Methylation Series Pt1: The Why, The How, and The When.

Introduction to Methylation

In this section, the speaker introduces the topic of methylation and its importance in the body. They discuss the role of methylation in various physiological processes and highlight the need for nutrients and genetic factors for optimal methylation.

• Methylation is the transfer of one carbon atom and three hydrogen atoms from one substance to another.

• Methyl groups act as switches that regulate gene expression, mood, hormone detoxification, energy production, and healthy aging.

• Methylation occurs billions of times per second in every cell of the body.

• Nutrients from diet, such as vitamins, minerals, and amino acids, are essential for proper methylation.

• Genetic factors and oxidative stresses can affect methylation pathways.

• Methylation plays a role in various processes including creatine production, DNA synthesis, hormone regulation, detoxification, neurotransmitter production, fat metabolism, immune function, cognitive function, and cardiovascular health.

The Folate Cycle

This section focuses on the folate cycle within methylation. The speaker explains how folates are converted into active forms and emphasizes the importance of 5-methyltetrahydrofolate (5-MTHF).

• Folate exists as polyglutamate in the diet and needs to be converted into folate monoglutamate in the small intestine for absorption.

• Synthetic folic acid is more easily absorbed than dietary folates but doesn't necessarily mean it's better or should be supplemented with.

• Neither dietary folate nor folic acid is biologically active until converted to tetrahydrofolate (THF).

• The end product of the folate cycle is 5-MTHF or 5-methyltetrahydrofolate.

Methionine Cycle and Transulfuration Pathway

This section covers the methionine cycle and transulfuration pathway, which are important processes in methylation. The speaker explains the interplay between these pathways and their role in maintaining homeostasis.

• The methionine cycle involves the conversion of methionine to S-adenosylmethionine (SAM), which is a major methyl donor.

• SAM is involved in various methylation reactions throughout the body.

• Homocysteine is produced as a byproduct of SAM metabolism.

• Homocysteine can be remethylated back to methionine or converted to cysteine through the transulfuration pathway.

• The transulfuration pathway produces glutathione, an important antioxidant.

• Imbalances in these pathways can disrupt overall methylation status.

Clinical Relevance of Methylation Testing

In this section, the speaker discusses the clinical relevance of methylation testing and identifies symptoms that may indicate methylation imbalances.

• Methylation testing can be valuable for certain patients.

• Symptoms such as mood disorders, hormone imbalances, detoxification issues, cognitive dysfunction, immune dysfunction, and cardiovascular problems may point to methylation imbalances.

• Methylation testing helps clinicians identify underlying imbalances and guide treatment decisions.

Importance of Methylation

This section emphasizes the importance of methylation in various physiological processes and highlights its role as a ubiquitous biochemical process.

• Methylation plays a crucial role in regulating mood, hormone detoxification, energy production, healthy aging, neurotransmitter production, fat metabolism, immune function, cognitive function, and cardiovascular health.

• Nutrients from diet are necessary for optimal functioning of methylation pathways.

• Genetic factors and oxidative stresses can impact methylation processes.

Methylation Processes

This section explains how methylation involves the transfer and donation of methyl groups between molecules, leading to changes in their structure and function.

• Methylation is a biochemical process that occurs billions of times per second in every cell of the body.

• Methyl groups act as switches that turn genes on or off.

• Methylation processes require nutrients such as vitamins, minerals, and amino acids from the diet.

• Methylation impacts various physiological processes including mood regulation, hormone production, detoxification, energy production, aging, neurotransmitter metabolism, fat metabolism, immune function, cognitive function, and cardiovascular health.

Webinar Introduction

The speaker introduces themselves as a clinician educator at Genova Diagnostics Europe. They outline the purpose of the webinar series to increase understanding of methylation pathways and interpretation of Genova's methylation panel. Case study examples will be covered in the next installment.

• The webinar aims to increase understanding of methylation pathways and imbalances.

• Genova's methylation panel combines genomic and functional analytes related to methylation pathways.

• Case study examples will be discussed in the next installment.

New Section

This section discusses the conversion of folate or folic acid into biologically active forms and the role of various enzymes and cofactors in the folate cycle.

