mthfr - SpectraCell Laboratories

Transcription

MTHFR Mutational Effects
• Relationship to:
• subtype and elevated
homocysteine
• ADD/bipolar/PMS/ chronic
fatigue/fibromyalgia
• Tongue/fingernails/thin hair
• vericose veins/spider veins
• hypercoagulopathy/infertility
• elevated testosterone in
men/PCOS in women
• Acne, fibroids, endometriosis,
menometorrhagia
• depression when on OCPs
• Exacerbation of menopausal
symptoms on BHRT
• elevated uric acid and
elevated ammonia levels
• gut barrier and dysbiosis
• CANCER
• estrogen dominance and
obesity (estrogens effects on
insulin and the insulin
receptor and adiponectin)
Folate cycle and glutathione
MTHFR
• irreversibly reduces 5,10-methylenetetrahydrofolate
to 5-methyltetrahydrofolate/5-MTHF.
• 5,10-methylenetetrahydrofolate is used for de novo
Thymidine synthesis.
• 5-Methyltetrahydrofolate is used to convert
Homocysteine to Methionine by the enzyme
methionine synthase using methylcobalamine.
Betaine-Homocysteine
Methyltransferase
• Homocysteine can be
converted to Methionine by
a folate-independent
enzyme, BHMT.
• Zinc metallo-enzyme that
catalyzes the transfer of a
methyl group from betaine
to homocysteine to produce
dimethylglycine and
Methionine respectively
MTHFR contains a Flavin cofactor and uses
NAD(P)H as the reducing agent
FAD
NAD
Ribbon diagram of the active site of E. coli MTHFR. The flavin cofactor
(top) is shown interacting with the bound substrate NADH
Frequency of MTHFR Polymorphisms
• 30-40% of Americans are found to have either a single or
double polymorphism of either C677T or A1298C
• There is ethnic variability in the frequency of the T allele frequency in Mediterranean/Hispanics > Caucasians >
Africans/African-Americans)
Worldwide distribution of a common methylenetetrahydrofolate reductase mutation. Am J
Hum Genet 62 (5): 1258–60
• Higher frequency in chronic disease: Autoimmune diseases,
diseases of the gut, Fibromyalgia, Chronic Fatigue, Chronic
Retroviruses, Cancer, Hormone dysregulation, Mood issues,
Cardiometabolic patients.
Transsulfuration is dependent on
healthy methylation
Folate cycle nutrients, homocysteine
and Transsulfuration
• Transsulfuration is the conversion of Homocysteine with the
co-factors B6 and Serine to Cystathionine.
• Cystathionine is then converted to Cysteine which is one of the
three amino acids that compose Glutathione.
• Polymorphisms in Cysta beta-synthase may affect the rate of
conversion of Cystathionine to Cysteine.
• Cysteine is thought to be the rate limiting step in the formation
of Glutathione.
Folate cycle and glutathione
MTHFR
• MTHFR activity may be inhibited by binding of dihydrofolate
(DHF) and S-adenosylmethionine (SAM, or AdoMet)
• The enzyme is coded by the gene with the symbol MTHFR on
chromosome 1 in humans.
• There are genetic polymorphisms associated with this gene.
• In 2000 a report brought the number of polymorphisms up to
24. Two of the most investigated are C677T and A1298C single
nucleotide polymorphisms.
C677T Single Nucleotide
Polymorphism
• The MTHFR nucleotide at position C677T encodes a
thermolabile enzyme with reduced activity.
• Individual with two copies of 677C (677CC) have the “normal”
or “wildtype” genotype.
• 677TT individuals (homozygous) are said to have mild to
moderate MTHFR deficiency.
• The degree of enzyme thermolability is much greater in 677TT
individuals (18-22%) compared with 677CT (56%) and 677CC
(66-67%).
C677T Single Nucleotide
Polymorphism
• These C667T mutants are at a higher risk for errors in transsulfuration of amino acids and hyperhomocysteinemia (high blood
homocysteine levels)
• Low folate intake affects individuals with the 677TT genotype to a
greater extent. C677TT mutants lose the FAD cofactor three times
faster than the wild type protein.
• 5-Methyl-THF slows the rate of FAD release in both the wild-type
and mutant enzymes.
• Mutations in the MTHFR gene could be one of the factors leading to
increased risk of developing chronic diseases and psychiatric
problems.
“Effects of the methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism on
executive function in schizophrenia.” Schizophr Res 92 (1–3): 181–8
A1298C Single Nucleotide
Polymorphism
• 1298AA is the “normal” homozygous, 1298AC the
heterozygous, and 1298CC the homozygous for the “variant.”
• In studies of human recombinant MTHFR, the protein encoded
by 1298C cannot be distinguished from 1298A in terms of
activity, thermolability, FAD release, or the protective effect of
5-methyl-THF.
• The C mutation does not appear to affect the MTHFR protein.
It does not result in thermolabile MTHFR and does not appear
to affect homocysteine levels.
“Effects of common polymorphisms on the properties of recombinant human
methylenetetrahydrofolate reductase.” Proc. Natl. Acad. Sci. U.S.A. 98 (26): 14853–8
Severe MTHFR deficiency
• Severe MTHFR deficiency is rare (about 50 cases worldwide)
and caused by mutations resulting in 0-20% residual
enzyme activity.
Characterization of six novel mutations in the methylenetetrahydrofolate reductase
(MTHFR) gene in patients with homocystinuria. Hum Mutat 15 (3): 280–7
• These patients exhibit developmental delay, motor and gait
dysfunction, seizures, and neurological impairment and have
extremely high levels of homocysteine in their plasma and
urine as well as low to normal plasma methionine levels.
Folic Acid/5-mTHF Cycle
Methionine Synthase (MTR)
• Enzyme that in humans is encoded by the MTR gene
(5-methyltetrahydrofolate-homocysteine methyltransferase
• Enzyme is responsible for the regeneration of methionine
from homocysteine. Methionine synthase forms part of the
S-adenosylmethionine (SAMe) biosynthesis and
regeneration cycle.
• Methionine synthase contains the cofactor methylcobalamin
(MeB12) and uses the substrates N5-methyl-tetrahydrofolate
(N5-mTHF) and homocysteine
Methionine Synthase
• The enzyme works in two steps in a ping-pong reaction.
• First, methylcobalamin is formed by a methyl group transfer from
N5-mTHF with formation of MeB12 and tetrahydrofolate (THF)
• In the second step, MeB12 transfers this methyl group to
homocysteine, regenerating the cofactor cobalamin and releasing
the product methionine.
• Methionine synthase is the only mammalian enzyme that
metabolizes 5-mTHF to regenerate the active cofactor THF.
Deficiency in methionine synthase function may be due to genetic
mutations, reduced levels of its cobalamin cofactor B12, or decreased
levels of the enzyme (methionine synthase) reductase which is
required for the sustained activity of MS.
Methionine Synthase Mutations
• Mutations in the MTR gene have been identified as the
underlying cause of methylcobalamine deficiency.
• This may lead to megaloblastic anemia as seen in elevated
MCV in the patients CBC.
• Patients may actually have high levels of B12 in the serum as
the Cobalamine is not converting to Methylcobalamine.
• May compound mutations in MTHFR
Methionine
S-ame
Homocysteine
SAH
Cystathionine
Cysteine
The reaction reaction catalyzed by
Methionine Synthase
Tetrahydrofolate
Homocysteine
5methytetrahydrofolate
Methionine
Methylcobalamin
Methylcobalamin is a cobalamin used in the treatment of
peripheral neuropathy, diabetic neuropathy, and as a
preliminary treatment for amyotrophic lateral sclerosis.
Role in biomethylation
• Methylcobalamin plays an important role in the environment.
It is produced preferentially by some bacteria. In the
environment, it is responsible for the biomethylation of certain
heavy metals. For example, the highly toxic methylmercury is
produced in this way.
Methylcobalamin
has been studied in
conjunction with
sleep-wake rhythm
disorders, where it
appears to yield
benefits, but at a
low or inconsistent
level.
Psychiatry Clin Neurosci. 1999 Apr;53(2):2113.
Double-blind test on the efficacy of
methylcobalamin on sleep-wake rhythm
disorders.
Mega Dosages of Me-B12
• Supplementation with
megadoses of MeB12 has
been advocated to protect
the cognitive function of
patients suffering from:
• Chronic Fatigue
• Stroke
• Depression
• Alzheimer’s disease
Neurological diseases
Me-B12 reduces
Glutamate
Methylcobalamin has been
used to reduce
neurotoxicity and lower
excess glutamate levels,
resulting in the reduction of
fatigue, stabilization of
mood, improvement of
memory, and executive
function.
Methyl Vitamin B12, but not Methylfolate,
rescues a motor neuron-like cell line from
Homocysteine-mediated cell death.
•
Motor Neuron Cell Biology Group, Department of Neurology, Carolinas Neuromuscular/ALS-MDA Center, Carolinas
Medical Center, Charlotte, NC 28203, USA. [email protected]
•MeCbl
Abstract in the micromolar range was able to provide
andacid
neurorescue…
against
millimolar
•neuroprotection…
Homocysteine is an excitatory amino
implicated in multiple diseases
including
amyotrophic lateral sclerosis
(ALS). Information on the toxicity of homocysteine in motor neurons is limited and few studies have examined how
homocysteine
exposure.
hadneuron-neuroblastoma),
no effect on
this toxicity can be modulated.
In NSC-34DIn
cellscontrast,
(a hybrid cell lineMTHF…
derived from motor
homocysteine induces apoptotic cell death in the millimolar range with a TC₅₀ (toxic concentration at which 50% of
homocysteine-mediated
death.
maximal cell death is achieved) of 2.2 mM,cell
confirmed
by activation of caspase 3/7. Induction of apoptosis was
independent of short-term reactive oxygen species (ROS) generation. Methyl Vitamin B12 (MeCbl) and methyl
Toxicol
Appl Pharmacol.
2011clinically
Mar 15;251(3):217-25.
Epub 2011
Janwere
13 tested for their ability to
tetrahydrofolate
(MTHF), used
to treat elevated homocysteine
levels,
reverse homocysteine-mediated motor neuron cell death. MeCbl in the micromolar range was able to provide
neuroprotection (2 h pretreatment prior to homocysteine) and neurorescue (simultaneous exposure with
homocysteine) against millimolar homocysteine with an IC₅₀ (concentration at which 50% of maximal cell death is
inhibited) of 0.6 μM and 0.4 μM, respectively. In contrast, MTHF (up to 10 μM) had no effect on homocysteinemediated cell death. MeCbl inhibited caspase 3/7 activation by homocysteine in a time- and dose-dependent
manner, whereas MTHF had no effect. We conclude that MeCbl is effective against homocysteine-induced cell death
in motor neurons in a ROS-independent manner, via a reduction in caspase activation and apoptosis. MeCbl
decreases Hcy induced motor neuron death in vitro in a hybrid cell line derived from motor neuron-neuroblastoma
and may play a role in the treatment of late stage ALS where HCy levels are increased in animal models of ALS.
Serine Hydroxymethyltransferase
• Serine Hydroxymethyltransferase (SHMT) is an enzyme which
catalyzing the conversions of L-serine to Glycine and
Tetrahydrofolate to 5,10-Methylenetetrahydrofolate.
• Glycine is a critical amino acid in the formation of the tripeptide, Glutathione. Glycine production is supported by
L-serine supplementation
A Ferritin-responsive internal
ribosome entry site regulates folate
metabolism.
•
Woeller CF, Fox JT, Perry C, Stover PJ.
