cbd oil for undermethylation

Undermethylation vs. Overmethylation: Causes, Symptoms, Treatments

While there are a number of potential factors that may cause mental illness, one that many people don’t consider is that of methylation. It is believed that genetic mutations may lead to abnormalities in the methylation process. The methylation cycle is a specific biochemical pathway that is responsible for influencing a variety of critical biological processes including: the immune system, DNA maintenance, production of energy, and detoxification.

During the methylation process, neurotransmitters are created, giving rise to certain emotional states. If there is problems with the methylation process, it is speculated that a number of different psychiatric symptoms may arise. These speculative symptoms are thought to be specifically correlated with whether someone is considered to be stuck in a state of “overmethylation” vs. “undermethylation.”

Undermethylation vs. Overmethylation

There isn’t anything wrong with being overmethylated nor undermethylated, but it may cause some sort of mental distress. Too much methyl (overmethylation) results in a unique set of symptoms that are different than those resulting from too little methyl (undermethylation). While undermethylation is generally considered less problematic than overmethylation, you may want to consider “methylation” and its potential to cause mental illness.

Undermethylation (Histadelia) : Too little methyl (a carbon group with three hydrogen atoms). Essentially they have low levels of SAM-e, which donates methyl. This can lead to perfectionism, high accomplishment, and high achievement. Now why would this be problematic? 15-20% of these people experience problems as a result of the undermethylation. This can lead to low levels of serotonin, making them susceptible to depression.

Overmethylation (Histapenia) : Too much methyl (a carbon group with three hydrogen atoms). It is very active in the brain, and too much leads to “too much of a good thing.” This causes an overproduction of serotonin, norepinephrine, and dopamine in the brain. In many cases, high serotonin levels can cause psychological problems including reduced motivation, reduced libido, weight gain, and confusion.

MTHFR Gene Polymorphisms Causes Undermethylation or Overmethylation

There are a number of theories regarding the causes of overmethylation as well as undermethylation. The most obvious cause is that of MTHFR genetic polymorphisms. If you possess a polymorphism of the MTHFR gene, this may be directly associated with poor methylation or highly efficient methylation. Fortunately you can detect whether you have a mutation of the MTHFR gene by getting a simple blood test.

MTHFR is an acronym for “Methylenetetrahydrofolate Reductase.” MTHFR is an enzyme and the MTHFR gene is necessary for the creation of the enzyme which regulates the methyl cycle. When the gene is considered abnormal, the methyl cycle may function abnormally. There are a variety of possible mutations that may occur to the MTHFR gene as well as its nucleotides.

If the gene’s nucleotide is abnormal, it’s called a SNP (Single Nucleotide Polymorphism). With the MTHFR gene, there are a variety of nucleotide polymorphisms (SNPs) that may occur, some are considered heterozygous (on one strand of DNA) while others can be homozygous (on both strands of DNA). Upon diagnosis, a professional will be able to explain whether you have homozygous or heterozygous MTHFR and the specific SNP location.

  • Source: http://www.ncbi.nlm.nih.gov/books/NBK6561/

Biochemistry of Undermethylation

The undermethylation can lead to a number of deficiencies in nutrients throughout the body. Some researchers believe that if a person is a poor methylator, they are likely to have high levels of histamine, reduced zinc, and a high basophil count.

  • High histamine: It is thought that a metabolic imbalance occurs when a person isn’t capable of sufficient methylation, leading to abnormally high histamine (e.g. 70 ng/ml).
  • Low zinc: Some individuals who are chronically undermethylated tend to have zinc deficiencies.
  • Low copper: Those with poor methylation ability tend to have low levels of copper.
  • High Basophil count: This is often detected in blood tests and is associated with infections and allergic reactions. If levels exceed 50 cells/cu mm, there’s a better chance of undermethylation.
  • High homocysteine: The greater the degree of undermethylation, the higher you can expect your homocysteine levels.
  • High heavy metals: Proper methylation is thought to help detoxify the body of heavy metals and toxins. If a person is undermethylated, some speculate that they may have increased build-up of heavy metals.

Undermethylation Symptoms

Keep in mind that if you are suffering from undermethylation, you may not experience every symptom on this list. There are different genetic polymorphisms of the MTHFR gene as well as other factors that may dictate your experience. Below is a collective list of symptoms that people with undermethylation tend to exhibit.

  • Addictions: Those who are considered undermethylators may be more likely to battle addictions and/or have addictive personalities.
  • Competitive: It is believed that many undermethylators are extremely competitive in sports, business, and other facets of life. Competition is a notable signal that a person may not have a sufficient methylation process.
  • Concentrative endurance: Some individuals who are suffering from undermethylation may have a difficult time maintaining focus for prolonged periods of time. In other words, their concentration ability may wane quicker than average.
  • Delusions: Certain individuals that fall into the undermethylation diagnosis may experience delusions or beliefs that aren’t based in reality. While these generally are not severe, then can interfere with the accuracy of a person’s perception of reality.
  • Headaches: Some researchers believe that undermethylation may cause physical symptoms such as headaches.
  • High achievement: One characteristic (rather than symptom) of people with low levels of methylation is that of accomplishment and achievement. Many individuals considered top athletes, CEOs, and professionals may be fueled in part by undermethylation.
  • High libido: A person may be highly interested in sex and/or have a higher than average “drive” compared to others.
  • Obsessive compulsive: Undermethylation may provoke symptoms of OCD or other obsessive tendencies. In fact, someone may actually get diagnosed with obsessive-compulsive disorder as a result of their methylation deficiency.
  • Oppositional defiance: Another common finding is that those displaying signs of oppositional defiant disorder (ODD) tend to also have undermethylation. While this isn’t a very common diagnosis, it is thought to be related to undermethylation.
  • Inner tension: While a person who is an undermethylator may appear to exhibit a calm demeanor, they may be filled with inner tension.
  • Low pain tolerance: Individuals with undermethylation tend to have a poor tolerance to any sort of pain.
  • Perfectionism: Another trait of undermethylators is that of perfectionism. They aren’t satisfied unless tasks are completed in accordance to their specific methodology. They may be intolerant to less-than-perfect outcomes.
  • Phobias: Certain phobias or irrational fears may be caused in part by undermethylation.
  • Ritualistic behavior: Those who are undermethylated may engaged in ritualistic behavior with rigid schedules. They may have specific daily rituals to which they must adhere.
  • Seasonal allergies: It has also been suggested that allergies may be stronger among undermethylators, particularly during seasonal transitions. This may be related to naturally elevated levels of histamine, leading to more pronounced reactions.
  • Self-motivated: A person who is undermethylated may be highly self-motivated in both school and work functions. They may not need any outside inspiration or encouragement to complete their work, they are fuelled internally by themselves.
  • Social isolation: Some individuals with undermethylation may isolate themselves from others and prefer to be left alone, especially during the completion of work.
  • Strong willed: Another characteristic that is found among those who are undermethylated is that of a strong will.

Undermethylation and Serotonin : Researchers speculate that undermethylation is associated with abnormally low levels of serotonin in the brain. While the mainstream media has lead most people to believe that low serotonin is always bad, clearly not all outcomes are poor. However, the fact that undermethylators tend to have low serotonin, it can make some of these individuals more prone to developing depression. It is estimated that 15% to 20% of people dealing with undermethylation tend to find it problematic; likely as a result of serotonergic deficits.

Undermethylation Treatment

Assuming you are suffering from undermethylation, there are numerous supplements you can consider for treatment. Generally treatment doesn’t yield drastic improvement overnight, and in some cases it can take 8 to 12 months before a person feels noticeably better. However, it is also important to realize that nutritional intervention can be highly effective and successful over the long-term.

Since undermethylation results in low levels of calcium, magnesium, methionine, and Vitamin B6 – it’s important to consider these for supplementation. Additionally it may be important to avoid folic acid as levels may be abnormally high throughout neurons. Correcting nutritional imbalances is considered an important step towards improving undermethylation symptoms.

Supplements for Undermethylation

Most people aren’t fans of taking pharmaceutical medications once they have pinpointed the problem of undermethylation. If you know for a fact that undermethylation is the problem, taking a targeted nutritional approach is your best bet. Work with a professional to determine what quantities of vitamins and/or “stack” you should be taking.

  • Choline
  • Calcium
  • Magnesium
  • Methionine
  • Omega-3 fatty acids
  • SAM-e
  • Vitamin B12
  • Vitamin C

Medication Outcomes

If you plan on taking a medication, some researchers have noted that outcomes among individuals with undermethylation are considered specific based on the type of drug utilized.

