Tuesday, 31 January 2012

Drinking alcohol and MS

Epub: Foster et al. Associations of moderate alcohol consumption with clinical and MRI measures in multiple sclerosis. J Neuroimmunol. 2012 Jan.

OBJECTIVE: To examine the associations of alcohol consumption patterns with disability and brain injury in MS'ers.

DESIGN: This study included 423 subjects (272 MS'ers, 151 healthy controls) participating in a study of clinical, environmental and genetic risk factors in MS. Disability was assessed with the EDSS and the MS Severity Scale (MSSS). Brain injury was assessed using MRI. Information related to alcohol-consumption patterns was obtained with standardized questionnaires. 

RESULTS: The frequency of MS'ers who did not consume alcohol after the diagnosis of MS (19.4%) was higher than the frequency before MS (p<0.001). The EDSS and MRI parameters exhibited a non-linear dependence on duration of alcohol consumption after MS onset.

CONCLUSION: The duration of alcohol consumption is associated with disability and MRI measures in MS.

"This study is saying that alcohol consumption is bad for MS and is associated with a worse outcome clinically and on MRI."

"Until these results are confirmed I wouldn't take these result as fact. Alcohol consumption is associated with reversible shrinkage of the brain in normal people. May be the same effect is happening in MS'ers?"

Monday, 30 January 2012

Are you on natalizumab (Tysabri)? Do you know your JCV status?

Epub ahead of printFox & Rudick. Risk stratification and patient counseling for natalizumab in multiple sclerosis. Neurology. 2012 Jan 25.  

Natalizumab has demonstrated efficacy in multiple sclerosis (MS), but is associated with increased risk of progressive multifocal leukoencephalopathy (PML), a potentially fatal brain infection caused by the JC virus. At the time of natalizumab's reapproval in 2006, the estimated risk of PML was 1:1,000 (95% confidence interval 0.2-2.80) over 17.9 months of treatment. Since then, 3 risk factors for natalizumab-related PML have emerged: 

  1. Cumulative exposure to natalizumab, with risk increasing up to 3 years, after which risk appears to plateau. 
  2. Previous treatment with cytotoxic or immunosuppressive (IS) drugs
  3. Prior history of JCV infection, as indicated by the presence of JCV antibodies. 
Using a new assay for anti-JCV antibodies ~55% of MS'ers are anti-JCV antibody positive; with false-negative rate of ~2.5% (unfortunately no laboratory assay is perfect). 

To date, 28 natalizumab-related PML cases had blood specimens available 1 year or more prior to the development of PML. Using this assay, all 28 patients were JCV seropositive, which is highly statistically significant (p = 3.21 × 10(-8)).

MRI of someone with PML
This assay is now clinically available in the United States and Europe.

See Stratify JCV website.

"The negative predictive value of this assay is important. In other words  if you are JCV seronegative (negative test) your risk of getting PML is very low. If you are either on natalizumab, or deciding to go onto natalizumab,  you may find this information very helpful."

Please see the latest post on this blog about risk stratification:

14 Jan 2012
Please note that if you have been on Natalizumab longer than 24 months, have previously been treated with immunosuppressive drugs, for example mitoxantrone, and you are JC virus positive (blood tests shows you have ...

CoI: Multiple

Research: Viral Trigger of Relapse

Multiple Sclerosis (MS) is considered to be an autoimmune disease with unknown cause and with immune system dysregulation. Among environmental factors, viruses are most often connected with the aetiology of MS. Human Endogenous Retroviruses (HERVs) constitute 5-8% of human genomic DNA and have been detected as transcripts and proteins in the central nervous system (CNS) and peripheral blood, frequently in the context of neuroinflammation.

The HERV-Fc1, which belongs to the HERV-H/F family, has received our attention largely because of the genetic association with MS. The expression of a capsid (Gag) coat protein of HERV-H/F origin by flow cytometry in blood cells from healthy controls and from MS patients with non-active or active disease.

There was a significant increase in HERV-H/F Gag expression in CD4+ (P less than 0.001) and CD8+ T lymphocytes (P less than 0.001) and in monocytes (P=0.0356*) in the blood from MS patients with active disease. Furthermore, we have undertaken the first rigorous qualitative PCR (were you amplify the genes so that they can be detected and quantitated) to quantify extracellular HERV-Fc1 RNA viral loads in the plasma (blood fluid) from MS patients and healthy controls.

A 4 fold increase in extracellular HERV-Fc1 RNA titres in patients with active MS was found compared with healthy controls (P less than 0.001). These findings strengthen the link between HERV-Fc1 and the pathology of MS. The cause and biological consequences of these differential expressions will be the subject of further investigation. HERV-Fc1 biology could be a compelling area for understanding the pathology of MS and possibly other autoimmune disorders.

If you managed to attend the Research Day, you will have heard about the Charcot Project, which aims to try and tackle the possibility of a viral cause of MS.

As you will be aware Prof G has been investigating the link of MS with Epstein-Barr Virus, but Prof G Down Under has ideas about other viruses and these may be HERVs. There has been studies suggesting that HERVs may be more common in MSers. This current study suggests these endogenous retrovirus may become active during active disease in MS, so is this the trigger? Is there a link with EBV? There is one way to find out. That is deal with these viruses to stop them becomiing activated and see what happens in MS.

Please see previous posts

HERVS you heard it here
Is MS caused by a retro virus
Do human endogenous retroviruses play a role in MS

Research: Virus Causing autoimmunity

Libbey JE, Cusick MF, Tsunoda I, Fujinami RS. Antiviral CD8(+) T cells cause an experimental autoimmune encephalomyelitis-like disease in naive mice. J Neurovirol. 2012 Jan 27. [Epub ahead of print]

Major histocompatibility complex class I-restricted CD8(+) cytotoxic T lymphocytes are involved in the pathogenesis of multiple sclerosis (MS) and both autoimmune, experimental autoimmune encephalomyelitis, and viral, Theiler's murine encephalomyelitis virus (TMEV) infection, animal models of MS.

