock Gene Expression in Obese Visceral Adipose Tissue Is Associated with Metabolic Syndrome

Clock gene expression was associated with different components of metabolic syndrome (MS) in human adipose tissue. However, no study has been done to compare the expression of clock genes in visceral adipose tissue (VAT) from lean and obese subjects and its clinical implications. Therefore, we studied in lean and obese women the endogenous 24 h expression of clock genes in isolated adipocytes and its association with MS components. VAT was obtained from lean (BMI 21–25 kg/m2; n = 21) and morbidly obese women (BMI >40 kg/m2; n = 28). The 24 h pattern of clock genes was analyzed every 6 hours using RT-PCR. Correlation of clinical data was studied by Spearman analysis. The 24 h pattern of clock genes showed that obesity alters the expression of CLOCK, BMAL1, PER1, CRY2 and REV-ERB ALPHA in adipocytes with changes found in CRY2 and REV-ERB ALPHA throughout the 24 h period. The same results were confirmed in VAT and stromal cells (SC) showing an upregulation of CRY2 and REV-ERB ALPHA from obese women. A positive correlation was observed for REV-ERB ALPHA gene expression with BMI and waist circumference in the obese population. Expression of ROR ALPHA was correlated with HDL levels and CLOCK with LDL. Obese subjects with MS exhibited positive correlation in the PER2 gene with LDL cholesterol, whereas REV-ERB ALPHA was correlated with waist circumference. We identified CRY2 and REV-ERB ALPHA as the clock genes upregulated in obesity during the 24 h period and that REV-ERB ALPHA is an important gene associated with MS.

Brain transcriptomic profiling in idiopathic and LRRK2-associated Parkinson's disease

LRRK2 mutations are the most common genetic cause of Parkinsons disease (PD). We performed a whole-genome RNA profiling of locus coeruleus post-mortem tissue, a histopathologically affected brain tissue in PD, from idiopathic PD (IPD) and LRRK2-associated PD patients. The differentially expressed genes found in IPD and LRRK2-associated PD are involved in the gene ontology terms of synaptic transmission and neuron projection. In addition, differentially expressed genes in the IPD group are associated with immune system related pathways. Specifically, the study performed highlights the presence of differential expression of genes located in the chromosome 6p21.3 belonging to the class II HLA. Our findings support the hypothesis of a potential role of neuroinflammation and the involvement of the HLA genetic area in IPD pathogenesis. Future studies are necessary to shed light on the relation of immune system related pathways in the etiopathogenesis of PD.

Does reduced [123I]-FP-CIT binding in Huntington's disease suggest pre-synaptic dopaminergic involvement?

OBJECTIVE To evaluate the usefulness of SPECT in assessing damage to the pre-synaptic dopaminergic system in Huntingtons disease (HD) using [(123)I]-FP-CIT (DaTSCAN), a selective radioligand with regulatory approval as the diagnostic test for investigating functional dopaminergic neuron loss in the striatum in Parkinsons disease. METHODS We studied twelve symptomatic HD patients using DaTSCAN/SPECT imaging. [(123)I]-FP-CIT caudate and putamen uptake levels were qualitatively and semi-quantitatively analyzed to assess pre-synaptic damage in the striatal dopamine system. Possible correlations were analyzed between HD severity on the Unified Huntingtons Disease Rating Scale (UHDRS), duration of clinical symptoms, and [(123)I]-FP-CIT/SPECT striatal uptake. RESULTS DaTSCAN/SPECT qualitative analysis showed reduced striatal uptake in eight patients. Semi-quantitative analysis revealed a significant reduction in four. Of these four, uptake reduction was at putamen level in all, and also at caudate level in one. Although we observed no linear correlation between HD severity and reduced striatal [(123)I]-FP-CIT uptake, the patients with the worst UHDRS scores had more severe reductions in radioligand uptake. CONCLUSION This is the first study to use in vivo [(123)I]-FP-CIT/SPECT imaging to confirm prior descriptions using PET of a pre-synaptic dopaminergic system defect in HD.

Progressive presynaptic dopaminergic deterioration in Huntington disease: a [123I]-FP-CIT SPECT two-year follow-up study.

To illustrate the potential of [I]-FP-CIT SPECT DaTSCAN® in investigating the progression of presynaptic dopaminergic degeneration in Huntington disease (HD), we performed a 2-year follow-up [I]-FP-CIT study on 4 HD patients, evaluating the SPECT imaging based on qualitative and semiquantitative analysis. The mean annual decline in [I]-FP-CIT uptake in caudate and putamen after 2 years of follow-up was 5.8% and 9.6%, respectively. Our findings suggest that [I]-FP-CIT SPECT is useful in investigating the progression of presynaptic dopaminergic degeneration in HD, and may be useful as a disease biomarker, providing an objective method for measuring the effectiveness of future neuroprotective therapies.