Conversion of Folate/Folic Acid

• Folate or folic acid enters the next stage of the folate cycle through various steps.

• The conversion process determines the production of five mthf, which provides a methyl group for the methionine cycle.

• Enzymes and cofactors such as B6, iron, B2, and B3 play important roles in propelling the conversion.

• Five mthf is the predominant circulating folate in serum, while other non-circulating forms are mainly found intracellularly.

New Section

This section focuses on folic acid supplementation and its potential drawbacks.

Folic Acid Supplementation

• Folic acid is better absorbed than dietary folate but has low and variable activity in the liver due to dhfr enzyme.

• High intake of folic acid can crowd out dietary folate by competing for dhfr binding sites.

• Elevated levels of unmetabolized folic acid in the bloodstream may have negative health consequences.

• Potential consequences include reduced natural killer cell activity, increased cancer risk, cognitive impairment among older adults, and lower cognitive development scores in children.

New Section

This section introduces phelanic acid as an alternative form of supplemental folate.

Phelanic Acid

• Phelanic acid can readily enter the folate cycle without being reduced by dhfr enzyme.

• It may be more effective for individuals with polymorphisms on the dhfr gene or sensitivity to methylfolate.

• Phelanic acid is considered gentler, especially for those with psychiatric conditions involving neurotransmitter imbalances.

New Section

This section explores the folate cycle and its role in methylation and nucleotide synthesis.

Folate Cycle

• Dehydropholate reductase converts folate into tetrahydropholate, which then gets metabolized by shmc enzyme using serine as a methyl donor.

• The direction of the conversion depends on methylation demand, either towards five mthf for methylation or pure nucleotide synthesis.

• Shmt enzyme, dependent on B6, iron, and Sam, prioritizes nucleotide synthesis over Sam synthesis.

• Disturbances in the methylation cycle and conditions like cancer can be linked to polymorphisms on the shmt enzyme.

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New Section

This section discusses the role of methyl transferase enzymes and their importance in various biological functions.

Methyl Transferase Enzymes

• Methyl transferase enzymes, such as DNMT, HNMT, COMT, PNMT, ASMT, TEMT, and GAMT, play a crucial role in various biological processes.

• These enzymes are involved in DNA methylation, histamine breakdown, detoxification of catechol compounds, conversion of noradrenaline to adrenaline, production of melatonin and phospholipids, and creatine production.

• Deficiencies in 5-MTHF and slow COMT enzyme can lead to symptoms related to excess estrogen and an amplified stress response.

New Section

This section explores the interconnection between the folate cycle and the methionine cycle in producing SAM.

Folate Cycle and Methionine Cycle Interconnection

• The purpose of the methionine cycle is to convert homocysteine into methionine which then becomes SAM through methylation.

• The MTR enzyme plays a key role in converting homocysteine to methionine using methylfolate as a methyl donor and methyl B12 as a catalyst.

• The MTRR enzyme maintains adequate levels of activated or methylated B12 for efficient homocysteine recycling. Snips or variations on this enzyme can impact homocysteine conversion.

• Insufficient activated B12 can lead to ineffective homocysteine recycling and potential backup in homocysteine levels.

• The BHMT pathway acts as a backup or salvage pathway for homocysteine conversion when methyl folate levels are low. Choline oxidation to betaine is a key step in this pathway.

• Zinc serves as a cofactor for the BHMT enzyme, and DMG, the product of the BHMT pathway, can inhibit BHMT through negative feedback.

New Section

This section emphasizes the importance of controlling homocysteine levels in methylation and highlights its role as a major branch point in the methylation pathway.

Homocysteine and Methionine Cycle

• Controlling homocysteine levels is crucial for proper methylation and serves as an indicator of methylation capacity.

• Homocysteine can be converted to methionine through either the methylfolate and methyl B12 dependent pathway or the BHMT or Salvage pathway.

New Section

This section discusses the importance of homocysteine as a biomarker and the risks associated with elevated levels.

Homocysteine and Methylation Capacity

• Elevated homocysteine can enhance vascular smooth muscle cell proliferation, increase platelet aggregation, and contribute to the development of atherosclerosis and cardiovascular diseases.

