GlyphosateandtheGut
StephanieSeneffMITCSAIL
Au2smOne2018
Outline• Au2smandGutIssues• PathogenOvergrowth• Diges2veEnzymeImpairment• Celiacdisease• An2bio2cResistance• BTBRMousemodelofAu2sm• ImpairedCYPenzymesandBileAcidSynthesis• ImpairedMyosinandPeristalsis• ClostridiaOvergrowthandAu2sm• Summary
Au2smandtheGut*
“Prospec2ve,controlledstudiessuggestthatasmanyas70%ofau2s2cchildrenexhibitchronicGI-relatedsymptoms[1,5,6]includingdiarrhea,laxa2ve-dependentcons2pa2on,abdominaldistension,failuretothrive,weightloss,feedingproblems,andabdominalpainrelatedtoextremeirritability,aggression,andself-injury.”
*SJWalkeretal.PLOSOneMarch2013;8(3):e58058.
GlyphosateandtheGut:PathogenOvergrowth
• Glyphosateisanan2microbialagentthatpreferen2allykillsbeneficialmicrobes,allowingpathogenstoflourishinthegut*
• Immunecellsinvadethegutandreleaseinflammatorycytokines– ThiscausesincreasedrisktoinflammatoryboweldiseasessuchasCrohn’sandulcera2vecoli2s
*SamselandSeneff.Entropy2013;15:1416-1463.
*PlotprovidedbyDr.Mar2nMichener
Clostridia Salmonella
PathogenOvergrowthinPoultryMicrobesExposedtoGlyphosate*
Bifidobacteria
GlyphosateandtheGut:Diges2veEnzymes
• Glyphosatehasbeenfoundasacontaminantindiges2veenzymestrypsin,pepsinandlipase*
• Trypsinimpairmentpreventsproteinslikegluteninwheatfrombeingdigested
• Undigestedproteinsinducereleaseofzonulinwhichopensupgutbarrier**
• Zonulinlingersbecausetrypsinisdefec2ve
*ASamselandSSeneff.JBiolPhysChem2017;17:8-32**JJGildeaetal.JClinNutrDiet.2017,3:1.
Trypsin,PepsinandLipaseareallcontaminatedwithglyphosate*
*ASamselandSSeneff.JournalofBiologicalPhysicsandChemistry2017;17:8-32
Enzyme Glyphosate(PPB)
Pepsin(ELISA) <40Pepsin(GC-MS) 430Pepsin(HPLC-MSMS) 290Trypsin(ELISA) 62Lipase(ELISA) 24
H
GlycineGlyphosate
Glyphosateisanon-codingaminoacidanalogueofglycine
H
Glyphosate
Glyphosateisanon-codingaminoacidanalogueofglycine
Glyphosatesubs2tu2onbymistakeforglycine
duringproteinsynthesiscouldexplainglyphosatecontamina2onintrypsin
Trypsin,PepsinandLipaseareallcontaminatedwithglyphosate*
*ASamselandSSeneff.JournalofBiologicalPhysicsandChemistry2017;17:8-32
Trypsin’sac2va2ondomaincontainsfourcrucialglycinerichsubdomains:*
N-terminustoGly19Gly142toPro152Gly184toGly193Gly216toAsn223
AScenarioforGlutenIntolerance
ZonulinTrypsindefec2ve
Zonulinreleased
LeakybarrierGliadin
Leakybrainbarrier
Glyphosateinwheat
Autoimmuneneurologicaldisease
Systemicimmuneresponseinducesmul2plecomplexsymptoms
*SamselandSeneff,InterdiscipToxicol.2013;6(4):159–184.