TLR-activated conventional DCs promote γ-secretase-mediated conditioning of plasmacytoid DCs

Cooperative events between DC subsets involve cell contact and soluble factors. Upon viral challenge, murine pDCs induce cDC cooperation through CD40‐CD40L interactions and IL‐15 secretion, whereas in humans, the same effect is mediated by IFN‐α. Conversely, during bacterial infections, pDC maturation may be induced by activated cDCs, although no mechanisms had been described so far. Here, we investigate how human pDCs are “conditioned” by cDCs. Blood‐borne DC subsets (cDCs and pDCs) were sorted from healthy donors. IL‐3‐maintained pDCs were cocultured with LPS‐activated, poly (I:C)‐activated, or control cDCs [cDCLPS, cDCP(I:C), cDCCTRL]. Coculture experiments showed that cDCLPS‐conditioned pDCs up‐regulated maturation markers, such as CD25 and CD86, whereas SNs contained higher amounts of IL‐6 and CCL19 compared with control conditions. Gene‐expression analyses on sorted cDCLPS or cDCP(I:C) conditioned pDCs confirmed the induction of several genes, including IL‐6 and CCL19 and remarkably, several Notch target genes. Further studies using the γ‐secretase/Notch inhibitor DAPT and soluble Notch ligands resulted in a significantly reduced expression of canonical Notch target genes in conditioned pDCs. DAPT treatment also hampered the secretion of CCL19 (but not of IL‐6) by cDCLPS conditioned pDCs. These results reveal the involvement of γ‐secretase‐mediated mechanisms, including the Notch pathway, in the cell contact‐dependent communication between human DC subsets. The resulting partial activation of pDCs after encountering with mature cDCs endows pDCs with an accessory function that may contribute to T cell recruitment and activation.

Non-targeted metabolomic biomarkers and metabotypes of type 2 diabetes: A cross-sectional study of PREDIMED trial participants

AIM To characterize the urinary metabolomic fingerprint and multi-metabolite signature associated with type 2 diabetes (T2D), and to classify the population into metabotypes related to T2D. METHODS A metabolomics analysis using the 1H-NMR-based, non-targeted metabolomic approach was conducted to determine the urinary metabolomic fingerprint of T2D compared with non-T2D participants in the PREDIMED trial. The discriminant metabolite fingerprint was subjected to logistic regression analysis and ROC analyses to establish and to assess the multi-metabolite signature of T2D prevalence, respectively. Metabotypes associated with T2D were identified using the k-means algorithm. RESULTS A total of 33 metabolites were significantly different (P<0.05) between T2D and non-T2D participants. The multi-metabolite signature of T2D comprised high levels of methylsuccinate, alanine, dimethylglycine and guanidoacetate, and reduced levels of glutamine, methylguanidine, 3-hydroxymandelate and hippurate, and had a 96.4% AUC, which was higher than the metabolites on their own and glucose. Amino-acid and carbohydrate metabolism were the main metabolic alterations in T2D, and various metabotypes were identified in the studied population. Among T2D participants, those with a metabotype of higher levels of phenylalanine, phenylacetylglutamine, p-cresol and acetoacetate had significantly higher levels of plasma glucose. CONCLUSION The multi-metabolite signature of T2D highlights the altered metabolic fingerprint associated mainly with amino-acid, carbohydrate and microbiota metabolism. Metabotypes identified in this patient population could be related to higher risk of long-term cardiovascular events and therefore require further studies. Metabolomics is a useful tool for elucidating the metabolic complexity and interindividual variation in T2D towards the development of stratified precision nutrition and medicine. Trial registration at www.controlled-trials.com: ISRCTN35739639.

Integrative Analysis to Select Genes Regulated by Methylation in a Cancer Colon Study

Methylation is a regulatory mechanism known to be associated with tumour initiation and progression. Finding genes regulated by methylation is a first step to develop therapies that target these genes, for instance to inhibit tumor development. This study addresses this problem by comparing two methods, one based on mutual information, and a new one based on clustering the coefficients of fitted curves. The methods are tested on a Cancer Colon study and the biological analysis of the resulting lists suggests that at least some of the genes selected are indeed genes regulated by methylation, opening the door to an automatic mining method.

Influence of early neurological complications on clinical outcome following lung transplant

Background Neurological complications after lung transplantation are common. The full spectrum of neurological complications and their impact on clinical outcomes has not been extensively studied. Methods We investigated the neurological incidence of complications, categorized according to whether they affected the central, peripheral or autonomic nervous systems, in a series of 109 patients undergoing lung transplantation at our center between January 1 2013 and December 31 2014. Results Fifty-one patients (46.8%) presented at least one neurological complication. Critical illness polyneuropathy-myopathy (31 cases) and phrenic nerve injury (26 cases) were the two most prevalent complications. These two neuromuscular complications lengthened hospital stays by a median period of 35.5 and 32.5 days respectively. However, neurological complications did not affect patients’ survival. Conclusions The real incidence of neurological complications among lung transplant recipients is probably underestimated. They usually appear in the first two months after surgery. Despite not affecting mortality, they do affect the mean length of hospital stay, and especially the time spent in the Intensive Care Unit. We found no risk factor for neurological complications except for long operating times, ischemic time and need for transfusion. It is necessary to develop programs for the prevention and early recognition of these complications, and the prevention of their precipitant and risk factors.