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Presentationtips Talks • 10minutes- CoSBBI…... iBRIC 10-12minutes • 1slide/minute • NEVERGOOVER. • Knowyouraudience. • Samedata/projectcanbepresentedinverydifferentways dependingonyouraudience.Tailoryourtalktowhatyour audienceknowsorwantstoknow.Beengaging. • Tellastory • Breatheandrelax SNHG7isanIGF1regulatedlong non-codingRNAnecessaryfor proliferation DavidBoone,PhD AdrianLee,PhD Donotallowyourbackgroundor texttobedistracting • Contrastisimportant dfaasfdafd Asdfasdfas asdfasdfasd Noryouranimations IGF1signaling Forallofyou,themajorityofyourtalk maybebackground/describinga problem,yourhypothesis/goal,and howyouattemptedtotacklethe problem IGF1 IGF1R PI3K mTOR Growth Survival Useimagesand minimaltext…evenfor backgroundinfo lncRNAs? miRNAs? piRNAs? AKT MAPK Proliferation Fold >1.5; P<0.01 Creighton etal.2008.JCO Don’tforgetcitations! Longnon-codingRNAs(LncRNAs) • 85%ofthegenomeistranscribed • Only2%ofgenomeiscodingregions • ~20000proteincodinggenes • ~16000lncRNAs • Diversebiologicalfunctions. • Chromatinremodeling • Transcriptionalco-activatororco-repressor • Proteininhibition • Dysregulated inhumancancer. • XIST– downregulated inbreastandovarian. • HOTAIR– upregulated inbreastcancerandpromotesmetastisis. Morris &Mattick.Theriseofregulatory RNA.NRG(2014) Goals • Furtherunderstandthemolecularmechanismsof IGF1’sfunctionsbyexaminingIGF1-regulatedlncRNAs. • LeveragetheIGF1systemtoidentifyandcharacterize functionallycriticallncRNAs inbreastcancer. 1) DoesIGF1regulatetheexpressionoflncRNAs? 2) AreIGF1-regulatedlncRNAs importantforIGF1mediatedbiology? HaveaGoalsorHypothesisslide!Everyoneshouldalwaysknowwhatyourdoingandwhy Outlineslidesaregood,butdonotjusthaveintro,methods,results,conclusions. Havedescriptiveheadersforeach.Tellthemwhatyouaregoingtotellthem,tell them,thentellthemwhatyoutoldthem. Outline • Researchinterests • • • • • IGFandlncRNA background IGFregulatesSNHG7andotherlncRNAs SNHG7isnecessaryforproliferation IGF/SNHG7feedback/fine-tuningloop Future • UPCIAcademy/CoSBBI/iBRIC • Description • MentoringandTeaching • Details,Expectations,andOpportunities • PreliminaryOutcomes • Longitudinal • Surveysandawardsin2015 IGF1signalingalterstheexpressionofa subsetoflncRNAs Flowcharts>text IGF1signalingalterstheexpressionofa Titlesshouldbeyour subsetoflncRNAs takeawaymessage fromtheslide RNAseq RelativeFC RT-qPCR validationofRNAseq 4 3.5 3 2.5 2 1.5 1 0.5 0 ctl3hr IGF3hr ctl8hr IGF8hr Blue- IGF1-regulated lncRNAs at3hrsFDR<0.05 Red– IGF1-regulated lncRNAs genesat8hrsFDR <0.05 RNAseq FC8hrs-1.742.9-3.5-2 *lnc annotationsfromBodyMap ANDUCSCknowngene Interactwithyourfiguresand slides!Explainx/ygraphs.Explain methodifyoualreadydidn’t,etc SmallNucleolar HostGene7 (SNHG7) • A classoflncRNAs thatcontainsnoRNAs intheir introns.Ex.Gas5 Makesuretextisn’ttoosmalland donotputanythinginyourtalkor onposterthatyouaren’tprepared totalkabout Predictedsecondarystructure Don’tbeafraidtothrowinmoreintrohere ratherthanatthebeginning.Tellastory! • ThesesnoRNAs areNOTregulatedbyIGF1 RNAFold 1.4 1.2 1 0.8 0.6 0.4 0.2 0 SNHG7expression SNHG7expression ctl IGF1 RelativeFC RelativeFC IGF1decreasesSNHG7stabilityviatheMAPKpathway 1.2 1 0.8 0.6 0.4 0.2 0 ctl IGF 4hrs 8hrs 12hrs 24hrs 48hrs TimeafterIGF1stimulation IGF1R PI3K mTOR AKT MAPK Itisnicetoreconnecttointro. RelativeFC IGF1 1.4 1.2 1 0.8 0.6 0.4 0.2 0 SNHG7expression ctl IGF 3hrs 8hrs DMSO 3hrs 8hrs MEKinhibitor SNHG7regulatesproliferationinadose-dependentmanner 12 0.8 siCtl 0.6 siSNHG7 0.4 0.2 CellCycle Proliferation 10 8 siCtl 6 siSNHG7 4 2 0 0 1 3 5 7 9 Daysaftertransfection SNHG7 Keepcolors andaxes