21st century breakthroughs in methylation...
TRANSCRIPT
21st Century Breakthroughs in Methylation
Biochemistry for Clinicians
Presenter:
Benjamin Lynch, ND
Methylation Summit
Chicago, IL
April 12, 2014
“Clinicians will be central to helping consumer-patients use genomic information to make health
decisions.” – NEJM
1(c) 2014: Benjamin Lynch, ND
Disclaimer & Disclosures
The information presented here is for informational and educational purposes only. Seeking Health,
LLC and Benjamin Lynch will not be liable for any direct, indirect, consequential, special, exemplary, or
other damages arising from the use or misuse of any materials or information published.
I am President and CEO of SeekingHealth.com, SeekingHealth.org and founder of MTHFR.Net
I am compensated for this presentation.
I am on the Xymogen Board of Advisors.
My kids aren’t perfect.
2(c) 2014: Benjamin Lynch, ND
3
4
(c) 2014: Benjamin Lynch, ND 5
6(c) 2014: Benjamin Lynch, ND
Why?
7(c) 2014: Benjamin Lynch, ND
What is Methylation?
The addition of a single carbon group with three hydrogens onto a compound
8(c) 2014: Benjamin Lynch, ND
Methyl Donors support Methylation
Methyl Donors assist S-adenosylmethionine (SAM) production1. Methylfolate2. Methylcobalamin3. Serine4. Glycine5. Choline6. Betaine (TMG)7. DMG
9(c) 2014: Benjamin Lynch, ND
(c) 2014: Benjamin Lynch, ND 10
11
S-Adenosylmethionine (SAM): Functions
• Cofactor for Methyltransferases• COMT, PEMT, HMT, PNMT, GNMT, GAMT, DNMT…
• Binds to CpG Sites and Islands to block gene transcription• Maintains normal liver function• Induces apoptosis in liver cancer cells• Modulates iNOS synthesis• ↑ viral latency• ↓ NFk-B• ↑ CBS cysteine, hydrogen sulfide, taurine, pyruvate and glutathione
12(c) 2014: Benjamin Lynch, ND
Dietary Choline and Betaine Intakes and Risk of Cardiovascular Diseases: Review of Epidemiological Evidence and http://emergency.doctorsonly.co.il/wp-content/uploads/2011/03/SAMe-therapy-in-liver-disease-J-HEP-11.12.pdf
13
What should you be asking??
S-Adenosylmethionine (SAM): Functions
Biosynthesis
• Serotonin Melatonin
• Norepinephrine Epinephrine
Degradation
• Histamine
• Dopamine
• Catechol estrogens
• Epinephrine
• Arsenic
14(c) 2014: Benjamin Lynch, ND
(c) 2014: Benjamin Lynch, ND 15
S-Adenosylmethionine (SAM): Pain Reduction
Dopamine
• ↑ Dopamine ↓ Pain Tolerance
• SAM = cofactor for COMT
16(c) 2014: Benjamin Lynch, ND
Dietary Choline and Betaine Intakes and Risk of Cardiovascular Diseases: Review of Epidemiological Evidence and http://emergency.doctorsonly.co.il/wp-content/uploads/2011/03/SAMe-therapy-in-liver-disease-J-HEP-11.12.pdf
(c) 2014: Benjamin Lynch, ND 17
Functions of Methylation
Several Functions of Methylation:1. Turn on and off genes (gene regulation)2. Process chemicals, endogenous and xenobiotic compounds
(biotransformation)3. Build neurotransmitters (norepinephrine epinephrine, serotonin
melatonin)4. Metabolize neurotransmitters (dopamine, epinephrine)5. Process hormones (estrogen)6. Build immune cells (T cells, NK cells)7. DNA and Histone Synthesis (Thymine aka 5-methyluracil)8. Produce energy (CoQ10, carnitine, creatine, ATP)9. Produce protective coating on nerves (myelination)10. Build and maintain cell membranes (phosphatidylcholine)
18(c) 2014: Benjamin Lynch, ND
“Methylation is a common but generally minor pathway of xenobiotic transformation.” – Toxicology, 8th ed.
How is Methylation Disturbed?
