BOC Midterm 2 PDF

BIOL380 : The Biology of Cancer Lecture #6 -- chapter #4 Paper presentation #1 Session 2 Chapter 4 – Growth factor Chapter 5 – Cell Cycle Chapter 6 – Tumor Suppresor Chapter 7 – Apoptosis Chapter 8 – Cancer Stem Cell. Session 3 : Chapter 9-13. Growth factor signaling and oncogenes Over...

BIOL380 : The Biology of Cancer Lecture #6 -- chapter #4 Paper presentation #1 Session 2 Chapter 4 – Growth factor Chapter 5 – Cell Cycle Chapter 6 – Tumor Suppresor Chapter 7 – Apoptosis Chapter 8 – Cancer Stem Cell. Session 3 : Chapter 9-13. Growth factor signaling and oncogenes Overview EGFR signaling Oncogenes Kinase as drug targets Only a selected subset of our cells are being replaced, to sustain tissue architecture.  Only a selected subset of our cells are being replaced, process transferring to sustain tissue architecture. of signal and yhlFromdueintred time parteris of requires 4types growth HOW? of in transduction factorsigned gothfactor fetters transdwers intracellularsignal transcription mulcar factors  TF Transcription Proliferation genes Proliferation genes factors Intracellular Signal transducers Growth factors receptors Growth factors  intracellularsignaltransdmen overier externalsignalGutt fat Tiggerchainreartin Gutfont receptor atintionofgenesthatdive transcription for tu cellputt insidenucleus activated puters transdurers 4 93ʳᵈ mulear directlyinterantofDNAtoturnon Transcription politeration genes TF Proliferation genes factors spinf Genes Taiusion respite middleplayers represented byprimepieres passsignalfromrecept cellsurface Intracellular unseus of a refytmetgh.tt Signal transducers receptoractivated Growth factors receptors P EditEnamel father signalling Growth factors molecules likekeysthatinitiate bind  sugarthelow Fftolook on cles key outline waygutfators montfty me thatadd phosphate group Overview funATPGTP EGFR signaling tospentAA in putin Oncogenes Kinase as drug targets phosphate gaptotyrosine othertargetserine threonine phosphate afineiienihngheaii.at a j thisplusphylationstepcurialforrelayingsignalsinsidethecell  EGF signaling: a paradigm EGFR tyrosinekinase four growth creatiered extracellular blind domain extend utilisation dwain EGFR: Y kinase. Substrate: conformational change meetsEstimating EGFR  EGF signaling: overview steps 1 Y 1 GF binding mppmmm.mpnpn.ee residues earthestyrosine 2 dimerization 5 Activation of intracellular 3 4 transducers Recruitment of proteins with 6 S/T kinase t.in athm SH2/SH3 cascade state domains cell tyrosineAA 7 8 Target gene TF activation regulation  EGF signaling: overview Steps5 8 1 GF binding signaltransdivers I tentethitained 2 dimerization farnesylation 5 Activation of intracellular 3 4 transducers Recruitment of proteins with 6 S/T kinase SH2/SH3 cascade of activation transcription factorin mulens muters domains kinasecascade RAF MEK MAPK In on genescivolved in cellgunthand 7 serieskinaseoutwation of eachkinaseactivateshe TF activation 8 Target gene division regulation nextinline GF binding dimerization Auto phosphorylation sheetsintertitles bingingtogether autophosphylationactivates thereceptorcomplex  Auto phosphorylation (inside the cell) w/o EGF EGF treated Phospho-EGF receptor T.it Translocation of SOS (RAS activator) leads to Ras (membrane-bound) activation iEiE Phthaleinother ftp.hgteettgsia warpaint SoSbiyngrear membrane RASITTembram En.IE RAs Raus mtsignalling forcellguth signal SosfamilitatesactivationofRAS SH2: bind to Y-p SH3: bind to proline and hydrophobic aa exchange bypromoting of attractpatentwithSH2Src Sosbuding GRBS antevideexchange guanine farterCAF 2 domains RAS  bind to phosphylatedtyloids GRBshassH3domainsthatbind to RASactivated tethered GRP2hasSHzdomains prolinerichregionsofSos anching to membrane inactiveGTP itnearcellmembranewhereRASis boundforminterartof downstreameffectors  MAP kinase cascade justaparallelsignallingpathwaydifferRAS PI3K AKT mTOR