Archive for November, 2018

Immune Study Adds to Evidence Of Body-Wide Problems With Energy Production in Chronic Fatigue Syndrome (ME/CFS)

Increased expression of CD24 …could thus reflect abnormalities in maintaining appropriate ATP generation (in ME/CFS). The authors

Numerous studies suggest problems with energy production exist in chronic fatigue syndrome (ME/CFS). The huge energy needs exertion places on the muscles and brain suggest they’re an obvious place for energy production problems to show up. Ramping up to fight off pathogens also places extreme demands on energy production in immune cells. Now comes a study which suggests that energy production problems in a subset of B-cells could be setting people with ME/CFS up for problems with inflammation and autoimmunity.

B-cells and ME/CFS

The first couple of successful Rituximab trials brought renewed interest in the immune cells most effected by the drug – the B-cells. The last, large Rituximab trial unfortunately failed – the drug does not work in ME/CFS – but the Rituximab effort succeeded in other ways.

First off, it brought two creative and dedicated researchers, Oystein Fluge and Olav Mella to the field and energized ME/CFS research in Norway.

B-cell

A B-cell producing antibodies to fight pathogens

Secondly, the B-cells – which play a huge role in immunity (and autoimmunity) – finally got some study in ME/CFS.  Keeping with ME/CFS’s time-honored tradition of falling between the cracks in medicine, several studies found no indication of altered levels of “classical B-cell markers”.

Something unusual did, however, pop up, in an extended analysis, which went well beyond the classical markers usually explored. In 2015 a study found that a molecule called CD24 was highly expressed in a group of B-cells.

CD24 is an adhesion molecule which turns on various signaling networks – it basically tells cells what to do. It is most highly expressed on early stage or transitional B-cells as they emerge from the bone marrow.

During the normal transition from immature to mature metabolically active B-cells, early B-cells are tested again and again for evidence that they may be turning into autoantibody producing cells and many are removed.  Over time, as these cells transform themselves into mature B-cells, the CD24 molecule gradually disappears from their surfaces. High levels of these molecules in people with ME/CFS suggested that a problem with B-cell maturation might be present.

Since antibody producing B-cells play a major role in fighting off infections, having a bunch of immature B-cells hanging around the immune systems of ME/CFS patients could constitute a problem.

Plus there’s evidence that the CD24 molecule plays a role in several diseases.  CD24 polymorphisms (genetic variants of CD24) have been associated with increased risk for and accelerated progression of autoimmune diseases including multiple sclerosis, rheumatoid arthritis, and systemic lupus erythematous. CD24 can also be over‐expressed in many cancers, including B cell lymphomas.

In short, it’s not a molecule you want to ignore.

Energy Production Problems in the Immune System

Front Immunol. 2018 Oct 22;9:2421. doi: 10.3389/fimmu.2018.02421. eCollection 2018. CD24 Expression and B Cell Maturation Shows a Novel Link With Energy Metabolism: Potential Implications for Patients With Myalgic Encephalomyelitis/Chronic Fatigue Syndrome. Mensah FFK1, Armstrong CW2, Reddy V1, Bansal AS3, Berkovitz S4, Leandro MJ1, Cambridge G1.

In a 2018 study, a UK and Australian group took B-cells from ME/CFS patients and healthy controls, stimulated them and then monitored what happened.  As before, they found an increased frequency of CD24+ B-cells in ME/CFS patients.

What made the increased frequency of these naïve CD24 packed B cells in ME/CFS so interesting, though, was their mode of energy production.

Examining the metabolism of the ME/CFS patients’ cells, the researchers found a “strong(ly) positive” association between the amount of glycolysis and lactate produced and the expression of CD24 molecules on ME/CFS patients’ B-cells. Put another way, the more glycolysis was used to produce energy and the greater the lactate production (a by-product of glycolysis) – the more the CD24 molecule showed up on ME/CFS patient’s B-cells.

energy production me-CFS

Findings suggesting that a body-wide problem with energy production are present in ME/CFS are piling up.