•
Source
Cytoplasmic
serineMolecular
Hydroxymethyltransferase
• Graduate Field of Biochemistry,
and Cellular Biology, Cornell University, Ithaca,(cSHMT)
New York 14853, USA.
enzyme
levels are elevated by the expression of the heavy
• Abstract
chain
Ferritin (H ferritin) cDNA in cultured cells without
• Cytoplasmic serine hydroxymethyltransferase (cSHMT) enzyme levels are elevated by the expression of the heavy chain ferritin
(H ferritin) cDNA in cultured cells without corresponding changes in mRNA levels, resulting in enhanced folate-dependent de
corresponding
changes in mRNA levels, resulting in enhanced
novo thymidylate biosynthesis and impaired homocysteine remethylation. In this study, the mechanism whereby H ferritin
regulates cSHMT expression was determined. cSHMT translation is shown to be regulated by an H ferritin-responsive internal
folate-dependent
de novo Thymidylate biosynthesis and
ribosome entry site (IRES) located within the cSHMT mRNA 5'-untranslated region (5'-UTR). The cSHMT 5'-UTR exhibited IRES
activity during in vitro translation of bicistronic mRNA templates, and in MCF-7 and HeLa cells transfected with bicistronic
impaired
Homocysteine remethylation.
mRNAs. IRES activity was depressed in H ferritin-deficient mouse embryonic fibroblasts and elevated in cells expressing the H
ferritin cDNA. H ferritin was shown to interact with the mRNA-binding protein CUGBP1, a protein known to interact with the
alpha and beta subunits of eukaryotic initiation factor eIF2. Small interference RNA-mediated depletion of CUGBP1 decreased
IRES activity from bicistronic templates that included the cSHMT 3'-UTR in the bicistronic construct. The identification of this H
ferritin-responsive IRES represents a mechanism that accounts for previous observations that H ferritin regulates folate
metabolism.
J Biol Chem. 2007 Oct 12;282(41):29927-35. Epub 2007 Aug 16.
Serine hydroxymethyltransferase
Methylation Support:
Vesselcare 1 qd
Somnolin qhs
Ceralin forte 3 day
Folapro 1-3 a day
Intrinsi B12/Folate 1-2day
Estrofactors 3 day
Symphora 1-2 day
Lilly
93 yo CF c/o fatigue, IBS, recently
dx with elevated TSH, high ferritin
found incidentally
• Hemochromatosis HFE double
mutation
• MTHFR mutation A1298C+
A1298C+
• Hypothyroidisms
• Initial Labs: Ferritin 1022, iron
total 195, iron Sat% 83.55%
Lilly:
• TIBC 234 L
• Homocysteine 14.2
• Treatment:
• Vesselcare 1 qam
• Somnolin 1 qhs
• Metalloclear 3 bid
• Outcome: Ferritin 759, total iron
132, iron sat 49%, TIBC 266
Hank
• 46yo HM hx of BetaThalassemia, Gout, Diabetes
type II, Cirrhosis and endstage liver disease, on
transplant list. Fatigue, poor
stamina, weight loss.
• High Ferritin HFE -/• Low testosterone, Diabetes
well controlled
• MTHFR C677T+/A1298C+
Hank cont.
Treatment: Vesselcare 1 bid,
Alapars 1 bid,
Methylcobalamine 1000mcg
IM injection q 3 days,
Ferritin: 1054> 675>414>359
nl <400
Tot iron: 283> 138
Sat: 71.81% > 65%
Tot test 319> 433 (250-750)
Free Test: 23.6> 30.1
Patient did so well he was
removed from the Transplant
List!
Nadia
• Treatment:
• Infertility: 6 miscarriages,
3 failed IVFs
• Estrofactors 3 day
• 36yo MF G7P1 Beta Thalassemia • 28 day detox Ultraclear Renew
Hgb/Hct 9.5/31.7
• EPA/DHA
• Recurrent fetal loses at 2-6 wks
EGA
• High Ferritin 949
• Plt 521, WBC 17, Tot Bili: 5.4
• Mthfr A1298+ heterozygote, HFE
negative
• Intrinsi B12/Folate
• Topical progesterone days 14-28
of cycle
Nadia – Cont’d
• Achieved pregnancy in 1st
month after detox without
IVF!! Beta hcg quant.
doubled every 48 hours;
Pregnant and doing well
taking Intrinsic B12 and
5MTHF with prenatal
vitamins. Has required less
frequent blood transfusions
since starting intrinsic factor
with B12/5MTHF
• Last Ferritin 532; last TB 3.2
Stephen
• 56 yo CM hx of
• Htn
• Abdominal discomfort/IBS
• Paresthesias in extremities
• Muscle pain
• Rash on upper/lower arms
• Fatigue
• Mouth ulcers/ Gingivitis
Stephen
• Labs:
• HgA1c 6.1
• Total Chol 220, TG 206, Hdl 31
Ldl 152 pattern B
• CPK 289 U/L (24-204) Ferritin
1004 (<400)
• HFE ++ mutation
• AST 70,ALT 127, GGT 79
• MTHFR C677T/A1298T
• HLA DQ2/HLA DQ8 -/-
Stephen con’t
• Treatment: new diagnosis of
Diabetes, Hyperlipidemia,
NAFLD
• Metformin for to treat
Diabetes II
• Simcor: tx hyperlipidemia
• Folapro 1 po TID
• Symphora for Cortisol DHEA
support (improved energy,
but…activated skin/gums)
Stephen
• Symphora was switched to
Vesselcare 1 am,
• Somnolin 1 pm
• Alapars bid for NAC and
Niacinamide support of
glutathione
• Advaclear bid
• Magnesium Glycinate 200mg
qhs.
Stephen – Cont’d
• Response: Skin and gums
improved remarkably! Skin
lesions on his arm completely
resolved. Fatigue and muscle pain
resolved, improved moods and
abdominal discomfort. Better
Sleep, feeling more rested.
• Labs: GGT:79>62>50>49>42>41
• Ferritin:
1002>860>901>687>668>582>563
• HgA1c 6.1>5.8
• CPK: 74
Ricardo
• 44yo HM hx of
• Hyperlipidemia
• Hypertension
• Hypothyroidism
• Chronic Exhaustion
Ricardo
• Serum Testoterone 233>534
(280-800)
• TSH: 7.1 (.450-4.5) >1.25 ( on
Synthroid 75mcg)
• Homocysteine: 12.6 (<15)>8.1
• Crp: 1.01 (<3.0)
• MTHFR:C677T+- HFE -/• Glutathione
739>616>691>685
• GGT 30
Uric acid: 6.7
Treatment:
• Pre-provocative: Serum heavy
metal: wnl, 24hr Urine heavy
metal (Arsenic 35, Mercury 2)
• Post-provocative: Serum
Mercury 2.2>0, Serum Arsenic
0> 5>0, 24hr Urine Arsenic
50>42>10, Mercury 0.
• Iron 123>182>144,Iron sat%
34%>56%>43%,Ferritin 761>
662>592>519>458
Ricardo
• Patient had initial labs and
was placed on
• 28 day detox program with
Ultraclear renew
• Advaclear 2 bid
• Vesselcare 1 am
• Ceralin forte 3 day
Ricardo
• Patient reported
dramatic improvement
in energy, and stamina
• Remarkable drops in his
bp to 120-130/70-80s
while on the same bp
meds and thyroid
medications.
Inhibition of angiogenesis by
Methionine
•
Sahin M, Sahin E, Gümüşlü S, Erdoğan A, Gültekin M.
•
Source
•
Health Sciences Research Centre, Faculty of Medicine, Akdeniz University, 07070 Antalya, Turkey.
[email protected]
•
Abstract
•InMetastasis
is a leading our
cause ofstudy
mortality suggests
and morbidity inthat
cancer. One
of the steps in metastasis process is the
conclusion,
Methionine
formation of new blood vessels. Aberrant DNA methylation patterns are common in cancer cells. In recent studies,
Supplementation
useful
against
angiogenesis
as effects
a on some
S-adenosylmethionine (SAM),may
which isbe
a DNA
methylating
agent, has been
found to have inhibitory
carcinoma cells in vivo and in vitro. In the present study, we have used SAM to investigate whether it is effective
natural
bio-product
against angiogenesis
in vitro. Our results have shown that SAM can reduce the formation and organization of
capillary-like structures of endothelial cells in tumoral environment. Besides, we have found SAM can block
endothelial cell proliferation and the migration of cells towards growth factors-rich media. In conclusion, our study
suggests that SAM may be used against angiogenesis as a natural bio-product.
Biochem Biophys Res Commun. 2011 Apr 29;408(1):145-8. Epub 2011 Apr 2
S-adenosylmethionine (AdoMet)
supplementation for treatment of
chemotherapy-induced liver injury
•
Santini D, Vincenzi B, Massacesi C, Picardi A, Gentilucci UV, Esposito V, Liuzzi G, La Cesa A, Rocci L, Marcucci F, Montesarchio V,
Groeger AM, Bonsignori M, Tonini G
•
Source
Conclusion
The results of our study clearly demonstrate a protective effect
of AdoMet in cancer chemotherapy-induced liver toxicity.
Further large phase III studies are required to assess the real
clinical benefit associated with AdoMet supplementation.
•
Medical Oncology, Center for Biomedical Research (CIR), Laboratory of Internal Medicine and Hepatology, Campus Bio-Medico University, Rome, Italy.
•
Abstract
•
BACKGROUND:
•
Liver toxicity can be observed during treatment with most chemotherapic agents, and represents one of the principal causes of dose reduction or chemotherapy delays. SAdenosylmethionine (AdoMet) plays a critical role in the synthesis of polyamines and provides cysteine for the production of glutathione (GSH), the major endogenous hepatoprotective
agent. Our study was aimed at assessing the protective effect of AdoMet supplementation in cancer chemotherapy-induced liver toxicity.
•
PATIENTS AND METHODS:
•
Fifty cancer patients who developed, for the first time, anticancer chemotherapy-induced liver toxicity were studied. Enrolled patients received oral AdoMet supplementation.
•
RESULTS:
•
AST, ALT and LDH levels recorded at the moment of the recognition of liver toxicity were significantly reduced after one week of AdoMet therapy (respectively p: 0.009, 0.0005 and 0.012).
AST, ALT and LDH decrease was confirmed after two weeks of treatment. Furthermore, the effect on these enzyme levels persisted in the following chemotherapy courses, permitting our
patients to perform the scheduled chemotherapy courses with a minimal number of dose reductions or administration delays. The efficacy of AdoMet supplementation was not influenced
by the presence of liver metastases, and no appreciable side-effects were recognized.
•
CONCLUSION:
•
The results of our study clearly demonstrate a protective effect of AdoMet in cancer chemotherapy-induced liver toxicity. Further large phase III studies are required to assess the real
clinical benefit associated with AdoMet supplementation.
Anticancer Res. 2003 Nov-Dec;23(6D):5173-9
S-adenosylmethionine inhibits the growth of
cancer cells by reversing the hypomethylation
status of c-myc and H-ras in human gastric cancer
and colon cancer
•
Luo J, Li YN, Wang F, Zhang WM, Geng X.
•
Source
•
Department of biochemistry, Tianjin Medical University, Tianjin, 300070 China.
•
Abstract
S-adenosylmethionine
treatment inhibited cell
• A global DNA hypomethylation might activate oncogene transcription, thus promoting carcinogenesis and tumor
development.in
S-Adenosylmethionine
(SAM) serves
as a major
methylcolon
donor in biological
transmethylation
growth
gastric cancer
cells
and
cancer
cells,events.