• Elevated homocysteine induces inflammatory cytokines and contributes to disease progression.

• High homocysteine levels can impair bone health by interfering with osteoclast activity.

• Elevated homocysteine increases central nervous system phosphorylated tau, leading to increased neurofibrillary tangle formation seen in Alzheimer's dementia.

• Homocysteine can interfere with methyl transferase enzymes related to neurotransmitter synthesis and have neurotoxic properties.

New Section

This section explores the causes of high homocysteine levels and factors that contribute to its accumulation.

Causes of High Homocysteine Levels

• Lack of cofactor nutrients such as methylfolate, methylcobalamin, B vitamins, zinc, choline, and iron can lead to high homocysteine levels.

• Single nucleotide polymorphisms (SNPs) may downregulate the activity of certain enzymes involved in methylation pathways, such as SHMT enzyme, MTHFR enzyme, MTRR enzyme, and BHMT pathway.

• Lifestyle factors like high alcohol intake, tobacco use, and caffeine consumption can also contribute to elevated homocysteine levels.

New Section

This section discusses the potential issues associated with low homocysteine levels and factors that can contribute to its decrease.

Low Homocysteine Levels

• Low homocysteine levels are not well represented in literature, but they may lead to issues such as reduced glutathione production.

• Factors contributing to low homocysteine levels include low dietary intake or poor absorption of protein and upregulation in the transulfuration pathway.

• Low homocysteine levels result in lower levels of methionine and S-adenosylmethionine (SAM), which are important for various biological processes.

New Section

This section explains the conversion of homocysteine to methionine and the regulation of SAM levels.

Conversion of Homocysteine to Methionine

• Homocysteine is converted to methionine via the MAT enzyme, which depends on ATP, magnesium, and potassium. Depletion of these compounds or nutrients can downregulate SAM production.

• SNPs in the MAT enzyme can also downregulate its activity, leading to a depletion of SAM.

Regulation of SAM Levels

• Elevated SAM acts as a negative feedback to MTHFR and BHMT enzymes, inhibiting their activity to maintain homeostasis and prevent hypermethylation.

• High levels of SAM have been associated with adiposity and obesity, although the mechanism is still being studied.

New Section

This section discusses the conversion of excess S-adenosylmethionine (SAM) to S-adenosylhomocysteine (SAH) and its role in maintaining homeostasis.

Conversion of SAM to SAH

• The enzyme responsible for this conversion is GNMT.

• GNMT catalyzes the methyl group transfer of fonsam to glycine, ultimately forming sarcosine (SAR).

• SAR plays an important role in removing excess SAM.

• This removal process is down-regulated when there are low levels of methyl folate and SAM.

New Section

This section explains the hydrolysis of homocysteine to complete the methionine cycle and the impact of elevated homocysteine on SAH production.

Hydrolysis of Homocysteine

• Homocysteine is hydrolyzed to homocysteine when there are low levels of methyl folate and SAM.

• This reaction completes the methionine cycle.

• The dynamics strongly favor the production of SAH, leading to an increase in SAH with elevated homocysteine levels.

New Section

This section highlights the inhibitory effect of SAH on methylation reactions and its significance as a marker for certain diseases.

Inhibition by SAH

• SAH acts as a potent feedback inhibitor for methotransferase enzymes within various tissue components, including DNA, RNA, and phospholipids.

• Plasma levels of SAH have been shown to be more sensitive markers for clinical cardiovascular disease, renal disease, and Alzheimer's disease than plasma homocysteine levels.

New Section

This section emphasizes the importance of SAM removal and the role of AHCY enzyme in methylation reactions.

SAM Removal and AHCY Enzyme

• Methylation reactions depend on the removal of SAH.

• The functioning of the AHCY enzyme is crucial for SAH removal.

• The availability of vitamin B3 is essential for the functioning of the AHCY enzyme.

New Section

This section introduces the transsulfuration pathway and its connection to methylation, mitochondrial energy production, and glutathione biosynthesis.

Transsulfuration Pathway

• Transsulfuration is the main route for irreversible homocysteine disposal.