Lower levels of mitochondria in these cells suggested one reason ME/CFS patients’ B-cells may be stuck in this mode.  Not surprisingly, the authors found this more prominent B-cell subset in ME/CFS was also largely unresponsive to stimulation.

That suggested that not only do the energy problems in ME/CFS extend to the immune cells but that they could be impacting immune functioning – in this case by keeping ME/CFS B-cells in a naïve state – that may be associated with disease.

Plus increased levels of the CD24 molecule have also been associated with a damaging state called “senescence”.  Instead of undergoing a process called autophagy during which a cell’s contents are safely recycled, during senescence – which is often associated with aging –  damaged mitochondria cause cells to slowly deteriorate while producing scads of pro-inflammatory factors.

An exercise physiologist, Graham Salmun, recently reported that his exercise study results suggest senescence is indeed occurring in ME/CFS.  He believes problems with aerobic energy production are a) impairing ME/CFS patients ability to produce energy and b) creating a senescent state that is causing chronic inflammation.

Anaerobic Thresholds, Fatty Acid Problems and Autophagy: Dr. Klimas’s Exercise Study

Conclusion

This study provided an intriguing metabolic snapshot of the immune system. The fact that increased expression of the CD24 molecule has been associated with autoimmune disorders and cancer makes the CD24 finding in ME/CFS interesting, but the metabolic connection the researchers found may be more important.

Their findings suggest that the same problems producing energy found elsewhere in ME/CFS may also be occurring in their immune cells.  Plus the findings suggest that a state of senescence, chronic inflammation and cellular unresponsiveness may be present as well. The possible penalties of having an immune system with a broken aerobic energy production system could go beyond fatigue and pain and extend to problems with autoimmunity, and perhaps in rare cases, cancer.

Studies finding an increased reliance on anaerobically produced energy in the muscles, the brain, in neutrophils and now in some of the B-cells all suggest that a body-wide disruption in energy production may be present in ME/CFS.

 

Could “Junk DNA” Be Causing Chronic Fatigue Syndrome / Myalgic Encephalomyelitis?

Carmen Scheibenbogen

Dr. Scheibenbogen continues to successfully plough new ground

It seems like every time you turn around another part of the genome pops up. It’s amazing how far our knowledge of the human genome has progressed since the Human Genome Project was completed just 15 years ago. Thankfully the small band of researchers involved in chronic fatigue syndrome / myalgic encephalomyelitis (ME/CFS) seem to be keeping up with the latest findings.

Dr. Scheibenbogen seems intent on ploughing new ground. First she re-energized the search for autoantibodies in ME/CFS. Then she examined the effectiveness of a promising treatment called immunoadsorption. With her latest study she and her colleagues at Institute for Medical Immunology, Charité-Universitätsmedizin in Berlin became the first in this disease to examine a peculiar part of our genome called long non-coding RNA’s.

Hope for an ME/CFS Autoimmune Subset: A German Researcher Steps Forward

When most people think of RNA they’re probably thinking of the messenger RNA (mRNA) which carries the genetic code from the DNA in the nucleus of our cells to ribosomes. The ribosomes then translate the mRNA into proteins – which then do the work of our cells.

pre mRNA

pre-mRNA -(Wikipedia—httpsupload.wikimedia.orgwikipediacommonsthumbaa4Pre-mRNA-1ysv-tubes)

Not all RNA produces proteins, however. Formerly termed “junk DNA“, non-coding RNA’s – RNA’s which do not encode proteins –  make up a substantial part of our genome.  (They lack the “reading frames” necessary for the process of translation from RNA to protein to begin). Long non-coding RNA’s (lncRNA) are particularly long pieces of RNA (>200 nucleotides long) which do not encode proteins.

While their presence has been known for decades, it wasn’t until the 1990’s that the first hint of the role they play in regulating gene expression and epigenetics  appeared. Research since then has shown that lncRNA’s play an important role modulating the activity of transcription factors which turn the expression of our genes on and off.

