Combatting Brain Calcification in Some Autism (and Bipolar and Schizophrenia) and Osteoprotegerin (OPG) as a potential biomarker, implicating Cav1.2

In today’s post there is more supposition than normal, but plenty of anecdotal evidence.  It follows on from the previous post that suggested calcification might be an issue in some types of autism.  As we know, many unrelated biological dysfunctions can lead to autism, but there do seem to be some commonly affected pathways.

This subject is definitely worthy of much more detailed study than my post, which is based on an initial review of the science.  Some leading researchers, like Persico and Courchesne are fully aware of the issue.  I am not sure who would undertake such a study.  There is no physician specialty dedicated solely to osteoporosis, so we are lacking experts.  The bone-vascular axis is worthy of more study, as much for heart disease as autism.

A variety of medical specialists treat people with osteoporosis, including internists, gynecologists, family physicians, endocrinologists, rheumatologists, physiatrists, orthopaedists, and geriatricians.  If you do not know what a physiatrist is, I also had to look it up.  Physical Medicine and Rehabilitation (PM&R) physicians, also known as physiatrists, treat a wide variety of medical conditions affecting the brain, spinal cord, nerves, bones, joints, ligaments, muscles, and tendons.

 

Overview

There is more support for the potential use of calcium channel blockers that affect Cav1.2, via its effect on calcification by modulating Osteoprotegerin (OPG).  OPG is known to be elevated in autism and its two older brothers schizophrenia and bipolar.

It appears that in some people with severe brain calcification, that shows up on CT scans, biphosphanate drugs can be helpful, but do not actually shrink the calcification, perhaps they stop it growing.

Biphosphanate drugs used to treat osteoporosis are not without side effects in some people.

Some people have disturbed calcium homeostasis as a result of drugs they are taking, for example antiepileptic drugs.

So-called “chelation” using powerful intravenous antioxidants has been shown in the TACT clinical trial to reduce future heart problems, but only in people with diabetes. Diabetics are known to have disturbed calcium homeostasis leading to calcification, heart disease and osteoporosis.

In some counties intravenous antioxidants have long been given to people with diabetes to treat its main side effects but not to clear calcification.  In those countries this is seen as perfectly safe and routine. Preventative care for diabetics is actually rather poor in the UK and US.

Vitamin K plays a key role in calcium homeostasis and in some people just giving large amounts of this vitamin has the required therapeutic effect.  Unless given alongside blood thinning drugs, it is claimed that high dose Vitamin K does not have side effects.

Perhaps the most common osteoporosis therapy, calcium plus vitamin D is shown in some trials to be of no value whatsoever.  This therapy would most likely be ill advised in autism.

Osteoprotegerin (OPG)

Osteoprotegerin (OPG) is a cytokine involved in calcification and inflammation.

Osteoprotegerin has been used experimentally to decrease bone resorption in women with postmenopausal osteoporosis.

 It has been particularly related to the increase in cardiovascular risk in patients suffering from diabetes

Interestingly it has been shown that the L type calcium channel Cav1.2 regulates Osteoprotegerin (OPG) expression and secretion.

A NASA space shuttle flight in 2001 tested the effects of osteoprotegerin on mice in microgravity, finding that it did prevent increase in resorption and maintained bone mineralization.  Space flight is not good for your bones.

Osteoprotegerin levels are elevated in people with bipolar and schizophrenia.

Osteoprotegerin levels in patients with severe mental disorders

Severe mental disorders are associated with elevated levels of inflammatory markers. In the present study, we investigated whether osteoprotegerin (OPG), a member of the tumour necrosis factor receptor family involved in calcification and inflammation, is elevated in patients with severe mental disorders.

Methods

We measured the plasma levels of OPG in patients with severe mental disorders (n = 312; 125 with bipolar disorder and 187 with schizophrenia) and healthy volunteers (n = 239).

The mean plasma levels of OPG were significantly higher in patients than in controls (t₅₃₁ = 2.6, p = 0.01), with the same pattern in bipolar disorder and schizophrenia. The increase was significant after adjustment for possible confounding variables, including age, sex, ethnic background, alcohol consumption, liver and kidney function, diabetes, cardiovascular disease, autoimmune diseases and levels of cholesterol, glucose and C-reactive protein.

Conclusion

Our results indicate that elevated OPG levels are associated with severe mental disorders and suggest that mechanisms related to calcification and inflammation may play a role in disease development.

As shown in the study below, many inflammatory cytokines are elevated in autism, just look at those insulin-like growth factor binding proteins.  Osteoprotegerin is a modest 500% of what it might be expected to be in non autism.

Neuroglial Activation and Neuroinflammation in the Brain of Patients with Autism

Chelation

Because of the continuing non-debate in scientific terms about vaccines and autism, it is unlikely that there will ever be any study about calcium chelation and autism.  Rather than admit that in a small number of cases vaccination may trigger mitochondrial disease and result in autism, there is complete denial, at least in public. In private it is an open secret.

The planned chelation trial in autism was banned, on “safety grounds”.

It looks to me that the enemy is not mercury or other heavy metals, the problem is much less exotic. 

Oxidative Stress

Most people with autism have oxidative stress, which should be improved by any potent antioxidant.  Agents used to chelate metals have to be potent antioxidants.