Following TMEV infection, certain T cell hybridomas, generated from cloned (single identical cells) TMEV-induced CD8(+) T cells, were able to produce clinical signs of disease (flaccid hind limb paralysis) upon adoptive transfer into naive mice. Dual T cell receptors (TCR) are present on the surface of these cells as both Vβ3 and Vβ6 were detected by polymerase chain reaction (PCR) screening and flow cytometry and multiple Vα mRNAs were detected by PCR screening.

This is the first demonstration of antiviral CD8(+) T cells having more than one TCR initiating an autoimmune disease in the natural host of the virus. We hypothesize that this is a potential mechanism for virus-induced autoimmune disease initiated by CD8(+) T cells.

Theiler's murine encephalomyelitis virus is a natural pathogen of mice, which sometimes gets into the brain and causes a demyelinating disease, that may generate the development of autoimmunity by a number of different mechanisms such as molecular mimicry (sequence similarities between foreign(virus) and self-peptides are sufficient to result in the cross-activation of autoreactive T or B cells by pathogen derive peptides. So viral directed immune responses are mis-directed to attack the nervous tissue) or deteriminant spread (where damage generated in response to the immune attack of virus, triggers the development of a immune responses to the damaged protiens and thus trigger autoimmune attack. This study suggests another mechanism.

The T cell receptor is the stucture that allows the T cell to see its target. This is anchored in the cell membrane and consists of two halves, which form a pair of protein chains. The halves are called the alpha (α) and beta (β) fragments (in γ/δ T cells, the halves are gamma (γ) and delta (δ) fragments). Each fragment is divided in turn into a constant (C) and variable (V) region. These are constructed by the action of joining the gene products of a variable number of constant, joining, diversity and variable T cell receptor gene. There are about 50 variable genes for the beta chain and over 70 for the alpha.

So in this cell there were two variants (3 and 6) of the beta chain variable reion and many different alpha genes. Normally only one alpha and beta are present and can recognise one target if there is an extra beta then it could recognise a different target. T cells that reacted with the virus were cloned (so all the cells are identifical) and so the T cell receptor would react with the virus. So that they have a source of immortal cells to do reach with they fused the T cell clone with a tumor that lacks T cell receptor. They found that the T cell clone must have contained many different T cell receptor and when they put one of these tumours in mice it caused neurological problems suggesting that some of the T cell receptors were reacting with a nerve component. So by having more than one set of T cell receptors viral specific cells could also react with nerve proteins and trigger autoimmunity.

This concept whereby cells have dual (two) T cell receptors has been demonstrated many years ago, particularly with cells containing extra alpha chains in the context of of other viral infection or autoimmune conditions. This is yet another way that autoimmunity could develop following viral infection.

Sunday, 29 January 2012

Barts and London Research Day

A Big Thank You
All the People that Attended the Research Day Yesterday

Hope you All Had a Good Day and a Safe Journey Home

January. Unrelated Blogger Comments 4

Sometimes You want to post a comment that is Unrelated to the thread.

Therefore I have Created this Spot for You
It jumps around so that it is visible

Research: Monitoring Progression

Gnanapavan S, Grant D, Pryce G, Jackson S, Baker D, Giovannoni G. Neurofilament a biomarker of neurodegeneration in autoimmune encephalomyelitis.Autoimmunity. 2012 Jan 26. [Epub ahead of print]

Monitoring neuroaxonal loss in multiple sclerosis is an important objective to study the pathogenesis and response to treatment of the disease. The release of neurofilaments is a potential surrogate biomarker of neurodegeneration. One route to explore this aspect further is through the use of animal models that have well-defined, and predictable, disease courses.

The release of neurofilaments into plasma (blood) across the course of relapsing-remitting and secondary progressive experimental autoimmune encephalomyelitis in mice was assessed.

It was found that there was an immediate release in neurofilaments into the blood following the initial paralytic attack. Neurofilament release was continuous in relapse and remission but returned towards baseline in chronic disease, as animals entered the slowly developing post-relapsing progressive phase of the disease.

In contrast neurofilament levels increased dramatically as neurodegeneration and clinical disease occurred in the G93A SOD1 transgenic model of amyotrophic lateral sclerosis.

These data further support neurofilament levels as a good surrogate measure of neurodegeneration and their potential use as a surrogate endpoint in neuroprotective studies.

Green colour shows the neurofilament as an internal protein of the nerve. Red = glial cells

Progression of clinical disease occurs too slowly to undertake lots of clinicalt trials based on clinical features. Therefore we have we have been seeking indicators of disease worsening that can change quicker than clinical disability and so we could use those outcomes as surrogates to show disease activity. One thing we have found is neurofilament.

This is an internal part of the nerve and as nerves get damaged the nerves content is released first in to the cerebrospinal fluid and then into the blood. This study looked at two models of nerve damage, one that is known to occur by attacks of the immune system and another that is caused by neurodegenerative effects independent of autoimmunity. This study shows that with immune attack neurofilament can even be detected in the blood and tells us that stopping relapsing disease is a good thing. We know that each attack is indeed associated with nerve damage and that we can pick break down products of nerve attack in the blood supports this.

Next it says that in progressive MS models that slow progression that is independent of autoimmunity, such as secondary and primary progressive disease was not picked up in the blood in one model of neurodegeneration but was found in the blood in another model of neurodegeneration in a model that resembles motor neuron disease.

This tells us two things that measuring neurofilament release into biological fluids such as cerebrospinal fluid and the blood can give us an indication of the damage that is occuring in the central nervous system and that we may have to look in the cerebrospinal fluid rather than the blood.

Therefore if you have not watched the video and answered the survey on having three lumbar punctures as a method of detecting nerve damage and its control in a new trial disease design that will speed getting drugs to progressive MSers. Please watch the video and do the survey on the left. If you will not do it is just as important to know as if you will do it. These is no point in developing an idea that can not get off the ground

Are you up for having lumbar puncture

CoI: This is the work of Team G as part of Promse 2010

Saturday, 28 January 2012

Bad Science:Interferons after Mitoxantrone

Today is our research day and I have to deliver Bad Science. When you read papers you have to digest whether they actually tell us anything. You do not always believe things you read. Importantly is it worth reading them in the first place?