Celiacdisease
GlyphosateandCeliacDisease*
GlyphosateUseonwheat
CeliacDisease,GlyphosateandNonHodgkin’sLymphoma
• Glyphosatepreferen2allykillsBifidobacteria*• Bifidobacteriaaredepletedinceliacdisease**• CeliacdiseaseisassociatedwithincreasedrisktononHodgkin’slymphoma***
• GlyphosateitselfisalsolinkeddirectlytononHodgkin’slymphoma****
*A.A.Shehataetal.,CurrMicrobiol.2013Apr;66(4):350-8.**M.Velasquez-Manoff,NYTimesSundayReview,Feb.23,2013.***C.Catassietall,JAMA.2002Mar20;287(11):1413-9.****M.Erikssonetal.,IntJCancer.2008Oct1;123(7):1657-63.
*Figure20,NLSwansonetal.JournalofOrganicSystems9(2),2014,p.25.
GlyphosateInducesAn2bio2cResistance*• Ac2nobacteriaproduceafreeradicalscavengerinresponsetoglyphosatethatprovidesresistancetoawiderangeofan2bio2cs,includingpenicillin
• E.coliexposedtoglyphosatedevelopan"effluxpump"thatincreasesresistancetothefluoroquinoloneCiprofloxacinandtheaminoglycosideKanamycin.– SameeffectobservedinSalmonellaexposedtoglyphosate
*AHCVanBruggenetal.ScienceoftheTotalEnvironment2018;616-617:255–268.
GlyphosateUsageandPapersonAn2bio2cResistance*
*AHCVanBruggenetal.ScienceoftheTotalEnvironment2018;616-617:255–268.
ABTBRMouseModelofAu2sm*Thesemicehadallthemousefeaturesofau2smTheywerefed“standardrodentchow”–glyphosatecontaminated?Somefeaturesinthegut:• Reducedlevelsofbileacids(duetoimpairedCYP7A1ac2vityinthe
liver)• Furtherreducedlevelsofsecondarybileacids(impairedmetabolismby
gutmicrobes)• ReducedlevelsofLactobacillusandBifidobacteria(microbesthat
metabolizebileacids)– Thesemicrobesarepreferen2allykilledbyglyphosate
• Serotonindeficiency(dueinparttotryptophanconversiontokynureninetofightinfec2on)– Serotoninisderivedfromtryptophan,aproductoftheshikimatepathway
whichglyphosatedisrupts
*AVGlubevaetal.EBioMedicine.2017Oct;24:166-178.
GlyphosateDisruptsCytochromeP450(CYP)Enzymes*
• GlyphosatehasbeenshowntoseverelysuppressCYPenzymesinratliver
• CYPenzymeshaveauniqueFXXGXRXCXGmo2fwithtwoandsome2mesthreecri2calglycineresidues**
*ASamselandSSeneff.Entropy2013;15:1416-1463.**KSyedandSSMashele.PLOSONE2014;9(4):|e95616.
GLYCINES
GlyphosateDisruptsCytochromeP450(CYP)Enzymes*
• GlyphosatehasbeenshowntoseverelysuppressCYPenzymesinratliver
• CYPenzymeshaveauniqueFXXGXRXCXGmo2fwithtwoandsome2mesthreecri2calglycineresidues**
*ASamselandSSeneff.Entropy2013;15:1416-1463.**KSyedandSSMashele.PLOSONE2014;9(4):|e95616.
GLYCINES
CYPenzymesareneededtoproducebileacidsfordiges2ngfats,toac2vatevitaminDandto
detoxifymanyenvironmentaltoxicants
BTBRmicehavelowacetate,andglyphosatedisruptsacetatesynthesisingut*
Childrenwithau2smhadonly3.5mg/mlacetateinstoolsamplescomparedto5.1incontrols.**
BTBRmice
*LNNielsenetal.EnvironmentalPollu2on2018;233:364e376.**Adamsetal.BMCGastroenterology2011;11:22.
BTBRmicehavelowacetate,andglyphosatedisruptsacetatesynthesisingut*
Childrenwithau2smhadonly3.5mg/mlacetateinstoolsamplescomparedto5.1incontrols.**
BTBRmice
*LNNielsenetal.EnvironmentalPollu2on2018;233:364e376.**Adamsetal.BMCGastroenterology2011;11:22.