• It involves transferring a sulfur group to various molecules, such as cystathionine, cysteine, glutathione, pyruvate, and taurine.

• The pathway is upregulated during oxidative stress when there is a higher demand for glutathione.

• It also contributes to energy production when glutathione demand is low.

New Section

This section focuses on CBS enzyme's role in connecting homocysteine to cystathionine in the transsulfuration pathway.

CBS Enzyme and Cofactor Nutrients

• CBS (cystathionine beta synthase) enzyme interconnects homocysteine to cystathionine.

• CBS requires three cofactor nutrients: B6, iron, and serine.

• Excess methionine and SAM upregulate CBS activity, leading to increased conversion of homocysteine to cystathionine.

• Insufficiencies in B6 can lead to backup in this pathway and potentially insufficient glutathione production.

New Section

This section discusses the impact of CBS enzyme variations and other enzymes on glutathione synthesis.

Impact of CBS Enzyme Variations

• Variations in the CBS enzyme can lead to upregulation or downregulation of homocysteine removal via transsulfuration.

• Downregulation can result in a backup of homocysteine and potential lack of glutathione production.

• Other enzymes, such as CTH and GSS, also play a role in glutathione synthesis.

• Insufficiencies in B6 can affect this pathway and potentially lead to insufficient glutathione production.

New Section

This section highlights the importance of B6 and the often overlooked nutrient's impact on glutathione production.

Importance of B6

• B6 is a cofactor nutrient for enzymes involved in the transsulfuration pathway and glutathione synthesis.

• Insufficiencies in B6 can lead to backup in the transsulfuration pathway, potentially affecting glutathione production.

• Many reports on metabolomics and nutritional panels highlight insufficient B6 levels in individuals.

New Section

This section emphasizes the significance of glutathione as an outcome of the transsulfuration pathway.

Importance of Glutathione

• Glutathione plays a crucial role in neutralizing hydrogen peroxide and lipid peroxides through GSH peroxidase reactions.

• It is involved in phase two detoxification by conjugating hormones, toxins, and xenobiotics to make them water-soluble for excretion.

New Section

This section addresses when to test methylation status and which clients or patients would benefit from such testing.

When to Test Methylation

• Methylation plays a role in various processes, including creatine production.

• The decision to test methylation status depends on the specific client or patient and their health concerns.

New Section

This section discusses the implications of methylation in various biological processes and conditions such as DNA and RNA synthesis, gene regulation, hormone regulation, detoxification of environmental toxins, neurological conditions, mental health conditions, and cardiovascular disease.

Methylation Implications

• Methylation is highly implicated in conditions like cancer .

• Methylation plays a role in hormone regulation and detoxification of environmental toxins .

• Hormone imbalances and symptoms like estrogen dominance can be related to methylation defects .

• Methylation is important for cell membrane repair and myelination in neurological conditions like Parkinson's disease .

• Mental health conditions such as depression, anxiety, bipolar disorder, schizophrenia, and mood disorders may have methylation defects affecting neurotransmitter production .

• Methylation plays a role in endothelial function and nitric oxide production related to cardiovascular disease .

New Section

This section explores common barriers to optimal methylation that healthcare providers should consider when offering methylation testing. These barriers include GI dysbiosis, oxidative stress, stress from childhood abuse or trauma, exposure to environmental toxins, nutrient deficiencies, sleep deprivation, and medication use.

Common Barriers to Optimal Methylation

• GI dysbiosis can impact DNA methylation due to microbiome imbalances and their effect on folate production .

• Increased oxidative stress affects the activity of enzymes responsible for regulating DNA methylation .

• Early life stress like childhood abuse has lasting effects on methylation into adulthood .

• Exposure to environmental toxins can lead to aberrant changes in epigenetic pathways and hypomethylation .

• Specific environmental chemicals like heavy metals and organic pollutants can directly reduce enzymatic activity responsible for DNA methylation .

• Nutrient deficiencies, especially those related to cofactors for methylation enzymes, can impact methylation capacity .

• Sleep deprivation, both acute and chronic, affects DNA methylation patterns in genes involved in metabolism and circadian rhythm .