Calcification

In some yet to be determined percentage of people they potentially have disturbed calcium homeostasis resulting in some calcium deposits in the brain.  Those chelating to remove, most likely non-existing, “toxins” may sometimes be reducing harmful calcification.

Fortunately there has been a very large study, called TACT, on de-calcification (calcium chelation) in Coronary Heart Disease.

One large group of people at risk from low bone density are those with diabetes.

Patients with diabetes, who made up approximately one third of the 1,708 TACT participants, had a 41 percent overall reduction in the risk of any cardiovascular event; a 40 percent reduction in the risk of death from heart disease, nonfatal stroke, or nonfatal heart attack; a 52 percent reduction in recurrent heart attacks; and a 43 percent reduction in death from any cause.

   

Chelation for Coronary Heart Disease

§  Patients with diabetes, who made up approximately one third of the 1,708 TACT participants, had a 41 percent overall reduction in the risk of any cardiovascular event; a 40 percent reduction in the risk of death from heart disease, nonfatal stroke, or nonfatal heart attack; a 52 percent reduction in recurrent heart attacks; and a 43 percent reduction in death from any cause. In contrast, there was no significant benefit of EDTA treatment in participants who didn't have diabetes.

Source:  https://my.americanheart.org/idc/groups/ahamah-public/@wcm/@sop/@scon/documents/downloadable/ucm_446204.pdf

From the Mayo Clinic:-

          Results of trial to assess chelation therapy (TACT) study presented

 A further review from TACT just looking at patients with diabetes:- 

The Effect of an EDTA-based Chelation Regimen on Patients with Diabetes and Prior Myocardial Infarction in TACT

Patients with diabetes:-

Patients without diabetes (no benefit over placebo):-

Treatment

The 10 component 500 mL intravenous solution in TACT consisted of 3 g of disodium EDTA, adjusted downward based on estimated glomerular filtration rate; 7 g of ascorbic acid; 2 g of magnesium chloride; B-vitamins, and other components (eTable 4). The placebo solution consisted of 500 mL of normal saline and 1.2% dextrose (2.5 g total). The solution was infused over at least 3 hours through a peripheral intravenous line weekly for 30 weeks and then biweekly to bimonthly to complete 40 infusions.

Background

The Trial to Assess Chelation Therapy (TACT) showed clinical benefit of an ethylene diamine tetraacetic acid (EDTA-based) infusion regimen in patients 50 years or older with prior myocardial infarction (MI). Diabetes prior to enrollment was a pre-specified subgroup.

Methods and Results

Patients received 40 infusions of EDTA chelation or placebo. 633 (37%) had diabetes (322 EDTA, 311 placebo). EDTA reduced the primary endpoint (death, reinfarction, stroke, coronary revascularization, or hospitalization for angina) [25% vs 38%, hazard ratio (HR) 0.59, 95% confidence interval (CI) (0.44, 0.79), p<0.001] over 5 years. The result remained significant after Bonferroni adjustment for multiple subgroups (99.4% CI (0.39, 0.88), adjusted p=0.002). All-cause mortality was reduced by EDTA chelation [10% vs 16%, HR 0.57, 95% CI (0.36, 0.88) p=0.011], as was the secondary endpoint (cardiovascular death, reinfarction, or stroke) [11% vs 17% HR 0.60, 95% CI (0.39, 0.91), p=0.017]. After adjusting for multiple subgroups, however, those results were no longer significant. The number needed to treat to reduce one primary endpoint was 6.5 over 5 years (95% CI (4.4, 12.7). There was no reduction in events in non-diabetics (n=1075, p=0.877), resulting in a treatment by diabetes interaction (p=0.004).

Conclusions

Post-MI diabetic patients age 50 or older demonstrated a marked reduction in cardiovascular events with EDTA chelation. These findings support efforts to replicate these findings and define the mechanisms of benefit. They do not, however, constitute sufficient evidence to indicate the routine use of chelation therapy for all post-MI diabetic patients.

Effect of the Polypill on Calcification

Oral antioxidants like NAC and Alpha lipoic Acid given daily will have both a direct and indirect “chelating” effect.

Alpha-Lipoic Acid Promotes Osteoblastic Formation in H2O2 -Treated MC3T3-E1 Cells and Prevents Bone Lossin Ovariectomized Rats.

 

Alpha-lipoic acid (ALA), a naturally occurring compound and dietary supplement, has been established as a potent antioxidant that is a strong scavenger of free radicals. Recently, accumulating evidences has indicated the relationship between oxidative stress and osteoporosis (OP). Some studies have investigated the possible beneficial effects of ALA on OP both in vivo and in vitro; however, the precise mechanism(s) underlying the bone-protective action of ALA remains unclear. Considering this, we focused on the anti-oxidative capacity of ALA to exert bone-protective effects in vitro and in vivo. In the present study, the effects of ALA on osteoblastic formation in H(2)O(2) -treated MC3T3-E1 pre-osteoblasts and ovariectomy (OVX)-induced bone loss in rats were investigated. The results showed that ALA promoted osteoblast differentiation, mineralization and maturation and inhibited osteoblast apoptosis, thus increasing the OPG/receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL) ratio and leading to enhanced bone formation in vitro and inhibited bone loss in vivo. Further study revealed that ALA exerted its bone-protective effects by inhibiting reactive oxygen species (ROS) generation by down-regulating Nox4 gene expression and protein synthesis and attenuating the transcriptional activation of NF-κB. In addition, ALA might exert its bone-protective effects by activating the Wnt/Lrp5/β-catenin signaling pathway. Taken together, the present study indicated that ALA promoted osteoblastic formation in H(2)O(2) -treated MC3T3-E1 cells and prevented OVX-induced bone loss in rats by regulating Nox4/ROS/NF-κB and Wnt/Lrp5/β-catenin signaling pathways, which provided possible mechanisms of bone-protective effects in regulating osteoblastic formation and preventing bone loss. Taken together, the results suggest that ALA may be a candidate for clinical OP treatment.

