ISGAIN-OF-FUNCTIONRESEARCHUNLOCKINGTHE SECRETSOFHIDDENDANGEROUSVIRUSES?
Dr RaghavendraRaoM.V*,Dr MahendraKumarVerma,Dr M.M.Karindas ,Dr IlieVasiliev, 5Dr.DilipMathai
1Scientist-Emeritus,ApolloInstituteofMedicalSciencesandResearch,JubileeHills,Hyderabad,TS,India(*CorrespondingAuthor)
2AssistantProfessor,AmericanUniversitySchoolofMedicine,Aruba,NetherlandsAntilles
3Professor,DepartmentofOncology,WorldAcademyofMedicalSciences,Netherlands.
4Professor,DepartmentofInternalMedicine,WorldAcademyofMedicalSciences,Netherlands
5DistinguishedProfessorandAdvisor,TheApolloUniversity,Chittoor,AndhraPradesh,India
ABSTRACT
Gain-of-functionexperimentsmayhelpmedicalresearcherstotestscientifictheories,developnewtechnologiesandfindtreatmentsforinfectiousdiseases.Whenthe original SARS-CoV outbreak occurred in 2003, scientists developed a method to study the virus in the laboratory, by growing the virus in mice. This endeavour showed a model for researching the virus and testing potential vaccines and treatments. Inadequate and insufficient research on viruses will authorize unaware of futurepandemics.Buttoolittleconsciousnessofsecuritythreatswillmultiplythehazardsthatanexperimentvirusmayescapealabthroughanaccidentandcausean outbreakofitsown.Avirusgeneratedinthelabbygeneticallymodifieditsgeneproducts,alteringanorganism,andifaccidentallyreleased,couldresultinmillionsof deaths.
Gain-of-Functionresearchisfollowedinscientificstudiestocautiouslyunderstandthepossibilityoftheemergenceofinfectiousvirusesinnature.Gain-of-Function experimentsincludeengineeringinthelabgeneticalterationsthatsimulateanincreaseinvirulence.Theorganismmodifiedthiswayistestedonlaboratoryanimals likeferrets,mice,andguineapigs.Thesetestsprovideinsightthatisutilizedtodevelopvaccines,testoutnewantivirals,understandthehost-immuneresponse,and comprehendthedisease-causingabilityofthevirus.Inthisway,wecanremainastepaheadofviralevolutionbydevelopingsuitablemedicalcountermeasures.Gainof-functionresearchistheserialpassagingofmicroorganismstoincreasetransmissibility,virulence,immunogenicity,andhosttropism.
KEYWORDS:Gain-of-Function,mice,andguineapigs,immunogenicity,Dual-useresearchofconcern(DURC)
Introduction
GOFRisasubsetof“dual-useresearch”—i.e.,researchthatcanbeusedforboth beneficialandmalevolentpurposes(1,2)
Dual-useresearchofconcern(DURC)isoneofthemosthotlydebatedscience policy issues during the 21st century, with controversy surrounding a series of published experiments with potential implications for biological weaponsmaking.(3)
Gain-of-function research (GoF research or GoFR) is medical research that geneticallyaltersanorganisminawaythatmayenhancethebiologicalfunctions ofgeneproducts.(4)
Introducing a mutation that would allow influenza B to infect rabbits in a controlledlaboratorysituationwouldbeconsideredagain-of-functionexperiment, asthevirusdidnotpreviouslyhavethatfunction(5)
Thattypeofexperimentcouldthenhelprevealwhichpartsofthevirus'sgenome correspondtothespeciesthatitcaninfect,enablingthecreationofantiviralmedicineswhichblockthisfunction(6)