  • Antihistamines: Since those with undermethylation tend to have high levels of histamine, using an antihistamine may result in favorable effects.
  • Benzodiazepines: People taking benzodiazepines like Xanax tend to have unfavorable responses if they are suffering from undermethylation.
  • SSRIs: Those dealing with undermethylation tend to respond well to selective-serotonin reuptake inhibitors. These are antidepressant drugs that specifically increase the amount of extracellular serotonin. These are considered helpful among undermethylators because they have low levels of serotonin.
  • Vitamin B Complex: Those taking Vitamin B Complex may experience adverse effects. This is because the person already has high levels of folic acid (Vitamin B9) and the complex serves to further elevate it.

Biochemistry of Overmethylation

Overmethylation is the polar opposite of undermethylation. If you are overmethylated, you will likely have elevated levels of serotonin, low levels of histamine, and likely a low count of absolute basophils. Additionally you may discover high levels of copper, but low presence of zinc and histamine throughout the body.

  • High copper: Abnormally high levels of copper are generally present among those who exhibit overmethylation.
  • Low basophil count: In cases of overmethylation, basophil count tends to be depressed, meaning theres less likelihood of an allergic reaction.
  • Low histamine: Overmethylation results in the reduction of histamine, making it completely unnecessary to use an over-the-counter antihistamine.
  • Low zinc: Another common finding among those who are overmethylators is low levels of zinc.

Overmethylation Symptoms

Below is a list of possible symptoms that you may experience during overmethylation. Understand that overmethylation will not necessarily cause every single symptom on the list and that severity of each symptom is highly subject to individual variation.

  • ADHD: Those who are overmethylators may have attention deficits and may have been previously diagnosed with ADHD.
  • Anxiety: In some cases, the overmethylation can lead to increased levels of anxiety and susceptibility to panic attacks.
  • Artistic: Some sources have suggested a link between artistic and musical pursuits and overmethylation. Whether this is accurate is subject to debate.
  • Depression: Those with overmethylation may become depressed as a result of high levels of serotonin, lack of motivation, and accomplishment.
  • Dry skin: It has also been speculated that in those who are overmethylated tend to be more susceptible to bouts of dry skin.
  • Food sensitivities: If you are an overmethylator, you may notice that you are highly sensitive to certain foods and/or chemicals.
  • Frustration: Another sign among overmethylators is rampant frustration or becoming easily frustrated in seemingly benign situations.
  • High pain threshold: Unlike those who are poor methylators, overmethylation is associated with a higher pain tolerance.
  • Low libido: Another symptom of overmethylation is that of a decreased sex drive. They may be considerably less interested in sex than average.
  • Low motivation: The general tendency among overmethylators is that of deficient achievement in workplace or competitive settings. This is often a direct result of the motivational deficit that may be a byproduct of overmethylation.
  • Nervousness: This ties into the generalized anxiety that a person may experience as a result of the overmethylation.
  • Obsessions: Some have suggested that individuals with overmethylation may demonstrate clear obsessions, but they aren’t usually accompanied by compulsions.
  • Overweight: Those who are overmethylated may be more likely to pack on weight in part due to motivational deficits and depression.
  • Paranoia: Among those who are strongly affected by overmethylation, paranoid thoughts, and possibly auditory hallucinations may emerge.
  • Restless legs: Another possible symptom that you may experience if you’re dealing with too much methylation is restless legs.
  • Self-imposed isolation: Those who isolate themselves from others may do so in part as a result of depression and or anxiety from overmethylation.
  • Self-harm: Researchers believe that among those who commit acts of self-harm and mutilation, overmethylation tends to occur.
  • Sleep disorders: Those with sleep problems may be more likely to have overmethylation.

Overmethylation Treatment

If you are dealing with overmethylation, many experts will recommend a specific supplement regimen over the course of 3 to 6 months. Some believe that it is easier to offset the effects of overmethylation than undermethylation. While you probably won’t notice major benefit overnight, you may start to feel gradually better over the course of several months.

To properly cope with overmethylation, it is recommended to work with a professional to construct a specific regimen of supplements. These supplements work to correct nutritional imbalances and should improve symptoms of the condition.

Supplements for Overmethylation

Working with a professional will help you come up with a supplement regimen that effectively offsets the overmethylation. The goal is to gradually reduce the amount of methylation that occurs with targeted nutritional interventions. Below are some supplements that someone may take if they are overmethylated.

  • DMAE
  • Folic Acid (Folate)
  • Niacinamide
  • Omega-3 fatty acids
  • Vitamin B6
  • Vitamin C
  • Vitamin E
  • Zinc

Medication Outcomes

If you are taking a medication and have a mutation of MTHFR that leads to overmethylation, below are some likely reactions.

  • Antihistamines: Since you already have low levels of histamine, you are going to respond poorly to any antihistamine drug. People who are overmethylated tend to have less allergic responses than usual as a result of the low endogenous histamine.
  • Benzodiazepines: Favorable responses have been noted among those with overmethylation to taking benzodiazepines.
  • Lithium: The mood stabilizing agent Lithium has also been thought to yield noticeable improvement among those who are overmethylated.
  • Oestrogen therapy: Should you engage in oestrogen therapy as an overmethylator, you are likely to experience an adverse reaction.
  • SAM-e: This supplement should be avoided by any individual that is dealing with overmethylation. Adverse reactions are likely to occur as this will further increase methylation.
  • SSRIs: In general, people who are overmethylated already have high levels of serotonin and don’t require an SSRI. Further increasing serotonin levels may be problematic and may yield unwanted side effects or adverse reactions.

Does treating undermethylation or overmethylation cure mental illness?

Assuming you are a person who has a polymorphism of the MTHFR gene that is causing undermethylation or overmethylation, balancing the methylation process could drastically improve your mood or psychiatric symptoms. While many people preach that treating “over” or “under” methylation with supplement stacks will “cure” you of your psychitric illness, it is important to realize that this isn’t often the case.

Addressing the methylation imbalances could significantly improve your symptoms and/or overall health. However, it does NOT mean that you will be eventually cured of your severe anxiety or depression. A considerable number of people do improve once they address their MTHFR mutations with nutritional-based therapy, but don’t isolate methylation as the sole cause of your mental illness. Your condition may be a result of a collective number of other genetic and environmental factors.

Are you suffering from overmethylation or undermethylation?

It is important to avoid assuming that you are an overmethylator or undermethylator based on the symptoms listed above. While you may be one or the other, self-diagnosing based off of information you’ve read online may end up doing more harm than good. If you are serious about learning whether you truly have a methylation problem, get the MTHFR gene tested and have the results analyzed.

Determining whether you have an MTHFR mutation is the best way for you to get proof that supports your hypothesis of being an overmethylator or undermethylator. Using a treatment protocol for overmethylation or undermethylation when you have no actual methylation dysfunction may be more problematic than beneficial. If you have had your MTHFR gene tested and successfully treated your methylation dysfunction, feel free to share your experience in the comments section below.

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Hi. I am suffering from chronic tension type daily headache since 1997. I used almost all types of antidepressants for years without any benefit. SSRI, SNRI etc make my mind very active and create insomnia. In 2016 I stoped using antidepressants and started taking GNC products.

The following vitamins/ minerals work for me:

-Niacin
-Chromium Picolinate (reduces headache 80%)
-Methyl Folate 400mcg
-Magnesium 500m
-B12 worked initially but now it makes me more sleepy.

Also calcium and copper increase restlessness. In the morning I feel tired and there is lack of motivation and in the evening I feel more energy. I need help in adjusting the proper dosage of the vitamins.

I was tested 6 months ago and am an overmethylator. My doctor put me onto a vitamin B and folate mix and I have also been to a naturopath. I have been feeling really great until 10 days ago when I had a cortisone injection.

It occurred to me this may be what has re-caused the symptoms I was experiencing last year. My question is whether cortisone can have this effect, particularly in overmethylators.

Please can you clarify, does having an MTHFR genetic mutation make you overmethylated? So in my case I have MTHFR C77T +/-….would this lead to overmethylation? (as opposed to undermethylation). I have also low histamine, low basophils, high copper, low zinc. I have a SAM/SAH ratio of 4.3 and high levels of SAH but my Doctor and I aren’t clear if this means overmethylation? Or just normal methylation. Thank you.

Yes, you are overmethylated, at least you were at the time you took the test. Your low histamine, low basophil count, high copper, and high SAH, plus a high SAM to SAH ratio (SAM should be no more than 80 percent of SAH) are all indicative of overmethylation.

(Zinc is low in most MTHFR persons and both under- and overmethylators). You should also find low homocysteine levels, if your doctor may have checked them at the same time. You will find research of the above levels indicating overmethylation documented all over Google Scholar, if you wanted to look up to show your doc.