The object of this study is to explore the influence of SAM on the status of methylation at the promoter of the
oncogenes
H-ras and tumor-suppressor
gene p16 (INK4a),
well as its inhibitory effecthigher
on cancer cells. The
and
thec-myc,
inhibition
efficiency
wasassignificantly
results indicated that SAM treatment inhibited cell growth in gastric cancer cells and colon cancer cells, and the
inhibition
efficiency
significantly
higher thancells
that in the normal cells. Under standard growth conditions, C-myc
than
that
inwasthe
normal
and H-ras promoters were hypomethylated in gastric cancer cells and colon cancer cells. SAM treatment resulted in
a heavy methylation of these promoters, which consequently downregulated mRNA and protein levels. In contrast,
there was no significant difference in mRNA and protein levels of p16 (INK4a) with and without SAM treatment.
SAM can effectively inhibit the tumor cells growth by reversing the DNA hypomethylation on promoters of
oncogenes, thus down-regulating their expression. With no influence on the expression of the tumor suppressor
genes, such as P16, SAM could be used as a potential drug for cancer therapy.
Int J Biol Sci. 2010 Dec 6;6(7):784-95
S-adenosylmethionine inhibits the growth of
cancer cells by reversing the hypomethylation
status of c-myc and H-ras in human gastric cancer
and colon cancer
•
Luo J, Li YN, Wang F, Zhang WM, Geng X.
•
Source
•
Department of biochemistry, Tianjin Medical University, Tianjin, 300070 China.
•
Abstract
SAM
can effectively inhibit the tumor cells growth by reversing the
• A global DNA hypomethylation might activate oncogene transcription, thus promoting carcinogenesis and tumor
S-Adenosylmethionineon
(SAM)
serves as a major of
methyl
donor in biological transmethylation
events.
DNAdevelopment.
hypomethylation
promoters
oncogenes,
thus downThe object of this study is to explore the influence of SAM on the status of methylation at the promoter of the
oncogenes c-myc,
H-rasexpression.
and tumor-suppressor
gene p16
(INK4a),
as well as its inhibitory
on cancer cells. The
regulating
their
With
no
influence
on theeffect
expression
results indicated that SAM treatment inhibited cell growth in gastric cancer cells and colon cancer cells, and the
inhibition
efficiency
was significantly higher
than that in the normal
cells. Under
C-myc
of the
tumor
suppressor
genes…SAM
could
be standard
usedgrowth
as aconditions,
potential
and H-ras promoters were hypomethylated in gastric cancer cells and colon cancer cells. SAM treatment resulted in
druga heavy
formethylation
canceroftherapy.
these promoters, which consequently downregulated mRNA and protein levels. In contrast,
there was no significant difference in mRNA and protein levels of p16 (INK4a) with and without SAM treatment.
SAM can effectively inhibit the tumor cells growth by reversing the DNA hypomethylation on promoters of
oncogenes, thus down-regulating their expression. With no influence on the expression of the tumor suppressor
genes, such as P16, SAM could be used as a potential drug for cancer therapy.
Int J Biol Sci. 2010 Dec 6;6(7):784-95
Glutaclear helps to support
healthy glutathione production
and detoxification while
supporting Selenomethionine
Glutathione and
Detoxification
Support
Glutaclear 2 pill twice a
day
Selenomethionine
N-acetyl Cysteine
Broccoli seed extract
Niacinamide
Utilization of Seleniomethionine for
Glutathione Support
• Se-met is activated initially
by adenosylation,
demethylated and converted
to Se-cys via
Selenohomocysteine and
Selenocystathionine in
analogy to Met and without
involving Se-met–specific
enzymes
• Se-met prevented the
decline of plasma Se and
glutathione peroxidase
activity
• Se-met oxide is easily
reduced back to Se-met by
glutathione and oxidative
damage to Se-met is
reversible. On the basis of
this observation, Se-met and
GSH were suggested to act
as an antioxidant system,
protecting cells against
oxidants such as
peroxynitrite
•
(Journal of Nutrition. 2000;130:1653-1656.)
•
© 2000 The American Society for Nutritional Sciences
Catechol-O-Methyltransferase
• Involved in phase II
metabolism of hydroxyestradiols
• Involved in metabolism of
xenobiotics
chocolate
caffeine by-products
(catechins)
excitatory neurotransmitters
• S-adenosylmethionine and
magnesium dependent
• Linked to estrogen
imbalance disorders
• Is involved in
hyperhomocystinemia in
Parkinson patients on L-dopa
• Is linked to psychiatric
disorders
E
S
T
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o
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e
s
Susan
• 49 yo CF recent dx of Stage II
Breast Ca hormone Rec +,s/p
partial mastectomy,
radiation, irregular periods,
shingles, intolerant of
Fereston due to Headaches,
constipated. Hx of fibroids,
heavy periods, fibrocystic
breast. G2P2
• nl vitals, Estradiol
26.8pg/ml (0-32.2
menopause range)
• Sed rate 22 (0-20)
Susan Cont.
• Cbc, chem, thyroid panel
cholesterol wnl
• Vit D 153 (30-100)
• GGT 15 Uric Acid 3.2
• Glutathione 992
• MTHFR C677+/A1298C+
• Cytokine: base Il-6++, IL-17+,
IL-12--,IFN gamma ++, TNF-a
++, IL-4++, IL-5--, IL-10++, IL8++, G-CSF++
Susan Cont.
Susan Cont.
• Treatment: 28 day Ultraclear
Renew with extra meal
supplementation due to low
body weight.
• Co-Q10 support
• Meta I3C for estrogen
balance
• High concentrate EPA/DHA
Magnesium Glycinate 200mg
qhs for alkaline, mineral, and
bowel support.
• Vesselcare bid methylation
support
• Somnolin with 5HTP and
methyl-B12 to improve sleep
and pain control.
• Glutaclear 2 bid to increase
glutathione
• Ceralin Forte for anti-oxidant
support.
• Ultraflora IB
• Exercise daily, check urine ph
goal> 6.6. Sauna at the gym,
alkaline diet, Xenoestrogen
awareness!
Susan
Response: “I feel the
best that I have felt in
years…This is
Miraculous…I sleep
deeper than ever, no
hot flashes, I’m
exercising because I
have ENERGY. My
husband says I’m back
again b/c I’m SASSY
again!”
Xenobiotic Effects on COMT
• In humans, 2-Oh and 4-Ohestradiol (catechol estrogens) are
rapidly O-methylated to form monomethyl ethers catalyzed by
COMT and S-adenosyl-L-methionine.
• Xenobiotics may strongly inhibit COMT-mediated
O-methylation of catechol estrogens by xenobiotics and may
facilitate the development of estrogen-induced tumors.
• Xenobiotics may therefore deplete intermediates in the Folate
cycle. Environmental burden of Xenobiotics may create a
higher need for methylation support.
COMT genotype, micronutrients in
the folate metabolic pathway and
breast cancer risk
•
Goodman JE, Lavigne JA, Wu K, Helzlsouer KJ, Strickland PT, Selhub J, Yager JD.
•
Department of Environmental Health Sciences, Johns Hopkins University, Bloomberg School of Public Health, Baltimore, MD 21205, USA.
•
Abstract
•
Catechol-O-methyltransferase (COMT) catalyzes the O-methylation of catechol estrogens (CEs), using S-adenosylmethionine (SAM) as a
methyl donor. Several studies have indicated that the val108met COMT polymorphism, which results in a 3-4-fold decrease in activity, is
associated with increased breast cancer risk. Folate, whose intake levels have also been associated with breast cancer risk, and other
micronutrients in the folate metabolic pathway influence levels of SAM and S-adenosylhomocysteine (SAH), a COMT inhibitor generated
by the demethylation of SAM. Because these micronutrients have been shown to alter SAM and SAH levels, we hypothesized that they
could also affect COMT-catalyzed CE methylation. Although measurements of SAM and SAH were not initially collected, a secondary
analysis of data from two nested case-control studies was performed to examine whether serum levels of folate, vitamin B12 (B12),
pyridoxal 5'-phosphate (PLP), cysteine and homocysteine, in conjunction with COMT genotype, were associated with breast cancer risk.
COMT(HH) (high activity COMT homozygote) breast cancer cases had statistically significantly lower levels of homocysteine (P = 0.05) and
cysteine (P = 0.04) and higher levels of PLP (P = 0.02) than COMT(HH) controls. In contrast, COMT(LL) (low activity COMT homozygote)
cases had higher levels of homocysteine than COMT(LL) controls (P = 0.05). No associations were seen between B12, COMT genotype,
and breast cancer risk. An increasing number of COMT(L) alleles was significantly associated with increased breast cancer risk in women
with below median levels of folate (P(trend) = 0.05) or above median levels of homocysteine (P(trend) = 0.02). These findings are
consistent with a role for certain folate pathway micronutrients in mediating the association between COMT genotype and breast cancer
risk.
These findings are consistent with a role for certain folate
pathway micronutrients in mediating the association between
COMT genotype and breast cancer risk.
Carcinogenesis. 2001 Oct;22(10):1661-5
Association of genetic polymorphisms of
COMT gene with psychiatric disorders
•
Gao LB, Zhong SR, Jing Q.
•
Source
•
Department of Forensic Medicine, Kunming Medical College, Kunming, Yunnan, P.R. China.
•
Abstract
The enzyme catechol-O-methyltransferase (COMT) transfers a
• The enzyme catechol-O-methyltransferase (COMT) transfers a methyl group from S-adenosylmethionine to the benzene ring of
methyl
group
S-adenosylmethionine
to the This
benzene
ringin the
catecholamines
includingfrom
the neurotransmitters
dopamine, epinephrine and norepinephrine.
methylation results
of catecholamines. The involvement of the COMT gene in the metabolic pathway of these neurotransmitters has
ofdegradation
catecholamines
including
thedisorders.
neurotransmitters
dopamine,
made it an attractive candidate gene
for many psychiatric
This review focuses on the association
between the
genetic polymorphisms of COMT gene and psychiatric disorders.
epinephrine and norepinephrine. This methylation results in
the degradation of catecholamines.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2010 Dec;27(6):650-3
Inhibition of human liver
catechol-O-methyltransferase by tea catechins
and their metabolites: structure-activity
relationship and molecular-modeling studies
•
Department of Chemical Biology, Ernest Mario School of Pharmacy, Rutgers University, 164 Frelinghuysen Road, Piscataway, NJ 08854,
USA.
• Abstract
(Epigallocatechin-3-gallate
(EGCG) is the major polyphenol
• (Epigallocatechin-3-gallate (EGCG) is the major polyphenol present in green tea. We previously demonstrated that EGCG was both a
substrate andin
potent
inhibitor oftea.
human liver
cytosolic
catechol-O-methyltransferease
(COMT). We now report
the structure-activity
present
green
We
previously
demonstrated
that
EGCG
relationship for the inhibition of COMT-catalyzed O-methylation of catecholestrogens in human liver cytosol by tea catechins and some of
their metabolites. The most potent inhibitors were catechins with a galloyl-type D-ring, including EGCG (IC(50)=0.07 microM), 4''-Owas
both(IC(50)=0.10
a substrate
and potent
inhibitor
of microM),
human
liver
methyl-EGCG
microM), 4',4''-di-O-methyl-EGCG
(4',4''-DiMeEGCG)
(IC(50)=0.15
and (-)-epicatechin-3-gallate
(ECG)
(IC(50)=0.20 microM). Catechins without the D-ring showed two to three orders of magnitude less inhibitory potency. Enzyme kinetic
analyses revealed that EGCG behaved as a mixed inhibitor, whereas 4',4''-di-O-methyl-EGCG exhibited competitive kinetics for the Scytosolic
catechol-O-methyltransferease
(COMT).
nowa new
report
adenosylmethionine
(SAM), and noncompetitive kinetics for the catechol binding site.
These compoundsWe
may represent
type of
COMT inhibitor. In silico molecular-modeling studies using a homology model of human COMT were conducted to aid in the
the catalytic and inhibitory mechanisms.