Todays story is an an example. NO LINK PROVIDED, READING VALUE IS LOW

Objective: The objective of this study was to assess the effect of treatment with interferon (IFN) β-1a, 44 µg subcutaneously (sc) three times weekly (tiw), on clinical and magnetic resonance imaging (MRI) outcomes in patients with relapsing multiple sclerosis (MS) following mitoxantrone therapy.

Methods: This was an open-label, randomized, multicentre, rater-blinded, 96-week observational study. Clinically stable patients with relapsing forms of MS, who had discontinued mitoxantrone treatment 1-6 months before study entry, were randomized to IFN β-1a sc 44 µg tiw, or no treatment. The primary endpoint was time to first relapse. Secondary endpoints included the number of relapse-free patients, disease activity assessed by MRI and time to 3-month confirmed Expanded Disability Status Scale (EDSS) progression, all at week 96.

Results: A total of 30 patients were randomized (intent-to-treat population: 14 IFN β-1a, 15 untreated; one patient from the safety population discontinued the study after 25 days owing to an adverse event and without providing any postbaseline efficacy data, and was thus excluded from the intent-to-treat population). Overall, 71.4% (10/14) of patients in the IFN β-1a group remained relapse free over 96 weeks, versus 46.7% (7/15) in the untreated group (p = 0.26 THIS ACTUALLY MEANS THERE WAS NO DIFFERENCE DESPITE THE INFERENCE). IFN β-1a delayed the time to first relapse versus no treatment (p = 0.14. THIS ACTUALLY MEANS THERE WAS NO DIFFERENCE IN TIME TO RELAPSE DESPITE THE INFERENCE); time to first relapse (25th percentile) was 95.4 (IFN β-1a) versus 46.0 weeks (no treatment). Confirmed EDSS progression was observed in five patients in each treatment group. Mean change in EDSS score was 0.3 in both groups (p = 0.79 THIS ACTUALLY MEANS THERE WAS NO DIFFERENCE). Changes in the number or volume of T1 and T2 lesions at week 96 were not significantly different between treatment groups (THEY ACTUALLY TELLL US THAT THERE WAS NO DIFFERENCE). There were no new or unexpected adverse events related to IFN β-1a treatment.

Conclusions: Several endpoints appeared to show a benefit of IFN β-1a treatment, but no significant differences could be detected owing to the small sample. THEY TELL US NOTHING WAS SHOWN Therefore, these data only permit, at best, tentative conclusions (THEY DO NOT PERMIT CONCLUSIONS AS THE DATA COULD EASILY BE DUE TO CHANCE AND MEAN NOTHING) about the disease course in patients with MS after de-escalation from mitoxantrone and continuation with or without IFN β-1a. Larger confirmatory studies are required.

While we want to know if after you stop mitoxantrone (you can only have so many doses because of risks to the heart) can beta interferon be of benefit in halting relapses. But we have to digest the information and ask, is there anything of real importance occuring here?

If one looked at more people would there be benefit to be seen? However, as the results stand you have to conclude that they show nothing concrete. This is a reason why it is important that studies are properly powered (that is contain a lare enough number of samples to provide meaningful information) so that you get a result that informs otherwise you have to do it all again. The authours should have waitied until they had enough data to say something.

Bad Refereeing: New model for Progression

Normally I do not report on animal models of MS but as Mouse Doc is doing "Bad Science Day" at our research Day and the media have found that "Diabetic mice have provided a surprising breakthrough for MS research 1:5:2012" todays new breakthrough extends this further.

We have been talking about the publication process. Part of this is the independent review process. The peer review process of publication aims to see if work is original and whether the claims made are supported by the data shown and whehter the data presented is put into context of the exisitng knowledge. Sometimes this works sometimes it is not so good.

Anderson AC, Chandwaskar R, Lee DH, Sullivan JM, Solomon A, Rodriguez-Manzanet R, Greve B, Sobel RA, Kuchroo VK. A Transgenic Model of Central Nervous System Autoimmunity Mediated by CD4+ and CD8+ T and B Cells. J Immunol. 2012 Jan [Epub ahead of print]

Experimental autoimmune encephalomyelitis (EAE) is a widely used model of multiple sclerosis. Non obsese diabetic (NOD) mice that can develop type I diabetes, develop EAE as a relapsing-remitting disease that transitions to a chronic progressive disease, making the NOD model the only mouse model that recapitulates the full clinical disease course observed in most multiple sclerosis patients. [Sack the peer-reviewers.........as they do not know how to do a literature review to see if this is not the ONLY model. Maybe they could have found others maybe one a few or many years ago...likewise do a literature review and wonder if it is really a progressive model or maybe a relapsing neurodegenerative model see figure 1]. If you just look at diagrams with out knowing the course in individual animals it is easy to say that studies show that disease is progressive, when it is degenerative, relapsing-remitting disease that is not occurring in a asychronous fashion (See below).

A T cell receptor (TCR) transgenic mouse that expresses the α and β-chains of a myelin oligodendrocyte glycoprotein (MOG) 35-55-reactive TCR (1C6) on the NOD mouse (shown earlier to be rather resistant to the development of EAE) genetic background was made and called 1C6 TCR transgenic mice. These spontaneously generate both CD4(+) and CD8(+) T cells that recognize MOG and produce proinflammatory cytokines, allowing for the first time to our knowledge the simultaneous examination of myelin-reactive CD4(+) and CD8(+) T cells in the same host. [Check out studies of which there are loads, in normal mice with CD4 and CB8 -specific antibodies]

