MoniqueMendez

August12,2017
LairsonLab
TheScrippsResearchInstitute

InternshipAbstracts

Forthepastseveralweeks,IhadtheopportunitytointernattheLairsonlabwitha
higherfocusonbiochemistry.Throughoutthemonth,myresearchprojectprimarilyfocused
onunderstandingtheregulationofthecGAS-STINGpathway,howita ectedthosewith
lupusandhowourlabismostimportantlyattackingthisdisablingdisease.
TheLairsonlabhastheirfocusofresearchinawidearrayofbiomedical elds;from
biochemistrytoorganicchemistry,drugdiscoverytoregenerativemedicine,theirresearch
touchesbasewithnearlyalldisciplines.Whilethelabdoesfocusinotherareasofresearch,
theyspecializeinhigh-throughputscreeningforsmallmoleculeactivatorsandinhibitors.
Currently,theyarescreeningforsmallmoleculeinhibitorstoblockcGAS-STINGsignaling
inpatientswithLupus.
TheLairsonlabfocusesonLupusbecausewhenthecGAS-STINGpathwayis
activated,itisessentiallyactivatingtheinterferonin ammatoryresponse.Whenwereally
getdowntoit,STINGstimulatesthetranscriptionfactorIRF3,causingtheactivationofthe
interferonpathway,leadingtohigherlevelsofIFNandIFNexpression.Whenthe
in ammatoryresponseisactivated,muchofanindividualscardiovascular,nervous,
hematopoieticandmusculoskeletalsystemisa ectedwith,ofcourse,in ammation..

 (Figure1) (Figure2)

Forour rstsetofresults,wewereabletoidentifyoptimizedenzymaticcGAS
reactions.WewereabletooptimizereactionconditionsinordertomeasurechangesinATP
concentrationandalsotodetecttheappearanceof23-cGAMPthroughuseofHPLC
machine.

(Figure3) (Figure4)

Forthesecondportionofmyresults,wewereabletooptimizeanassayinorder
changesinATPconcentrationasanoutputforcGASenzymaticactivity.Inour rstchart,
ourdatashowsthatwewereabletooptimizecGASenzymaticactivitythroughanATP
standardcurvewithKinaseGloMax.WemeasuredtheproductionofcGAMPinordertosee
ifitwouldbindtoSTING.UsingadilutionseriesofATP,wewereabletogeneratealinear
standardcurveasdemonstratedinF
igure3andgatheranR2 of.99.InF
igure4,isour
controls,anditillustrateshowmuchourATPlevelsvaried.

Throughoutmyinternship,Iwasabletolearnnumerousskills.ComparedtohowI
feltatthestartofmyinternship,Ifeelmorecon dentinmyscienceandlabknowledgenow.
Ifnotformylabmate,fellowpeers,mentor(s)andP.I.,IdonotknowhowIcouldhave
survivedthroughthis.IbelievethatwhileImayhavegainedmanyskills,Istilllackvery
muchinnumerousothers.
WhileIbelievethatIde nitelybito morethanIwasabletochewthistimearound,
IfeelthatIovercameobstaclesthroughsimpleactions.Iwasmostfortunateenoughtohave
beenintroducedtonovelconceptsandtechniques.AndwhileIfeelthatImayhave
struggled,mymentorshelpedkeepmea oat,answeringallofmyquestionsandbeingmy
biggestcheerleader.
Lookingbackonmyoverallexperience,IstillfeelthatIhaveastrongconnectionto
thescience eld.Iunderstandthatwecannotextendourinternshiportimewithourlab,
however,itfeelsasifthereisun nishedworkstilllefttobedone.IunderstandthatIwas
de cientinmanyofthenecessaryskillsrequired,butIfeelasthoughthereisstillastrong
possibilityformetoworkinthescience eldhavingnowcompletedtheLifeSciences
SummerInstituteinternship.