Statins are known to promote bone health.

Statins and osteoporosis:new role for old drugs.

Osteoporosis is the most common bone disease, affecting millions of people worldwide and leading to significant morbidity and high expenditure. Most of the current therapies available for its treatment are limited to the prevention or slowing down of bone loss rather than enhancing bone formation. Recent discovery of statins (HMG-CoA reductase inhibitors) as bone anabolic agents has spurred a great deal of interest among both basic and clinical bone researchers. In-vitro and some animal studies suggest that statins increase the bone mass by enhancing bone morphogenetic protein-2 (BMP-2)-mediated osteoblast expression. Although a limited number of case-control studies suggest that statins may have the potential to reduce the risk of fractures by increasing bone formation, other studies have failed to show a benefit in fracture reduction. Randomized, controlled clinical trials are needed to resolve this conflict. One possible reason for the discrepancy in the results of preclinical, as well as clinical, studies is the liver-specific nature of statins. Considering their high liver specificity and low oral bioavailability, distribution of statins to the bone microenvironment in optimum concentration is questionable. To unravel their exact mechanism and confirm beneficial action on bone, statins should reach the bone microenvironment in optimum concentration. Dose optimization and use of novel controlled drug delivery systems may help in increasing the bioavailability and distribution of statins to the bone microenvironment. Discovery of bone-specific statins or their bone-targeted delivery offers great potential in the treatment of osteoporosis. In this review, we have summarized various preclinical and clinical studies of statins and their action on bone. We have also discussed the possible mechanism of action of statins on bone. Finally, the role of drug delivery systems in confirming and assessing the actual potential of statins as anti-osteoporotic agents is highlighted.

Verapamil via the effect on OPG should have positive effect on bones and reduce vascular calcification.

Use of Biphosphanate Drugs to Treat Brain Calcification

Primary brain calcification in patients undergoing treatment with the biphosphanate alendronate

Brain calcification might be associated with various metabolic, infectious or vascular conditions. Clinically, brain calcification can include symptoms such as migraine, Parkinsonism, psychosis or dementia. The term Primary Brain Calcification was recently used for those patients without an obvious cause (formerly idiopathic) while Primary Familial Brain Calcifications was left for the cases with autosomal dominant inheritance. Recent studies found mutations in four genes (SLC20A2,PDGFRB, PDGFB and XPR1). However, these genes represent only 60% of all familial cases suggesting other genes remain to be elucidated. Studies evaluating treatments for such a devastating disease are scattered, usually appearing as single case reports. In the present study, we describe a case series of 7 patients treated with Alendronate, a widely prescribed biphosphanate. We observed good

tolerance and evidence of improvements and stability by some patients. No side effects were reported and no specific symptoms related to medication. Younger patients and one individual continuing a prescription (prior to study commencement) appeared to respond more positively with some referred improvements in symptoms. Biphosphanates may represent an excellent prospect for the treatment of brain calcifications due to their being well tolerated and easily available. Conversely, prospective and controlled studies should promptly address weaknesses found in the present analysis.

Patient 3. A 43-year-old man, one of seven children born to the same mother (described below as Patient 4), presented with rapid progression of parkinsonism. In the last 5 years, a progressive presentation of general bradykinesia, rigidity, and paresis in the right arm had developed. He had previously been an active individual with regular employment. Prior to recruitment, this patient had been on carbidopa/levodopa, which was continued throughout the duration of the present study. Genetic screening identified a SLC20A2 mutation (c.1483 G > A)3, and the patient was placed on alendronate therapy.

Patient 4. This 84-year-old woman presented with mild depression, late-stage parkinsonism, and large calcifications (10.85 cm3) in the basal ganglia and cerebellum. She is the mother of Patient 3 and carries the same SLC20A2 mutation. This patient had been taking alendronate for 10 years due to a diagnosis of osteoporosis. Intriguingly, she presented with fewer symptoms than her son, despite being 41 years old older.

We chose alendronate due to its availability, safety, and comfortable dosing schedule (oral administration, once a week). Etidronate probably works via a different mechanism (bulk action binding to hydroxyapatite) than

the newer amino bisphosphonate alendronate (inhibition of osteoclasts). This might explain why the effects seen in our series were less dramatic than those seen in patients treated with etidronate. Thus, while alendronate has a more convenient dosing schedule and, possibly, fewer side effects, a larger clinical trial should consider the choice of bisphosphonate carefully.

To date, there is no specific treatment for primary brain calcification; the main goal is symptom management.