1C6 CD8(+) T cells alone can induce optic neuritis and mild EAE with delayed onset; however, 1C6 CD4(+) T cells alone induce severe EAE and predominate in driving disease when both cell types are present. [As shown previously about 15-20 years earlier with monoclonal antibodies] When 1C6 mice were crossed with mice bearing an Immunoglobulin heavy chain (antibody) specific for MOG, the mice develop spontaneous EAE with high incidence, but surprisingly the disease pattern does not resemble the neuromyelitis optica-like disease observed in mice bearing CD4(+) T cells and B cells reactive to MOG on the C57BL/6 background. [Most EAE models show optic nerve and spinal cord disease without significant brain involvement.....so crappy reviewers to papers in the past. This is not neuromyelitis optica and the brain lesions found in mice in this study in the cerebellum of mice without typical neurological disease have been reported previously and again ignored in the paper. The authors have spun their story and the reviewers have let them get away with it. The peer-reviewers should retire from the review process due to incompetence] Collectively, the data show that although myelin-reactive CD8(+) T cells contribute to disease, disease is primarily driven by myelin-reactive CD4(+) T cells and that the coexistence of myelin-reactive T and B cells does not necessarily result in a distinct pathological phenotype. [As shown previously in many other mouse studies]

The graph from Team G shows the results of the development of disease in a group of animals with EAE. Disability is on the scale on the left so 3 is more disabled than 1 and time is days after the disease was induced. After the first attack is this progressive disability? Well it looks like it. However this is the result of relapsing disease with neurodegeneration that occurs as a consequence of the relapse. Disease worsening is occuring in a step wise fashion with each attack in each individual animal but apart for the first attack the relapses are not synchronised. Therefore sometimes you need more information to know what is going on than simply looking at line graphs

Is this study interesting "yes it is", is it good quality work.."yes it is", are these mice going to be useful "yes they probably are" Can I decipher the good stuff in the paper? Yes I can, but does it over-egg the importance "yes it does". Just an example of you don't have to believe everything you read or you have to decipher what is being said. This is what we are trying to do on the blog. Obviously we have our own bias when we review the studies. Any critism being made is supposed to be constructive... as this is an interesting study.

Friday, 27 January 2012

Research Beta Interferon does not stop secondary Progressive MS

La Mantia L, Vacchi L, Di Pietrantonj C, Ebers G, Rovaris M, Fredrikson S, Filippini G. Interferon beta for secondary progressive multiple sclerosis.Cochrane Database Syst Rev. 2012 Jan;1:CD005181.

BACKGROUND: Therapy with either recombinant beta-1a or beta-1b interferons (IFNs) is worldwide approved for Relapsing Remitting Multiple Sclerosis (RRMS). A major unanswered question is whether this treatment is able to safely reverse or retard the progressive phase of the disease.

The main objective was to verify whether IFNs treatment in Secondary Progressive Multiple Sclerosis (SPMS) is more effective than placebo in reducing the number of patients who experience disability progression.

SEARCH METHODS: Cochrane Multiple Sclerosis Group's Trials Register (1995 to 15 February 2011), the reference lists of relevant articles and conference proceedings were searched. Regulatory agencies were used as additional sources of information. All randomised, double or single blind, placebo-controlled trials (RCTs) evaluating the efficacy of IFNs versus placebo in SPMS patients were included.

DATA COLLECTION AND ANALYSIS: Two review authors independently assessed all reports retrieved from the search. They independently extracted clinical, safety and MRI data, using a predefined data extraction form, resolving disagreements after discussion with a third reviewer. Risk of bias was evaluated to assess the quality of the studies. Treatment effect was measured using Risk Ratio (RR) with 95% confidence intervals (CI) for the binary outcomes and Standard Mean Difference with 95% CI for the continuous outcomes.

MAIN RESULTS: Five RCTs met the inclusion criteria, from which 3122 (1829 IFN and 1293 placebo) treated patients contributed to the analysis. Included population was heterogeneous in terms of baseline clinical characteristics of the disease, in particular the percentage of patients affected by secondary progression with superimposed relapse ranging from 72% to 44%. IFN beta 1a and 1b did not decrease the risk of progression sustained at 6 months (Relative risk, 95% CI: 0.98, [0.82-1.16]) after three years of treatment. A significant decrease of the risk of progression sustained at 3 months (RR, 95% CI: 0.88 [0.80, 0.97]) and of the risk of developing new relapses at three years (RR 0.91, [0.84-0.97]) were found. The risk of developing new active brain lesions decreased over time but this data was obtained from single studies on Magnetic Resonance Imaging (MRI), performed in subgroups of patients; in spite of no effect on progression, the radiological data supported an effect on MRI parameters. The safety profile reflects what is commonly reported in MS IFN-treated patients.

AUTHORS' CONCLUSIONS: Well designed RCTs, evaluating a high number of patients were included in the review. Recombinant IFN beta does not prevent the development of permanent physical disability in SPMS. We were unable to verify the effect on cognitive function for the lack of comparable data. This treatment significantly reduces the risk of relapse and of short -term relapse-related disability. Overall, these results show that IFNs' anti-inflammatory effect is unable to retard progression, when established. In the future, no new RCTs for IFNs versus placebo in SPMS will probably be undertaken, because research is now focusing on innovative drugs. We believe that this review gives conclusive evidence on the clinical efficacy of IFNs versus placebo in SPMS.
The Cochrane Collaboration is an international not-for-profit and independent organization, dedicated to making up-to-date, accurate information about the effects of healthcare readily available worldwide. It produces and disseminates systematic reviews of healthcare interventions and promotes the search for evidence in the form of clinical trials and other studies of interventions. The Cochrane Collaboration was founded in 1993 and named after the British epidemiologist, Archie Cochrane.

Recombinant IFN beta does not prevent the development of permanent physical disability in Secondary Progressive MS. This is old news

Research: IL-1 inhibits Nerve Damage

Rossi S, Furlan R, De Chiara V, Motta C, Studer V, Mori F, Musella A, Bergami A, Muzio L, Bernardi G, Battistini L, Martino G, Centonze D. Interleukin-1β causes synaptic hyperexcitability in multiple sclerosis. Ann Neurol. 2012; 71:76-83. doi: 10.1002/ana.22512.

The frequency of inflammatory episodes in the early stages of multiple sclerosis (MS) has been correlated with late neurodegeneration, but the mechanism by which inflammation gives rise to delayed neuronal damage is unknown. Increased activity of the neurotransmitter glutamate is thought to play a role in the inflammation-driven neurodegenerative process of MS, and therefore we tested whether inflammatory cytokines released during acute MS attacks have the property of enhancing glutamate-mediated transmission and excitotoxicity in central neurons.