consistent whenever possible RelativeFC 1.2 12 SNHG7Expression 1.0 10 0.8 8 0.6 0.4 RFU(10^5) RelativeFC 1 Cell Number(10^4) 1.2 6 4 0.2 2 0.0 0 Percentageofcells SNHG7Expression 100% 80% 60% 40% 20% 0% Proliferation(day5) G2/M S G0-G1 SNHG7overexpression enhancesproliferationofMCF7 cellsinthepresenceofserum Proliferation 3.5 12 3 10 SNHG7_p2 8 SNHG7_p1 6 vector_p1 2.5 2 1.5 1 0.5 0 RelativeProliferation RelativeExpression SNHG7overexpression vector_p2 4 2 0 1 3 5 Days 7 SNHG7andIGF1signalingsimilarlyregulate theexpressionofmanygenes. Egr3;Lif IGF1 SNHG7 Irs2;Socs2 RelativeFCtomatchedCtls RNAseq Expression(RT-qPCR) 6 Ctl 5 IGF 4 SNHG7 3 IGF Ctl siSNHG7 1.3FC siSNHG7 2 -1.3FC 1 0 Lif Egr3 Irs2 Socs2 Useanimationtoyouradvantage SNHG7regulatesthetranscriptlevelsof membersoftheIGF1signalingpathway. IGFSignalingTranscripts RNAseq siCtl siCtl siSNHG7 siSNHG7 IGFBP4 IGF1R 2239 IPA 1.5FC 1659 -2FC 2FC AnnotatedgenesFDR<0.05 IGFBP3 IGFBP5 -1.5FC IGFBP2 IRS2 Model SteadyState Inducedgenes IGF1signaling SNHG7 IGF1signaling SNHG7 Repressedgenes Inducedgenes Steadystate Proliferation Itisnicetohaveagraphical representationofyourconclusions IGF1pulse Deregulated Inducedgenes Inducedgenes IGF1signaling SNHG7 Repressedgenes IGF1signaling SNHG7 Repressedgenes HyperProliferation Proliferation Breastcancerpatientswithtumorsthathavehighly expressedSNHG7havedecreasedoverallsurvival. SNHG7AmplifiedorUpregulated in49(6%)ofcases. Logrank testp-value:0.0395 100 UnalteredSNHG7 Upregulated SNHG7 %Surviving 80 60 40 20 0 0 50 100 150 MonthsSurvival 200 Summary Endwithasummaryandthenfuturedirectionsor whatyouwouldve donewithmoretime • IGF1signalingdecreasesSNHG7stabilitythroughtheMAPK pathway. • SNHG7regulatesproliferationinadose-dependentmanner. • Identifiedafine-tuningmechanismofIGF1-regulatedgene expressionandproliferation. • BreastcancerpatientswithelevatedSNHG7expression havedecreasedoverallsurvival. Donotsaysomethingdifferentthanthetext.Peoplecan’treadandlistenatthesame time Acknowledgements Neverforgettothankeveryone includingfunders(askyour mentors) AdrianLee,PhD SteffiOesterreich,PhD RyanHartmaier,PhD AmirBahreini Dani Hochbaum,PhD CenterforSimulationandModeling(SAM) ChandraBathula,PhD MikeBarmada,PhD YanGonghong,PhD Itisokaytonotknowtheanswertoquestions!Itisokaytosaythatis SusanFarabaugh,PhD agoodquestionandIdonotknow…orweshouldlookintothat..etc Rekha Gyanchandani,PhD Youarenotbeingquizzed,youarehavingadiscussionwithanother WhitneyCrosson,MS scientist. AlisonNagle NolanPriedigkeit Younowknowmoreaboutthisdomainthanmostpeopleintheworld. BethKnapick Chen(Linda)Jingci McGillUniversity Rangos ResearchCenter– CHP William Horne Funding: Posters • Size36x44isacommonsize. • SubmittoVickie • https://osticket.dbmi.pitt.edu:444/index.php • [email protected] • Atleast3daysbeforeyouneedit(Tuesday) • Sametipsastalks…knowyouraudienceandtella story.Prepareforpeoplewithdifferent backgrounds.Beabletoexplainyourworkin2 minutes,5minutes,10minutes. • Engagetheperson.Askthemquestions.Makeeye contact • Interactwithyourposter. Posters • Limittext • Somesymposiumwantyourabstracttobeincluded.Other thanthisyoushouldneverhaveblocksoftext.Usebullet pointsifnecessarybutpicturesarebest. • Whitespace.Don’tcram. • Makefontvisiblefrom5feetaway(size12iswaytoo small) • Figuresarewelllabeledandeasytounderstand. • Itisokaynottoknowsomething. • Havefunandlearn! • Example CoSBBI – AMIA • https://www.amia.org/amia2016/high-schoolscholars • 10minutetalk/presentation • Abstract/shortpaperdifferentthisyear.Lookat requirements • Recommendationfromyourmentordescribing yourcontributions(GIVETHEMTIME!Several weekspreferably). • DUESept8th