Clinicians should make sure that the idiopathic/primary profile is accurately defined to rule out any underlying organic cause, e.g., in non-idiopathic basal ganglia calcification caused by abnormal calcium regulation, such as in primary endocrine disorders.

Bisphosphonates represent the only effective (although still anecdotal) treatment that could have wider applications in basal ganglia calcification. Prospective, controlled studies should be conducted to address the weaknesses of the present manuscript and establish a definitive analysis of bisphosphonate therapy for primary brain calcification. Furthermore, the excellent tolerability profile of alendronate in primary brain calcifications suggests that a trial in asymptomatic patients could help address the potential benefit of this strategy to control symptoms in younger patients.

Conclusion

Bisphosphonates may be applicable, safe and change the natural progression of primary brain calcifications, especially in younger patients and across prolonged periods. Nevertheless, future studies with adequate design should answer remaining questions.

Metabolic Bone Diseases

There are numerous things that can affect the bone-vascular axis including various  metabolic diseases.  This is rather beyond the scope of an autism blog, but if you are interested here is a link.

Imaging Findings and Evaluation of Metabolic Bone Disease

Conclusion

Unless you have evidence of osteoporosis, or a brain scan showing calcification, it might be rather extreme to take a biphosphanate drug like Fosamax.

If you already take oral NAC , ALA or L-carnitine you have a pretty potent therapy which would target any calcification, if indeed it existed.  Intravenous ALA, as used my Monty’s Grandad for years, should be even more effective as it is for diabetic neuropathy.

Those using verapamil appear to have another layer of protection against calcification. I did suggest to Agnieszka that elevated OMG might indeed be the biomarker needed for the use of verapamil in Autism. Remember to contact her to participate in her study.

Verapamil use in Autism – Request for Case Reports from Parents

Vitamin K2 is claimed to be extremely safe unless you are taking a blood thinning drug like Warfarin, that are Vitamin K antagonists.

Some studies claim great results from K2, while some others are more mixed.  It is likely that depending on what underlying dysfunction exists, high dose K2 may help or do nothing.  It is clear that low amounts of K2 are damaging.

So K2 would seem worthwhile trialing.  It is found in the not so pleasant tasting Natto.  Vitamin K (more K1 than K2) is found in broad-leafed vegetables.  The excellent Linus Pauling Institute reviewed all the vitamin K evidence and concluded people should:-

 “eat at least one cup of dark green leafy vegetables daily”

This brings me back to where I started the previous post with the Mediterranean diet, rich in dark green leafy vegetables.

Intravenous infusion of antioxidants looks like a very good idea for people with diabetes.  Where we live this has been standard practice for years, where Monty’s grandad goes twice a year for 10 days of ALA infusion, the rest of the year he is prescribed oral ALA.  This is given to control diabetic neuropathy, but clearly a side effect is that it will reduce the likelihood of a heart attack or stroke.

I have no doubt IV infusion of ALA would be beneficial for some with autism, but I think they might get sufficient benefit from oral ALA or indeed NAC.

I wish the FDA would permit the “chelation” autism trial in the US, I have no doubt it would show a positive effect, but not for the reasons put forward by DAN doctors and the chelation cults. 

The TACT chelation trial in older people showed that the therapy was very well tolerated.  IV ALA therapy is also well tolerated.

Public health officials should not fear the truth.  In the long run the truth is the best policy and when given all the facts the public are not stupid.  If vaccination is in the interest of their child, enough parents will happily cooperate. The Herd Immunity Threshold (HIT) is the percentage of people who need to be vaccinated.  HIT is 95% for measles.  Therapies used at Johns Hopkins exist to minimize the possible damaging effect on mitochondria and never give paracetamol/acetaminophen to children after a vaccination.

Activated Microglia and Inflammation in Autism

There have been yet more autism studies recently, highlighting neuroinflammation and the role of cells called microglia.  The result is this rather long post; but there is film to watch, if it gets heavy going.

Glia derives from a Greek word for glue. The original thought was that the glial cells “glued” the neurons together.

It turned out that glial cells do very much more and might be better thought of as “resident immune cells”.  They have other functions including synaptic pruning, which appears to have gone awry in autism.  They also form myelin, and when this goes wrong, big problems follow.

Microglia are inside the blood brain barrier and one of their jobs is to swallow up any foreign bodies that should not be there, before they can do damage.  It appears that this process is mainly modulated via potassium channels.  The majority of research focuses on the calcium-activated K⁺ channels, particularly KCNN4/KCa2 and 3.1, and ATP-sensitive K⁺ channels (K_ATP).  Administration of diazoxide, a classic K_ATP channel activator, is shown to reduce microglial activation and is neuroprotective in a variety of models involving neuroinflammation. 

However, K_v 1.3 and K_v 1.5 are also involved in activated glia.  We have seen in earlier posts, that blocking K_v 1.3 can be effective in autism (remember those TSO worms).

Immunomodulatory Therapy in Autism - Potassium Channel Kv1.3, Parasitic Worms, and their ShK–related peptides

For the scientists among you:-

Microglial Ion Channels as Potential Targets for Neuroprotection in Parkinson’s Disease

Protective Roles for Potassium SK/KCa2 Channels in Microglia and Neurons

Synaptic pruning

A very small Acer Palmatum

Synaptic pruning could itself be the subject of an entire blog.  I will just use the analogy of a different kind of pruning.