• Common medications like antacids, non-steroidal anti-inflammatory drugs, corticoids, and methotrexate can inhibit DNA methylation .

New Section

This section summarizes the types of patients who can benefit from methylation panel testing. Methylation defects can have a broad range of symptoms associated with them.

Patients Who Can Benefit from Methylation Panel Testing

• Patients with questionable detoxification or a risk of cancer may benefit from methylation panel testing .

• Patients with cardiovascular disease or a family history of cardiovascular disease may benefit from methylation panel testing .

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End of Presentation and Questions

The presenter concludes the presentation and invites participants to ask questions. They provide instructions on how to submit questions through the webinar control panel or via email. They also mention that a recording of the webinar will be provided, along with the slides in PDF format.

Questions and Feedback

• Participants are encouraged to write their questions in the question tab on the go-to-webinar control panel or contact the team via email. Feedback is also appreciated.

Recording and Slides

• The presenter informs participants that there will be a recording of the webinar available for those who asked about it. Slides will also be sent in PDF format.

• The methylation support guide can be found in the handouts section on the control panel, while slides will be sent separately via email.

Question about CBS Enzyme and Cysteine Levels

A participant asks about high cysteine levels, potential upregulation of CBS enzyme, and suggestions for reducing sister thinine (homocysteen) while increasing cysteine.

• The junction between sister thinine and cysteine is regulated by the CBS enzyme, which depends on B6 availability, iron, and serine as cofactors.

• If sister thinine is elevated but cysteine levels are low, it suggests a backup in the pathway due to lower availability of cofactor nutrients for proper enzyme function.

• This backup can affect glutathione production.

Preparing Clients for Metabolomics Plus Test

A participant asks about preparing clients for the metabolomics plus test, which is a urine test.

• The presenter suggests checking the page on their website dedicated to the metabolomics plus test for preparatory guidelines.

• Alternatively, participants can send an email for more information.

Ammonia Increase with CBS and BHMT SNPs

A participant asks about the potential increase in ammonia when individuals have CBS and BHMT SNPs and whether this puts pressure on the urea cycle, resulting in BH4 deficiency.

• The presenter explains that theoretically, upregulation of CBS may lead to increased ammonia levels, impacting BH4 production and putting pressure on the urea cycle.

• High ammonia symptoms could be related to GI dysbiosis and overproduction of ammonia by dysbiotic bacteria.

• It's important to consider factors beyond methylation when addressing high ammonia symptoms.

Using Methylation Panel for Chronic Fatigue Syndrome

A participant asks how the methylation panel can help clients with chronic fatigue syndrome.

• The presenter mentions various methyl transferase enzymes involved in energy production, cellular membrane health, nervous system health, and creatine production.

• If a patient with chronic fatigue syndrome shows hypomethylation (lower levels of SAM), supporting them through proper cofactor nutrients for folate and methionine cycles can be beneficial.

• Identifying any SNPs that may slow down or downregulate SAM production is also important.

Test Procedure for Methylation Panel

A participant asks about how the methylation panel test is taken.

• The test involves a combination of a blood draw and a buccal swab to analyze functional analytes in the blood sample and SNPs related to methylation.

New Section

This section discusses the impact of methylation on different parts and analytes, as well as the importance of immediate centrifuging for certain tests.

Impact of Methylation on Analytes

• Methylation can have an impact on certain analytes, but it varies depending on which parts of the methylation cycle are affected.

Immediate Centrifuging for Testing

• The test requires immediate centrifuging, so it is important to consider this when scheduling a phlebotomy appointment for a client or patient.

New Section

This section addresses questions related to testing for Mast Cell Activation Syndrome and supplementation of folic acid.

Testing for Mast Cell Activation Syndrome

• It is recommended to test methylation for clinical presentations related to Mast Cell Activation Syndrome due to its connection with histamine detoxification and metabolism.

Supplementation of Folic Acid

• There is no upper limit for dietary folic acid intake as it does not negatively impact the dhfr enzyme. However, supplemental synthetic folic acid can downregulate this enzyme. The upper limit for supplemental folate or folic acid is 1000 MCG.

New Section

This section discusses supplementation options for improving glutathione levels and evaluating the sum to SAR ratio.