Invirology,gain-of-functionresearchisusuallyemployedwiththeintentionof better understanding current and future pandemics. In vaccine development, gain-of-function research is conducted in the hope of gaining a head start on a virusandbeingabletodevelopavaccineortherapeuticbeforeitemerges(7)
Theterm"gainoffunction"istorefertoapathogentoreplicatemorequicklyor causemoreharminhumansorotherclosely-relatedmammals8)
Thetermfirstgainedawidepublicaudiencein2012,aftertwogroupsrevealed that they had tweaked an avian influenza virus, using genetic engineering and directedevolution,untilitcouldbetransmittedbetweenferrets(9)
Theexperimentsdidmakethevirusamenabletoresearch,however,andtheteam sharedbothitandtheengineeredmicewithothers.Itledtoplentyofnewfindings. (10)
ResponseswithSARS-CoV-2,suggestingthatifinterferonisprovidedasatreatment,itshouldbeearlyinthecourseofthedisease(11)
Researchersalsousedthemouse-adaptedMERS-CoVtotestnewvaccinesand treatments. The Iowa team's collaborators tested a vaccine that is a hybrid of
parainfluenzaviruswiththeMERS-CoVversionofthespike.Thevaccinewasn't very effective when injected, but it did protect DPP4-expressing mice from MERS-CoVquitewellwhenprovidedthroughthenose(12)
AlthoughMERSoutbreakshaven'tledtosustainedtransmission,thisinformation has proved valuable in the COVID-19 pandemic: a vaccine with the same design,butagainsttheSARS-CoV-2spike,worksinmiceandferretsandisnow undergoingearlyclinicaltrials(13)
Parainfluenzavirustype5(PIV5)isanegative-strandedRNAvirusinthefamily Paramyxoviridaethathasbeenevaluatedasavaccinevectorforinfluenza,respiratorysyncytialvirus(RSV),rabies,andavarietyofotherpathogens(14,15).
Inanimalmodels,PIV5issafeandisnotassociatedwithanydisease,withthe exceptionofkennelcoughindogs(16).
IntranasallyadministeredkennelcoughvaccinescontaininglivePIV5havebeen used for more than four decades with an excellent safety record. Dogs immunizedwithkennelcoughvaccinescanshedPIV5forupto5days,andithasbeen safetohumansinclosecontactwithimmunizedanimals(17,18).
InJanuary2020,theNationalScienceAdvisoryBoardforBiosecurityconvened anexpertpaneltorevisittherulesforgain-of-functionresearchandprovidemore clarityinhowsuchexperimentsareapproved,andwhentheyshouldbedisclosed tothepublic(19,20)
TheoriginsofGOFR
Theterm"gainoffunction" refersto"researchwhichcouldenableapandemicpotential pathogen to replicate more quickly or cause more harm in humans or otherclosely-relatedmammals."
Whatdoesgainoffunctionmean?
Anyorganismcanacquireanewabilityorproperty,or“gain”a“function.”
Gainoffunctioncanalsobeusefulforenvironmentalreasons,suchasmodifying E.colisothatitcanconvertplasticwasteintoavaluablecommodity
InthecurrentdebatearoundSARS-CoV-2,thevirusthatcausesCOVID-19,gain offunctionhasamuchnarrowermeaningrelatedtoavirusbecomingeasierto move between humans, or becoming more lethal in humans. It is important to remember,though,thattheterm“gainoffunction”byitselfcoversmuchmore
Copyright©2023,IERJ.Thisopen-accessarticleispublishedunderthetermsoftheCreativeCommonsAttribution-NonCommercial4.0InternationalLicensewhichpermitsShare(copyandredistributethematerialinany mediumorformat)andAdapt(remix,transform,andbuilduponthematerial)undertheAttribution-NonCommercialterms.
thanthistypeofresearch.
Anyorganismcanacquireanewabilityorproperty,or“gain”a“function.”