You might know that MTHFR SNPs more often result in undermethylation than over, as the mutation disables your ability to convert folic acid into folate, impeding one of the major pathways to methylation.

Your less common overmethylation may be due to a number of other factors, including other genetic mutations, nutritional imbalances, chronic or acute trauma or stress, metal toxicities, radiation exposure, various medications – particularly antihistamines and antidepressants, which both lower histamine and increase methylation, – food allergies or sensitivities, and food and other environmental chemical exposures.

Please help me. I am an overmethylator. I recently bought niacin, folic acid and b12 and I felt relief 10 minutes after I took my dose. my headache went away, my eyes relaxed. This stuff is real. Is this enough to help me with my overmethylation problem or do I need to supplement with more stuff? I’m thinking of upping my dose to 2000mg niacin, 2mg b12 & 5mg folic acid. Is this too much?

You shouldn’t be taking Folic acid. Folate is the one you can take.

I don’t think that is correct… UNDERmethyators are the ones that have problems with Folic Acid, not OVERmethylators. Best to ask your Doctor or naturopath.

I have a son who is suffering. I cannot tell whether he is an under or an over-methylator. His B6 is on the lower side (13). He has very high zinc (though still within the high normal range), but very low copper (but still at the edge of the low normal range). He has very low homocysteine (5.7), and good B12 levels (about 800).

He flushes dramatically with only 50mg of Niacin (as nicotinic acid). He has lots of anxiety (compulsive irrational worries and ruminations). He would love to move on with his life. He has tried multiple SSRIs in the past with only a few helping a bit, but the effect is very short-lived. No SSRI’s effect has lasted longer than 2.5 weeks, no matter how high the dose.

It has been almost 2 years since he tried medication now, and because he is so sensitive to everything, he is afraid to go down that road again. So, he has just tried various supplements. He cannot tolerate even the slightest amount of methylfolate (it makes his head tight and ramps up his anxiety). In fact, he cannot tolerate even the slightest stimulant (sugar, caffeine, scary movies, intense video games, etc).

These things make his head tight, give him heart-palps, agitation, massive sweating, itching everywhere, etc. He cannot even tolerate the folic acid that is in breakfast cereal because it is slightly stimulating. Even B vitamins (even those without the folate and B12) help him feel better temporarily (for the first 12 hours), but then leave him with even more anxiety and agitation than he had to begin with.

At one point he tried TMG (trimethylglycine, the precursor to SAM-e), pouring out all but about 1/12th of a 175mg capsule, because he reacts to things so strongly. It was such a small amount we could barely see it at the bottom of the capsule (about 15mg). He reacts extremely to even that much. For the first 8 hours after taking even that much, his anxiety and agitation is very intensified, but then he has a period where he feels good–very calm and ok– for the next 6 hours.

Then it all returns to normal anxiety again. Does that mean he needs more? But the period where it increases his anxiety initially, before it helps, is so intense, he is afraid to take more. We’re also aware that TMG lowers homocysteine and his is already so low, at 5.7, so he probably can’t afford that. So what does that mean?

He has been to every doctor (GP, neurologist, endocrinologist, immunologist, psychiatrist, nutritionist, wellness coach, etc). It has been such a long time since he has felt completely well, and no one seems to know what to do to help him. He is so nervous about trying any additional supplements at this point. He just wants to give up.

Jen, my teenage son shares some of the symptoms your son suffers from, although not debilitating. I highly recommend Trudy Scott’s website and book (also Julia Ross’s). Per Trudy’s recommendations I started my son on inositol and magnesium. Over the course of a year, they nearly eliminated his palpitations, anxiety and obsessions.

The itching your son experiences sounds like his histamine levels are high, which can have an effect on mental state, so perhaps that is another avenue of research/exploration for you. Was his copper/zinc tests SERUM tests? Read up on copper at Mensah Medical website. Has he tried an elimination diet?

Gluten sensitivity (not necessarily just celiac) can have amazingly severe mental symptoms! Look at Dr. Tom O’Bryan’s website for info on that. Also, I would not rely on this article too heavily for advice as it seems to have a number of inaccuracies. Hope some of these help. Best of luck!

Hi Jen. This is exactly my son. Did you find out whether your son is an over or undermethelator? Ilana

Niacinamide may really help the anxiety and other mental issues… I’d start low, maybe 50-100mg… and ramp it up… it won’t cause the flushing.

See also  hempworx cbd oil what is it good.for

I have disabling OCD/tics; CBT does not work and medication caused tinnitus and hyperacusis so am unable to return to that route. I am getting worse and my life is a living hell. I have other issues, immune related: chronic fatigue, psoriasis and the arthritis, asthma, allergies, believe I have fibromyalgia and M.E. though awaiting diagnoses. I am fighting suicidal depression because of how hard my life is; it is a real battle and there is no hope for me.

I learned about leaky gut/brain and realized this could be why all these supplements have no helped. I want to detox the gut and brain, heal the leakage and infuse with the nutrition it needs to balance my brain. I am very poor, on benefits and live in Manchester UK. I am willing to partake in studies, tests or any trials that may be going but nowhere seems to be taking anyone on at this time.

I have asked all over and getting nowhere. Can you suggest anything please? I can’t bear this anymore, and don’t know how long I can hold on. I am incredibly strong but have reached my limit to what I can stand. I am reaching out to anyone, anywhere for help.# Best Regards, Miss Armstrong

Your best bet to treat a gut problem is the GAPS diet. Get hold of Dr Natasha Campbell-McBride’s book Gut and Pyschology Syndrome. She is a neurologist with a masters in nutrition. It is the best gut healing diet, but there are several. What they all have in common is – cut out sugar, cut out starch (all grain, potatoes). GAPS is based on another healing diet which you can find details and food lists on the internet called SCD (specific carbohydrate diet), check out SCD website breakingtheviciouscycle.info. Good luck!

Anne, So sorry to hear this. I recommend the book Dirty Genes. Good information! Do the AIP diet.

I have the MTHFR C677T mutation and methylation problems. I’ve been trying to treat it with supplements but my symptoms flip back and forth every few days from overmethylation to undermethylation and back. I can’t stabilize it! My biggest symptom for overmethylation is an extreme insomnia (

90 minutes of sleep per night) and the opposite when undermethylated (

9-12 hours sleep).

I use TMG and benfotiamine (methylated B1) to increase methylation and niacin to decrease it. I also take a lot of other supplements and have researched each one to determine how it affects methylation levels.

I’m not sure why I’m commenting. I guess I’m just throwing this out there in case it helps someone suffering similarly. It seems no one has definite answers.

I need to have my daughter tested for methylation problems. We live in the Philippines. Would anyone know where we can get this test done?

I am based in the UK and would like to get tested for Methylation and other renamed tests, but don’t know where to start. Any advice greatly appreciated.

I’m still waiting for credible scientific evidence supporting the idea of undermethylation or overmethylation and the scientific evidence that vitamin therapy can cure mental disorders. Vivian

Lots of research done on niacinamide for mental problems up to and including schizophrenia… Google it.

I have had the MTHFR testing done. It came back positive for c677t. How can I tell if I am over or under methylating?

I was diagnosed with Scoliosis, Fibromyalgia, Sjogrens, AMD, heterozygous for MTHFR and have had severe anxiety all my life. Taking the natural form of FOLATE has been of great help. My AMD (eyes) are presently stable and my anxiety is calming. Family members with severe depressive disorders, Tourettes, Obstinate Defiant Disorder, etc.

Perhaps natural Folate and no folic acid would be of help to those experiencing some of these issues. My Fibromyalgia is gone after stopping Folic Acid and taking natural Folate: along with Magnesium. The MTHFR factor being diagnosed was significant as a turning point in my health.

I am trying to understand why folate is bad for undermethylators? Is both FOLIC ACID and L-MethylFOLATE supplements bad for undermethylaors? What is the reason?

Most websites say the opposite and most integrative doctors treat undermethylators with methylfolate. However my undermethylating son had a massive emotional & hallucinatory reaction to a tiny amount so I can see why there is confustion.

Some can help methylation, but too much folic acid, folinic acid or methylfolate could be a problem for some undemethylators (if serotonin activity is low), and could also result in depression (reduce serotonin/dopamine neurotransmission) if they get too much.

Isn’t it easier to blood test for histamine to know if you have methylation problems?

I’d say no, given most websites say under methylators have high homocysteine but this page says they have low homocysteine. So I guess it can’t be used as a diagnostic test.