Either D-ring or
B-ringthe
of EGCGinhibition
could be accommodated
the substrate binding
theunderstanding
structure-activity
relationship
for
of toCOMTpocket of human COMT. However, the close proximity (2.6A) of 4''-OH to the critical residue Lys144, the higher acidity of the hydroxyl
groups of the D-ring, and the hydrophobic interactions between the D-ring and residues in the binding pocket greatly facilitated the
interaction of theO-methylation
D-ring with the enzyme, and resulted
in increased inhibitory potency. These results
provide mechanistic
insight into the
catalyzed
of catecholestrogens
in human
liver
inhibition of COMT by commonly consumed tea catechins.
cytosol by tea catechins and some of their metabolites
Biochem Pharmacol. 2005 May 15;69(10):1523-31.
Mechanisms of ethanol-drug-nutrition
interactions.
• Alcohol Research and Treatment Center, VA Medical Center, Bronx, NY 10468.
• Abstract
• Mechanisms of the toxicologic manifestations of ethanol abuse are reviewed.
Hepatotoxicity of ethanol results from alcohol dehydrogenase-mediated excessive
hepatic generation of nicotinamide adenine dinucleotide and acetaldehyde. It is now
recognized that acetaldehyde is also produced by an accessory (but inducible) pathway,
the microsomal ethanol-oxidizing system, which involves a specific cytochrome P450. It
generates oxygen radicals and activates many xenobiotics to toxic metabolites, thereby
explaining the increased vulnerability of heavy drinkers to industrial solvents,
anesthetics, commonly used drugs, over-the-counter medications and carcinogens. The
contribution of gastric alcohol dehydrogenase to the first pass metabolism of ethanol
and alcohol-drug interactions is now recognized. Alcohol also alters the degradation of
key nutrients, thereby promoting deficiencies as well as toxic interactions with vitamin A
and beta-carotene. Conversely, nutritional deficits may affect the toxicity of ethanol and
acetaldehyde, as illustrated by the depletion in glutathione, ameliorated by S-adenosyl-Lmethionine. Other supernutrients include polyenylphosphatidylcholine, shown to correct
the alcohol-induced hepatic phosphatidylcholine depletion and to prevent alcoholic
cirrhosis in non-human primates. Thus, a better understanding of the pathology induced
by ethanol has now generated improved prospects for therapy.
ETOH generates oxygen radicals and activates many
xenobiotics to toxic metabolites, thereby explaining
the increased vulnerability of heavy drinkers to
industrial solvents, anesthetics, commonly used drugs,
over-the-counter medications and carcinogens.
Jackie
• 32 yo women presented after
discharge from the hospital in San
Diego.
• Reported while out to dinner she
had a single glass of Chardonnay,
after which she went home a took a
Tylenol PM as a sleep-aid.
• Woke up in the middle of the night
with acute abdominal pain. Found
to have fulminant hepato-renal
failure. Emergency liver transplant
was performed.
Mechanisms of ethanol-drug-nutrition
interactions.
• Alcohol Research and Treatment Center, VA Medical Center, Bronx, NY 10468.
• Abstract
• Mechanisms of the toxicologic manifestations of ethanol abuse are reviewed.
Hepatotoxicity of ethanol results from alcohol dehydrogenase-mediated excessive
hepatic generation of nicotinamide adenine dinucleotide and acetaldehyde. It is now
Alcohol
also alters
the degradation
of key nutrients,
thereby
recognized
that acetaldehyde
is also produced
by an accessory
(butpromoting
inducible) pathway,
the microsomal
ethanol-oxidizing
system, which
involves aAspecific
cytochrome P450. It
deficiencies
as
well
as
toxic
interactions
with
vitamin
and
beta-carotene.
generates oxygen radicals and activates many xenobiotics to toxic metabolites, thereby
Conversely,
deficits
may affect
the
toxicity
of ethanol
and
explainingnutritional
the increased
vulnerability
of heavy
drinkers
to industrial
solvents,
anesthetics, as
commonly
usedby
drugs,
medications and
carcinogens.byThe
acetaldehyde,
illustrated
the over-the-counter
depletion in glutathione,
ameliorated
contribution of gastric alcohol dehydrogenase to the first pass metabolism of ethanol
S-adenosyl-L-methionine.
Other
supernutrients
include
and alcohol-drug interactions
is now
recognized. Alcohol
also alters the degradation of
key nutrients, therebyshown
promoting
deficiencies
well as toxic interactions
phosphatidylcholine,
to correct
the as
alcohol-induced
hepaticwith vitamin A
and beta-carotene. Conversely, nutritional deficits may affect the toxicity of ethanol and
phosphatidylcholine
depletion
anddepletion
to prevent
alcoholicameliorated
cirrhosis inbynonacetaldehyde, as illustrated
by the
in glutathione,
S-adenosyl-Lmethionine.
Other supernutrients include polyenylphosphatidylcholine, shown to correct
human
primates.
the alcohol-induced hepatic phosphatidylcholine depletion and to prevent alcoholic
cirrhosis in non-human primates. Thus, a better understanding of the pathology induced
by ethanol has now generated improved prospects for therapy.
Inhibition of human catechol-O-methyltransferase
(COMT)-mediated O-methylation of catechol
estrogens by major polyphenolic components
present in coffee
•
Zhu BT, Wang P, Nagai M, Wen Y, Bai HW.
•
Source
•
Department of Pharmacology, Toxicology and Therapeutics, School of Medicine, University of Kansas Medical Center, Kansas City, KS
66160, USA. [email protected]
•
Abstract
• In the present study, we molecular
investigated the inhibitory
effect of three catechol-containing
polyphenols,that
chlorogenic acid, caffeic acid
Computational
modeling
analysis coffee
showed
and caffeic acid phenethyl ester (CAPE), on the O-methylation of 2- and 4-hydroxyestradiol (2-OH-E(2) and 4-OH-E(2), respectively)
catalyzed by the cytosolic catechol-O-methyltransferase (COMT) isolated from human liver and placenta. When human liver COMT was
used as the enzyme,
chlorogenic
and caffeic acidacid
each inhibited
O-methylation
2-OH-E(2) in a soluble
concentration-dependent
chlorogenic
acid
andacidcaffeic
canthebind
to ofhuman
manner, with IC(50) values of 1.3-1.4 and 6.3-12.5 microM, respectively, and they also inhibited the O-methylation of 4-OH-E(2), with
IC(50) values of 0.7-0.8 and 1.3-3.1 microM, respectively. Similar inhibition pattern was seen with human placental COMT preparation.
COMT
active
siteacidinforainhibiting
similar
manner
as but
the
catechol
CAPE at
had athe
comparable
effect as caffeic
the O-methylation
of 2-OH-E(2),
it exerted
a weaker inhibition of the Omethylation of 4-OH-E(2). Enzyme kinetic analyses showed that chlorogenic acid and caffeic acid inhibited the human liver and placental
COMT-mediated O-methylation of catechol estrogens with a mixed mechanism of inhibition (competitive plus noncompetitive).
estrogen
substrates…
coffee
polyphenols
have
Computational
molecular modeling analysis
showed that
chlorogenic acid and caffeic
acid canhigher
bind to humanbinding
soluble COMT at the active
site in a similar manner as the catechol estrogen substrates. Moreover, the binding energy values of these two coffee polyphenols are
lower than
of catechol
estrogens, which
means that
coffee
polyphenols have
higher binding affinity for the enzyme than the natural
affinity
forthatthe
enzyme
than
the
natural
substrates.
substrates. This computational finding agreed perfectly with our biochemical data.
J Steroid Biochem Mol Biol. 2009 Jan;113(1-2):65-74. Epub 2008 Nov 2
Catechol-O-methyltransferase (COMT)-mediated
metabolism of catechol estrogens: Comparison
of wild-type and variant COMT isoforms
•
Dawling S, Roodi N, Mernaugh RL, Wang X, Parl FF.
•
Source: Department of Pathology, Vanderbilt University Medical Center, Nashville, Tennessee 37232, USA.
•
Abstract
•
The oxidative metabolism of 17beta-estradiol (E2) and estrone (E1) to catechol estrogens (2-OHE2, 4-OHE2, 2-OHE1, and 4-OHE1) and
estrogen quinones has been postulated to be a factor in mammary carcinogenesis. Catechol-O-methyltransferase (COMT) catalyzes the
methylation of catechol estrogens to methoxy estrogens, which simultaneously lowers the potential for DNA damage and increases the
concentration of 2-methoxyestradiol (2-MeOE2), an antiproliferative metabolite. We expressed two recombinant forms of COMT, the
wild-type (108Val) and a common variant (108Met), to determine whether their catalytic efficiencies differ with respect to catechol
estrogen inactivation. The His-tagged proteins were purified by nickel-nitrilo-triacetic acid chromatography and analyzed by
electrophoresis and Western immunoblot. COMT activity was assessed by determining the methylation of 2-OHE2, 4-OHE2, 2-OHE1, and
4-OHE1, using gas chromatography/mass spectrometry for quantitation of the respective methoxy products. In the case of 2-OHE2 and 2OHE1, methylation occurred at 2-OH and 3-OH groups, resulting in the formation of 2-MeOE2 and 2-OH-3-MeOE2, and 2-MeOE1 and 2OH-3-MeOE1, respectively. In contrast, in the case of 4-OHE2 and 4-OHE1, methylation occurred only at the 4-OH group, yielding 4MeOE2 and 4-MeOE1, respectively. Individual and competition experiments revealed the following order of product formation: 4-MeOE2
> 4-MeOE1 >> 2-MeOE2 > 2-MeOE1 > 2-OH-3-MeOE1 > 2-OH-3-MeOE2. The variant isoform differed from wild-type COMT by being
thermolabile, leading to 2-3-fold lower levels of product formation. MCF-7 breast cancer cells with the variant COMT 108Met/Met
genotype also displayed 2-3-fold lower catalytic activity than ZR-75 breast cancer cells with the wild-type COMT 108Val/Val genotype.
Thus, inherited alterations in COMT catalytic activity are associated with significant differences in catechol estrogen and methoxy
estrogen levels and, thereby, may contribute to interindividual differences in breast cancer risk associated with estrogen-mediated
carcinogenicity.
Thus, inherited alterations in COMT catalytic activity are
associated with significant differences in catechol estrogen
and methoxy estrogen levels and, thereby, may contribute to
interindividual differences in breast cancer risk associated
with estrogen-mediated carcinogenicity.
Cancer Res. 2001 Sep 15;61(18):6716-22
N-acetylcysteine blocks formation
of cancer-initiating estrogen-DNA
adducts in cells
•
N-acetylcysteine blocks formation of cancer-initiating estrogen-DNA adducts in cells.
•
Zahid M, Saeed M, Ali MF, Rogan EG, Cavalieri EL
•
Source: Eppley Institute for Research in Cancer and Allied Diseases, College of Public Health, University of Nebraska Medical Center,
Omaha, NE 68198, USA.
•
Abstract
This
finding suggests that NAcCys, a common dietary
• Catechol estrogens, especially 4-hydroxylated metabolites of 17beta-estradiol (E(2)), are responsible for estrogen-induced carcinogenesis.