Epigenetics –  heritable changes in gene expression that do not involve changes in the underlying DNA sequence – is all the rage now. The ability of infections and other stressors to turn genes on or off via epigenetics presents an intriguing explanation for how an infection could result in ME/CFS. Because lncRNA’s can regulate the epigenetic process, they could provide even more basic insights into how ME/CFS began.

LncRNA’s can also catalyze biological reactions and respond to cellular signals. Their extreme flexibility allows them to interact with proteins, DNA and RNA to affect many physiological processes. One review stated they, “can impact almost all physiological functions.” Another review called them, “a new and crucial layer of biological regulation”.

They’re certainly providing a fresh look at complex diseases. Search for long non-coding RNA’s in PubMed and you’ll get a long list of diseases they may be implicated in. A recent review of the role lncRNA’s may play in cancer called them, “new players in the old battle against cancer”. Some regulate mitochondrial synthesis and energy production.  Pathogens can induce the production of lncRNA’s in humans that then promote viral survival.

It’s become increasing apparent that they can play a role in fundamental developmental processes that can produce chronic disease states. One review called them “arguably the hottest area of RNA research” today.  Still much remains to be learned about the roles they play.

“Remarkable” Finding

The expression signature of very long non‑coding RNA in myalgic encephalomyelitis/chronic fatigue syndrome. Chin‑An Yang1,2,3,4 , Sandra Bauer5, Yu‑Chen Ho3, Franziska Sotzny5, Jan‑Gowth Chang1,3,4† and Carmen Scheibenbogen. Transl Med (2018) 16:231 https://doi.org/10.1186/s12967-018-1600-x

In this study – the first of its kind in ME/CFS – Dr. Scheibenbogen examined the expression of ten very large lncRNAs (> 5 kb) involved in immune regulation, or which influence genes involved in the stress response and/or metabolic and neurologic processes.

The authors were clearly surprised by their findings.  After all, they’d simply taken ten lncRNA’s that they thought, based on findings in other diseases, might play a role in ME/CFS.

In a finding they called “remarkable”, the expression of those ten lncRNA’s was enough to distinguish ME/CFS patients from healthy controls. In fact, the expression of any two of three of these lncRNA’s (NTT, MIAT and EMX2OS) was all that was needed to pick out most ME/CFS patients.

Diagnositic accuracy

The elevated expression of just three lncRNA’s was enough to identify most ME/CFS patients

Then seeking to understand if the chronic illness state present in ME/CFS could be turning these lncRNA’s on by exposing them to biological stressors known to be present in ME/CFS. They found that punishing cell lines with oxidative stress did, in fact, increase the expression of the lncRNA’s. Subjecting the cell lines to a viral stressor also increased expression of one of them. Doing it increased the expression of a gene associated with chronic inflammation and blood vessel dysfunction – two problems that are believed to be present in ME/CFS.

ME/CFS seems to revel in producing findings that baffle researchers and this study was no expression. The enhanced expression of one lncRNA (EMX2OS) was something of a mystery as it is rarely found in the cells (peripheral blood mononuclear cells) they examined. It is, however, increased in brain hypoxia; i.e. low oxygen levels in the brain – which Dr. Shungu’s studies suggest may be present in ME/CFS.

Study Suggests “Bad Energy” is Core Problem in Fibromyalgia and Chronic Fatigue Syndrome (ME/CFS)

Finally, the authors noted that it took just three lncRNA’s (NTT, MIAT, and EMX20s) to do what it took Kerr 88 genes and Naviaux 8-13 metabolites to do – differentiate ME/CFS patients from healthy controls. One wonders if Scheibenbogen’s examination of lncRNA’s is getting at some basic components of ME/CFS.

Treatment Implications

No direct treatment implications were mentioned. In the short-term they’re viewed more as providing excellent diagnostic biomarkers. For instance, lncRNA’s are now being used to one of most difficult diagnostic scenarios of all – prostate cancer. They’re now being used in men with high PSA levels and negative biopsy results.