He said histamine, not Hcy. Hcy can blur the interpretation of over or under methylation and is not definite indicator, while everyone almost invariably agrees that elevated all blood Histamine level is the indication of under methylation. Other primary indications are:
-Low (serum) copper
-High Basophil count
-High heavy metals
(see Walsh Nutrient Power: p. 60, 80, 122)

Yes, and LOW whole blood Histamine level is the indication of OVER methylation… best to get copper and zinc levels checked too.

I found that increasing my Magnesium intake to 400mg+/day REALLY helped… not sure if this is related.

I believe I am an overmethylator… waiting for the Whole Blood Histamine test results…

I started getting paranoia, OCD, and severe anxiety after taking SAM-e. There is nothing good about being an overmethylator.

According to this website low homocysteine is an indicator of undermethylation. According to other websites it’s just the opposite, the rationale being people can’t convert homocysteine to methionine. The contradictory information is very confusing.

Exactly my thoughts. I found this article while trying to find info on undermethylators with Low homocysteine (and high blood folate), which is the category by son falls into. Most sources just assume high homocysteine with undermethylation and recommend methylfolate, as did his integrative doctor.

Despite his blood tests showing low homocysteine, she put him on methylfolate which apparently reduces homocysteine because that is ‘what you do’ for undermethylators. But even a tiny amount turned him into an emotional wreck – he put his foot through the wall and started hallucinating! I haven’t given it to him again and stopped seeing that doctor. I rather liked this article as at last he seemed to ‘fit’ the diagnosis, but it doesn’t make sense.

Too much conflicting info. I can only conclude that the science of methylation simply is not fully understood yet and some people with methylation problems do not fit neatly into “over” or “under” methylators.

I want to make sure I am reading this correctly. There are 3 duplications that can be used for either methylators.

Undermethylation
Calcium
Magnesium
Methionine
Omega-3 fatty acids <—
SAM-e
Vitamin B6 <—
Vitamin C <—

Overmethylation
Choline
DMAE
Folic Acid (Folate)
Niacinamide
Omega-3 fatty acids <—
Vitamin B12
Vitamin B6 <—
Vitamin C <—
Vitamin E
Zinc

This is the best article I have read and the most accurate. Thanks, Steve

I found out that I was MTHFR heterozygous when I lost my first pregnancy. I had to take extra folic acid and a daily aspirin during my next two pregnancies to carry to full term. That was ten years ago. Anyway I currently have terrible headaches and recurring sinus infections. I have severe hair loss and long time history of depression. I take Wellbutrin XL and brintellix, working okay. Doctor added deplin, I don’t know if it’s helping or not. My zinc is low but homocysteine is normal. Have terrible brain fog and zero motivation. What should I take/not take?

My DD& has the MTHFR C677T homozygous mutation. We have test results which also show very low homocysteine, low basophils, and high B6 and B12. I am confused as to whether she is over or undermethylating. I can see symptons in both. Namely, her symptons are:
`Moody, quick to anger, has rages, lashes out.
`Depressed and sad a lot. Cries a lot but doesn’t know why
`Unmotivated, socially isolates herself.
`Compulsive, almost addicted, to certain things/activities.
`often doesn’t feel well, some nausea or diarrhea.

She was tested positive for coeliac disease by biopsy. I try to limit dairy, but otherwise no other food sensitivities. Eats a clean unprocessed diet, Mostly paleo. I’m interested in helping her with her mood swings, and depression. Do you think that SAMe would help? Thanks, 🙂

It’s best to get tested before trying supplements, but what you’re describing sounds a lot like me and I overmethylate. I actually thought I was an undermethylator and supplemented antihistamines and methionine (which gave me fragrance sensitivities, brain fog, heart palpitations, panic attacks, more anxiety, etc…) before I finally got the whole blood histamine test and came back at “13” – pretty severe overmethylation.

I have low basophils, high b6 and b12 too and can definitely relate to the sentimentality and isolation- spent so many nights crying myself to sleep and not even knowing why. Also the lack of motivation – a strong sense of inner direction leaves me unaffected by extrinsic status/labels, and addictive personality – everything is intense for me, everything becomes an obsession because I ruminate unendingly, which has also made it hard for me to take practical steps (do sh#t) and manage time well.

Anyway just thought I’d offer myself as a glimpse into the mind of an overmethylator haha, hope she finds healing soon. 🙂

She may have the celiac like problems because she has SIBO – some guy bacteria produce folate. SIBO mimics celiac disease. Might want to talk to your doctor about a SIBO program. Nicholas

The best thing any of this information provides is temporary hope. The fact is medical doctor or not – no one has a clue how this stuff works. The orthomolecular doctors who put these theories together were looking for solutions to problems that are still beyond our scientific understanding. Most of it is based on anecdotal observation and ultimately the people who tend to benefit the most are the naturopaths perpetuating nonsense and charging $200 for BS tests.

Don’t get me wrong I’m all for alternative medicine and not all docs are full of it – most are just as blind as the majority of people searching for solutions. But answer me this if methylation, candida, adrenal, Lyme’s, heavy metals, toxicities, deficiencies, etc, etc, were the issue and were solved by taking supplements – then why aren’t there thousands of people testifying that they’re cured?

In some 20 years of obsessively reading as much as I could on alternative mental recovery I can only recall about half a dozen people out of a potential 50,000 who’ve ever solved their problem. Most people it seems do better for a day or two, week or two and then something swings the other way. I just hate how ideas are perpetuated and people get swept up in it spending fortunes on random theories. Anyway good luck.

Good comment. I’m thinking of all the supplements I’ve tried, and I can’t think of any that helped my fatigue/lack of motivation/anxiety/depression. So far, not much change. I’m embarrassed at the thousands of hours and dollars I’ve spent researching this and buying supplements.

What has helped is tyrosine, mixture of T3/T4 for hypothyroid, gf/cf diet. Reduction in starches, elimination of processed food. Probiotics, psyllium, and lots of water with a little salt added have healed the gut, and this is something I must be diligent about every day. Prescription drug NuVigil really helps with energy and focus, but it costs over $700/month, which I cannot afford, and insurance won’t pay for.

Thanks for putting things in perspective.

I’m also taking lots of supplements, and my son is too. I’ve never felt like they have helped with much, except for Glucosamine and Chondroitin for joint pain. Good food seems to work best, especially green smoothies (with extra flaxseed oil).

But one thing I’ve found that’s really helpful with my son’s anxiety and paranoia is CBD oil (cannabidiol). I’m hoping that it will keep him from having another psychotic episode also. The Mayo Clinic has even published a dosing guide for cannabidiol to treat schizophrenia.

RMC has read about 50,000 people. Did RMC read the Dr Walsh book? True, Dr Walsh only amassed information on 30,000 people and we should throw that out the window? My MEDICAL doctor works with the Walsh method and we are lucky to have a few DOCTORS in the Chapel Hill area who also subscribe to and work with the Walsh method, along with other approaches.

It is true not one thing works for EVERYONE. The blood tests may not suit everyone but it is worth the time and effort. Blood tests are reasonable on lifeextension… RMC already says he is obsessive and it is obvious he is a naysayer.

Hi RMC, I’d like to counter your comments with my own positive experience of the Dr Walsh protocol. After battling with ME, depression and anxiety for years, I now finally feel normal and able to function in the world. The ‘mad’ feeling I experienced that I couldn’t describe or make sense of diminished after a few days of limiting folinic acid from my diet.

A few weeks later my mood elevated from L-Methionine supplementation and zinc and B6 has helped reduce anxiety and improve energy levels. Yes I’m not 100 % perfect – I’m still a perfectionist! But I can go about my daily life and have set up my own business. Thanks to Margo Goldspink the only Walsh practitioner in the UK!

What precisely do you mean by limiting folinic acid from your diet? Did you limit food sources of folate?

As a mother of two undermethylated sons who are also affected by pyrrole disorder, one with schizophrenia and the other with depression, I would like to add that I think that the reason that treatment takes so long is that most people have too much folate in their diet, even if they are not supplementing with any folate. Any folate, including methylfolate and folinic acid, make my sons worse. They are now on a limited folate diet. Having pyrrole disorder along with undermethylation makes limiting folate even more important.

Hi Kim, I too have the same diagnosis as your sons. Pyrrole disorder and undermethylation. Are your sons on a current supplement regime and if so would you be open to sharing?

This was a great article, but the recommendations to get the MTHFR test as a sole measure or indicator of methylation status is problematic. There are numerous factors involved in the methylation cycle which can lead to imbalance not just the MTHFR gene. Your best bet is to get a whole blood histamine test. See the work of Dr. William Walsh (book: Nutrient Power) or Dr. Albert Mensah (many videos on this topic on his website – Mensah medical) for better information on how to get tested and for a good breakdown of how the methylation cycle works. It is also possible to have high copper and be undermethylating – if your zinc levels are low already than other metals like copper and lead can be high.