4-Hydroxyestradiol (4-OHE(2)), a major metabolite of E(2) formed preferentially by cytochrome P-450 1B1, is oxidized to E(2)-3,4supplement,
used
as adducts
a potential
chemo-preventive
quinone, which can reactcould
with DNA tobe
yield the
depurinating
4-OHE(2)-1-N3Ade
and 4-OHE(2)-1-N7Gua. The apurinic sites
generated by the loss of these depurinating adducts induce mutations that could lead to cancer initiation. In this study, we have
agent
block
the initial
in the
genotoxicity
caused
byepithelial cell
evaluatedto
the effects
of N-acetylcysteine
(NAcCys)step
on the metabolism
of two
cell lines, MCF-10F (a normal
human breast
line) and E6 (a normal mouse mammary epithelial cell line), treated with 4-OHE(2) or its reactive metabolite, E(2)-3,4-quinone. Extensive
HPLC with electrochemical
detectionquinones.
and UPLC-MS/MS analyses of the cell media demonstrated that the presence of NAcCys very
catechol
estrogen
efficiently shifted the estrogen metabolism toward protective methoxylation and conjugation pathways in multiple ways, whereas
formation of depurinating DNA adducts was inhibited. Protection by NAcCys seems to be similar in both cell lines, irrespective of their
origin (human or mouse) or the presence of estrogen receptor-alpha. This finding suggests that NAcCys, a common dietary supplement,
could be used as a potential chemopreventive agent to block the initial step in the genotoxicity caused by catechol estrogen quinones.
Free Radic Biol Med. 2010 Aug 1;49(3):392-400. Epub 2010 May 31.
Jan
• 61 yo CF hx Breast Cancer at
age 53, dysmetabolic
syndrome, obesity, myalgia,
restless leg syndrome, IBS,
depression, insomnia, lower
extremity swelling with
limited mobility
• Sulfa allergic
• PMHx: infertility, hx of
endometriosis with TAH at
age 32
• Vesselcare am
• MTHFR A1298+/A1298+
sedrate: 26 mm/Hr
• Treatment:
• UltraInflamX360
• Somnolin pm
• Ceralin forte TID
Jan– Cont’d
• Progress: Patient
reports shocking
improvement in lower
extremity swelling and
improved stamina due to
resolved lower extremity
pain. Improved mood,
sleep, hot flashes, and
energy, abdominal
discomfort resolved.
Tammy
• 45 yo CF with hx of
hypothyroidism on
• Armour Thyroid, reports
ongoing fatigue, poor
stamina, low mood and
desire to lose weight.
Financial concerns were
discussed.
• Treatment:
• UltraClear Renew for 28
days
• Advaclear 2 bid for
bifunctional liver
support, lost 14 lbs but
could not keep the
weight off when she
resumed her gluten
free/dairy free diet.
Tammy Cont.
Could not afford a lot of
supplements.
• Glutaclear 2 bid
• Vesselcare 1 bid
• Response: patient lost 8 lbs
in 4 weeks, effortlessly!
Bi-functional
Liver Support
Advaclear
Choline
N-Acetylcysteine
5-mthf/formly
B12
Niacin
100mg
66mg
267mcg
30mcg
23mg
Selenomethionine 50mcg
EGCG
Vesselcare for Methylation Support
• Riboflavin 5mg
• Folate (folic acid and L-5mthf)
• B6 25mg
• B 12 500mcg (Methyl)
• Zinc 5mg
• Trimethylglycine 500mg
• Choline 100mg
• Intrinsic factor 20mg
Estrogen and
Menopausal Balance
Vit. A
2500 IU
Vit. D
200IU
Vit. E
200IU
Vit. K
40mcg
B6
50mg
Folate Blend
800mcg
B12 Blend
30mcg
TMG
200mg
Chrysin
90mg
Isoflavones
100mg
Turmeric Extract
210mg
Rosemary Extract
200mg
Resveratrol
2mg
Epigenetics and autoimmunity, with
special emphasis on methylation
•
Epigenetics and autoimmunity, with special emphasis on methylation.
•
Renaudineau Y, Youinou P.
•
Source
•
Laboratory of Immunology, Brest University Medical School Hospital, Brest, France.
Once a gene promoter has been demethylated, the
gene recovers its capacity to be transcribed, e.g. genes
for cytokines, activation receptors on cells, and
endogenous retroviruses.
•
Abstract
•
Epigenetics signifies stable and heritable changes in gene expression without changes in the genetic code. There is a wealth of emerging
evidence for such processes in promoting autoimmunity. The first clue is that inhibition of DNA methyl transferases (DNMTs) induces
systemic lupus erythematosus (SLE) in animals. Similar immune-mediated disorders have been generated by injecting normal T cells
incubated with DNMT inhibitors into healthy mice. Further, monozygotic twins display differences in DNA methylation that parallel
discordances in SLE. Moreover, defects in DNA methylation characterize lymphocytes from SLE, synoviocytes from rheumatoid arthritis,
and neural cells from multiple sclerosis patients. It has also been shown that DNA hypomethylation of T and B cells correlates with
reduced DNMT efficacy and histone acetylation in SLE. Once a gene promoter has been demethylated, the gene recovers its capacity to
be transcribed, e.g., genes for cytokines, activation receptors on cells, and endogenous retroviruses. This outcome has been associated
with a blockage of the Erk pathway and/or a growth arrest at the G0/G1 interface of the cell cycle. Of importance is the fact that these
changes can be reversed. For example, blockade of the interleukin-6 autocrine loop in SLE B cells restores DNA methylation status, thus
opening new perspectives for therapy.
Keio J Med. 2011 Mar;60(1):10-6.
Differential methylation of Epstein-Barr virus
latency promoters facilitates viral persistence in
healthy seropositive individuals
•
Paulson EJ, Speck SH.
•
Source
•
Department of Pathology, Washington University School of Medicine, St. Louis, Missouri, USA.
•
Abstract
•
Epstein-Barr virus (EBV) establishes a life-long infection in humans, with distinct viral latency programs predominating during acute and
chronic phases of infection. Only a subset of the EBV latency-associated antigens present during the acute phase of EBV infection are
expressed in the latently infected memory B cells that serve as the long-term EBV reservoir. Since the EBV immortalization program elicits
a potent cellular immune response, downregulation of viral gene expression in the long-term latency reservoir is likely to facilitate
evasion of the immune response and persistence of EBV in the immunocompetent host. Tissue culture and tumor models of restricted
EBV latency have consistently demonstrated a critical role for methylation of the viral genome in maintaining the restricted pattern of
latency-associated gene expression. Here we extend these observations to demonstrate that the EBV genomes in the memory B-cell
reservoir are also heavily and discretely methylated. This analysis reveals that methylation of the viral genome is a normal aspect of EBV
infection in healthy immunocompetent individuals and is not restricted to the development of EBV-associated tumors. In addition, the
pattern of methylation very likely accounts for the observed inhibition of the EBV immortalization program and the establishment and
maintenance of a restricted latency program. Thus, EBV appears to be the first example of a parasite that usurps the host cell-directed
methylation system to regulate pathogen gene expression and thereby establish a chronic infection.
EBV appears to be the first example of a parasite
that usurps the host cell-directed methylation
system to regulate pathogen gene expression
and thereby establish a chronic infection.
J Virol. 1999 Dec;73(12):9959-68
Epigenetic and immune function
profiles associated with post-traumatic
stress disorder
•
Epigenetic and immune function profiles associated with posttraumatic stress disorder.
•
Uddin M, Aiello AE, Wildman DE, Koenen KC, Pawelec G, de Los Santos R, Goldmann E, Galea S.
•
Source
•
Department of Epidemiology, University of Michigan School of Public Health, Ann Arbor, MI 48109, USA.
•
Abstract
•
The biologic underpinnings of posttraumatic stress disorder (PTSD) have not been fully elucidated. Previous work suggests that alterations
in the immune system are characteristic of the disorder. Identifying the biologic mechanisms by which such alterations occur could
provide fundamental insights into the etiology and treatment of PTSD. Here we identify specific epigenetic profiles underlying immune
system changes associated with PTSD. Using blood samples (n = 100) obtained from an ongoing, prospective epidemiologic study in
Detroit, the Detroit Neighborhood Health Study, we applied methylation microarrays to assay CpG sites from more than 14,000 genes
among 23 PTSD-affected and 77 PTSD-unaffected individuals. We show that immune system functions are significantly overrepresented
among the annotations associated with genes uniquely unmethylated among those with PTSD. We further demonstrate that genes whose
methylation levels are significantly and negatively correlated with traumatic burden show a similar strong signal of immune function
among the PTSD affected. The observed epigenetic variability in immune function by PTSD is corroborated using an independent biologic
marker of immune response to infection, CMV-a typically latent herpesvirus whose activity was significantly higher among those with
PTSD. This report of peripheral epigenomic and CMV profiles associated with mental illness suggests a biologic model of PTSD etiology in
which an externally experienced traumatic event induces downstream alterations in immune function by reducing methylation levels of
immune-related genes.
Externally experienced traumatic events
induces downstream alterations in immune
function by reducing methylation levels of
immune-related genes.
Proc Natl Acad Sci U S A. 2010 May 18;107(20):9470-5. Epub 2010 May 3
Gail
• 72 yo CF hx of
Hypothyroidism,
Hypertension,
Hyperlipidemia, Menopausal
symptoms, Atrophic Vaginitis
with lichensclerosis,
Recurrent Shingles, Muscle
and Joint pain. Hx of total
abdominal hysterectomy age
28 for Cervical Cancer.
Reported some memory and
brain fog.
• Initial Labs: Ferritin 1034,
HFE heterogenous mutation
• Mthfr C677T+/C677T+
Homocysteine 17.9
• Vap test: tot cholesterol 287,
hdl 37, ldl 187 pattern B
• Lp(a), LpPla2, d-dimer,
fibrinogen all high
Gail Cont.
• Treatment: Bioidentical
hormone cream
• Ceralin Forte 3day (5mthf
500mcg, mB12 500mcg, NAC
600mg, B3 250mg, Acetyl-LCarnitine 600mg)
• Referred to FLT LGI diet
• Plant Sterols 3 grams/day
• RYR 2400mg a day/Niatain
500mg qhs
• Ferritin: dropped to 456,
• no further herpetic
outbreaks,
• cholesterol dropped by
50pts,
• pt reported amazing energy!
• subtype and elevated
homocysteine
• ADD/bipolar/PMS/ chronic
fatigue/fibromyalgia
• Tongue/fingernails/thin hair
• vericose veins/spider veins
• hypercoagulopathy/infertility
• elevated testosterone in
men/PCOS in women
• Acne, fibroids, endometriosis,
menometorrhagia
• depression when on OCPs
• Exacerbation of menopausal
symptoms on BHRT
• elevated uric acid and
elevated ammonia levels
• gut barrier and dysbiosis
• CANCER
• estrogen dominance and
obesity (estrogens effects on
insulin and the insulin
receptor and adiponectin)
Relationship to
• chronic low glutathione
• leaky gut and food
allergies
• Chronic viruses
• high serum B12 levels
• low serotonin and
melatonin
• high serum ferritin and
HFE mutation
Relationship to
• Thalassemia and
hepatorenal failure
• FSG and renal failure
• autoimmunity frequency
in women
• chronic dysbiosis/
chronic yeast infections
• estrogen dominant
cancers
Genetics of homocysteine metabolism
and associated disorders
•
Genetics of homocysteine metabolism and associated disorders.
•
Brustolin S, Giugliani R, Félix TM.
•
Source
•
Serviço de Genética Médica, Hospital de Clínicas de Porto Alegre, Porto Alegre, RS, Brasil.
•Hyperhomocysteinemia
Abstract
is observed in approximately 5% of the
•general
Homocysteine
is a sulfur-containingand
amino acid
from the metabolism
methionine,
an essential amino
and is
population
is derived
associated
withof an
increased
riskacid,for
metabolized by one of two pathways: remethylation or transsulfuration. Abnormalities of these pathways lead to
hyperhomocysteinemia.