METHODS:We compared the effect of cerebrospinal fluid (CSF) from active and quiescent MS patients on glutamate-mediated excitatory postsynaptic currents (EPSCs) and excitotoxic damage in rodent brain slices. We also measured CSF concentrations of tumor necrosis factor-α, of interleukin-1β (IL-1β), and of IL-1 receptor antagonist (IL-1ra, an IL-1 blocker), and correlated cytokine levels with cortical excitability assessed in MS patients by means of paired-pulse transcranial magnetic stimulation (TMS).

RESULTS: CSF from MS patients with enhanced brain lesions at magnetic resonance imaging was able to increase spontaneous EPSC frequency and glutamate-mediated neuronal swelling in cell culture through a mechanism dependent on enhanced IL-1β signaling and increased glutamate α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor stimulation. Furthermore, IL-1β/IL-1ra ratio was significantly higher in the CSF of active MS subjects, and correlated with intracortical facilitation, a measure of glutamate transmission. Finally, we identified transient receptor potential vanilloid 1 channels as essential intermediates for the synaptic action of IL-1β on central glutamatergic synapses.

INTERPRETATION: Our results provide compelling evidence of the synaptic mechanism linking inflammation and excitotoxic neurodegeneration in MS.

Glutamate is the major neurotransmitter (chemical that triggers nerve impulses) that stimulates nerves. However if a nerve gets too much stimulation it can trigger the cells to commit suicide which is known as excitotoxicity. Using mouse brain tissue the electrical activity of nerves was measured before and after cerebrospinal fluid was used to bathe the brain tissue. This was taken when MS was active and when it was non-active. It was found that when disease was active there were chemicals released that could cause nerve damage. Using this approach one of the damaging molecules was identified and away that it can stimulation of glutamate through AMPA glutamate receptors. Glutamate receptor anagonists could be a potential treatment that could be suggested from this work, but have the potential to cause side-effects or Vannilod receptor antagonists could be an alternative new avenue for treatment.

CoI: G. Martino from Milan is Part of Team G

Research: MicroRNA in Multiple Sclerosis

Paraboschi EM, Soldà G, Gemmati D, Orioli E, Zeri G, Benedetti MD, Salviati A, Barizzone N, Leone M, Duga S, Asselta R. Genetic association and altered gene expression of mir-155 in multiple sclerosis patients.Int J Mol Sci. 2011;12:8695-712.

Multiple sclerosis (MS) is a complex autoimmune disease of the central nervous system characterized by chronic inflammation, demyelination, and axonal damage. As microRNA (miRNA)-dependent alterations in gene expression in hematopoietic cells are critical for mounting an appropriate immune response, miRNA deregulation may result in defects in immune tolerance.

In this frame, we sought to explore the possible involvement of miRNAs in MS pathogenesis by monitoring the differential expression of 22 immunity-related miRNAs in peripheral blood mononuclear cells of MS patients and healthy controls, by using a microbead-based technology.

Three miRNAs resulted greater than 2 folds up-regulated in MS vs controls, whereas none resulted down-regulated. Interestingly, the most up-regulated miRNA (mir-155; fold change = 3.30; P = 0.013) was previously reported to be up-regulated also in MS brain lesions. Mir-155 up-regulation was confirmed by qPCR experiments. The role of mir-155 in MS susceptibility was also investigated by genotyping four single nucleotide polymorphisms (SNPs) mapping in the mir-155 genomic region. A haplotype of three SNPs, corresponding to a 12-kb region encompassing the last exon of BIC (the B-cell Integration Cluster non-coding RNA, from which mir-155 is processed), resulted associated with the disease status (P = 0.035; OR = 1.36, 95% CI = 1.05-1.77), suggesting that this locus strongly deserves further study.

Recently we introduced you to the concept of microRNA and that they may influence the effects of proteins, Mir 155 has been implicated in controlling the immune response, which is aletered during MS. Animal studies have suggested that Mir55 can promote autoimmune inflammation by enhancing inflammatory T cell development Mice that lack Mir55 develop less MS-like disease than mice that produce Mir155. That Msers have elevated levels of Mir55 may not be a good thing, but it suggests that by inhibiting the development of Mir55 there may be benefit to be had.

Thursday, 26 January 2012

Interleukin 7 and T cell function

Kreft KL, Verbraak E, Wierenga-Wolf AF, van Meurs M, Oostra BA, Laman JD, Hintzen RQ. The IL-7Rα Pathway Is Quantitatively and Functionally Altered in CD8 T Cells in Multiple Sclerosis. J Immunol. 2012 Jan [Epub ahead of print]

The interleukin 7 receptor alpha chain (IL-7Rα) single nucleotide polymorphism rs6897932 is associated with an increased genetic risk for multiple sclerosis (MS). IL-7Rα is a promising candidate to be involved in autoimmunity, because it regulates T cell function, proliferation, and anti-apoptotic signaling. However, the exact underlying mechanisms in the pathogenesis of MS are poorly understood.

We investigated whether CD4 and CD8 lymphocyte subsets differed in IL-7Rα expression and functionality in 78 MS patients compared with 59 healthy controls. A significantly higher frequency of IL-7Rα(+) CD8 effector memory (CD8EM) was found in MS. Moreover, IL-7Rα membrane expression was significantly increased in MS in naive and memory CD8 (all p < 0.05) with a similar trend in CD8EM (p = 0.055).

No correlation was found between the expression level or frequency of IL-7Rα(+)CD8(+) and rs6897932 risk allele carriership.

Upon IL-7 stimulation, MS patients had stronger STAT5 (a transcription factor) activation in CD8EM compared with Healthy controls. IL-7 stimulation had a differential effect on both mRNA and protein expression of granzyme A and granzyme B (a protein that punches holes in its target. Imagine a pin prick to a balloon this is what granzymes can do to virally infected cells) between MS and HC. Stainings of different lesions in post-mortem MS brain material showed expression of IL-7 and CD8(+)IL-7Rα(+) in preactive, but not in active, demyelinating MS lesions, indicating involvement of IL-7Rα(+) lymphocytes in lesion development.