Direct treatments based on lncRNA’s are probably years away but should be noted that lncRNA’s are a very active area of research for a good reason: turning down their expression could turn off basic processes that cause disease. The author of a recent study which found that LncRNA’s play a role regulate fat metabolism reported:

“We are still in the early stages of figuring out how lincRNAs function in human disease, but what used to be considered ‘junk’ in the genome may actually point us towards the jackpot of developing effective therapeutic approaches for cardiometabolic diseases,” Jennie Lin, MD, MTR

This new exploration of ME/CFS patient’s genomes further substantiates the notion that ME/CFS is a disease of immune dysregulation. Given the study’s strong results surely more lncRNA study in ME/CFS is on the way.

 

Could Crippled Herpesviruses Be Contributing to Chronic Fatigue Syndrome (ME/CFS) and Other Diseases?

We provide evidence…. that herpesviruses dUTPases…(have) unique immunoregulatory functions that can alter the inflammatory microenvironment and thus exacerbate the immune pathology of herpesvirus-related diseases including myalgic encephalomyelitis/chronic fatigue syndrome, autoimmune diseases, and cancer. Williams et. al.

Most people are exposed to herpesviruses such as Epstein-Barr virus (EBV) early in their lives and carry the viruses in latent form in their B cells. Sometimes – particularly when the body is under stress – the immune system slips a bit and the viruses reactivate, causing anything from no symptoms at all to – more rarely – being associated with such devastating disorders as autoimmune diseases and cancer.  One study suggests that glucocorticoids released during stress tell EBV to come out of hibernation.

EBV virions leukemia

EBV infected (green) leukemia cells

Herpesviruses have a long enough history in ME/CFS for the disease to have been referred to as chronic Epstein-Barr virus syndrome by some in the 1980’s. However, over thirty years later, the role herpesviruses play in ME/CFS is unclear. Are they simply a common trigger of ME/CFS or do they play a more fundamental role? Several studies have found no evidence of herpesvirus reactivation while others suggest immune problems exist that could allow the virus to wreak havoc in some patients.

The Ohio State University team lead by Maria Ariza and Marshall Williams believes researchers have missed an obvious possibility. They don’t believe the virus per se is the problem. (If they’re right, you can basically throw out all the viral load studies.)

It’s not that the virus is reactivating; in fact, they believe the virus may be most dangerous in ME/CFS when it fails to reactivate properly and produces kind of a very low-level, smoldering infection. Even as the immune system in people with ME/CFS is mostly smothering EBV, the virus is producing a protein that’s causing harm.

“Surprisingly, none of these studies have approached the possibility that virus encoded proteins, rather than the viruses themselves, may act as drivers of/contribute to the pathophysiological alterations observed in a subset of patients with ME/CFS.” Authors

It turns out that in herpesviruses a failure to replicate produces something called “abortive lytic replication”.  As it does that, it produces proteins that get ejected into the blood stream or get inserted in vesicles called exosomes, which then travel through the blood. These exosomes are now believed to play important roles in cell to cell communication.  (Maureen Hanson is now studying exosomes in ME/CFS).

The protein released during abortive lytic replication is an enzyme called deoxyuridine triphosphate nucleotidohydrolase or EBV-dUTPase. The unusual herpesvirus dUTPase saga at Ohio State University began way back in 1985 with a Williams/Glaser study. It gathered force in the mid-2000’s with a series of papers suggesting the protein might be a good target for chemotherapy, produced “sickness behavior” in mice, and triggered pro-inflammatory cytokine production.

In 2010 Ronald Glaser won an NIH grant to study the protein titled STRESS EFFECTS ON VIRUS PROTEIN INDUCED INFLAMMATION AND SICKNESS BEHAVIOR and the hunt was on to determine dUTPase’s effects in ME/CFS.  (This long standing grant continues today under Ariza and Williams’ name.)