Methylation is often disturbed by various mechanisms1. Lack of cofactors/substrate driving methylation forward (zinc, B2, magnesium, cysteine, B6,
methylcobalamin)2. Medications (antacids, methotrexate, metformin, nitrous oxide)3. Specific nutrients depleting methyl groups (high dose Niacin)4. Environmental toxicity, heavy metals, chemicals (acetylaldehyde, arsenic, mercury, high copper)5. Excessive end product (feedback inhibition – eg. DMG, SAMe, Methylfolate)6. Genetic mutations/polymorphisms (MTHFR, GSTM1, PEMT, MAT, GAMT, CBS)7. Mental state (stress, anxiety, lack of sleep, ‘can’t do it’, pessimist, optimist)
19(c) 2014: Benjamin Lynch, ND
Methylation Genetics
Folate• DHFR• MTHFD• MTHFR• SHMT• FOLR• TYMS• SLC19A1
B12• TCN2 and TCN3• MMAB (aka cblB)
Methionine Cycle• MTR/MTRR• MAT1• AHCY• CBS
Methyltransferases• COMT• PEMT• BHMT• GAMT• HNMT• GNMT• PNMT• DNMT
Glutathione• GSTM1• GCS• GPX1• GR
Detoxification• Cyt P450’s• SULT• NAT
Additional:• PON1• SOD2• DAO• G6PD• NOS1, 2 and 3
http://www.ornl.gov/sci/techresources/Human_Genome/project/info.shtml
Pathways Supporting Methylation(not comprehensive)
• Folate• B12• Methionine• ROS• Biotransformation
20(c) 2014: Benjamin Lynch, ND
(c) 2014: Benjamin Lynch, ND 21
Diet
DietDiet
DietDiet
& CpG sites
Mitochondrial Genetics
Mitochondrial• SOD• CAT• NDUF• ATP• COX
http://cshperspectives.cshlp.org/content/5/5/a012641.full.pdf and http://www.nature.com/scitable/topicpage/mtdna-and-mitochondrial-diseases-903 and http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3753274/pdf/pone.0074513.pdf
Mitochondrial DNA (mtDNA)• Inherited only from Maternal side (family hx Important)• Majority of ATP produced in mitochondria• Require importing nDNA gene products to function• Mitochondrial dysfunction cell dysfunction
methylation dysfunction• SNPS/mutations in mtDNA may be pathological
• Cancer• Diabetes• Cardiovascular Diseases• Neurodegenerative Diseases• Aging• Degenerative Diseases
• mtDNA copy number ↑ cell survival and function
22(c) 2014: Benjamin Lynch, ND
Genetics: Mitochondria
Acton B et al. Mol. Hum. Reprod. 2004;10:23--‐32 European Society of Human Reproduction and Embryology
Mitochondrial DNA (mtDNA)• Sperm – 700 molecules of mitochondria
• Oocytes – 200,000 molecules of mitochondria
23(c) 2014: Benjamin Lynch, ND
Cell Division and Mitochondria:• Mitochondria ‘float’ in cytoplasm
• Lack of Spindles
• Randomized
Cell Division and Replication of ‘Sick’ Cells?
24(c) 2014: Benjamin Lynch, ND
Stimulate DNA Bases and ↑ Cell Proliferation• “New” cells created:
• ↓ Glutathione
• Oxidized cell membrane
• ↓ Potassium
• ↑ ROS
• ↑ Cell death
Flare of Patient Symptoms with addition of Folate / B12.
Necessitates Treatment Flow
Epigenetics
“As an organism grows and develops, carefully orchestrated chemical
reactions activate and deactivate parts of the genome at strategic times
and in specific locations.
Epigenetics is the study of these chemical reactions and the factors that
influence them.”
25(c) 2014: Benjamin Lynch, ND
http://learn.genetics.utah.edu/content/epigenetics/ and Epigenetics and the
developmental origins of inflammatory bowel diseases.
“Epigenetic changes are environmentally responsive mechanisms that can
modify gene expression independently of the genetic code.”
Epigenetics 1st Identify Causation of Dysfunction
• Environment
• Lifestyle
• Diet
• Pathogens
• Heavy Metals
• Xenobiotics
• Oxidative Stress
• Nutrient Deficiencies
• Nutrient Excess
• SNPs
26(c) 2014: Benjamin Lynch, ND
System-Wide Dysfunction
27
Most stable epigenetic alteration at CpG islands
(c) 2014: Benjamin Lynch, ND
Epigenetic Example: Inflammatory Bowel Disorders
High monozygotic twin discordant rates in Crohn disease and UC.70+ loci associated with CD. 40+ for UC = epigenetic control.
28(c) 2014: Benjamin Lynch, ND
Source: Epigenetics and the developmental origins of inflammatory bowel diseases.
High monozygotic twin concordant rates in Celiac disease.Single HLA locus linked to 40% of heritability = genetic control
Epigenetic Control in Crohn’s Disease and UC
29(c) 2014: Benjamin Lynch, ND
Epigenetics in Action
a) Lab Setting b) Environment
Folic Acid GMH
MTHFR increasing in the population.