activeRASativates phosphyates antivatesMEK ERK phspmlateiftgh.nlaedputeintmase entermiles paypal.lt 7 phophylated7F AP-1 (JUN/FOS) likeAPTintichareJNNFOS  proteins bindto DNA activepartin transcription 2 majorclassifications untatedgenesthat outline of cause saver Incogenes Overview antenna version EGFR signaling ftp.edtiyttd Oncogenes unfatedgenesthat promotecarerby causing adsto gunanddivide Kinase as drug targets to wh dominant only 1 mutatedallele enough to haveeffort The first oncogene v-src returims containRNAgenome caninsert a Putcopy int a host chromosome  Discovery of Rous Sarcoma Virus (RSV) unsescancausecaner sanoma in dickens andfreefitrate fundueter sarone induecour in healthychicken Dr. Peyton Rous fundamentalprimple of cancerbiology almostallfouronagers forms mural are altered of genes a partonogens  SRCunmallyinactivestate TailfingEntitled binds to phosphate guy closed shape 7453 or removesoff switch keepingin excessive keepskinase domain partof alwaysalive Ske thatactivatesotherproteins cellsignalling H EGFR 1 Fishingtins i as i removing Tyr530 n regulatory site v-SRC = Always active, EEm truncation amplify the signaling adly Other examples of viral oncogenes mins PPKF Iguth fat thatpantescellgirth wands instead ofbeing stays incell incytoplasm secreted stays v-SIS: truncated version Growth factors keep PDEF of PDGF (growth factor) Estimative v-ERBB: truncated mutated version of EGFR missingexternalbindingpart mutated receptalwaysone causingcells todividewithout needingguthsignal  mutationsinRASfound in 307 love Ras ofhumantumors ofmost anon anogenes Mutation in aa 12, 13, and 61 Constitutively active Ras unallyswitchesfor ATP GDP G inactivate mutationprevents this keepingRASinstantly Raf htddba t kepsgmthsgnals oNinadls.igwig B-Raf, commonly mutated in melanoma PE.PEhesitantflat 3has Constitutively active kinase Another example of Oncogene – fusion protein after chromosomal translocation ABL-nuclear tyrosine kinase involved in DNA damage CML Excessive cell proliferation. I Reduced apoptosis (programmed cell death). Accumulation of immature white Philadelphia chromosome blood cells in the bone marrow and bloodstream. t(9:22) 95% of Chronic myelogenous in care chromosomes 9 22 snap leukemia (CML) plans called Philadelphiachromosome Another example of Oncogene – fusion protein after chromosomal translocation ABL-nuclear tyrosine kinase involved in DNA damage (activated by ATM) BCR-ABL fusion iMiifffJ iiopg fffgpgg mmi Philadelphia chromosome Dimerization, autoP  activation in t(9:22) cytoplasm  Novel substrates, interfering with 95% of Chronic myelogenous normal signal transduction leukemia (CML)  Abnormal proliferation Transcription factors i Instantiates Shesitating signalsforall guith JUN/FOS upregulation CHP1 transciptinfatorinvolvedof complex (truncation of 3’ UTR  increased stability; glad deletion/mutation in the promoter region; genomic amplification) Tree girth MYC mIttg intestinal chromosomal translocation t(8,14), under strong promoter of IgG leadingto highMYexpression HighMYCactivity drivesuncontrolledcelldivision camer  tonallyactivategenesthatinhibitgouth p Thyroid hormone receptor ERBA ---- target genes: inhibit growth Dominant negative mutation - bind to DNA & block access of wild-type receptors III Iii DNAbutblooksawe's ofviralcontdtypereceptors tonal katana mutationsinthyroidhormone survey neptor v ERBA preventional thyroid hormonesignalling byblockingfurtimalactivators untilledcellguith ecDNA. ----- Non mendelian inheritance leading to amplification circularDNAoutside of duplicatedindependentlyleadingtogene mainchromosomes amplification that_templifchtimal Chromatin accessibility is higher DNA movearesible IItattmendtieitita.ua transcription comes agility maintained incelldivision Fae higherexpression cDNAgenus of  Experiment designed to identify some of the oncogenes spentDNAfragment thatantainanges thrillingthan arrogenes