The intralesional production of IL-7 in combination with the lower threshold for IL-7-induced cytotoxicity in MS may enhance the pathogenicity of these CD8 T cells. This is of special interest in light of the established demyelinating and cytotoxic actions of granzyme A.

Variants in Interleukin seven receptor are associated with an increase risk of developing MS. We know that interleukin 7 is associated with the function of the immune system but how does the variant affect MS?. This study does not answer this question as the expression of IL-7R on T cells did not correlate with the variant of IL-7R genes, but it does suggest that cells expressing IL-7R are part of the disease process. There was more IL-7R on T cells from people with MS compared with healthy people and cells expressing this molecule accumlate in early MS lesions.

What Story Did You Make Up?

Thinking of brain training how did you remember this. Make up a story and it is easy to remember. Mine worked. The one on the left is a VV

Brain Training: The results

Can you remember the figures on the brain training picture from last week?

Could not be bothered.....

or did you remember:

A guy with shaded eyes; a fish; a guy with Big Teeth; Two one cent coins; a middle Eastern Man; a light bulb with a dollar sign and a newspaper.

Wednesday, 25 January 2012

Research: Human Stem cells

The recent introduction of technologies capable of reprogramming human somatic cells into induced pluripotent stem (iPS) cells offers a unique opportunity to study many aspects of neurodegenerative diseases in vitro that could ultimately lead to novel drug development and testing.

Here, we report for the first time that human dermal fibroblasts from a patient with relapsing-remitting Multiple Sclerosis (MS) were reprogrammed to pluripotency by retroviral transduction using defined factors (OCT4, SOX2, KLF4, and c-MYC).

The MSiPS cell lines resembled human embryonic stem (hES) cell-like colonies in morphology and gene expression and exhibited silencing of the retroviral transgenes after four passages. MSiPS cells formed embryoid bodies that expressed markers of all three germ layers by immunostaining and Reverse Transcriptase (RT)-PCR.

The injection of undifferentiated iPS cell colonies into immunodeficient mice (so that they do not reject the human cells) formed teratomas (an encapsulated tumour), thereby demonstrating pluripotency. The MSiPS cells were successfully differentiated into mature astrocytes, oligodendrocytes and neurons with normal karyotypes. Although MSiPS-derived neurons displayed some differences in their electrophysiological characteristics as compared to the control cell line, they exhibit properties of functional neurons, with robust resting membrane potentials, large fast tetrodotoxin-sensitive action potentials and voltage-gated sodium currents. This study provides for the first time proof of concept that disease cell lines derived from skin cells obtained from an MS patient can be generated and successfully differentiated into mature neural lineages. This represents an important step in a novel approach for the study of MS pathophysiology and potential drug discovery.

The abstract says it all and is encouraging for stem cell technology but there will be be more twists and turns as these technologies are developed towards the clinic, but a step in the right direction. Furthermore, this types of technologies can provide access to cells to study human biology that may lead to other benefits.

Research:Biomarkers for MS

The complex pathogenesis of multiple sclerosis, combined with an unpredictable prognosis, requires identification of disease-specific diagnostic and prognostic biomarkers.Objective: To determine whether inflammatory proteins, such as neurofilament light chain (marker of nerves), myelin oligodendrocyte glycoprotein and myelin basic protein (markers of myelin), and neurodegenerative proteins, such as tau (distressed nerves) and glial fibrillary acidic protein (active astrocytes), can serve as biomarkers for predicting the clinical subtype and prognosis of MS.

Methods: Cerebrospinal fluid and serum samples were collected from patients with a diagnosis of clinically isolated syndrome (n = 46), relapsing-remitting MS (n = 67) or primary-progressive MS (n = 22) along with controls having other non-inflammatory neurological disease (n = 22). Western blot analyses were performed for the listed proteins. Protein levels were compared among different clinical subtypes using one-way analysis of variance analysis.

Results: The results showed that each of tau, GFAP, MOG and NFL protein concentrations differed significantly (p less than 0.001) in multiple sclerosis clinical subtypes compared with the controls. Levels of the proteins also differed between the multiple sclerosis clinical subtypes, which may be associated with the underlying disease process. Classification studies revealed that these proteins might be useful for identifying multiple sclerosis clinical subtypes.

Biomarkers We showed that select biomarkers may have potential in identifying multiple sclerosis clinical subtypes. We also showed that the predictive value of the prognosis increased when using a combination of the proteins versus using them individually.

Radar diagram of the value of differnt proteins in predicting relapisng remitting primary progressive non converting clinically isolated sydromes and CIS converting to MS and other neurological controls.

Although many of the markers were higher in the MS groups compared to control samples when they the results from analysis of 4 different proteins were compared they had a 95% chance of detecting MS from other neurological condidtions and analysis of three of them together had a 95% chance of predicting whether you are PP MS or RRMS. It this can be repeated then it may be of value in selecting MSers for treatments that may work verses those that do not work on PPMS for example.

Research: BG12 targeting Nrf2 to help save nerves

Scannevin RH, Chollate S, Jung MY, Shackett M, Patel H, Bista P, Zeng W, Ryan S, Yamamoto M, Lukashev M, Rhodes KJ. Fumarates Promote Cytoprotection of Central Nervous System Cells Against Oxidative Stress via the Nrf2 Pathway.J Pharmacol Exp Ther. 2012 Jan 20. [Epub ahead of print]

Oxidative stress is central to the pathology of several neurodegenerative diseases, including multiple sclerosis (MS), and therapeutics designed to enhance antioxidant potential could have clinical value. The objective of this study was to characterize the potential direct neuroprotective effects of dimethyl fumarate (DMF) and its primary metabolite monomethyl fumarate (MMF) on cellular resistance to oxidative damage in primary cultures of central nervous system (CNS) cells and further explore the dependence and function of the Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) pathway in this process. Treatment of animals or primary cultures of CNS cells with DMF or MMF resulted in increased nuclear levels (levels within the cell nucleus) of active Nrf2, with subsequent upregulation of antioxidant target genes. DMF-dependent upregulation of antioxidant genes in vivo was lost in mice lacking Nrf2 (Nrf2-/-). DMF or MMF treatment increased cellular redox potential, glutathione, ATP levels and mitochondrial membrane potential in a concentration-dependent manner. Treating astrocytes or neurons with DMF or MMF also significantly improved cell viability after toxic oxidative challenge in a concentration-dependent manner. This effect on viability was lost in cells that had eliminated or reduced Nrf2.