With ornamental trees, to obtain the perfect form, pruning is very important.  You have to clear away the dead wood and encourage growth in particular areas to achieve the optimal shape.  You need to know when to cut, where to cut and how much to cut.

The human brain develops with far too many synapses and they too need pruning.  The weak ones need to give way for the strong ones to prosper.  Too many synapses lead to poor brain function.  This process is going on from childhood to early adulthood.  Microglia are heavily involved in this pruning process, as you will see in the video shortly.

We know that synaptic pruning is implicated in autism and very likely in its big brother, schizophrenia.

Impaired Synapse 'Pruning'Linked to Autism

Activation of Microglia

Microglia can be in either a resting or activated state. In the activated state, for no good reason, they can do damage.  They can also react with mast cells to produce more inflammation.

(here is a link for the mast cell followers of Theoharides; they know who they are)

Dysregulated Brain Immunity and Neurotrophin Signaling in Rett Syndrome and Autism Spectrum Disorders

The subject is very complex.  For those with an hour to spare there is an excellent presentation by Beth Stevens from Harvard.  Click on the link below to go to the SFARI website and the video.

http://sfari.org/sfari-community/community-blog/webinar-series/2013/webinar-beth-stevens-discusses-brain-immunity-and-wiring

By a bizarre coincidence, there is another B Stevens researching glial cells and autism.  This time it is Bruce Stevens, in Florida.

His paper is interesting because he is using a known anti-oxidant (alpha lipoic acid, ALA) to affect brain glial cells.

One of the odd things is that we know in autism there is both oxidative stress and neuro-inflammation; they are a self-perpetuation combination.  There are numerous effective anti-oxidants; almost too many.  There is, however, a paucity of effective, safe, anti-inflammatory drugs.  In fact the best anti-inflammatory drug is probably an anti-oxidant.  So called Reactive Oxygen Species (ROS) are among the biggest causes of neuroinflammation.  With anti-oxidants you can neutralize the ROS, and thereby you take a big bite out of the neuroinflammation.

  

Alpha-lipoic acid effects on brain glial functions accompanying double-stranded RNA antiviral and inflammatory signaling.

Abstract

Double-stranded RNAs (dsRNA) serve as viral ligands that trigger innate immunity in astrocytes and microglial, as mediated through Toll-like receptor 3 (TLR3) and dsRNA-dependent protein kinase (PKR). Beneficial transient TLR3 and PKR anti-viral signaling can become deleterious when events devolve into inflammation and cytotoxicity. Viral products in the brain cause glial cell dysfunction, and are a putative etiologic factor in neuropsychiatric disorders, notably schizophrenia, bipolar disorder, Parkinson's, and autism spectrum. Alpha-lipoic acid (LA) has been proposed as a possible therapeutic neuroprotectant. The objective of this study was to test our hypothesis that LA can control untoward antiviral mechanisms associated with neural dysfunction. Utilizing rat brain glial cultures (91% astrocytes:9% microglia) treated with PKR- and TLR3-ligand/viral mimetic dsRNA, polyinosinic-polycytidylic acid (polyI:C), we report in vitro glial antiviral signaling and LA reduction of the effects of this signaling. LA blunted the dsRNA-stimulated expression of IFNα/β-inducible genes Mx1, PKR, and TLR3. And in polyI:C treated cells, LA promoted gene expression of rate-limiting steps that benefit healthy neural redox status in glutamateric systems. To this end, LA decreased dsRNA-induced inflammatory signaling by downregulating IL-1β, IL-6, TNFα, iNOS, and CAT2 transcripts. In the presence of polyI:C, LA prevented cultured glial cytotoxicity which was correlated with increased expression of factors known to cooperatively control glutamate/cysteine/glutathione redox cycling, namely glutamate uptake transporter GLAST/EAAT1, γ-glutamyl cysteine ligase catalytic and regulatory subunits, and IL-10. Glutamate exporting transporter subunits 4F2hc and xCT were downregulated by LA in dsRNA-stimulated glia. l-Glutamate net uptake was inhibited by dsRNA, and this was relieved by LA. Glutathione synthetase mRNA levels were unchanged by dsRNA or LA. This study demonstrates the protective effects of LA in astroglial/microglial cultures, and suggests the potential for LA efficacy in virus-induced CNS pathologies, with the caveat that antiviral benefits are concomitantly blunted. It is concluded that LA averts key aspects of TLR3- and PKR-provoked glial dysfunction, and provides rationale for exploring LA in whole animal and human clinical studies to blunt or avert neuropsychiatric disorders

The obvious question is whether other antioxidants have the same effect.  Most likely nobody knows.  I did ask both B Stevens #1 and B Stevens #2 for their thoughts on this – so far no answer.

Brain inflammation a hallmark of autism, according to large-scale analysis

Finally to the subject of this post, the recent Johns Hopkins study that shows inflammation in the autistic brain.

This is the press release from Johns Hopkins so it is quite readable.

While many different combinations of genetic traits can cause autism, brains affected by autism share a pattern of ramped-up immune responses, an analysis of data from autopsied human brains reveals. The study, a collaborative effort between Johns Hopkins and the University of Alabama at Birmingham, included data from 72 autism and control brains. It was published online today in the journal Nature Communications.