Hyperhomocysteinemia
is observed in approximately
5% of the general population and is associated
many
disorders,
including
vascular
and
neurodegenerative
with an increased risk for many disorders, including vascular and neurodegenerative diseases, autoimmune disorders, birth
defects, diabetes,
renal disease, osteoporosis,
neuropsychiatric birth
disorders, and
cancer. We review
here the correlation
between
diseases,
autoimmune
disorders,
defects,
diabetes,
renal
homocysteine metabolism and the disorders described above with genetic variants on genes coding for enzymes of
homocysteineosteoporosis,
metabolism relevant to clinical
practice, especially common variants
of the MTHFRand
gene, 677C>T
and 1298A>C.
disease,
neuropsychiatric
disorders,
cancer
We also discuss the management of hyperhomocysteinemia with folic acid supplementation and fortification of folic acid and
the impact of a decrease in the prevalence of congenital anomalies and a decline in the incidence of stroke mortality.
Rxs of 5-mTHF
Brands
• Oral doses of levomefolic acid have been developed in Germany and are marketed under the
brand name Metafolin.
• Cerefolin NAC (5.6 mg) – Nutritional requirements of individuals under a physician's treatment
for neurovascular oxidative stress or hyperhomocysteinemia, with particular emphasis for
those individuals diagnosed with or at risk for mild to moderate cognitive impairment, vascular
dementia, Alzheimer's disease or recurrent or ischemic.
• Deplin (7.5 mg or 15 mg) – Dietary management of suboptimal folate levels in depressed
patients.
• Metanx (2.8 mg) – Dietary management of endothelial dysfunction in patients with diabetic
peripheral neuropathy.
• Néevo and NeevoDHA (1 mg) – Dietary management of those women under a physician's
treatment for vitamin deficiency throughout pregnancy, postnatal and the lactating periods.
Néevo is specifically indicated for: patients with high risk pregnancies, older OB patients, and
patients unable to fully metabolize folic acid.
• Zervalx (1 mg) – Dietary management of the following patient conditions: preconception
hyperhomocysteinemia; intermediate to higher-risk pregnancies that require an increased
dietary folate intake; hemolytic, megaloblastic or sickle cell anemia complicated by folate
deficiency; and methotrexate induced plasma hyperhomocysteinemia.
Methylenetetrahydrofolate reductase
(MTHFR) genetic polymorphisms and
psychiatric disorders
•
Methylenetetrahydrofolate reductase (MTHFR) genetic polymorphisms and psychiatric disorders: a HuGE review.
•
Gilbody S, Lewis S, Lightfoot T.
•
Source: Department of Health Sciences, Alcuin College, University of York, York, United Kingdom. [email protected]
•
Abstract
•
The authors performed a meta-analysis of studies examining the association between polymorphisms in the 5,10methylenetetrahydrofolate reductase (MTHFR) gene, including MTHFR C677T and A1298C, and common psychiatric
disorders, including unipolar depression, anxiety disorders, bipolar disorder, and schizophrenia. The primary
comparison was between homozygote variants and the wild type for MTHFR C677T and A1298C. For unipolar
depression and the MTHFR C677T polymorphism, the fixed-effects odds ratio for homozygote variants (TT) versus
the wild type (CC) was 1.36 (95% confidence interval (CI): 1.11, 1.67), with no residual between-study heterogeneity
(I(2) = 0%)--based on 1,280 cases and 10,429 controls. For schizophrenia and MTHFR C677T, the fixed-effects odds
ratio for TT versus CC was 1.44 (95% CI: 1.21, 1.70), with low heterogeneity (I(2) = 42%)--based on 2,762 cases and
3,363 controls. For bipolar disorder and MTHFR C677T, the fixed-effects odds ratio for TT versus CC was 1.82 (95%
CI: 1.22, 2.70), with low heterogeneity (I(2) = 42%)-based on 550 cases and 1,098 controls. These results were
robust to various sensitively analyses. This meta-analysis demonstrates an association between the MTHFR C677T
variant and depression, schizophrenia, and bipolar disorder, raising the possibility of the use of folate in treatment
and prevention.
This meta-analysis demonstrates an association between the
MTHFR C677T variant and depression, schizophrenia, and
bipolar disorder, raising the possibility of the use of folate in
treatment and prevention.
Am J Epidemiol. 2007 Jan 1;165(1):1-13. Epub 2006 Oct 30
Methylenetetrahydrofolate reductase
gene polymorphisms in children with
attention deficit hyperactivity disorder
•
Methylenetetrahydrofolate reductase gene polymorphisms in children with attention deficit hyperactivity
disorder.
•
Gokcen C, Kocak N, Pekgor A
•
Source
•
1. Department of Child and Adolescent Psychiatry, Medicine Faculty of Gaziantep University, Gaziantep, Turkey.
Conclusions:
Preliminary data imply a possible relationship
• Abstract
between
and5,10the
ADHD
• Objective: TheA1298C
purpose of thisMTHFR
study was to polymorphisms
evaluate the relationship between
methylenetetrahydrofolate
reductase (MTHFR) polymorphisms and Attention Deficit Hyperactivity Disorder (ADHD) in a sample of Turkish
children.Study Design: MTHFR gene polymorphisms were assessed in 40 patients with ADHD and 30 healty controls.
Two mutations in the MTHFR gene were investigated using polymerase chain reactions and restriction fragment
length polymorphisms.Results: Although there were no statistically significant differences in genotype distributions
of the C677T alleles between the ADHD and the control groups (p=0,678) but the genotypic pattern of the
distributions of the A1298C alleles was different between the ADHD patients and the controls
(p=0,033).Conclusions: Preliminary data imply a possible relationship between A1298C MTHFR polymorphisms and
the ADHD.
Int J Med Sci. 2011;8(7):523-8. Epub 2011 Aug 30
Rachel
16 yo female c/o migraines,
neck pain and stiffness, PTSD
after a cheerleader fell on
her, dx with cervicogenic
headache syndrome, hx of
ADD and cannot get out of
bed unless she takes her
Adderall.
Rachel Cont.
Seasonal allergies, allergic to
gluten, eggs, and dairy,
abdominal pain for years,
dysmenorrhea and PMDD
using progesterone cream 2
weeks prior to cycle, h/o PCOS
and ovarian cysts, Severe
anxiety and mood swings.
Rachel Cont.
• Labs: Double MTHFR
mutation
(C677T/A1298C), DHEAS
466, Testosterone 87
c/w PCOS!, Food allergy
test: +gluten, wheat,
dairy, eggs, yeast….Leaky
Gut Barrier, US 4x3cm
ovarian cyst
ADD Teen Rachel – Cont’d
Treatment
• PCOS: Estrofactors 3 day
• Insomnia/Anxiety: 5HTP/LTheanine, Somnolin QHS
• Headaches: Magnesium and
5-htp (5HTP/L-Theanine)
• Magnesium glycinate 200mg
Qhs
• Trancor 2 Bid for Nac and
Theanine support
• Error in Methylation/
Transsulfuration:
Methylation support product
in am, Ceralin forte tid
Rachel
Progress: Sleep and
moods are
dramatically
better!
Headaches are
resolved, periods
and acne have
improved, better
energy and fresh
motivation!
Autism and Impaired Methylation
• An increased vulnerability to oxidative stress and a decreased
capacity for methylation may contribute to the development and
clinical manifestation of autism.
• …the children with autism had significantly lower baseline plasma
concentrations of methionine, SAM, homocysteine, cystathionine,
cysteine, and total glutathione and significantly higher
concentrations of SAH, adenosine, and oxidized glutathione
• consistent with impaired capacity for methylation (significantly lower
ratio of SAM to SAH) and increased oxidative stress (significantly
lower redox ratio of reduced glutathione to oxidized glutathione)
Metabolic biomarkers of increased oxidative stress and impaired methylation capacity in children with
autism 1 From the Department of Pediatrics, University of Arkansas for Medical Sciences, and the Arkansas
Children‘s Hospital Research Institute, Little Rock, AR (SJJ, SM, and SJ); Niagara Falls, NY (PC); Colden, NY
(LJ); Gaylor and Associates, LLC, Eureka Springs, AR (DWG); and Edison, NJ (JAN)2 Rep
Chris
• PMhx: 6 yo HM hx of severe
anxiety, unable to perform in
class, crying, restless,
insomnia, ADD, dyslexia.
Mother brought him in for
medication for anxiety and
referral to special education
program. Mom was
considering home schooling
due to severity of anxiety.
Small bumps on cheeks and
upper arms mom describes
as acne.
Chris Cont.
• Labs: HLA Dq2/ Hla Dq8
negative
• Food allergy test + gluten,
wheat, dairy, honey, yeast,
and some other minor
antibodies (leaky gut).
• MTHFR: c677T+/A1298C+
Chris – Cont.
• Treatment: Elimination diet 6
weeks followed by no gluten
or cow dairy.
• EPA/DHA liquid
• Multigenics Chewable
• Folapro
• Ceralin forte
• Somnolin 1 qhs
• Response: all anxiety and
sign of ADD completely
resolved!!
minor issues with dyslexia
continued, integrated back
into school after mom took
him out of school and home
schooled for a year until she
could control his diet.
Methyl-folate and methylB12 were added this past
year in Somnolin which helps
energy and focus
Glutaclear was added to help
support detoxification in
Phase II of liver as well as to
support weight loss.
• INTERVENTION 1: Supplementation with folinic acid and
betaine
• Although supplementation was effective in normalizing the
methionine cycle metabolites to the concentrations in the
control subjects, the intervention significantly improved but
did not normalize tGSH or GSSG concentrations or tGSH:GSSG.
• INTERVENTION 2: Supplementation with folinic acid and
betaine, and Methyl vitamin B12 (intervention 2)
• The addition of injectible methylcobalamin (intervention 2) did
not alter the mean concentrations of methionine, SAM, SAH,
or homocysteine beyond the alterations induced by the
intervention with folinic acid and betaine. However, relative to
intervention 1, the addition of injectible methylcobalamin
further decreased the concentrations of adenosine and GSSG
and further increased the concentrations of methionine,
cysteine, and tGSH and SAM:SAH and tGSH:GSSG.
Antisense inhibition of
methylenetetrahydrofolate reductase
reduces cancer cell survival in vitro and
tumor growth in vivo
•
Stankova J, Shang J, Rozen R.
•
Source
•
Departments of Human Genetics, Pediatrics and Biology, Research Institute, McGill University-Montreal Children's Hospital, 4060 Saint Catherine Street West, Montreal, Quebec, Canada
H3Z 2Z3.
•
Abstract
Conclusions:
Our results confirm that MTHFR inhibition decreases tumor
growth and suggest that inhibition of MTHFR by antisense or
small molecules may be a novel anticancer approach.
•
PURPOSE:
•
Many cancer lines are methionine dependent and decrease proliferation when methionine supply is limited. Methylenetetrahydrofolate reductase (MTHFR) generates the folate derivative
for homocysteine remethylation to methionine. We investigated the effect of antisense-mediated inhibition of MTHFR on survival of human cancer cells.
•
EXPERIMENTAL DESIGN:
•
We examined the in vitro and in vivo anticancer effects of a combination of MTHFR antisense and standard cytotoxic drugs.
•
RESULTS:
•
Specific antisense against MTHFR (EX5) showed significant inhibitory effects on growth of human colon, lung, breast, prostate, and neuroblastoma tumor cells in vitro compared with that of
the control oligonucleotide. Cytotoxic drugs (5-fluorouracil, cisplatin, or paclitaxel) potentiated the effect of EX5. In vivo, antisense alone or in combination with cytotoxic drugs inhibited the
growth of human colon and lung carcinoma xenografts. In comparison with control oligonucleotide, treatment with EX5 inhibited growth of colon tumors and lung tumors by 60% and 45%,
respectively. EX5 with 5-fluorouracil decreased growth of colon tumors by an additional 30% compared with EX5 alone, and EX5 with cisplatin decreased growth of lung tumors by an
additional 40% compared with cisplatin alone. Growth inhibition by EX5 was associated with decreased amounts of MTHFR protein and with increased amounts of an apoptosis marker.