These data suggest that DMF and MMF are cytoprotective (protrcts the cell) for neurons and astrocytes against oxidative stress-induced cellular injury and loss, potentially via upregulation of an Nrf2-dependent antioxidant response. These data also suggest DMF and MMF may function through improving mitochondrial function. The clinical utility of DMF in
multiple sclerosis is being explored through phase 3 trials with BG-12, which is an oral therapeutic containing DMF as the active ingredient.

Yesterday we were exploring ways that mtiochondrial energy depletion in astrocytes....and based on past posts nerves......may lead to nerve damage.
You wondered if there were an know causes and whether anything has been done about this. Well this study shows a way that something is being done about it.

There is a new oral drug called BG12 that has been found to be effective at inhibiting relapses (reduction of around 50%) in MSers in the phase III studies, but the question is whether it does other things that may be beneficial in progressive MS also?

This study suggests that there may be some potential. This drug may be working via a molecule called Nrf2 which acts to control oxidative stress that represents an imbalance between the production and manifestation of reactive oxygen species (damaging in high concentrations) and a biological systems ability to readily detoxify the reactive intermediates or to repair the resulting damage. But simplified means that it has anti-oxidant properties that will block some of the molecules that are known to block mitochondrial function that depletes the cells energy that can lead to nerve damage, as reported yesterday. Mice that lack Nrf2 do not do well when they are given an MS-like disease so a drug that can stimulate Nrf2 has the potential to be beneficial in MS. BG12 is based on a drug used to treat psorias for many years and so hopefully will have a good safety profile. Time will tell is there is any bnefit to be had in progressive MS, but there is some logical biology behind this hope.
CoI: This work was undertaken by Biogen Idec, the producers of BG12

Tuesday, 24 January 2012

Another Idea about the Causes of Progression

White-matter astrocytes, axonal energy metabolism, and axonal degeneration in multiple sclerosis. Cambron M, D'Haeseleer M, Laureys G, Clinckers R, Debruyne J, De Keyser J. J Cereb Blood Flow Metab. 2012 Jan [Epub ahead of print]

In patients with multiple sclerosis (MS), a diffuse axonal degeneration occurring throughout the white matter of the central nervous system causes progressive neurologic disability. The underlying mechanism is unclear.

This study describes a number of pathways by which dysfunctional astrocytes in MS might lead to axonal degeneration.

White-matter astrocytes in MS show a reduced metabolism of adenosine triphosphate-generating phosphocreatine, which may impair the astrocytic sodium potassium pump and lead to a reduced sodium-dependent glutamate uptake.

Astrocytes in MS white matter appear to be deficient in β(2) adrenergic receptors, which are involved in stimulating glycogenolysis and suppressing inducible nitric oxide synthase. Glutamate toxicity, reduced astrocytic glycogenolysis leading to reduced lactate and glutamine production, and enhanced nitric oxide levels may all impair axonal mitochondrial metabolism, leading to axonal degeneration.

In addition, glutamate-mediated oligodendrocyte damage and impaired myelination caused by a decreased production of N-acetylaspartate by axonal mitochondria might also contribute to axonal loss. White-matter astrocytes may be considered as a potential target for neuroprotective MS therapies.

We have described the energy deficit pathway before in nerves, this hypothesis claims the this occurs in astrocytes

Research: Loss of Citrullization of energy making protein is associated with disease RNA transporter

Deimination refers to conversion of protein-bound arginine into citrulline as a post-translational modification after DNA, is transcribed to form RNA and translated into the production of proteins. An mRNA carrier, RNA binding export factor (REF), present on mitochondria undergoes loss of deimination with impaired ATP5b mRNA transport in ND4
mice (model of multiple sclerosis) compared with the controls. We present evidence of (1) reduced ATP5b mRNA binding strength of non-deiminated REF compared with deiminated REF, (2) impaired ATP5b mRNA transport in ND4 mice and (3) reduced mitochondrial ATP synthase activity on inhibition of deimination in PC12 cells. Impaired deimination of REF and defect in mitochondrial mRNA transport are critical factors in mitochondrial dysfunction in ND4 mice.

The conversion of arginine into citrulline can have important consequences for the structure and function of proteins, since arginine is positively charged at a neutral pH, whereas citrulline is uncharged. This increases the hydrophobicity (fat solubility) of the protein, leading to changes in protein folding (gives 3D structure). Myelin basic protein is citrulated as are other proteins.

ND4 mice contain 70 copies (normally it would be two copies of genes, one from your mum and one from your dad) of the transgene encoding DM20, a myelin proteolipid protein. They appear clinically normal up to 3 months of age. By 8-10 months, they show tremors, unsteady gait, and die shortly thereafter. This is associated with the development of demyelination

This study shows that mRNA carrier, RNA binding export factor (REF), present on mitochondria undergoes loss of deimination in ND4 mice. This leads to loss of energy supply to the mitochondria (the powerhouses of cells) in nerve cells. As we have reported previously loss of energy supply in nerves that have been demyelinated can lead to progressive nerve loss. Therefore, it will be of interest to determine whether this process occurs in mutliple sclerosis as there is increasing evidence for a failure of mitochondrial function in the pathogenesis of MS.

When you know the cause you are nearer to a cure of the problem

Monday, 23 January 2012

Edible Science

Two of the "5 minute meal" films, that Alison Thomson made last year with members of the team will be shown in Dublin's Science Gallery from next month. 
 "How the eye is affected in Multiple Sclerosis." By Katie Lidster

5 minute meal - Katie Lidster, PhD student from somehow related on Vimeo.

"How nerve impulses are controlled by cannabis (synaptic neurotransmission)." By Professor David Baker . 