Gotta agree with the other replies here – genetic testing alone is known for being mis-leading, as you might have SNPs that cancel each other out. Functional testing (blood and urine) gives you a picture of what is actually happening in the body, not just a theory of what might be happening. I would only work with someone trained in William Walsh’s methods – he’s worked with 30,000+ patients, and has tried many methods and found what actually works.

I recently learned that I have a MTHFR genetic defect and cannot utilize Folic Acid. My doctor prescribed a methylated B complex. My homocysteine level is High. I don’t know if I am an under or over methylator. So don’t know what supplements to take. For example, should I take Sam-e? Any suggestions appreciated.

I totally agree with the last two posters. When you are high copper you are low zinc. You can’t be low in both. High copper can cause psychiatric symptoms. Undermethylators are high histamine people. Antihistamines are helpful for undermethylators. So, there are inaccuracies in this article but overall it is helpful.

Also, don’t put so much weight on the MTHFR gene. My daughter does NOT have the MTHFR gene but she is an under methylator. Dr. Walsh says 23andme is not helpful to determine methylation status. Check into his website and look up possible practitioners that can help you with this. They’ve been trained by Walsh himself: http://www.walshinstitute.org/clinical-resources.html

If a person has pyrrole disorder that is draining their zinc, yes, they can be low in both zinc and copper, but the zinc will probably be lower than the copper. I am speaking from experience with one of my sons because we have had to supplement his zinc like crazy and gradually his copper has become low, even though I give him a little copper, too.

Antihistamines might be helpful for some undermethylators, but where do you get the idea that it is helpful for undermethylators in general except maybe as a sleep aid? High histamine is a marker for undermethylation, but in general it isn’t the cause of the symptoms.

Thank you, Mom in Chicago! The link to practitioners is super helpful!! –another Mom in Chicago 😉

I think there is a lot of buzz surrounding the MTHFR test. Many of the sources I’ve been checking say that the MTHFR test alone is not enough to diagnose over or undermethylation. There are numerous other SNPs that could influence methylation status, not nust this one. Dr. Mensah (Mensah medical website), does a really good job of breaking down this topic and how to get tested. See the video section for a thorough discussion.

The other thing I’m finding is that it is possible to be an undermethylator with high copper. I don’t know what sources were used for this article but I believe there are some innacuracies. Zinc is recommended here for low copper?… that doesnt make sense. Zinc is a copper antagonist so taking more of it could worsen a copper deficiency.

I agree with Vanessa – my family has had good benefit from using Mensah Medical. Also check out the book “Nutrient Power” by William J Walsh PhD. He is clear that the ratio between methyl groups and folate is critical to consider – brilliant guy if you ask me!

Excellent suggestions. Walsh, author of nutrient power and Mensah Medical. There is so much here I would dispute but I would highly recommend these two. Turned my psych onto nutrient power and he uses it.

I am an undermethylator and high copper. We are trying to figure out the right mix of vits, etc. for me. My holistic psych tends to be distracted with other matters so I find her lack of follow thru disappointing but my naturopath has been more and more involved. Thank goodness. I added TMG to my mix to lower homocysteine but wonder how it will affect my higher copper.

To be short I was diagnosed with MTHFR 677 after a retinal blood clot and started taking Methyl Folate and B12 only at first and my symptoms of 10 years of Fibromyalgia were gone in four months! Although I think I am overmethylated now chronic fatigue and fibro fog can be cured. I had my family tested at 23andme.com $99. That is the best way to find all methylation mutation.

I have had 23AndMe analysis. How do I determine my MTHFR status?

With 23AndMe, you can buy the Strategene app (about $40). This will tell you which of your SNPs may affect your methylation. It does not give you a treatment plan, however. Dr. Ben Lynch, the designer of Strategene, says that properly managing stress, sleep, diet, exercise, and environment, is sometimes all the treatment you need for your body to work efficiently. He recommends getting a healthcare provider who understands MTHFR. (Some of them do telephone consultations.)

FWIW, I am about to have that testing done this week. By my doctor. She does it through a company called spectra cell. They do the methylation test and some vitamin deficiency testing.

Hi Helen, I live in Canada and was also thinking of getting the MTHFR gene analyzed and it says in the article that it’s possible through a blood test. I have my latest basic blood test results but don’t see anything I could relate to what I read here, so I think you’d have to specify to the general doctor before going for the test that you want to test that specific gene. Good luck!

Where can you get tested for the MTHFR gene and have it analyzed? I live in Auckland, NZ. I am also on hydrocortisone for adrenal fatigue but it is not effective anymore hence I am reducing it but I cannot tolerate (even small amounts of vitamins) which is my main frustration. Thank you for your reply. Most appreciated.

Hi Helen, I just saw your comment (you may have ready found somewhere). Most naturopaths can organize this test for you in NZ. If you get stuck go to
Organics Out West in West Auckland, we can sort it out for you. Take care.

I live in SE Auckland and would like to discuss MTHFR homogenous with you.

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Epigenetic Effects of Cannabis Exposure

The past decade has witnessed a number of societal and political changes that have raised critical questions about the long-term impact of marijuana (Cannabis sativa) that are especially important given the prevalence of its abuse and that potential long-term effects still largely lack scientific data. Disturbances of the epigenome have generally been hypothesized as the molecular machinery underlying the persistent, often tissue-specific transcriptional and behavioral effects of cannabinoids that have been observed within one’s lifetime and even into the subsequent generation. Here, we provide an overview of the current published scientific literature that examined epigenetic effects of cannabinoids. Though mechanistic insights about the epigenome remain sparse, accumulating data in humans and animal models have begun to reveal aberrant epigenetic modifications in brain and the periphery linked to cannabis exposure. Expansion of such knowledge and causal molecular relationships could help provide novel targets for future therapeutic interventions.

Introduction

Extensive political and societal debates are currently being waged at state and federal levels regarding the legalization of marijuana (Cannabis sativa), which remains today the most commonly used illicit substance in the United States and in many countries worldwide. As evident in Figure 1 , there has been a dramatic exponential increase of cannabis studies over the past two decades in response to the transformative implications resulting from the growing discussions and laws passed regarding legalization of recreational and medical marijuana use. Of the published studies to date, about 13% relate to the neurobiological effects of cannabis and approximately 27% is directed towards obtaining behavioral insights. Despite the perceived low health risk of cannabis use by the general public, there is growing clinical awareness about the spectrum of behavioral and neurobiological disturbances associated with cannabis exposure such as anxiety, depression, psychosis, cognitive deficits, social impairments, and addiction (1–7). The acute intoxication induced by cannabis consumption is strongly linked with concerns about its direct effects on cognition and motor function, but a central issue relates to its long-term impact especially when exposure occurs during critical periods of brain development. Key gaps of scientific knowledge pertain to the biological mechanisms that maintain persistent phenotypic and molecular alterations long after its acute use.

Number of publications in PubMed between 1960 and 2014 related to ‘cannabis’ research. The data shows the exponential increase in research studies over recent decades that coincides with changes in the legalization status (starting

1996) and debates of recreational and medical marijuana use. The drop in publications in the 1970s marks changes in state laws and local regulations banning possession or sale of cannabis and cannabis becoming a Schedule I drug (*).

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The major psychoactive cannabinoid within cannabis, Δ 9 -tetrahydrocannabinol (THC), targets the endocannabinoid (eCB) system, which plays a key role in the development of the brain and several other organs. In recent years, various human and experimental animal studies have evaluated the long-term impact of cannabis and cannabinoids on neurodevelopment, behavior and several biological systems such as immunological mechanisms and reproductive processes (reviewed in (7–10)). Moreover, behavioral abnormalities and molecular impairments in the brain have also been demonstrated to extend even into subsequent generations of offspring whose parents were exposed to cannabinoids before mating (11–15).

The epigenome provides a cellular fingerprint of environmental experiences, including drug exposure history, and thus is a highly relevant biological candidate expected to maintain persistent abnormalities and aberrant neuronal processing over time. The role of epigenetics in psychiatric disorders has been a major scientific focus during the past few years. According to the classic definition, “an epigenetic trait is a stably heritable phenotype resulting from changes in a chromosome without alterations in the DNA sequence” (as proposed by Conrad Waddington in the 1950s); this view implies heritability resulting in a phenotype. In the molecular biological era of recent years, “epigenetic” typically has been used to refer to mechanisms that modulate gene expression without altering the genetic code. Our article provides an overview of research endeavors relevant to cannabis-related epigenetic mechanisms that could shed light about the biological processes that establish the molecular platform that maintains marijuana’s protracted effects on gene expression and ultimately behavior.