combine w calciumphosphate helpsbutentercells in nextstep meetYamasee intestine III as cells Entome anyangenescantransform indicateangercaused thesecells abrumalguith Mechanisms: proto-oncogenes  oncogenes outline Overview EGFR signaling Y Oncogenes Kinase as drug targets  2 outside Intatterton Tyrosine kinase qpetitiveteestt.cat 1 Inhibitors (small molecules) RAS taegieatgai.is Kinase inhibitor (also bind to other targets) Binding reversible TBD Binding irreversible retstine covalentbonds B-RAF V600E 0 Target T790M (melanoma) (mutated in 50-60% of resistant cases) singentantionmEGFR B-RAF V600E (melanoma) Theater ihhtiskimdanmeitaier.atgirth developedtospecificallytarget inhibitABVBSRtyrosinekinase  r n wksbybindingto ATPbindingsite Development of imatinib – inhibit BCR-ABL fusion of BR AR blinking antithinylfallguth The value of early cancer detection CML has 3 phases Chronic Myeloid Leukemia Chronic 3-5 years. ----- 98% complete response. 9% relapse effativehere treatmentmust accelerated 3-9 months. 78% relapse --- 53% response datingEhmintertentive blast crisis 3-6 months --- 30% response EEEEI ei.ie haven'tdeveloped multiplyunontillably  all m I shapter4  BIOL380 : The Biology of Cancer Lecture #7 (chapter #5)  Chapter 5. Cell Cycle The 4 phases of cell cycle? I Which one is usually the shortest? 61 cellguns forPM replorationgyeongsangnam prepares fgymfy pggyyag j.my ganggpgygn.gg 1h18min forcelldivision m mitosis cell divides distributingchonsumes cytoplasm 1hr t.fm 2 daughtercells Hines attentive Infection  Mistheshutat Cell Cycle statage 1 fE.IE only forward moves mEiscins sure ifready Cell Cycle mostcellsintheadulthumanbodyare in whichphase Most cells in an adult: G0 Undifferentiated, quiescent cells can re- enter the cell cycle, induced by growth factors undifferientated notspecialized canbeinativebutreenterwell tugged bygutfators restingphase notactivelyengagedin CC  Cell Cycle R point, restriction point. Before R: cell division is dependent on mitogens (growth factors). After R: cells are irreversibly committed to progress through the cycle without the need for growth factors. antimethoughput synthesis Can.fiaimghhmtreedng sweads make sway whenconditionsarefaurable onlydivide resources areavailable butone committed onlyhavesmallwindowforsensing theyprovedindependentofexternalguithsignals preserve southfortus mitogens l Cell Cycle Different phases: regulated by CDK+cyclin  cyclins– regulatory subunits of their associated kinase (CDK) concentration varies in a cyclical fashion during the cell cycle fiit.EE B.DIIEiA The levels of CDK don’t fluctuate during the cell cycle attet CDKlevelsCOLAM cyclins– regulatory subunits of their associated kinase (CDK) concentration varies in a cyclical fashion during the cell cycle T The levels of CDK don’t fluctuate during the cell cycle  Mechanisms of CDK regulation Association with cyclin; Phosphorylation  activation conformation change Physphalted onephosphylatedkind regulatorstating Interfere with ATP binding I Croatiating kinase causesconformational changesmaking it CDK2, CDK1 active CDK4/6  Mechanisms of CDK regulation Association with cyclin; Phosphorylation  activation conformation change phosphylatedbyweet addinactivatingpurplates Phosphorylation  inactivation Interfere with ATP binding kinase T interferey p.EE een CDK4/6 CDK2, CDK1 Titillating tiratingCPK  Mechanisms of CDK regulation Association with cyclin; Phosphorylation  activation conformation change Association with CDK inhibitors Phosphorylation  inactivation p16, p15, p21 Interfere with ATP binding proteins likep16 PDetc canbindtoCDKor cookendinamplexes and invitthiratinty Fan 111 CDK4/6 CDK2, CDK1 K2 1  provenigrell f prog guard RB: a crucial regulator of Cell Cycle Entry Absence of DNA damage? Sufficient nutrition? periostitis's auditionsarentright if Rb A transcriptional regulator controlling cell cycle progression. A key checkpoint protein