“There are many different ways of getting autism, but we found that they all have the same downstream effect,” says Dan Arking, Ph.D., an associate professor in the McKusick-Nathans Institute for Genetic Medicine at the Johns Hopkins University School of Medicine. “What we don’t know is whether this immune response is making things better in the short term and worse in the long term.”

The causes of autism, also known as autistic spectrum disorder, remain largely unknown and are a frequent research topic for geneticists and neuroscientists. But Arking had noticed that for autism, studies of whether and how much genes were being used — known as gene expression — had thus far involved too little data to draw many useful conclusions. That’s because unlike a genetic test, which can be done using nearly any cells in the body, gene expression testing has to be performed on the specific tissue of interest — in this case, brains that could only be obtained through autopsies.

To combat this problem, Arking and his colleagues analyzed gene expression in samples from two different tissue banks, comparing gene expression in people with autism to that in controls without the condition. All told, they analyzed data from 104 brain samples from 72 individuals — the largest data set so far for a study of gene expression in autism.

Previous studies had identified autism-associated abnormalities in cells that support neurons in the brain and spinal cord. In this study, Arking says, the research team was able to narrow in on a specific type of support cell known as a microglial cell, which polices the brain for pathogens and other threats. In the autism brains, the microglia appeared to be perpetually activated, with their genes for inflammation responses turned on. “This type of inflammation is not well understood, but it highlights the lack of current understanding about how innate immunity controls neural circuits,” says Andrew West, Ph.D., an associate professor of neurology at the University of Alabama at Birmingham who was involved in the study.

Arking notes that, given the known genetic contributors to autism, inflammation is unlikely to be its root cause. Rather, he says, “This is a downstream consequence of upstream gene mutation.” The next step, he says, would be to find out whether treating the inflammation could ameliorate symptoms of autism.

The full study is here:-

Transcriptome analysis reveals dysregulation of innate immune response genes and neuronal activity-dependent genes in autism

What I liked about the study was the comment made by Arking, a specialist in genetics, that it did not seem to matter what the genetic cause was, all the brain samples exhibited the same inflammation.  So it does not matter which of millions of possible combinations of genetic dysfunction is present, one key physiological result is shared neuroinflammation.

Take home message:  Treat the neuroinflammation in people with Autism.

The question of course is how.

Since it seems easy to treat oxidative stress, a leading cause of neuroinflammation, we should go to extreme lengths to finish that job. 

I started it with NAC and recently added Sulforaphane/broccoli.  I suspect there are more “low hanging fruit” to be gathered here. Perhaps just an additional supplemental (exogenous) antioxidants, or perhaps something clever like increasing the amount DJ-1, which is needed to support Nrf2 which turns on the anti-oxidant genes. Early 2015 will see my oxidative stress therapy optimized.

Treating Neuroinflammation in Autism

There are lots of possible ways to treat neuroinflammation, some of which we have already covered in this blog.  Sometimes it gets called immunomodulatory therapy.

There are some natural options like quercetin and turmeric.  Turmeric is also possibly chemo-protective:-

“Currently there is no research evidence to show that turmeric or curcumin can prevent or treat cancer but early trials have shown some promising results.”

Cancer Research UK

Interestingly, people who eat a lot of curry (Indians) have a very low incidence of cancer.

1.     Steroids, like Prednisone

These are already used, particularly in regressive autism.  They are potent, but have side effects.

2.     Blockers of Potassium channel Kv1.3

This is a clever approach, since it appears that this potassium channel is involved in mediating the inflammatory response. By blocking these channels the response we have seen that the immune response can be moderated and in some people, there autism moderated.

3.     Activators of Potassium channel K_ATP

We learned earlier in this post about diazoxide

4.     Other Microglial Ion Channels

The various other potassium, calcium and sodium channels need to be considered.

5.     Ibuprofen

This common painkiller reduces inflammation and is used to reduce inflammation associated with autism secondary to mitochondrial disease.

Do not use acetaminophen/paracetamol/Tylenol.  These will increase oxidative stress, since it depletes GSH and also affect mitochondria.

6.     Leukotriene receptor inhibitors (i.e. montelukast, zafirlukast)

These are interesting because they are used to treat asthma and so are very widely used. They are not steroids and so do not have their side effects.  They are proved to have anti-inflammatory effects.

Montelukast/Zafirlukast is used to reduce inflammation associated with autism secondary to mitochondrial disease.

7.     Pregnenolone

I wrote a post a while back on Pregnenolone, which is interesting, since you do not need a prescription.  But does it work?

Well, after I wrote the post below, the results from a clinical trial in adults with autism was finally published.

Pregnenolone - an effective OTC anti-inflammatory therapy for autism?

Brief report: an open-label study ofthe neurosteroid pregnenolone in adults with autism spectrum disorder.