Supplementation for Glutathione Improvement

• If someone has low levels of glutathione and high levels of oxidative stress markers, it is recommended to supplement both NAC (N-acetylcysteine) and glutathione initially to increase levels. Once levels improve, switching to one may be sufficient.

Evaluating the Sum to SAR Ratio

• The sum to SAR ratio, which indicates the risk of hypo or hypermethylation, can be evaluated using a methylation panel. More information on this will be provided in the next week's presentation.

New Section

This section clarifies the relationship between cancer and methylation and discusses the safe upper limit for 5-methyltetrahydrofolate (5-MTHF).

Relationship Between Cancer and Methylation

• Snips on the shmt enzyme, involved in folate cycle, can impact DNA repair and regulation, potentially increasing the risk of cancer.

Safe Upper Limit for 5-MTHF

• The safe upper limit for 5-MTHF is 1000 MCG, similar to folic acid.

New Section

This section addresses questions regarding molybdenum supplementation for improving CBS pathway function and potential solutions for gnmt snips.

Molybdenum Supplementation for CBS Pathway

• Molybdenum is important for detox pathways, particularly sulfuration in the liver. It may help improve ammonia detoxification in cases of CBS snips. However, functional analytes should also be considered alongside genomic markers to understand their expression and interplay within pathways.

Potential Solutions for Gnmt Snips

• High sarcasin levels may indicate an upregulation in methylation due to an upregulation in gnmt. Further investigation is needed to determine potential solutions based on individual cases.

SAM Levels and Compensatory Mechanisms

The speaker discusses the significance of normal SAM (S-adenosylmethionine) levels as a compensatory mechanism to prevent hypermethylation. They emphasize the importance of checking both SAM and SAR (S-adenosylhomocysteine ratio) to determine if there are any issues related to methylation.

SAM Levels and Compensation

• Normal SAM levels may act as a compensatory mechanism to prevent hypermethylation.

• Checking both SAM and SAR is crucial in determining any issues related to methylation.

• The outcome of methylation depends on the levels of these two compounds.

Testing Children and Teenagers

The speaker addresses whether the testing can be performed on children or teenagers. They explain that the reference ranges provided are for an adult population, but adjustments can be made based on age. They also mention that sensitivity to having blood samples taken may hinder testing in younger individuals.

Testing Children and Teenagers

• Reference ranges provided are for adults aged 18 and above.

• Adjustments need to be made for children below 18 years old.

• Sensitivity to blood sample collection may affect testing in younger individuals.

Assessing Enzyme AHcy in DNA Add-On

The speaker responds to a question regarding why the enzyme AHcy is not assessed in their DNA add-on test. Although they do not have a definitive answer, they express their willingness to find out more about it.

Assessing Enzyme AHcy

• The reason for not assessing the enzyme AHcy in their DNA add-on test is unknown.

• Further investigation will be conducted to provide an explanation.

Recommended Supplements for MTHFR and COMT Enzyme SNPs

The speaker provides recommendations for supplements in patients with both MTHFR and COMT enzyme SNPs. They mention that magnesium, B2, zinc, and SAM are cofactor nutrients for COMT. However, the availability of 5-methyltetrahydrofolate (5-MTHF) may be affected by the MTHFR SNP, requiring supplementation with methylfolate.

Recommended Supplements

• Cofactor nutrients for COMT include magnesium, B2, zinc, and SAM.

• Supplementation with methylfolate may be necessary if there is a downregulation of 5-MTHF availability due to the MTHFR SNP.

• Testing function analytes can help determine the impact of SNPs on common enzyme activity.

• Elevated levels of anxiety and issues with stress response regulation may indicate a SNP on the common enzyme.

• Females with a SNP on the common enzyme may experience difficulties in estrogen detoxification.

Conclusion

The speaker concludes the webinar and expresses gratitude for participants' time. They inform attendees that they will receive a recording of the webinar along with slides. Additionally, they invite participants to join next week's session focused on methylation panels.

Conclusion

• The webinar concludes with appreciation for participants' time.

• Attendees will receive a recording of the webinar and slides.

• Next week's session will cover methylation panels.