controlling G1/S transition by modulating E2F-dependent transcription. RBcentresgone ELF whichisneeded activategenesforcellcycleprogression RBbindstoECF prevents EFfunactivating thesegenes prevent funmovingforward in CC RB activity is regulated by cyclin-CDK RB regulate E2F TF activity Moderate E2F partial- activation phosphorylation Note: textbook “text” part has typos in this section. Figure and figure legends are correct RB activity is regulated by cyclin-CDK RB regulate E2F TF activity transmptinreprenon the tt ntpitpiaidain artivatesCDK4 6 Moderate E2F activation ad partial- phosphorylation hyper- I mail.tt E.rEEE phosphorylation mme ies.int tieatne Note: textbook “text” part has typos in this section. Figure and figure legends are correct 4/6 2 2 sufficesantirate cycli ok complexorthat phphylate RB graduallytuing on ELF t allon Cuprogression Gi s  Structure and function of Rb – “pocket protein” HDA “pocket domain” there my modifyingchastinstature Sequesters E2F c PEEEEE EE.EE man genes Eif Cyclin A, thymidylate synthase, etc, Important for S phase EEE MAD2, mitosis  I G2 checkpoint: DNA damage response interphasetakeslonger bgi.mn rewgenes hetransabed ensureall deent pread to mitstsw damagedPNA  G2 checkpoint: DNA damage response indureskineseantivity issevere DNAdamage if P53activitycan prolonged Pathitiff P P leadtoapopt.si meditate DNAdamagedegradation phosphatase are putrepand v71 gapfun Negative P removes phosphate Feedback Loop p53tuning chekatting P imgapnphophteseCdkcdais normallyactivates this checkpoint txn Can be switched off Pfeifer.EE After DNA repair CyclinB removing by inhibitoryphphates whenideasinhibited Chsiption Binding adlaycleamttallo.fr “checkpoint recovery” Prtrepair byphphylation inhibitor fatorthatatientesdustream target i s The mitotic checkpoint: ensure correct mail.tk nt chromosomal segregation  Chromatids attached to mitotic spindle? mitotic spindle intact and functional? Titans preventerrorslike help distributionof uneven chromosomes Mutations in CDK and cyclins CDK4 mutation  blocks binding to cell cycle inhibitors mathfastate ok put a fraud areadidinson CDK6 overexpression. (leukemia, translocation) aerial addteams Cyclin overexpression. tartaric Tweeting Deletion of cell cycle inhibitors. allowCLto me forward canbeleveragedto develop allowcellstodividewithout summary therapentintreatments usualregulatingcontrols  Thmell Tflfallfuntion but Iii 4s gland homeostasis CDK4: dispensable for mammary but essential for breast cancer progression. I Breast cancer treatment: CDK4 inhibition + targeting Estrogen or Estrogen receptor ntgoodtoblokant aiEaaafhiea.is offer combined 4 estrgertargeting treatments orestgereceptor Simesomebreastcancersrelyon estrgerto go  G2 checkpoint: DNA damage response Make chemotherapy more effective (prevent cell cycle arrest, let more DNA damage accumulate, Leading to apoptosis) Negative Feedback Loop degradation P P phosphatase P I prevent 2 checkpoint fun stoppingcellcycletorepair DNA this checkpoint txn P bypreventingDNArepairchortherapy makes harder thecell Can be switched off it for After DNA repair avoidapoptosis Binding “checkpoint recovery”  BIOL380 : The Biology of Cancer Lecture #8 (chapter 6)  p53 couldabetherDart can getrepairedeffendy Rb Tumor suppressor genes Oncogenes  EXAMPLES of tumor suppressor genes – different processes Cell cycle Signal transduction Transcription regulation DNA repair Cell adhesion Etc. You are not required to memorize this table, but expected to understand the specific examples p53, BRCA (chapter2), PTEN, Rb. Rb two  M F MAP kinase cascade JEEThief PI3K AKT mTOR 2ndbranch ERK AP-1 (JUN/FOS) Lecture 6  PI3K-AKT-mTOR pathway andgarth matter sumal pathways You are not required to memorize this map!  