Epigenetic mechanisms

In a biological mechanistic context, knowledge of how gene expression is regulated by the cellular network of cis-acting elements and trans-acting factors has evolved substantially during the past decade. Generally, the interaction between genomic DNA elements (specific sequences with regulatory function), epigenetic modifiers and transcription factors determines the expression state of genes. This network of processes is tightly coordinated in space and time, in the specification of different cell, tissue and organ types, and throughout the lifespan of the individual (16–18).

Some of the most important ontogenetic regulatory decisions take place in early development, and thus have critical implications for drug exposure during this period. Epigenetic modifications that can regulate gene expression levels include DNA methylation, nucleosomal structure and positioning, post-translational modifications of nucleosomal histones, histone replacement, and small RNA molecules that influence protein production ( Figure 2A ). Mechanistic implications of the specific epigenetic processes that have thus far been linked to the effects of cannabis are briefly summarized below.

Several epigenetic mechanisms relevant to the effects of exogenous cannabinoids. (A) Gene expression is regulated by a network of DNA elements (e.g. promoters) and trans-acting factors (proteins that bind to the DNA) that interact physically and functionally to generate appropriate mRNA transcript levels from a gene. The resulting balance can be disrupted by drug exposure. Regulatory mechanisms include DNA methylation (Me), positioning and post-translational modifications of nucleosomes (small blue balls), recruitment of sequence-specific and basal transcription factors and RNA polymerase II, and non-coding RNAs. The DNA-protein structure forms three-dimensional structures (represented by the chromatin loop) that influence the expression of associated genes. (B) DNA methyltranserases (DNMT) generate 5-methylcytosine (pink stars) at CpG sites, facilitated by methyl-CpG binding domain (MBD)-containing proteins. Ten-eleven translocation (TET) proteins mediate the oxidation of 5-methylcytosine to 5-hydroxymethylcytosine (green stars), leading to demethylation of the DNA. (C) Modifications of nucleosomal histone tails such as methylation (Me) and acetylation (Ac) are mediated by histone methyltransferases (HMT) and histone acetyltransferases (HAT), respectively. Depending on modified amino acid residue, methylation can have either permissive (e.g. on lysine4, K4) or repressive (e.g. on lysine 9, K9) effects on transcription. Permissive modifications facilitate gene activation via the recruitment of the RNA polymerase II machinery. Acetylation is removed by histone deacetylases (HDAC) and can lead to transcriptional repression. (D) MicroRNAs are produced from specific genes and target protein-coding messenger RNAs (mRNA) for degradation, thereby prevention protein production.

DNA Methylation

The role of DNA methylation ( Figure 2B ) in the regulation of gene expression is still controversial and highly dependent on genomic location, developmental stage, cell type, or disease state. Historically, CpG methylation in promoter regions and transcriptional regulatory sequences has frequently been associated with gene silencing, whereas methylation within the gene body is less understood and may act as either positive or negative effectors (19, 20). Accumulating evidence now also indicates that DNA methylation in brain is reversible and its distribution changes throughout neuronal maturation and aging, in neurodevelopmental disorders, including addiction to drugs such as cocaine (21, 22). Mechanistically, DNA methylation (5-methylcytosine, 5mC) is generated by DNA methyltranserases (DNMTs). At promoter regions, 5mC is often associated with the binding of methyl-CpG binding domain (MBD)-containing proteins (e.g. Mecp2). The oxidation of 5mC to 5-hydroxymethylcytosine (5hmC) by ten-eleven translocation (TET) proteins can prevent access to DNMTs and thereby can maintain an unmethylated state of the promoter, leading to transcriptional activation (23). Interestingly, DNA methylation marks at specific gene loci have been shown to even persist during the maturation of germ cells (24, 25) and thus are interesting candidates for the propagation of the long-term effects of cannabis throughout multiple generations.

Histone modifications

On the protein level, the main epigenetic mechanism that has been implicated in neurobiological disturbances related to drug abuse is posttranslational modifications of nucleosomal histones ( Figure 2C ) which with the

146bp of DNA that encircle them comprise the basic unit of chromatin. Histones are subject to a variety of modifications including but not limited to, lysine acetylation, lysine and arginine methylation, serine and threonine phosphorylation, and lysine ubiquitination and sumoylation (26). These modifications occur primarily within the histone amino-terminal tails protruding from the surface of the nucleosome as well as on the globular core region, and have been shown to influence both the accessibility of genomic regions and the binding of trans-acting factors to the DNA (27). Changes in acetylation and phosphorylation in response to drug exposure are often transient and appear to be associated with the quick activation of genes rather than the maintenance of an altered transcription state (28). However, histone lysine methylation is known to maintain stable gene expression alterations, and it is also the nucleosomal modification that has been associated with the long-term effects of marijuana and different cannabinoids in neurons and other cell types (29–32).

Non-coding RNAs (ncRNAs)

These functional RNA molecules are transcribed from DNA but are not translated into proteins. Many ncRNAs regulate gene expression at the transcriptional and post-transcriptional level. Those ncRNAs that are known to be involved in epigenetic processes can be divided into two main groups — short ncRNAs (<30 nucleotides) and long ncRNAs (>200 nucleotides). The three major classes of short ncRNAs are microRNAs (miRNAs), short interfering RNAs (siRNAs), and piwi-interacting RNAs (piRNAs) (33). Of these, alterations in miRNA profiles have been associated with cannabinoid exposure in the mammalian brain, peripheral blood cells, and the gut ( Figure 2D ) (34–37). While the exact genomic targets of specific cannabinoid-affected miRNAs remain to be characterized, these observations are mechanistically intriguing given the variety of tissue-specific cellular and developmental processes that are influenced by miRNAs.

The endocannabinoid (eCB) system

Cannabis targets the eCB system, which contributes to organogenesis as well as neurogenesis and gliogenesis of the CNS. It is well documented that the eCB system controls neuronal hardwiring during prenatal ontogeny, relevant to the development of neural pathways such as the corticostriatothalamic circuit, which are implicated in addiction and psychiatric disorders (38, 39) During postnatal developmental, the eCB system is known to be a critical regulator of synaptic plasticity. In mammals, two cannabinoid receptors have been identified (CB1R and CB2R), along with two major endocannabinoids as their ligands, N-arachidonoy-lethanolamine (anandamide) and 2-arachido-noylglycerol (2-AG) (40). During development these endogenous cannabinoid transmitters act as signaling molecules via a primarily autocrine activation of CB1Rs colocalized in the same developing neurons, whereas in the mature brain, eCBs are synthesized by postsynaptic neurons and travel retrogradely across the synapse to inhibit presynaptic neurotransmitter release via CBRs (41). CB1R is the most abundant G-protein-coupled receptor in the adult brain and mediates in large part the neurobehavioral effects of THC ( Figure 3 ). Consistent with the known neurobiological and behavioral effects of the eCB system, CB1Rs are abundant in brain areas involved in learning and memory (e.g. hippocampus), motor function (e.g. basal ganglia, cerebellum), cognitive and emotional processes (e.g. striatum, amygdala, prefrontal cortex) (3), as well as the regulation of physiological and metabolic processes including feeding and stress response via the interaction of the Hypothalamic-Pituitary-Adrenal (HPA) and Gonadal (HPG) axes (42, 43). In neurons, CB1Rs are preferentially localized on the surface of presynaptic cells regulating both excitatory (glutamate) and inhibitory (GABA) transmission. Low expression of CB2Rs has recently been reported in the brain, frequently in association with inflammatory processes (44), and it has been detected in neurons within mesocorticolimbic brain regions relevant to cognition and motor function (45, 46). Despite its low abundance in brain, modulation of the CNS CB2R has been implicated in addiction-related behaviors (47, 48). Both CBRs are present in peripheral tissues, including the immune system, adipose tissue, liver, skeletal muscle and reproductive organs (49).

Biological processes affected by cannabinoid exposure. (A) The active compounds of cannabis target cannabinoid receptors (CB1R and CB2R; expression pattern in the body is indicated by green dots in the human figure). (B) Cannabinoid receptors are trans-membrane receptors of the G protein-coupled family. The CB1R (shown in red), the primary target of THC, is expressed most abundantly in the brain, but also in the lungs, liver, kidneys, immune system, gut, and in germ cells such as the sperm. The CB2R is present mainly in the immune system and in hematopoietic cells with low expression in brain. Cannabinoid receptors can be activated by endocannabinoids (eCBs, green polygons; retrograde signaling), THC, or synthetic cannabinoids (see also Table 1 ). In the adult brain, activation of the CB1R on the surface of pre-synaptic neurons modulates the release of neurotransmitters (orange dots) that bind to their specific receptors (light blue shapes) in the post-synaptic cell, thereby changing the communication between neurons.