Abstract

The objective of this study was to assess the tolerability and efficacy of pregnenolone in reducing irritability in adults with autism spectrum disorder (ASD). This was a pilot, open-label, 12-week trial that included twelve subjects with a mean age of 22.5 ± 5.8 years. Two participants dropped out of the study due to reasons unrelated to adverse effects. Pregnenolone yielded a statistically significant improvement in the primary measure, Aberrant Behavior Checklist (ABC)-Irritability [from 17.4 ± 7.4 at baseline to 11.2 ± 7.0 at 12 weeks (p = 0.028)]. Secondary measures were not statistically significant with the exception of ABC-lethargy (p = 0.046) and total Short Sensory Profile score (p = 0.009). No significant vital sign changes occurred during this study. Pregnenolone was not associated with any severe side effects. Single episodes of tiredness, diarrhea and depressive affect that could be related to pregnenolone were reported. Overall, pregnenolone was modestly effective and well-tolerated in individuals with ASD.

Trial doses were:-

Days 1-14: 100 mg

Week 1 and 2: 200 mg

Week 3 and 4: 350 mg

Week 5 and 6: 400 mg

Week 7 -12: 500 mg

So it was modestly effective, but the doses were huge.  It is a hormone and our endocrinologist did not much approve of the idea.

I will give this idea a miss.

8.     Statins

The current treatment for neuroinflammation in my Polypill is Atorvastatin.

I have already written a great deal about why statins may be effective in some people with autism; just make sure you do not have low cholesterol or mitochondrial disease.

Arthritis is another disease mediated by inflammation:-

Statin treatment for rheumatoid arthritis: a promising novel indication

To me it is no surprise that statins have therapeutic value in rheumatoid arthritis.

9.     NF-κB inhibitors

Because NF-κB controls many genes involved in inflammation, it is not surprising that NF-κB is found to be chronically active in many inflammatory diseases, such as inflammatory bowel disease, arthritis, sepsis, gastritis, asthma, atherosclerosis and others.

So perhaps NF-κB is for inflammation ,what Nrf2 is for oxidative stress, a force multiplier?

There are very many other inflammatory diseases like rheumatoid arthritis and so it is quite a well-trod path looking for inhibitors of NF-κB.

Before we get into that, a quick check on what we already know from research to schizophrenia (adult-onset autism).

The interaction of nuclear factor-kappa B (NF-κB) and cytokines is associated with schizophrenia. 

Abstract

BACKGROUND:

Many reports suggest that schizophrenia is associated with the inflammatory response mediated by cytokines, and nuclear factor-kappa B (NF-kappaB) regulates the expression of cytokines. However, it remains unclear whether the interaction between NF-kappaB and cytokines is implicated in schizophrenia and whether the effect of neuroleptics treatment for 4 weeks is associated with the alteration of cytokines.

METHODS:

Sixty-five healthy subjects and 83 first-episode schizophrenic patients who met DSM-IV criteria and who were never treated with neuroleptics previously were included. Serum levels of cytokines such as interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) were examined by using sandwich enzyme immunoassay (EIA). Peripheral blood mononuclear cell (PBMC) mRNA expressions of cytokines (IL-1beta, TNF-alpha) and NF-kappaB were detected by using semiquantitative reverse transcription polymerase chain reaction (RT-PCR). NF-kappaB activation was examined by using transcription factor assay kits.

RESULTS:

Schizophrenic patients showed significantly higher serum levels and PBMC mRNA expressions of IL-1beta and TNF-alpha compared with healthy subjects. However, treatment with the neuroleptic risperidone for 4 weeks significantly decreased serum levels and PBMC mRNA expressions of IL-1beta in schizophrenic patients. NF-kappaB activation and PBMC mRNA expression in patients were significantly higher than those in healthy subjects. Furthermore, PBMC mRNA expressions of IL-1beta and TNF-alpha were positively correlated to NF-kappaB activation in both schizophrenic patients and healthy control subjects.

CONCLUSIONS:

Schizophrenic patients showed activation of the cytokine system and immune disturbance. NF-kappaB activation may play a pivotal role in schizophrenia through interaction with cytokines.

It seems fair to conclude that NF-κB inhibitors are well worth investigating.

Interestingly, one of my new “pet” compounds, alpha lipoic acid appears to have another role here:-

Evidence that α-lipoic acid inhibitsNF-κB activation independent of its antioxidant function.

Abstract

OBJECTIVE:

α-Lipoic acid (LA) exerts beneficial effects in cardiovascular diseases though its antioxidant and/or anti-inflammatory functions. It is postulated that the anti-inflammatory function of LA results from its antioxidant function. In this study we tested whether inhibition of NF-κB by LA is dependent on its antioxidant function.

METHODS:

Human umbilical vein endothelial cells (HUVECs) were treated with tumor necrosis factor-α (TNFα) in the presence of various antioxidants, including LA, tiron, apocynin, and tempol. The activation of the nuclear factor-κB (NF-κB) signaling pathway was then analyzed.

RESULTS:

LA, but not other tested antioxidants, inhibited TNFα-induced inhibitor-kappaB-α (IκBα) degradation and VCAM-1 and COX2 expression in HUVECs. Although LA activated the phosphatidylinositol-3-kinase (PI3-kinase)/Akt pathway in HUVECs, inhibition of Akt by LY294002 did not affect inhibition of TNFα-induced IκBα degradation by LA. In transient co-transfection assays of a constitutively active mutant of IκB kinase-2 (IKK2), IKK2(EE), and a NF-κB luciferase reporter construct, LA dose-dependently inhibited IKK2(EE)-induced NF-κB activation in addition to inhibiting IKK activity in in vitro assays. Consistent with the effect on luciferase expression, LA inhibited IKK2(EE)-induced cyclo-oxygenase-2 (COX2) expression, suggesting that IKK2 inhibition by LA may be a relevant mechanism that explains its anti-inflammatory effects.