Some lab examples include creating more salt- and drought-resistant plants or modifyingdiseasevectorstoproducemosquitoesthatareresistanttotransmittingdenguefever Gainoffunctioncanalsobeusefulforenvironmentalreasons, suchasmodifyingE.colisothatitcanconvertplasticwasteintoavaluablecommodity
InthecurrentdebatearoundSARS-CoV-2,thevirusthatcausesCOVID-19,gain offunctionhasamuchnarrowermeaningrelatedtoavirusbecomingeasierto move between humans, or becoming more lethal in humans. It is important to remember,though,thattheterm“gainoffunction”byitselfcoversmuchmore thanthistypeofresearch.
Whywouldresearchersdogain-of-functionworkonpotentiallydangerous pathogens?
Gain-of-function experiments may help researchers test scientific theories, developnewtechnologiesandfindtreatmentsforinfectiousdiseases.Forexample,in2003,whentheoriginalSARS-CoVoutbreakoccurred,researchersdevelopedamethodtostudythevirusinthelaboratory
Oneoftheexperimentswastogrowthevirusinmicesotheycouldstudyit.This workledtoamodelforresearchingthevirusandtestingpotentialvaccinesand treatments.
Gain-of-function research that focuses on potential pandemic pathogens has beensupportedonthepremisethatitwillhelpresearchersbetterunderstandthe evolvingpathogeniclandscape,bebetterpreparedforapandemicresponseand developtreatmentsandcountermeasures.
Whataresomeexamplesofgain-of-functionresearch,andhowriskyisit?
PotentialBenefits
Somepotentialoutcomesofgain-of-functionresearchmayincludethecreation oforganismsthataremoretransmissibleormorevirulentthantheoriginalorganismorthosethatevadecurrentdetectionmethodsandavailabletreatments.
Otherexamplesincludeengineeringorganismsthatcanevadecurrentdetection methodsandavailabletreatments,orgrowinanotherpartofanorganism,suchas theabilitytocrosstheblood-brainbarrier
RiskandChallenges
Gainoffunctionexperimentsmakevirusesmoreinfectioustohumansinthelaboratory settings. This research look like gamble that civilization can't afford to risk.Thereisnosuchthingaszeroriskinconductingexperiments.Sothequestioniswhethercertaingain-of-functionresearchcanbeperformedatanacceptablelevelofsafetyandsecuritybyutilizingrisk-mitigationmeasures.
Arobust biosafety and biosecurity system, along with appropriate institutional response,helpstoensurethattheseincidentsareinconsequential.Thechallenge is to make sure that any research conducted – gain-of-function or otherwise –doesn'tposeunreasonableriskstoresearchers,thepublicandtheenvironment.
Functionresearchandglobalpandemic
Invirology,gain-of-functionresearchisusuallyemployedwiththeintentionof betterunderstandingcurrentandfuturepandemics.
In vaccine development, gain-of-function research is conducted in the hope of gainingaheadstartonavirusandbeingabletodevelopavaccineortherapeutic beforeitemerges.
Theterm"gainoffunction"issometimesappliedmorenarrowlytoreferto"research which could enable a pandemic-potential pathogen to replicate more quicklyorcausemoreharminhumansorotherclosely-relatedmammals.“
Someformsofgain-of-functionresearch(specificallyworkwhichinvolvescertain select agent pathogens) carry inherent biosafety and biosecurity risks, and arethusalsoreferredtoasdualuseresearchofconcern(DURC).
DuringtheCOVID-19pandemicanumberofhighlyspeculativetheoriesspread abouttheoriginoftheSARS-CoV-2virusandlinkstogain-of-functionresearch.
In January 2021, University of Saskatchewan virologist Angela Rasmussen wrote that one version of the information invoked previous gain-of-function workoncoronavirusestopromulgatetheideathattheviruswasoflaboratoryorigin
Gain-of-function (GOF) research involves experimentation that aims or is expectedto(and/or,perhaps,actuallydoes)increasethetransmissibilityand/or virulence of pathogens. Such research, when conducted by responsible scientists, usually aims to improve understanding of disease causing agents, their interactionwithhumanhosts,and/ortheirpotentialtocausepandemics.(21).
