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Inrecentyears,muchattentionhasbeenfocusedontheregulatoryactivityofaclassofRNAsknownasmicroRNAs(miRNAs).
AlthoughthereisnowwidespreadinterestinmiRNAs,thiswasn"talwaysthecase.Indeed,theseshort(~22bp)non-codingRNAsequenceswereonlydiscoveredin1993byVictorAmbrosandcolleagues.
ThefirstmiRNAprecursordiscovered,lin-4,wasfoundtoregulateworm(Caenorhabditiselegans)development(Leeetal.,1993).Consideredaninterestingbutworm-specificanomaly,theimportanceoflin-4’sdiscoverywasnotfullyappreciatedatthetime.
IttookseveralyearsbeforeresearchersnoticedthatmiRNAs,andthegenesthatprocessthem,werehighlyconservedacrossspeciesandworthyofmoreattention.OncethewidespreadnatureofmiRNAswasrealized,concertedeffortstodiscovermoremiRNAsandelucidatetheirfunctionbegan.
miRBase–adedicatedmiRNAannotationdatabase–counts28,645entriesinmorethan100speciestodate.Furthermore,recentresearchsuggeststhattherearemanymoremiRNAswaitingtobediscovered(Friedländeretal.,2014).
Figure1.ThenumberofmiRNAsinthemiRBasedatabasefrom2003to2014.
Decadeslater,weknowthathumansexpress1,900miRNAs(www.miRBase.org,2015),thattheyregulatethetranslationofmRNAandcansilencegenesattheDNAlevel.Unsurprisingly,suchfundamentalinfluenceoncellactivityhasimplicatedmiRNAsinanumberofdiseasesincludingcancer,neurologicaldisordersandcardiacfunction(Sayedetal.,2011).
miRNAsasbioMarkers
AnomalousexpressionofmiRNAshasbeenshowntobeahallmarkofspecificcancersand,asthesemoleculeshavebeenshowntocirculateintheblood,miRNAslooksettobecomepowerfulprognosticanddiagnosticbiomarkersintheyearsahead(Schwarzenbachetal.,2014).
ArecentCancerCellpaperhasillustratedwhatmaybeasignificantbroadeningofthemiRNAfield.Amulticenterresearcheffortdiscoveredthatbreastcancercellsshedmoreexosomesthanhealthycells,andthattheseexosomesareloadedwithmiRNAs,primedandreadytoregulatetheirtargetgenes(Meloetal.,2014).
Thismightbejustaninterestingquirk,afterallexosomesarejustsmallmembranebubblesregularlyshedbycells;however,thepaper’sauthorsdemonstratethatexosomespurifiedfromtheseraofcancerpatientscaninstigatetumorformationinhealthyepithelialcells.
Whileonepaperdoesn’tmakeafield,asimilarexosome-mediatedsheddingofmiRNAsandassociatedprocessingmachineryhasbeenobservedinHIVinfectedcells(Narayananetal.,2013)andincellsundergoinganoxicstress(Shenetal.,2013).ThisisjustoneofanumberofreportshighlightingopportunitiesforthedevelopmentofmiRNA-basedbiomarkersandtherapiesforcancerandanumberofotherhumandiseases(Sundarboseetal.,2013;VanRoosbroecketal.,2013);thisiscertainlyafieldtokeepacloseeyeon.
AnewmultiplextechnologytoprofilemiRNAs.
GiventheemergingroleofmiRNAsasbiomarkers,miRNAprofilinghastremendouspotentialforthediagnosisandprognosisofabroadrangeofdiseases.Usinginnovativetechnology,wehavedevelopedanassaythatenablesresearcherstodetectupto68miRNAsofchoiceeitherfrompurifiedRNAordirectlyfromcrudesamplesincludingserum,plasmaandexosomes.
WehavedemonstratedhowtheMultiplexCirculatingmiRNAAssaycanbeusedformiRNAprofilingwithPCRsensitivitydirectlyfromaslittleas10µLofserumorplasmawithnoneedforRNApurification,orfromlessthan100pgofpurifiedRNA.Theassayallowstheparallelprocessingof96samples,thusenablingthehigh-throughputanalysisrequiredforbiomarkervalidationstudies.