ElevatedpHlinkedtoglyphosateexposureresultsinsmallintes2nalbacterialovergrowth(SIBO)
*Figure2.IArgou-CardozoandFZeidán-Chuliá.Med.Sci.2018;6:29.
Glyphosate,Pathogens,Au2sm*
*Figure2.IArgou-CardozoandFZeidán-Chuliá.Med.Sci.2018;6:29.
Glyphosate,Pathogens,Au2sm*
AnewpaperInPressfromtheRamazziniIns2tuteinItalyshowedthatthegutmicrobiomewasdisrupted
inratsexposedto"safe"levelsofglyphosate.
EvidenceLinkingAu2smtoClostridiaOvergrowth*
• 14au2s2cchildrenwithgutdisordercomparedto21controls
• SignificantincreaseinClostridiaspeciesinthegutinau2s2cchildren
• Associatedwithreducedtryptophanlevelsandincreasedexpressionofinflammatorymarkers– Tryptophanisaproductoftheshikimatepathway,whichglyphosateblocks
– Macrophagesininflamed2ssuetakeuptryptophan,reducingbioavailabilitytothebrain
• Proposedroleforan2bio2cs– Glyphosateisapatentedan2microbialagent(2010)
*RALunaetal.,CellularandMolecularGastroenterologyandHepatology2017;3(2):218-230
• Triplets:twoboys,onegirl.Bothboyshaveau2smandgirlhasseizuredisorder
• Veryhighlevelsofglyphosateinurineinallthree• Clostridiaovergrowthduetoglyphosatedisrup2onofgutmicrobes– ClostridiaproducetoxinsHPHPAandp-cresol,whichblocktheconversionofdopaminetonorepinephrine.
– Damagetoneuronsinthebrainthroughoxida2vestress *W.Shaw.Integra2veMedicine2017;16(1);50-57.
*
MyosinintheGut• Myosinisamotorproteinfoundinhighlevelsinskeletalmuscles
• Myosinisalsoessen2alforgutmo2lity(peristalsis)andforthereleaseofbileacidsintotheupperintes2ne
• Myosincontainsahighlyconservedglycineatposi2on699*– Ifthisischangedtoalanine,theprotein’scontrac2leabilityisreducedtolessthan1%.
• Glyphosatehasbeenshowninfishstudiestosuppressmyosinexpression**
*FKinoseetal.TheJournalofCellBiology1996;134(4):895-909.**AnaPaulaRezendedosSantosetal.,Chemosphere2017;168:933e943.
MyosinintheGut• Myosinisamotorproteinfoundinhighlevelsinskeletalmuscles
• Myosinisalsoessen2alforgutmo2lity(peristalsis)andforthereleaseofbileacidsintotheupperintes2ne
• Myosincontainsahighlyconservedglycineatposi2on699*– Ifthisischangedtoalanine,theproteins’contrac2leabilityisreducedtolessthan1%.
• Glyphosatehasbeenshowninfishstudiestosuppressmyosinexpression**
*ACDukowiczetal.GastroenterolHepatol(NY)2007;3(2):112-122.
SIBO(SmallIntes2nalBacterialOvergrowth)isassociatedwithimpairedperistalsis*
Summary• Glyphosatecontamina2oninfoodproteinsmakesthemhardtobreakdown– Thisleadstoautoimmunedisease
• Diges2veenzymesarecontaminatedwithglyphosate– UndigestedproteinsinduceCeliacdiseaseandleakygut
• Glyphosateisakeyfactorintheemergenceofan2bio2cresistantpathogens
• TheBTBRmousemodelofau2smisconsistentwithglyphosatedamageinthegut
• GlyphosatepromotesClostridiaovergrowth– Thisinducesinflammatoryboweldisease,anepidemictoday– Au2smhasbeenlinkedtoClostridiaovergrowth– Clostridiareleasetoxinsthatinduceaninflammatoryresponse