The normal epigenetic control of the eCB system has recently been reviewed (50). In the currect article, we focus on how cannabis, THC, and other exogenous cannabinoid receptor modulators alter epigenetic mechanisms and developmental regulation ( Table 1 ). Briefly, however, various lines of evidence strongly suggest that the eCB anadamide and eCB signaling cascades mediated via CBRs regulate cellular functions in different tissues via epigenetic alterations in DNA methylation (e.g., cell differentiation in human keratinocytes, cells in the epidermis) (51), miRNA (regulating cells involved in interleukin production and inflammatory response) (36) and histone methylation (differentiation and inhibition of gliomagenesis) (29). These data highlight the role of the eCB system in regulating a repertoire of cellular functions in diverse tissues through multiple epigenetic modifications and suggest that exogenous modulation of these pathways with drugs may have long-lasting neurobiological impact.

Table 1

Asterisks indicate the examples where cannabinoids have been shown to affect epigenetic regulation in brain or neurons.

Cannabinoid Epigenetic alteration Biological target Associated effect or consequence References
Cannabis Increased CpG DNA methylation at promoter Human peripheral blood cells Negative correlation between CB1R methylation and mRNA levels in schizophrenic cannabis users (57)
Cannabis Met/Met COMT gene genotype and promoter CpG DNA methylation Human adolescent peripheral blood cells Less likely cannabis dependence and decreased risk of psychosis (61)
* THC H3K4me3, H3K9me2; Promoter, gene body Adult rat brain (NAc) Decreased Drd2 gene mRNA levels in response to in utero THC exposure (30)
* THC H3K9me2, H3K9me3; Promoter, gene body Adult rat brain (NAc shell) Increased Penk gene mRNA levels in response to adolescent THC exposure (31)
* THC CpG DNA methylation at promoters, intergenic regions, especially in gene bodies Adult rat NAc with parental THC exposure Altered methylation enriched in genes implicated in synaptic plasticity (15)
THC H3K4me3, H3K9me3, H3K27me3, H3K36me3; Promoters, intergenic regions, gene bodies Differentiating mouse lymph node cells Genome-wide alterations in histone modifications associated with dysregulated genes and non-coding RNAs (32)
THC Increased HDAC3 expression Human trophoblast cell line BeWo Gene dysregulation during placental development (70)
* THC DNA methylation at CpG islands; miRNAs Cerebellum and peripheral T cells of simian immunodeficiency virus-infected macaques Altered DNA methylation, mRNA and miRNA expression profiles (37)
THC miRNAs Mouse myeloid-derived suppressor cells Altered mRNA, miRNA, and differentiation profile (35)
THC miRNAs Intestine of simian immunodeficiency virus-infected macaques Altered miRNA profile and intestinal epithelial cell composition (34)
Exogenous anandamide Increased global DNA methylation Spontaneously immortalized human keratinocytes (HaCaT cell line) Decreased expression of differentiation-related genes and altered cell differentiation (51)
Exogenous anandamide miRNAs Mouse lymph node cells Altered interleukin production and inflammatory response (36)
HU-210, JWH-133 cannabinoid agonists H3K9me3; Global levels CB1R and CB2R-expressing human glioma stem-like cells (U87MG and U373MG lines) Induction of differentiation, inhibition of gliomagenesis (29)
* HU-210 cannabinoid agonist miRNAs Adolescent rat brain (entorhinal cortex) Altered miRNA profile (78)

Epigenetic mechanisms relevant to the long-term effects of cannabis

The study of epigenetics in relation to drugs of abuse has been a rapidly emerging field during the past several years, yielding important mechanistic revelations about different addictions and related neuropsychiatric disorders (52, 53). However, experimental data about epigenetic effects associated with cannabis exposure are still sparse in spite of the relatively easy accessibility and frequent use and abuse of this drug. Of the few published studies, various epigenetic regulatory mechanisms that have been associated with cannabinoid exposure are summarized in Table 1 . Epigenetic modifications have been shown to directly regulate the eCB system via targeting its individual components as well as downstream targets of eCB-associated pathways in a variety of cells types ( Figures 2 and ​ and3 3 ).

Human epigenetic studies

Of the different components of the eCB system, several investigations have focused on the epigenetic regulation of the CNR1 gene, which encodes the CB1R ( Figure 3 ). Specific genomic elements of the CNR1 gene have been shown to interact with trans-acting factors, some of which are implicated in methylation of CpG sites in the DNA and histone posttranslational modifications (54–56). A few of these studies have revealed that CB1R expression is dysregulated in different pathological conditions and upon exposure to drugs of abuse. For example, CB1R expression is increased in peripheral blood lymphocytes of schizophrenic patients with cannabis abuse and is inversely correlated to methylation of the CNR1 promoter ( Table 1 ) (57). However, that study had limitations in that most cannabis users also reported alcohol and cigarette use and were diagnosed with schizophrenia, making the direct delineation of any specific cannabis effect difficult. Nevertheless, CNR1 mRNA expression levels and promoter DNA methylation status detected in the blood was related to measures of cannabis craving, the severity of nicotine dependence and severity of cannabis (and alcohol) consumption that suggest a relationship to brain function. As such, lymphocyte CNR1 DNA methylation and CNR1 mRNA expression could potentially serve as peripheral biological marks. Clearly, a greater number of studies are needed to replicate these findings and to establish causal relationships in order to fully understand the functional relevance of peripheral epigenetic disturbances to neurobiological alterations induced by drug use. Moreover, whether such associations are evident in cannabis users without other comorbid neuropsychiatric conditions is also important to address.

One of the first gene x environment epigenetic associations described with cannabis use relevant to psychiatric vulnerability involved the COMT gene and schizophrenia risk. COMT (encodes catechol-O-methyltransferase that metabolizes catecholamine neurotransmitters such as dopamine) has also long been implicated in substance use. A well-known Val 108/158 Met COMT polymorphism increases COMT activity and thus levels of dopamine, which plays a critical role in reward, motivation, cognition and other behaviors linked to addiction. The Val allele has generally been associated with increased substance use disorder (58, 59) (but see meta-analysis in (60)). Recently, Val 108/158 Met genotype interaction with COMT DNA methylation status in blood was associated with non-daily cannabis use, which was not observed in either daily or non-users. Thus, adolescents with the Met/Met genotype in combination with high rates of COMT promoter methylation were less likely to be high-frequent cannabis users than adolescents with the Val/Val or Val/Met genotype (61). Given that the status of COMT DNA methylation depended on the frequency of cannabis use in active using adolescents, it remains unanswered whether such epigenetic alterations persist long after these individuals stop using the drug.

It is evident that a complex relationship exists between genetic and epigenetic interactions, and the relationship between peripheral epigenetic marks and methylation status in brain is still unknown. Despite the apparent associations of cannabis exposure with discrete molecular alterations in humans and the possibility to conduct studies on genetic associations, the specificity of the observed disturbances attributed to cannabis must be verified especially in the light of potential polysubstance exposure, which is common in humans. In addition, cannabis consists of over 60 cannabinoids, one of which is THC, and cannabis preparations can largely differ in amounts of these various cannabinoids, typically confounding clinical studies. Another important limitation is that given the low incidence of cannabis-related mortality that would allow postmortem brain molecular analyses, most human epigenetic studies can only be conducted in the periphery of live subjects and thus their relationships with brain changes remain unclear. Nevertheless, the accumulating data indicate epigenetic disturbances in human subjects relevant to cannabis use disorders that would predict the potential for long-term molecular alterations.