Thelast–andonly–foreignscientistintheWuhanlabspeaksout
Anexpertinbat-borneviruses,DanielleAndersonistheonlyforeignscientistto
haveundertakenresearchattheWuhanInstituteofVirology'sBSL-4lab,thefirst inmainlandChinaequippedtohandletheplanet'sdeadliestpathogens.Danielle Anderson was working in what has become the world's most notorious laboratory just weeks before the first known cases of Covid-19 emerged in central China.Yet,theAustralianvirologiststillwonderswhatshemissed.(22)
Covid-19 was a 'man-made' virus, claims scientist who worked at the Wuhanlab
Withmerelyanybiosafetyprovisions,China'sgain-of-functionexperiments— whichareconductedtogeneticallyalterorganismsinordertoenhancetheirbiological functions — led to a leak at the Wuhan lab.AUS-based scientist, who workedataresearchlabbasedinChina'sWuhanrevealedMondaythatCovid-19 wasa“man-madevirus”thathadleakedfromthefacility,accordingtoareportin TheNewYorkPost.(23)
TheLab-LeakTheory:InsidetheFighttoUncoverCOVID-19'sOrigins
As cities worldwide were shutting down to halt the spread of COVID-19, Demaneuf, 52, began reading up on the origins of SARS-CoV-2, the virus that causesthedisease.TheHuananwholesalemarket,inthecityofWuhan,isacomplexofmarketssellingseafood,meat,fruit,andvegetables.Ahandfulofvendors soldlivewildanimals—apossiblesourceofthevirus.(24)
NeoCovcoronavirusfoundinbatsmayposethreattohumansinfuture,scientistscaution
NeoCoviscloselyrelatedtotheMiddleEastrespiratorysyndrome,aviraldiseasefirstidentifiedinSaudiArabiain2012.Asinglemolecularchangeinthelab enabledacoronaviruscalledNeocovto“efficientlyinfect”humancellsusingthe samepathwaythattheSARS-CoV-2usestoinfecthumancells,researchersfrom WuhanUniversity,Wuhan,Chinasaidinareportthatisyettobepeer-reviewed.
Neocovhassofaronlybeenseeninbatsandnoinstanceshavebeenreportedin people, but being closely related to the Middle Eastern Respiratory Syndrome (MERS) coronaviruses --traditionally more lethal but less transmissible than Sarscov2--thestudyhasraisedconcernthatthistoomaylethallyproliferatein people Expertshoweversaythatsuchfearsareunwarranted.(25)
ApplicationofGofResearch
Currentmedicalcountermeasuresareofteninsufficientlargely Virusesdevelop resistance mechanisms that lead to “escape mutants,” that is, drug-resistant strainsandthecurrentmedicalcountermeasuresareofteninsufficientThereis,a needtodevelopnewantiviraldrugslike,immunotherapy,basedonneutralizing monoclonal antibodies The GoF studies, boostup viral yield and immunogenicity,arerequiredforvaccinedevelopment.Molecularmethodshelp withthecharacterizationofantigenicvariants,elucidatethebiologicalbasisfor adverse outcomes associated with vaccine candidates, and determine the basis forattenuationandstabilityofvaccinecandidates
AlternativeResearchMethodswithPotentiallyLessRisk
Ÿ Molecular dynamical modelling of influenza proteins and interactions withinhibitorsandreceptor
Ÿ Invitrostudiesofspecificpropertiesrequiredforhumanadaptation,using singleproteins
Ÿ Invitrostudiesofgeneticinteractionsbetweenlociinoneorseveralviral proteinsusingreplication-incompetentviruses–epistaticinteractions
Ÿ Sequencedatabasecomparisonsofgeneticpropertiesofhumanandavian adaptedviruses
Ÿ Comparisons of human seasonal isolates and zoonotic isolates from infectedhumansandavianisolates
'Notconvinced'Covid-19developednaturally,saysDrFauci --America'stopexpertoninfectiousdiseases,DrAnthonyFaucisaidheis"not convinced"thenovelcoronavirusdevelopednaturallyandhascalledforanopen investigationintotheoriginsofCovid-19virus.