CONCLUSIONS:

LA inhibits NF-κB activation through antioxidant-independent and probably IKK-dependent mechanisms.

 

This really makes ALA look very interesting.  It is cheap, widely available and well tolerated.

10.       Low Dose Naltrextone                       

Your local doctor will probably tell you that Low Dose Naltrexone (LDN) is a load of quack nonsense, partly because it is claimed to help so many unrelated disorders.

I would not have questioned that opinion, before I had started by investigation into the biology of the brain and seen how many apparently unrelated conditions are actually interrelated.  This can be established by science, not quackery.

First to note is that tiny doses of some substances do indeed sometimes have effects quite different to large doses.

We saw earlier how a tiny stimulation of the body’s nicotinic receptors produces a different effect to a large dose.

My own experience showed that a tiny, but specific, dose of Clonazepam has a marked effect, whereas conventional medical wisdom would say such a small dose would do absolutely nothing.  In this case, I was just following the clever idea of Professor Catterall, from the University of Washington.

I also found that tiny doses of a TRH analog had a positive effect and quite different to the “regular” dose.

The advocates of LDN suggest it for conditions including Crohn's disease, fibromyalgia and multiple sclerosis (MS).  As I mentioned earlier in this blog, some Fibromyalgia appears to be a condition that was almost autism; perhaps the final hit, in a multiple-hit process failed to occur.  Crohn’s is an immune disease and is a type of inflammatory bowel disease (IBD).  MS is an inflammatory disease in which the insulating covers of nerve cells in the brain and spinal cord are damaged.

Preliminary research suggests LDN may have an effect on inflammation. Naltrexone has an antagonistic effect on Toll-like receptor 4 (TLR4), which are found on microglia, which can modulate the body's response to inflammation. It has been hypothesized that LDN may have anti-inflammatory effects through this pathway.

  

Conclusion

The immediate conclusion is that there are plenty of ways, already existing, that might very well help reduce neuroinflammation in autism.  They just requires a little further thought and investigation.

The broader conclusion here is about the merit of genetic testing.

Undoubtedly, if you could analyze the entire genome in a person with autism and also measure the expression of those suspect genes in the brain, you would gain a great deal of information.  In a few cases, where there is a single gene causing the “autism”, you might well be able to figure out a therapy.

You cannot take brain biopsies from living people.  We did come across that clever Ricardo Dolmetsch, growing brain samples from skin cells.  He has now moved over to the private sector.

Future Science - Accurate Diagnosis of Autism Phenotype using Brain Biopsies grown from Skin Cells

So for the moment genetic testing will just generate a vast amount of data, that in many cases will not be of any immediate clinical relevance.

The good news, as pointed out by Dan Arking, from Johns Hopkins, is that many of these numerous, unrelated, genetic dysfunctions end up with the same biological manifestations.

There may be thousands, or even millions of combinations, of genetic dysfunctions that lead to autism with neuro-inflammation.

You can go ahead and treat the neuro-inflammation, without any knowledge of exactly which gene has which SNP (single nucleotide polymorphisms)  or who had what CNV (copy number variant).

For me, the identification of so-called autism genes like PTEN and BCL2 is interesting, as are the single gene causes of autism.  We can then see that a reduced expression of that gene might contribute to autism, caused by multiple gene dysfunction (multiple-hits).  For the great majority of people with ASD, they have had multiple-hits.

I read Ricardo Dolmetsch’s Stanford research into Timothy syndrome, which is caused just by one gene, albeit a very important one.  I considered that perhaps a partial dysfunction might occur, leading to disturbance in the protein expressed by this gene.  I had no idea whether in my son this dysfunction existed, whether it might be caused by a SNP (there are several known ones) or if a dysfunction was caused as a consequence of a metabolic disruption caused by autism, such as oxidative stress or neuroinflammation,  affecting the function of an undamaged gene.

It did not matter; I just carried on and did a little practical test.  This led me to include Verapamil in my Polypill.  No genetic testing was required.

It was suggested to me that genetic testing might help point me in the right direction.  I think it would likely point me in all directions.  We all carry many genetic errors, and most of us thrive regardless, so most genetic errors are irrelevant.

The clever future diagnostic tool is proteomics.

  

Clusters

From now, I will consider autism in terms of a manageable group of clusters.  Once you know, based on symptoms and some measurable biomarkers, which cluster you are in, you would have a good chance of predicting which drugs would be effective.

The underlying genetic causes may, or may not, overlap with other people in that cluster.

Some clusters may overlap. Note the case of siblings with autism, when one is early onset and the other is regressive.  Was the regressive one really symptom free early one? Or, was it just a second hit nudged him “over the edge” and then people noticed?

This would be a practical approach that could be used.  I think when people talk of phenotypes and autisms, they are thinking about very precise biological causes and then it just becomes too complicated to expect your local doctor to ever figure out.

90+% of people quite probably fit into a handful of clusters.  Then you just need a diagnostic flowchart leading to the relevant cluster and then a specific drug toolkit.

My Polypill is the drug toolkit for one cluster; and it is not a rare one.