PTEN antagonizes PI3K outfields'eteeper II.EE PiiiMxtaiItain Bettintitt's I ww Cell proliferation  tumor r Retinoblastoma --- Rb. twosuppner gene (initiated by loss of both RB1 alleles) eye fortumortoform botharytesneedtobeinactivatedmutated inheathyeyes RBIhelpsregular occurring most often in young children below 5 years 51T inrateadf.nl ~ 250-350 new cases occur every year in the USA inretinasawmonthlably twofination recognizedasthe gourdianofthe genome  P53 mutation landscape in cancers (cBioportal) reinforeprions observation agitatedwith p53  P53: Guardian of the Genome The first tumor suppressor identified. moststudiedhumangenome Highly connected to other signaling pathways. Tssprevent twor connectivity genomematters of byresponding development analogy flight map to DNAdamage chit Effite  P53: Guardian of the Genome Normal cells: low P53 level P53tendsusuallytwobecametheydon't custant need punters antiretim whenneededcanberegulated levelto repodfaster Sustain the expression of glutathione peroxidase 1 & sestrins antioxidanterymes 11 aft important Decrease ROS ( such as hydrogen peroxide H 2O2 ) P53 ByreduingRosdamage ROS can cause DNA oxidation, such as 8-oxoguanine. minimizes DNA DNA polymerase misrepairs this G, leading to G->T transverse mutation andmutationrisks  P53: Guardian of the Genome I restated justDNAdamage “danger signals”  increase P53 PROTEIN level Elijahaboral damaged or topment cells cellularhubthat our fun developinginto recievesmany funcells signals I at Ederases levels puntin interpretssignals stpmndtmffealhh.tt Tamans bloodresellformation Initwtitiont dangerouscells potentially_  ang Hardone forforSm experimentally mousemodel lotdon'there tur lantanase IIIE.IE spontaneouslydevelop Li-Fraumeni Syndrome Germline mutation of TP53 – increased risk of developing cancer at a young age similartoRb ypsyisthegenethatproducestheputenP53 4.9cm*7.0cm*5.9cm 4cm*5cm*3cm 32 month old boy 13 year old girl, Siblings with brain tumors, case report https://bmcpediatr.biomedcentral.com/articles/10.1186/s12887-021-03070-8 Tantalate'sEast 1 muchmorecopies comparedto humans just 1copy Churans p53 mutant mice display early ageing-associated phenotypes p53 Stabiye preventiging mulibank hearth leadstoconcludethat https://www.nature.com/articles/415045a Taste Wants getcamer  P53 protein: short half life of 6-20 min degrades suient tem.tnit MDM2: promotes degradation blocks transactivation 1nal majaty facilitate nuclear export happenshere  whereP53bindstoDNA to activateorrepress targetgenes P53 protein structure. Function as a tetramer Ei arrestapoptosis III MDM2: promotes degradation I 4FinofP53come blocks transactivation facilitate nuclear export togetherto regulate geneexpression  fullstablebioding tetramer inquirescomplete I interatofPutsegeme Bhai tie suboutsassembled Egulate allowto genesinward in SCarent apoptosis helpingpreventturn formation  p53 dominant negative mutations PETITE metathranded intetramer adding caninterferewithfuntionoftetramer majoritycurtaincurtants redmabilityofptstobad butand at mein atollingcell  g P53 protein structure. Function as a tetramer matinalhtspots gas MDM2: promotes degradation a ftp.t.fftfttF blocks transactivation facilitate nuclear export trigger p53iait downstream targetgenes  P53-MDM2 feedback loop negativefeedback loop helpsregulatep53 in comalcells PseettistsMore In normal cells Maintain low protein levels of p53, Until stress is sensed. ᵈSt p c 1EE den aurmfoteanisgesanlig maintaininglowlevels untilthere'sa streetsignal T.fi aaiai00 responrsditttyiesof Sequester Eitiyygmon's MDM2 to the nucleolus PIYARFisprdue hindgtmma.int.is tiastte Both ATM and CHK2 Both ATR and CKII stain lament Block MDM2 binding I Phosphorylate the NTD I phosphorylate p53 Disrupts MDM2-p53 binding initiate a apoptosis a terminsain Downstream effects after p53 activation EEE.EEEes Glycoproteins antiangiogenic Pro- apoptotic anti- Nucleotide apoptotic Excision repair  4 Repair or abandon? Initiation  additional canoran phphylationsites Post-translational modifications: S46 phosphorylation (DYRK2)  favors apoptosis of P53artication Short pulses (repair) vs sustained signal (apoptosis) I mad 11sFd Co-factors other and puteirs signalsKofanton caninfluencep53sdecisionmaking Repair or abandon? 