5 minute meal - Prof. David Baker from somehow related on Vimeo.

Edible: A taste of things to come. A free exhibition exploring the future of food. http://www.sciencegallery.com/ 

10th Feb - 6th April 2012
Tue - Fri 12:00 - 20:00
Sat & Sun 12:00 - 18:00

Science Gallery
Pearse Street
Trinity College
Dublin 2

"Well done Alison, we are proud of you!"

Fingolimod deaths and the EMA

Anxious? If you are on fingolimod (Gilenya) and have any queries please read the European Medicines Agency's press release, their Q&A paper or leave a comment on the blog which we will respond to.

Spinal fluid oligoclonal bands: are they the secret to uncovering the cause of MS?

Epub ahead of printKorn & Tumani. Patterns of intrathecal autoreactive antibodies in MS using antigen microarrays. Neurology. 2012 Jan 18.  

The role of autoreactive antibodies in MS has received intense attention since the discovery of oligoclonal immunoglobulin (Ig) bands in the spinal fluid of the majority (>95%) of MS'ers. However, many questions remain. We still do not know whether the production of Ig in the brain of MS'ers is an epiphenomenon of CNS autoimmunity, resulting from growth factor-driven expansion of long-lived B cells (the cells that make antibodies) in the meningeal* compartment or whether Ig production from antigen-driven proliferation of B-cell clones contributes to CNS pathology. CSF Ig from oligoclonal bands is produced by CSF B lineage cells and primarily recognizes epitopes of multiple neurotropic viruses and Epstein-Barr virus antigens.  However, antimyelin antibodies also occur in the serum and CSF of patients with MS.

*meningeal or meninges = the layer of membranes to covers the brain and spinal cord

"In my opinion the secret to the cause of MS lies in uncovering the antigen or proteins against which the antibodies are directed. In disease with known causes (e.g. viral encephalitis) that have oligoclonal IgG bands, the bands react to proteins from the causative agent."

"Cosimo Maggiore in our group is completing a PhD to try and delineate the antigens against which OCBs in MS'ers react. The project has a long way to run. Let's hope he gets there."

Research MSers react against myelin proteins

Epub ahead of print Quintana FJ, Farez MF, Izquierdo G, Lucas M, Cohen IR, Weiner HL. Antigen microarrays identify CNS-produced autoantibodies in RRMS.Neurology. 2012 Jan

Multiple sclerosis (MS) is characterized by the local production of antibodies in the CNS and the presence of oligoclonal bands in the CSF. Antigen arrays allow the study of antibody reactivity against a large number of antigens using small volumes of fluid with greater sensitivity than ELISA. We investigated whether there were unique autoantibodies in the CSF of patients with MS as measured by antigen arrays and whether these antibodies differed from those in serum.

METHODS: We used antigen arrays to analyze the reactivity of antibodies in matched serum and CSF samples of 20 patients with untreated relapsing-remitting MS (RRMS), 26 methylprednisolone-treated patients with RRMS, and 20 control patients with other noninflammatory neurologic conditions (ONDs) against 334 different antigens including heat shock proteins, lipids, and myelin antigens.

RESULTS: We found different antibody signatures in matched Cerebospinal fluid (CSF) and serum samples The targets of these antibodies included epitopes of the myelin antigens cyclic nucleotide phophodiesterase, myelin basic protein, myelin-associated oligodendrocyte basic protein, proteolipid protein (59%), Heat shock protein 60 KDa and heat shock protein 70 Kda (38%), and the 68-kDa neurofilament (3%).

The antibody response in patients with MS was heterogeneous; CSF antibodies in individual patients reacted with different autoantigens. These autoantibodies were locally synthesized in the CNS and were of the immunoglobulin G class. Finally, we found that treatment with steroids decreased autoantibody reactivity, epitope spreading (when the immune reaction changes from targeting the primary epitope to also targeting other epitopes. With time the number of targets expand) and intrathecal (within the space surrounding the brain and spinal cord) autoantibody synthesis.

CONCLUSIONS: These studies provide a new avenue to investigate the local antibody response in the CNS, which may serve as a biomarker to monitor both disease progression and response to therapy in MS.

Heterogeneous autoantibody reactivity in CSF antibodies from Relapsing remitting MS. Heatmap in which each column represents an RRMS patient, and each row shows the antibody reactivity to an antigen according to the colorimetric scale shown on the left.

The role of autoreactive antibodies in multiple sclerosis (MS) has received intense attention since the discovery of oligoclonal immunoglobulin (Ig) bands in the majority of patients with MS. However, many questions remain. It is still possible that the intrathecal production of antibody is an epiphenomenon of CNS autoimmunity, resulting from growth factor-driven expansion of long-lived B cells in the meningeal compartment. Alternatively antibody production from antigen-driven proliferation of B-cell clones could be a component of CNS immunopathology. Antibodies from oligoclonal bands is produced by B lineage cells within the CNS. These have been shown to recognizes epitopes of multiple neurotropic viruses and Epstein-Barr virus antigens. However, as this study shows anti-myelin antibodies also occur in the serum and CSF of patients with MS.

The array contained a large number of myelin protein epitopes that could be probed to see what antibodies were present in the blood and cerebrospinal fluid of MSers but these were largely linear peptide sequences of about 20 amino acids (proteins are often around 300-400 amino acids) and thus would fail to detect antibodies that detect conformational epitopes, which detect three-dimentional structures that have been found to be more important with regard to pathogenic-disease causing antibodies. There were some whole proteins and tissue extracts on this arrray.

As found previously in animal autoimmunity, there can be a wide diverisity of autoantibody (antibodies that react to the body) response and that this can broaden with time. There was alot of diversity seen between individuals and there was not one protein that everybody reacted to but it was evident that everyone with MS was reacting to at least one of the proteins. This occurred with relative high frequency for some of the epitopes such as over 60% of MSers vereses 0% in health individuals. However myelin is being damaged in MS and one wonders if this triggers an antibody response so it is secondary to the problem rather than being a primary problem.

If you get immunosuppression with your drug of interest, it may be hoped that the antibody responses would be reduced. This may be a new tool to detect this.