•
CONCLUSIONS:
•
Our results confirm that MTHFR inhibition decreases tumor growth and suggest that inhibition of MTHFR by antisense or small molecules may be a novel anticancer approach.
Adenosylmethionine produces and
inactive form of MTHFR
• Jencks DA, Mathews RG (February 1987). “Allosteric inhibition
of methylenetetrahydrofolate reductase by
adenosylmethionine. Effects of adenosylmethionine and
NADPH on the equilibrium between active and inactive forms
of the enzyme and on the kinetics of approach to equilibrium”.
J. Biol. Chem. 262 (6): 2485–93.
• NADPH promotes activation of MTHFR which has two flavin
moities. C677T polymorphisms tend to have a thermolabile
form of MTHFR and lose their B2 three times faster than the
wild type of MTHFR, decreasing it’s enzyme function.
Association of C677T gene polymorphism
of methylenetetrahydrofolate reductase
with insulin resistance among Kirghizes
•
Abstract
•
AIM:
•
To study an association of C677T gene polymorphism of methylenetetrahydrofolate reductase (MTHFR) and insulin resistance (IR) among
ethical Kirghizes.
•
METHODS:
Conclusion:
• 132 Kirghiz patients with IR according to HOMA index (n=132) and sex and age matched patients without IR, diabetes mellitus (DM) type
2 or metabolic syndrome (MS) (n=132) were included into this study. Measurements of blood pressure (BP), body mass index, waist and
In Kirghizes
ofinsulin,
T677
allele
gene
was
hip circumference, carriage
fasting blood sugar,
lipid parameters
and of
C677TMTHFR
gene polymorphism
of MTHFR
were performed in all patients.
associated
with IR, abdominal obesity, hypertriglyceridemia
• RESULTS:
• Frequency
CT and TT genotypes
was significantly higher in patients with IR than in controls (2 - 7.22, p - 0,027, OR - 1.68, 95%
and
low ofHDL-C
level.
confidence interval 1.13-2.5, p=0.027). T677 allele was also associated with obesity, hypertriglyceridemia and low level of high density
lipoprotein cholesterol (HDL-C).
•
CONCLUSION:
•
In Kirghizes carriage of T677 allele of MTHFR gene was associated with IR, abdominal obesity, hypertriglyceridemia and low HDL-C level.
Kardiologiia. 2011;51(3):58-62
Association of the C677T polymorphism in
the methylenetetrahydrofolate reductase
gene with sudden sensorineural hearing loss.
• Uchida Y, Sugiura S, Ando F, Shimokata H, Nakashima T.
• Source
• Department of Otorhinolaryngology, National Center for Geriatrics and Gerontology,
Our
results
suggest that the T allele of MTHFR C677T could be
Obu,
Japan. [email protected]
associated with susceptibility to SSNHL, and even imply that
• Abstract
this mutation could be a risk factor that is independent of
•blood
OBJECTIVES/HYPOTHESIS:
folic acid and homocysteine.
• To investigate the recently reported association of the C677T polymorphism in the
methylenetetrahydrofolate reductase (MTHFR) gene with sudden sensorineural hearing
loss (SSNHL), we analyzed data from a community-based Japanese population.
Laryngoscope. 2010 Apr;120(4):791-5
Andrew
• 68 yo HM presented to clinic
complaining of papular rash on
scalp started after visit to ER and
treatment with antibiotics for
severe diverticulitis. Hx of lower
extremity neuropathy, borderline
diabetes, fatigue, brain fog.
Reported sudden onset of loss of
hearing in the Rt. Ear and
treatment with injections of
steroid into ear by ENT.
• Cytokine blood test revealed
mycotoxicity, elevated hs crp,
sedrate, MTHFR C+/A+
polymorphism
Andrew Cont.
• Treated patient with Methylation
support product and 5HTP/LTheanine for methylation while
stool culture was pending, rash on
scalp improved. Patient was then
treated with diflucan, Lactobacillus
acidophilus NCFM/Bifodlbacterium
lactis, and Saccharomyces
boulardii combo product x 6
weeks. Patient was maintained on
Saccharomyces boulardii combo
product in am and candiBactin-AR
in pm for a total of three months
post Diflucan treatment.
• Partial Rt ear hearing
improvement!
Severe atherosclerosis in rheumatoid
arthritis and hyperhomocysteinemia:
Is there a link?
• El Bouchti I, Sordet C, Kuntz JL, Sibilia J.
• Source: Department of Rheumatology, Hautepierre Teaching Hospital, Strasbourg, France.
• Abstract
Rheumatoid arthritis (RA) is a chronic inflammatory joint disease
• Rheumatoid arthritis (RA) is a chronic inflammatory joint disease whose main complication is accelerated atheroma
whose
main
is accelerated
atheroma responsible
responsible
for high complication
rates of cardiovascular morbidity
and mortality. Hyperhomocysteinemia
is among the factorsfor
incriminated in RA-associated atheroma. We managed a 46-year-old patient with RA who required admission to
evaluate
severe of
arterial
and venous disease withmorbidity
involvement of theand
coronary,
renal, and peripheral arteries. She
high
rates
cardiovascular
mortality
had no laboratory evidence of rheumatoid vasculitis or conventional cardiovascular risk factors (diabetes and
hypercholesterolemia) and had never smoked. Her serum homocysteine level was elevated to 20.9 micromol/L as a
result of a homozygous C667T mutation in the methylenetetrahydrofolate (MTHFR) gene. Folate and vitamin B12
levels were normal. A circulating anticoagulant was identified. Hyperhomocysteinemia, which is defined as a
homocysteine level greater than 15 micromol/L, is a risk factor for arterial and venous disease.
Hyperhomocysteinemia is found in 20%-42% of patients with RA. Methotrexate therapy is the most common
causative factor. Other causes include MTHFR deficiency, vitamin B12 deficiency, renal failure, old age, and smoking.
Whatever the underlying cause, folic acid supplementation returns the homocysteine level to normal.
Hyperhomocysteinemia is found in 20%-42% of patients with RA.
Methotrexate therapy is the most common causative factor.
Other causes include MTHFR deficiency, vitamin B12 deficiency,
Joint
Bone
Spine. 2008
Epub 2008 May 23
renal
failure,
oldJul;75(4):499-501.
age, and smoking
Methotrexate
• Antimetabolite and antifolate drug
• Methotrexate competitively inhibits dihydrofolate reductase
(DHFR), an enzyme that participates in the tetrahydrofolate
synthesis. The affinity of methotrexate for DHFR is about one
thousand-fold that of folate. DHFR catalyses the conversion of
dihydrofolate to the active tetrahydrofolate.
• Folic acid is needed for the de novo synthesis of the nucleoside
thymidine, required for DNA synthesis. Also, folate is needed
for purine base synthesis, so all purine synthesis will be
inhibited. Methotrexate, therefore, inhibits the synthesis of
DNA, RNA, thymidylates, and proteins.
Methotrexate
• For the treatment of rheumatoid arthritis, patients should
supplement their diets with folate. In these cases, inhibition of
DHFR is not thought to be the main mechanism, but rather the
inhibition of enzymes involved in purine metabolism, leading
to accumulation of adenosine, or the inhibition of T cell
activation and suppression of intercellular adhesion molecule
expression by T cells.
"The anti-inflammatory action of methotrexate is not mediated by lymphocyte
apoptosis, but by the suppression of activation and adhesion molecules". Clinical
Immunology 114 (2): 154–63. doi:10.1016/j.clim.2004.09.001
Methotrexate (green) complexed into the active site of DHFR (blue)
Janeth
• 37 y/o Asian female, dx with RA a
few years ago, was being treated
with Methotrexate and Enbrel but
had a serious drug reaction and
developed meningitis. Joint pain
severe in all fingers, fatigue, worse
around her period, headaches,
insomnia, low mood.
• Labs: anti-ccp ab>250 MTHFR
double C677T SNP
• HLA DQ 2 ++/HLA DQ 8-+
Janeth
• Treatment:UltraInflamx360
medical food for inflammation
• EPA DHA 4g/day
• Kaprex for breakthrough days prior
to period
• Estrofacors 3 day
• Magnesium 400mg ghs
• Sam-e 200mg bid
• Somnolin qhs
• Ceralin forte
• Immucore for Immune support
Response: Feels as stable as
she did on meds without
medication side effects.
Inflammatory profile, age of onset, and the MTHFR
polymorphism in patients with multiple sclerosis
Sclerotic
patients also have elevated levels of plasma and CSF
• Inflammatory profile, age of onset, and the MTHFR polymorphism in patients with multiple sclerosis.
homocysteine.
• Alatab S, Hossein-nezhad A, Mirzaei K, Mokhtari F, Shariati G, Najmafshar A.
•
Source
•
Endocrinology and Metabolism Research Center, Tehran University of Medical Sciences, North Kargar Ave., 5th Floor, Dr. Shariati Hospital, Tehran
14114, Iran.
•
Abstract
•
Both genetic and inflammatory factors are suspected in the etiology of multiple sclerosis (MS). Of genetic factors, the methylenetetrahydrofolate
reductase (MTHFR) C677T polymorphism has been associated with increased levels of plasma homocysteine, a neuronal excitotoxic amino acid.
Sclerotic patients also have elevated levels of plasma and CSF homocysteine. In this study, the association between C677T polymorphism and MS was
tested by recruiting 230 healthy and 194 multiple sclerotic age- and gender-matched patients. The MTHFR C677T polymorphism and the serum levels
of inflammatory mediators IL-1β, TNFα, and CRP were measured. TNFα, CRP, and IL-1β levels were significantly higher in sclerotic patients. T allele was
1.7 times more present in this group. In patient's group, the levels of all inflammatory mediators were higher in T/T compared to two other genotypes.
Evaluation of the age of onset of disease revealed that subjects with T allele developed the MS disease, almost 4 years sooner than other genotype. We
concluded that having T allele of C677T in MS might be accompanied with higher levels of serum inflammatory mediators and a vulnerability to earlier
age of onset of disease. Further studies are needed to elucidate the underlying mechanisms.
TNFα, CRP, and IL-1β levels were significantly
higher in sclerotic patients. C677T allele was 1.7
times more present in this group.
We concluded that having T allele of C677T in MS might be
accompanied with higher levels of serum inflammatory mediators
and a vulnerability to earlier age of onset of disease.
J Mol Neurosci. 2011 May;44(1):6-11. Epub 2010 Dec 29
Hilda
• 34y/o HF Multiple Sclerosis patient
with weakness, fatigue, and
neurologic loss of function of
bladder, and extremities
• Swelling in knees, feet, worse with
Etoh
• Constipation, IBS, bloated
• Insomnia and easily aroused from
sleep
• Weight gain of 20 lbs in 6 months
• Anxiety and feels like she is going to
have “breakdowns”
Hilda Cont.
• RUQ pain under liver zone
• PMS: TAH 2006, BTL 2003 Hx of
Multiple Sclerosis
• Plan: refer to FLT for 28 day detox
and education of Elimination Diet,
then placed on UltraInflamX360
• Ultrafloa IB, Lactobacillus
acidophilus NCFM/Bifodlbacterium
lactis,
• avoid artificial sweetners
• S-adenosyl-L-methionine in pm,
gluten free diet,
• EPA/DHA
Hilda Cont.