When Dr Fauci was asked at an event whether he was still confident that the coronavirusdevelopednaturallyhesaid:"Iamnotconvincedaboutthat,Ithink weshouldcontinuetoinvestigatewhatwentoninChinauntilwecontinuetofind outtothebestofourabilitywhathappened."
"Certainly,thepeoplewhoinvestigateditsayitlikelywastheemergencefroman animalreservoirthattheninfectedindividuals,butitcouldhavebeensomething else,andweneedtofindthatout.So,youknow,that'sthereasonwhyIsaidI'm perfectlyinfavourofanyinvestigationthatlooksintotheoriginofthevirus,"Dr FauciwasquotedbyFoxNews.(26)
COVIDcrackedoutfromWuhanInstituteofVirology(WIV),laboratory
In what appears to be an astonishing revelation, a US-based scientist, who workedatacontroversialresearchlabinChina'sWuhan,hassaidthatCOVID19wasa"man-madevirus"thatleakedfromthefacility
COVID was leaked from Wuhan Institute of Virology (WIV), a state-run and fundedresearchfacility,twoyearsago,theNewYorkPostreported,quotingUSbasedresearcherAndrewHuff'sstatementintheBritishnewspaperTheSun.
Inhislatestbook,"TheTruthAboutWuhan,"epidemiologistHuffclaimsthatthe pandemic was caused by the US government's funding of coronaviruses in China.ExcerptsofHuff'sbookhavebeenoutintheUK-basedtabloidTheSun. AccordingtotheNewYorkPostreport,Huffistheformervicepresidentofthe EcoHealthAlliance, a non-profit organisation based in New York that studies infectiousdiseases(27)
OriginsofSARS-CoV-2:Whythelab-leakideaisbeingconsideredagain. Since the beginning of the COVID-19 pandemic, there have been various hypothesestoexplaintheoriginofSARS-CoV-2.
Sofar,noneofthesehypotheseshaveshowntheoriginsofthevirus.Oneofthe firstscenariosputforward—thattheWuhanseafoodmarketallowedforarapid dispersion of the virus — seems less certain now After a year of intensive research, the virus has still not been identified in any animals Animal-human transmission, however, remains one of the strongest hypotheses explaining the appearanceofthenewcoronavirus:thecoronaviruswastransmittedfrombatsto humans through an intermediate host. It would not be the first time this happened.InthecaseofMERS-CoV(MiddleEastRespiratorySyndrome),camels likelyservedastheintermediatehost.ForSARS-CoV-2,pangolins,scalyanteaterssoldillegallyintheWuhanmarket,maybethehost,althoughthishypothesis requiresmoreconvincingevidence.(28)
Conclusion
Invirology,gain-of-functionresearchisusuallyemployedwiththeintentionof betterunderstandingcurrentandfuturepandemics.Gain-of-Functionresearchis followed in scientific studies to cautiously understand the possibility of the emergence of infectious viruses in nature Gain-of-Function experiments include engineering in the lab genetic alterations that simulate an increase in virulence.Theorganismmodifiedthiswayistestedonlaboratoryanimalslike ferrets, mice, and guinea pigs. These tests provide insight that is utilized to developvaccines,testoutnewantivirals,understandthehost-immuneresponse, and comprehend the disease-causing ability of the virus. In this way, we can remain a step ahead of viral evolution by developing suitable medical countermeasures Gain-of-function research is the serial passaging of microorganisms to increase transmissibility, virulence, immunogenicity, and hosttropism.Invaccinedevelopment,gain-of-functionresearchisconductedin thehopeofgainingaheadstartonavirusandbeingabletodevelopavaccineor therapeuticbeforeitemerges.
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