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