• Labs: cbc, chem, iron,
ferritin, crp wnl
• Tot chol 210, TG 194, Hdl 60,
Ldl 111
• HgA1c 5.2, Uric Acid 5.8
• Estradiol 25, progesterone 6,
Free testosterone 53
• Vit. D 31.8, Vit B12 321,
• Glutatione 1050
• MTHFR C677T/C677T, Celiac
Panel
negative(HLADQ2/DQ8)
• Ammonia 107
• Spectracele: multiple vitamin
deficiencies
• Cytokines: elevated for
Candida4+++
• Homocysteine: 17
Hilda Cont.
Assessment:
• Disturbed Sulphur Amino
Acid Metabolism (270.4)
• Vesselcare +MethylB12
• Glutaclear 2 bid
• PCOS (246.4) Estrofactors 3
Qam
• Labs: hormones Q4weeks
• Multiple Sclerosis: Sadenyl methionine and
Methylation support
product to stimulate
methylation of DNA, and
activation of
Oligodendrocytes to
produce myelin
• Methylcobalamine
1000mcg IM twice
weekly
Hilda Cont.
• Vitamin D deficiency: add
5000IU/Day
• Insomnia: Somnolin 2 at
bedtime with methylators
• EPA/DHA high concentrate
4 grams a day, Wellness
Essentials (mvi, advaclear,
Epa/Dha)
• Progress: Decrease in
edema and lower
extremity swelling and
pain, insomnia resolved,
moods improved, appetite
improved, weakness
improved, abnormal
estrogen/testosterone
improved.
• Candida enteritis (112.84)
Diflucan 100mg x 6wks
• Ultraflora IB
• Proboulardii combo
product
• Candibactin-AR
Methods to Increase Glutathione
•
•
•
•
•
•
•
•
•
•
•
Methylation support
L-methionine
5HTP/L-Theanine
NAC/niacinamide/Glutaclear
Magnesium Glycinate
Ceralin forte
Estrogen Balance Support
product
Methylcobalamine
5MTHF
Cortisol support with DHEA
Choline
• GABA and Glutamate support
• Bifunctional detox support
product
• Heavy metal metabolism support
product
• 5MTHF
• Intrinsic B12/5MTHF
• Alpha Lipoic Acid
• Vitamin C
• L-Carnitine
• Vitamin E
• Trimethylglycine
MTHFR polymorphisms and low Sam-e
related to low Serotonin and Melatonin
deficiency
L-Tryptophan
B3
5-HTP
B6, VitC
Serotonin
5HIAA
S-adenyl-methionine
Melatonin
The conversion of 5-HTP to Serotonin with Vit B6 and then to Melatonin with
Sam-e as the co-factor is needed for healthy moods and sleep. Low
serotonin is associated with low mood, sleep difficulties, hot flashes, and an
increase appetite
S-adenosyl-L-methionine is a cofactor in the conversion
of norepinephrine to epinephrine to enhance energy
and motivation
L-Dopa
B6
Dopamine
Vit C
Norepinephrine
S-adenyl-methionine
Ephinephrine
S-adenosyl-L-methionine is the co-factor that helps to convert Norepinephrine
to epinephrine increasing excitatory neurotransmitters and energy as well as
the activator or COMT to digest catecholamines.
Tina Cont.
• Tchol 188, Ldl 112, Hdl 39,
Trig 185 Pattern B
• Apo B 109 (<90)
Homocysteine 14.2, lp-PlA2
164 (<200)
• Insulin 26 (<25) Fibrinogen
530 (<496) hs crp 7.3 (<1.0)
• MTHFR C677T+/C677T+
• Stool Cx 4+Candida albicans
Tina– Cont’d
• Treatment: Vesselcare bid
• Ceralin forte 1 bid
Pro-boulardii combo product
Candibactin-AR
• Advaclear bifunctional detox
support product 2 bid
• Repeat labs:
• stop Crestor & switch to
Simcor 20/500mg
(Simvastatin/Niacin)
• AST: 87>248>69
• arginine/magnesium 200mg
qhs
• We are trying to clean up
her NASH! No estrogen at
this time! Too risky.
• Diflucan 100mg x 10 days
• Ultraflora IB
• GGT: 491>473>477>79
• ALT: 67>111>103
Laura
• 61 yo CF abd. Pain, food allergies,
ibs constipation, bloating, rash skin
of neck and back when eats gluten
and dairy, blisters that itch and
burn, hyperlipidemia, 20lb wt loss
in 6 mo unintentionally, hair loss,
brittle nails, dry skin, uses tobacco
1 pk/dayx40yrs
• Labs: GGT: 18, Ferritin 273 (13150)
• Chol 350, Hdl 107, TG 119, Ldl 219
pattern A
Laura Cont.
• +Candida stool, + SIgA gliadin, soy,
dairy
• HLA DQ2-/DQ8+ MTHFR C677T-+
• Treatment: gluten, dairy, soy free,
Niatain 500mg 1 bid
• 28 day Ultraclear Renew Detox
• Somnolin qhs
• Vesselcare in am
• Ultraflora DF and Proboulardii,
• Candibactin-AR
Laura – Cont’d
• Response: patient reported
energy improved, sleep and
moods improved, felt less
achy and tender off gluten,
soy and dairy. IBS like
symptoms completely
improved, and pustular rash
on neck and back resolved.
• Tot Chol: 305>260>229>253
• Trigs: 72>114>67>105
• HDL: 92>88>88>72 (hdl2ab
dominant)
• Ldl: 199> 149>128>160
• (Note: bump in last lipid
panel was thought to be due
to patient using Diflucan
100mg a day for 6weeks
prior to final cholesterol lab
drawn)
Sally
• 39 yo Indian female, hx of slow
onset of papillary-like rash on
upper and lower extremities,
pruritic, creating terrible hyperpigmented scarring when
healing. Reported long hx of
endometriosis and
menometorrhagia, Total
Abdominal hysterectomy 6
months ago due to heavy
bleeding. Patient was placed on
synthetic hormone replacement
to help moods, sleep and hot
flashes. 6 weeks later is when
patient started to develop odd
skin papules.
Sally Cont.
• MTHFR mutation C677T/A1298C
• Treatment: synthetic hrt was
stopped, biest/progesterone
1.25mg/25mg topical cream was
rx’ed instead, methylation support
product am, 5HTP/L-Theanine pm,
Magnesium 400mg qhs,
NAC/niacinamide 2 bid
• Response: Rash resolved quickly
over 2-3 weeks and moods, sleep,
energy, and hormone symptoms
improved as well.
Chris
• 45 yo CM with hx of chronic
fatigue, chest pain, and
shortness of breath that started
after he developed
mononucleosis a year ago and
never recovered his energy. Hx
of recurrent oral herpes, cold
sores, shortness of breath and
chest pain that is worse if he
drinks Etoh, coffee, or sugar,
also notices that these make it
hard for him to fall asleep at
night. Skin lesions on his upper
arms… pimple like lesion he
Chris Cont.
he scratches the heads off… tend
to scar, and recur only around
upper and arms and elbows.
Works as a meter reader; around
gases
• Labs: MTHFR A1298+ + EBV IGG
6.5 EBV IGM –
• HSV IgM I/II + …chronic Herpes
Simplex
• MELISA + IgG Nickel
• Lymes: western blot• Lyme Immune Tolerance test:
positive
Chris – Cont’d
• Treatment:
• Symphora 2 qam
• Magnesium 400mg Qhs
• Somnolin in pm
• Advaclear 2 po bid
• Vit D3 5000 IU day
• Ceralin forte tid
• Progress: Patient states his
energy and fatigue are much
improved. His shortness of
breath and overall strength
feels normal daily unless he
over exerts himself with a
tough day on the job. His sleep
is much improved and he wakes
up feeling rested. He stated
that his skin lesions are
completely gone and his oral
cold sores are completely gone.
Diana
• 42 yo CF, reported hx of
partial hysterectomy last
year for endometriosis, with
recurrent candida enteritis
despite a yeast free and low
sugar diet, anxiety, low
mood, sleep difficulties,
recurrent oral cold sores,
fibrocystic breast, chronic
low energy.
• MTHFR A1298+/heterozygote
• Stool cx: no yeast, low levels
of lactobacilli
Diana– Cont’d
• Treatment:
• Vesselcare 1 QD
Methylcobalamine 1000mcg
qd
• Proboulardii 1 po qd
• Candibactin-AR
• Response: Patient stated she
could feel an immediate
difference in mood and
energy in a day, feels less
hormonal! Feels like all of
her IBS-like symptoms slowly
resolved.
Rob
• 80 yo CM hx of Parkinson’s
Syndrome, diarrhea, spinal
stenosis, takes Sinemet,
Azilect, Lodosyn, hx GERD, LE
neuropathy, fatigued and
unmotivated. Dental Crowns:
gold … “I want to try IV
glutathione”
• FMHX: brother and sister
Parkinson’s, Mother
Schizophrenia
Rob Cont.
Labs:
• MTHFR negative HLA DQ 2 /- HLA DQ 8 +/+
• Blastocystis hominis 4+
moderate infection in stool
cx
• B12 442 Homocysteine: 11.9
glutathione: 758
Rob – Cont’d
• Response: Patient reported
“ I am feeling much better... I
am eating gluten and dairy
free… less nausea, no more
diarrhea.” Stated he ran out
of GABA and Glutamate
support which was helping
with his anxiety and realized
how much it was really
helping him to feel more
calm. He noticed an increase
in energy and feeling of wellbeing once he finished 2
bottles of the heavy metal
detox support product. He is
continuing his Lactobacillus
acidophilus
NCFM/Bifodlbacterium
lactis, EPA DHA , Cortisol
support with DHEA and
methylators, GABA and
Glutamate support, Immune
support product. He has just
ordered NAC/niacinamide to
help increase his glutathione.
• Last glutathione: 836!!!!!
(544-1228uM)
Chris D.
• 9 yo CM bib mom with
report of behavioral
problems started very early
on, abusive of younger sister,
anger outbursts, poor coping
behavior at school,
abdominal pain, reflux,
burping, nausea, vomiting,
age 7 was found to have
precocious puberty with
bone age 2.5 years
accelerated and pubic and
axillary hair growth.
Chris cont,
Seen by Endo and placed on
meds to arrest puberty. Dry
scalp, joint and muscle pain
every night. Meds: Abilify,
Intuniv, Seroquel. “my son is
a zombie…he falls asleep
everywhere we go”
• Labs: Celiac panel negative,
MTHFR C677T+/A1298C+
• Glutathione: 501 (5441228uM)
Maria
• 27 y/o CF
Infertility/anovulation
• Autoimmune FSGS with renal
failure on kidney transplant
list
• Hypothyroidism with
Hashimoto’s thyroiditis
• Hyperlipidemia
• Labs: HLA DQ8++, MTHFR
C677T+, A1298T+
• Progesterone day 21: 7ng/ml
Maria Cont.
• TPO ab 304
• TREATMENT: Elimination diet/
gluten free
• Estrofactors 3 Qam
• Vesselcare bid
• Topical progesterone days 14-28
• Progesterone day 21: 18.5ng/ml
+Ovulation
• Kidney function is completely
normal now and she is not on
transplant list any longer.
Full Spectrum Liver/Kidney/Metal
Chelation/Alkaline Detox
10 or 28 Day Detox Plan:
Structured elimination diet food plan with slow
progressive up-regulation of Phase I and Phase
II detoxification systems.
Designed to Alkalinize the body to improve
Phase III Renal Elimination of Toxins
Also designed to up-regulate Metallothionine
to chelate and remove heavy metals from the
body.
Advise patients to take methyl-folate and
Methyl-B12 if clinical suspicion exists that the
patient is a slower methylator.
If patient complains of increase in headaches or
muscle aches, add vesselcare twice a day with
Methyl-B12 1000mcg a day.

mthfr - SpectraCell Laboratories

Transcription

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