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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
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contact
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• Example
CoSBBI – AMIA
• https://www.amia.org/amia2016/high-schoolscholars
• 10minutetalk/presentation
• Abstract/shortpaperdifferentthisyear.Lookat
requirements
• Recommendationfromyourmentordescribing
yourcontributions(GIVETHEMTIME!Several
weekspreferably).
• DUESept8th