• Folic acid fortification• ↑ Full-Term Pregnancies
• ↑ Folate SNPs
• ↑ Methylation SNPs
• ↑ Inferior SNPs
• ↑ Metabolic Issues
30(c) 2014: Benjamin Lynch, ND
↑ Susceptibility to Environmental Exposures
Natural DeSelection: Survival of the ‘Unfittest’
31(c) 2014: Benjamin Lynch, ND
(c) 2014: Benjamin Lynch, ND 32
(c) 2014: Benjamin Lynch, ND 33
(c) 2014: Benjamin Lynch, ND 34
(c) 2014: Benjamin Lynch, ND 35
SAM Deficiency via MATI/III Inhibitors
Oxidative Stress
• NO
• TNFα
• IL-6
Causes vicious cycle
↓ SAM ↓ CBS activity ↓ GSH, ↓ H2S & ↑ Hcy ↑ SAH
36(c) 2014: Benjamin Lynch, ND
http://emergency.doctorsonly.co.il/wp-content/uploads/2011/03/SAMe-therapy-in-liver-disease-J-HEP-11.12.pdf
Methionine intake may NOT ↑ SAM
(c) 2014: Benjamin Lynch, ND 37
Homocysteine
• Methionine = Methyl-Homocysteine
• Breakdown product of SAM SAH via AHCY
38(c) 2014: Benjamin Lynch, ND
Methionine
Homocysteine Kinetics
Methionine cycle low Km / high affinity for sulfur-containing substrate
• ↑ methionine (sulfur) ↓ methionine cycle
Betaine-Homocysteine Methyltransferase (BHMT) low Km for Hcy
Transsulfuration cycle high Km for sulfur-containing substrate
• ↑ methionine (sulfur) ↑ transsulfuration cycle • ↑ GSH
• ↑ H2S
• ↑ Taurine
• ↓ Hcy
• ↑ demand of P5P, cysteine and serine
39(c) 2014: Benjamin Lynch, ND
Homocysteine Metabolism
Tissue Specific
Plasma homocysteine is a ‘gross’ average.Not tissue specific.
40(c) 2014: Benjamin Lynch, NDRegulation of homocysteine metabolism and methylation in human and mouse tissues and http://www.procrelys.fr/telechargement/publications/hcy_fertil_steril.pdf
Ex. CBS• Brain • Liver• Pancreas• Muscle
Ex. BHMT• Liver• Oocyte• Kidney
Ex. MTR• Brain• Oocyte
Support ALL Routes
(c) 2014: Benjamin Lynch, ND 41
Low homocysteine ( < 6) is an important finding
Homocysteine Metabolism (Routes)
(c) 2014: Benjamin Lynch, ND 42
What did the doctor do? What happened here?
(c) 2014: Benjamin Lynch, ND 43
Here is the consequence. What happened? How to restore?
(c) 2014: Benjamin Lynch, ND 44
(c) 2014: Benjamin Lynch, ND 45
Low Alkaline Phosphatase• ↓ Phosphate• ↓ Magnesium• ↓ Zinc• Celiac• Malnutrition• Hypothyroid• Pernicious anemia
Manual of Laboratory and Diagnostic Tests, Fischbach
H2O2
CBS and CDO
46(c) 2014: Benjamin Lynch, ND
CBS• Cofactor: B6
• Requires: Serine or Cysteine & Hcy
• Inhibited by: NO
• Promoted by: SAM
• Product: H2S & Cystathionine
CDO• Cofactor: Iron & Zinc
• Requires: Cysteine
• Inhibited by: α-ketoglutarate, TNFα
• Promoted by: Sulfur/Cysteine
• Product: Cysteinesulfinate
http://www.wikigenes.org/e/gene/e/1036.html and http://www.ncbi.nlm.nih.gov/pubmed/11059810 and http://ajpendo.physiology.org/content/early/2011/06/15/ajpendo.00151.2011
CDO Inhibition may lead to Cysteine Excitotoxicity- Support CDO - Support COMT - Remove Pathogens- Support IDO - Support MAO - Remove Metals
Signs (sensitive to cysteine intake):
↑ Cysteine:Sulfate ↑ Cysteine:Taurine ↑ Sulfite:Sulfate ↑ Hydrogen Sulfide
(c) 2014: Benjamin Lynch, ND 47
(c) 2014: Benjamin Lynch, ND 48
Folates: Which are off?
49(c) 2014: Benjamin Lynch, ND
(c) 2014: Benjamin Lynch, ND 50
(Folinic acid)
(Methylfolate)
(Thymine)(5-FU site)
Testing Levels of Various Folate Forms
51http://www.hdri-usa.com/tests/methylation/
mislabeled
(c) 2014: Benjamin Lynch, ND
(c) 2014: Benjamin Lynch, ND 52
Keep to the Basics
• History• Lifestyle• Diet• Compliance• Motivation• Mindset• Environment• Water• Exercise
53(c) 2014: Benjamin Lynch, ND
When your patient feels good...Do NOTHING!
Look for the Apex of the “Bell Shape Curve”
54(c) 2014: Benjamin Lynch, ND
Stay Informed
Great ways to stay informed:• Newsletter Available at www.MTHFR.net• Facebook: https://www.facebook.com/drbenjaminlynch• Nutrigenomics Conference Part 1 & Part 2: www.SeekingHealth.org
55(c) 2014: Benjamin Lynch, ND
Thank You