Cannabinoid animal models and epigenetic factors

Animal models provide more controllable experimental strategies in which the protracted molecular consequences of long-term cannabinoid exposure can be better explored with regard to epigenetic mechanisms that could potentially maintain abnormal gene regulation and related behavioral disturbances. Such preclinical animal studies also facilitate the direct causal investigation of protracted effects in the brain as a consequence of developmental exposure to cannabinoid drugs. A number of early seminal animal studies demonstrated prenatal THC exposure on offspring behaviors and some suggested changes in gene expression (62, 63), confirmed by subsequent investigations (64–66). More recent research efforts into the developmental effects of THC directly described epigenetic alterations germane to addiction disorders. These studies focused in large part on the NAc, a critical neuroanatomical substrate underlying the pathophysiology of addiction (67–69). The CB1R is abundantly expressed on medium spiny neurons that represent the most abundant striatal cell-type and constitute the differential output pathways (striatopallidal and striatonigral) that regulate specific behaviors. Interestingly, exposure to low-to-moderate THC dosing paradigms has generally induced significant alterations of the dopaminergic D2 receptor (D2R) and the opioid neuropeptide proenkephalin (PENK) genes (9, 30, 31, 66), which are preferentially expressed on the striatopallidal neurons and have been linked with epigenetic impairments. The sensitivity of D2R gene (DRD2) and PENK to cannabis/THC exposure in both the human fetus and animal models is intriguing given the role of these genes in drug addiction vulnerability. Both human and animal postmortem studies have revealed specific disturbances in the expression of the PENK and DRD2 genes in the NAc of subjects exposed to THC during either prenatal or adolescent developmental periods that persists into adulthood (30, 31). Of the multiple epigenetic mechanisms, the regulation of histone modification is unique because methylation of distinct residues can have antagonistic effects on transcription ( Figure 2C ). Indeed, our previous studies revealed disturbances in the histone modification profile in the NAc of adult rats with prenatal THC exposure. These studies identified decreased levels of the trimethylation of lysine 4 on histone H3 (H3K4me3), a transcriptionally permissive mark, increased levels of dimethylation of lysine 9 on histone H3 (H3K9me2), a repressive mark, as well as decreased RNA polymerase II association with the promoter and coding regions of the gene in the NAc ( Table 1 ) (30). The combined epigenetic alterations were consistent with the observed reduction of the Drd2 gene expression and emphasize the enduring consequences of THC exposure following prenatal development. Similarly, persistent changes in repressive H3K9me2 and H3K9me3 were observed at the Penk locus in the NAc of adult rats following adolescent THC exposure in line with enduring upregulation of Penk mRNA levels (31). These findings emphasize an altered epigenetic landscape within the adult brain directly as a consequence of developmental cannabinoid exposure.

There is also evidence that THC exposure can affect the regulation of histone modification in other cell and tissue types during development. In differentiating mouse lymph node cells, alterations in H3K4me3, H3K9me3, H3K27me3, and H3K36me3 have been associated with dysregulated ncRNAs and mRNA genes (32). In addition, THC treatment dose-dependently increased the expression of HDAC3, a histone deacetylase, in a human trophoblast cell line indicating the possibility for cannabinoid exposure to affect placental development (70).

The studies discussed above highlight the long-term effects of cannabis exposure that influences the development of various cell and tissue types with functional and phenotypic consequences. Since these investigations so far have mainly been carried out at specific sets of candidate gene loci, rigorous future work will require comparisons between epigenomic and transcriptome alterations in order to address the mechanistic implications of these findings on the level of complex biological systems in different tissue types, and their dynamic regulation throughout development.

Multi-generational effects of cannabis

It has long been a subject of debates as to whether epigenetic disturbances that occurred during the lifespan of an individual are reprogrammed across most of the genome from parent to offspring, thereby establishing a new epigenetic “slate” for the next generation. Such concepts have been challenged in recent years by findings in various disease states where epigenetic aberrations that influence disease risk were shown to be inherited through the germline from parent to child (25, 71). More specifically, several cases of parent-child transmission regarding drugs of abuse have been published, describing both behavioral phenotypes and molecular disturbances in the offspring of parents that were exposed to drugs before mating (reviewed in (72)).

We have previously demonstrated that exposure of male and female adolescent rats before mating (“germline exposure”) leads to behavioral and molecular abnormalities in their unexposed offspring (11). Adult offspring of THC-exposed parents displayed increased work effort to self-administer heroin, with stereotyped behaviors during the period of acute heroin withdrawal. On the molecular level, parental THC exposure was associated with changes in the mRNA expression of cannabinoid, dopamine, and glutamatergic receptor genes in the striatum and altered synaptic plasticity in neurophysiological measures. In a more recent study and in line with the initial observations, DNA methylation disturbances were detected in the NAc of adult rats with parental germline THC exposure in an epigenome-scale investigation (15). The most significant finding was the identification of epigenetic alterations within an interaction network centered around the Dlg4 gene, encoding Psd-95, a membrane associated guanylate kinase scaffolding protein located in neural postsynaptic densities, involved in the regulation of dopamine-glutamate interactions. Psd-95 associates with the NMDA subtype of glutamate receptors and is required for synaptic plasticity associated with NMDA receptor function. A variety of genes involved in glutamatergic neurotransmission were also found to contain DNA methylation changes in the offspring of THC-exposed rats. Previously, epigenetic dysregulation of Dlg4 has been linked to abnormal glutamatergic transmission involved in morphine conditioning (73), consistent with the earlier observations of increased heroin self-administration in adult offspring with germline THC exposure (11). In other studies and in line with the above observations, adolescent female rats treated with the cannabinoid agonist WIN-55,212 before mating and pregnancy had progeny that exhibited increased morphine sensitivity (14, 74). These findings demonstrate that germline cannabinoid exposure can impact offspring phenotype, affect the molecular characteristics of the brain, and could possibly confer enhanced risk for addiction disorders.

Multi-generational epigenetic effects occur when an environmental trigger induces epigenetic changes that can be observed in at least one subsequent generation. The observations summarized above fit the classic concept of epigenetically inherited phenotypes. In-depth investigations are still needed to provide insights about epigenetic mechanisms underlying the transmission of cannabis effects through the germline. Moreover, important questions remain to be answered as to whether this represents a true transgenerational epigenetic transmission to subsequent generations (grandchildren and beyond) without direct germline exposure.

The eCB system plays important roles not only in the development of a variety of somatic cells and physiological systems, but also in reproduction. It is known that both male and female reproductive tissues express CBRs and eCBs and that in males, THC can disrupt gonadal functions (10, 75). Studies on the impact of cannabinoids on epigenetic changes in male fertility have been conducted in Cnr1 null mutant mice that displayed higher histone retention in germ cells compared to the wild type mice (76). In that study, CB1R expression was demonstrated to be necessary for spermiogenesis by controlling chromatin condensation in the developing sperm via the regulation of histone displacement during spermiogenesis, resulting in poor sperm quality. Adverse effects of cannabis use on the ovary of females have also been found to present a higher risk of primary infertility due to anovulation. Even when marijuana-using women undergo in vitro fertilization treatment, they produce poor quality oocytes and lower pregnancy rates (77). The effects of cannabis on the oocyte epigenome that could potentially lead to multi-generational transmission remain to be explored. Specifically, subsequent studies are required to assess how possible epigenetic processes (e.g. DNA methylation) are involved in the transmission of cannabinoid effects from parent to offspring.

Summary

Although still quite sparse in the number of studies and current mechanistic depth, there is solid scientific data that documents protracted effects of cannabinoids on brain as well as in other organs. Based on the current rapid growth in this scientific field, it is expected that significant developments in the near future will fill critical gaps of knowledge by focusing attention on long-term epigenetic processes and behavioral consequences of cannabis exposure.

The majority of addiction-related epigenetic neurobiological studies have targeted the adult brain. Even conceptually, very few studies have considered the potential lifelong or multi-generational epigenetic impact of cannabis. Although identifying mechanisms by which cannabis effects are maintained and transmitted is intriguing by itself, such explorations have potential far-reaching impact in the broader domain of developmental neurobiology since the identified epigenetic processes will no doubt be fundamental to transmission of other environmental insults across generations that bear on psychiatric vulnerability.

The mechanistic links between epigenetic modifications and gene expression impairments will require rigorous comparisons between epigenomic and transcriptome alterations. The overlay of results from approaches like RNA-sequencing, ChIP-sequencing and genome-scale DNA methylation studies in alignment to the genome will provide a unique potential to correlate epigenetic marks with the transcriptional regulation of neighboring genes. Moreover, the specific distribution and changes in 5-methylcytosine and 5-hydroxymethylcytosine (a demethylation intermediate, see Figure. 2B ) has not yet been studied in the context of cannabis, and will likely be an interesting direction for in-depth mechanistic investigations. Importantly, direct causal relationships will be gained through the use of genomic editing tools to determine the impact of specific epigenetic disturbances in relation to gene expression. Providing causal links between gene expression impairments and specific behavioral phenotypes using in vivo gene manipulations offers important mechanistic value and the potential for developing targeted therapeutic solutions.

Overall, the integration of information garnered from clinical populations with data emerging from animal models will provide innovative insights to guide future translational studies and better inform clinical treatment and prevention strategies for the long-term impact of cannabis and even for the growing use of synthetic cannabinoids.

Acknowledgments

This work was supported by grants from NIH/NIDA DA030359 and DA033660.

Footnotes

Financial Disclosure. All authors report no biomedical financial interests or potential conflicts of interest.

Publisher’s Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

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