A 2013 paper suggested dUTPase might provide a way to reconcile the studies which had not found herpesvirus reactivation in ME/CFS with others suggesting that the virus could be having profound effects. It found that even under conditions of low viral load, herpesvirus dUTPases were able to trigger a pro-inflammatory response strong enough to promote atherosclerosis and perhaps even precipitate a heart attack. In 2012, Williams, Ariza , Glaser and Martin Lerner and Lenny Jason produced the first direct evidence that dUTPases may be producing problems in ME/CFS. The small study found a prolonged antibody response to the protein in a large subset of ME/CFS patients.

A 2014 study indicated that during EBV’s last gasp while undergoing lytic replication, the virus was pouring enough dUTPase into exosomes to produce major immune effects that supported or promoted the establishment/maintenance of further EBV infections.

The 2017 ME/CFS Study

J Med Virol. 2017 Mar 17. doi: 10.1002/jmv.24810. [Epub ahead of print] Myalgic Encephalomyelitis/Chronic Fatigue Syndrome and Gulf War Illness patients exhibit increased humoral responses to the Herpesviruses-encoded dUTPase: Implications in disease pathophysiologyHalpin P1, Williams MV1,2, Klimas NG3,4, Fletcher MA3,4, Barnes Z3,5, Ariza ME1,2.

Then, in an expanded version of the 2012 study, the group in 2017 (which also included Nancy Klimas and Mary Fletcher) presented stronger evidence that herpesvirus produced dUTPases were present and could be causing harm in a subset of ME/CFS patients. The study looked for evidence that herpesvirus produced dUTPases were tweaking the immune systems of 74 ME/CFS patients – and found it.

The fact that antibodies to dUTPases produced by both EBV and HHV-6 were found in almost fifty percent of the ME/CFS patients in the study suggested that the two herpesviruses may be reactivating each other in ME/CFS – a feature also found in immune suppressed states such as organ transplant patients and drug induced hypersensitivity syndrome (DRSS).

Plus, for the first time, autoantibodies to the human dUTPases (humans produce a dUTPase as well) were found in ME/CFS – at much higher levels than in healthy controls (39% vs. 5%).

The authors suggested the Loebel’s 2014 study, which uncovered problems that ME/CFS patients’ T cell’s were having in suppressing EBV, could account for the evidence of multiple herpesvirus reactivations.

EBV I: A Deficient Immune Response, Increased Levels of Epstein-Barr Virus Opens Up EBV Question in Chronic Fatigue Syndrome Again

The immune system does ultimately jump in and suppress the virus in most people with ME/CFS, but it takes its time to do that. That delay appears to give herpesviruses the time they need to spill immune altering dUTPases into the bloodstream and slip them into exosomes to travel through the body.

EBV smoldering infection

EBV is halted from fully reactivating but the authors believe the smoldering infection present in ME/CFS could have significant consequences for some.

Besides the immune alterations possibly caused by herpesvirus produced dUTPases, they may be contributing to numerous symptoms including flu-like symptoms, fatigue, cognitive problems, anxiety, etc. in ME/CFS.

Plus, because failed herpesvirus reactivations commonly occur alongside actual herpesvirus reactivations, herpesvirus encoded dUTPases could end up being an excellent biomarker for herpesvirus reactivations.

This strange model of partial viral reactivation could end up playing a role in ME/CFS, Gulf War Syndrome and other diseases in several ways. It could be actually driving ME/CFS in a subset of patients, or it could, along with other possibly related immune issues, be exacerbating it.

Next Steps

However it all works out, it’s clear that the Ohio State University team’s long embrace of this novel protein is paying off. The more work they do with herpesvirus-encoded dUTPases, the more evidence they seem to find of its role in ME/CFS and other diseases. They have an 8-year continuing NIH grant under their belts – a grant that looks like it and the herpesvirus-dUTPase-ME/CFS saga will likely continue in the foreseeable future.

If the findings hold up, it may even provide a treatment option – the authors have published a paper alerting drug-makers to the potential this escaped protein may hold in treating herpesvirus infections.

Dr. Williams reported that the group has “some exciting data” concerning the potential role dUTPase plays in autoantibody production and the neurological effects the protein may be having in people with ME/CFS. The manuscripts are being written up now and will be submitted shortly.