1  example: MYC (oncogene) activation promotes p53’s regulations towards apoptosis ETigit.ieiitatin cellcyclearrest rathethanapoptosis Pstmpating can Apoptosis-stimulating iind it.tngp Proteins of p53 Enhance p53’s ability to selectively bind to apoptotic gene promoters Additional events Fiatient.is tYuYedP53ability bindgenesthattriggerapoptosis  M Mutations in the p53 pathway É CHK2 mutation (not activating p53) tmall.im iamage reakeringaellablityto damage Inf respond Predominantly MDM2 overexpression gat.tts missense mutations DNA binding domain ingusansexaminer degradepr3 reduits putativeeffortsoncell BAX/FASR inactivation  it Tlsnight and apoptsi inantine Viral proteins can inactivate p53 and RB EEYETA 45Eur bindto 183 blukits funtion appt blocking walhadtoRb releaseELF Eleftstrating allmcct.fmunacked uncontrollablecelldivision  cervical carcinoma 97% of these tumors – insertions of HPV genome fragments (oncogene) Human papillomavirus CintohumanDNA E2 negatively regulate E6 and E7 L1 capsid protein Targeting p53 pathway andfution p to restore folding Correct p53 mutation --- small molecule, helps (many point mutations result in aberrant protein conformation and interfere with DNA binding) P53 itmutated only suppresed Activate endogenous p53 (if p53 is not mutated. For example, when MDM2 is overexpressed) --- disruption p53-MDM2 interaction  stabilize endogenous protein disruptp53mouzbinding freep53t funtin Suppress endogenous p53 (temporary, reversible) --- reduce side effects of chemo/radio-therapy. (reduce apoptosis in hematopoietic organs, intestinal organs, hair loss) wouldhelppatent temporarysuppresin healthycellsduringtherapy  BIOL380 : The Biology of Cancer Lecture #9 Last lecture: Tumor suppressor genes p53, PTEN, Rb.  Overview --- apoptosis (chapter 7) Characteristics Molecular mechanisms P53 and apoptosis Relevance to cancer and chemotherapy Therapeutic strategies Which one is apoptosis? Sun burn Necrotic leg wound caused by a brown recluse spider bite apoptosis A highly regulated process of cell death. Active process of “cell suicide”. Swept clean by macrophages & neighboring cells “phagocytosis” www.mail Eliminates cells have extensive DNA damage & potential to lead to cancer me Defects in apoptosis also influence the effectiveness of conventional therapy (i.e. chemotherapy agents act primarily through DNA damage - apoptosis). waycanontreatments likechewtherapy workbycausing DNAdamage triggeringapoptosis ifapoptosisdoesn'tworktreatmentscanbelesseffective  damagedcells cartime survive gun Necrosis irreversible cell death caused by injury, infection, lack of blood flow, or toxins. cells die in an uncontrolled manner, Necrotic leg wound caused by a cells release their contents into the brown recluse spider bite surrounding tissue, often triggering inflammation. whichcouldlead further tissuedamage pain apoptosis Normal FAERIE ahdgm cods apoptosis Necrosis apoptosis injurytoxins T I canalsobitrggendhyextualwspenfnpathwa foutusfe.ge immunecellsattacking somatintissues  Detection of apoptosis sE hamapteti ladder ifeng.it Terminal deoxynucleotidyl transferase LymeLabel ( biotin, fluorescein) DNA ladder TUNEL assay detailsDNAbreaks Tdtaddslabeledmuletides ends fluoresceina biotin tothese3or allowusto visualizeunder  Caspases: central for apoptosis iii Caspase: cysteine-rich aspartate proteases titi Procaspase: ~2% of total Caspase activity. HarEEms Cleavage  full activity heomefllyatimatterthyondeaultns.to I initiallyinentive caspases are pronaspases mail.EE 5mYE casparesassade onearticatesthenext Chalet Patti Amplification of signal ensuresstrongresponse punishment salary caspasesinitiateanddivetheapoptosisprocess Extrinsic Intrinsic (signaling induced) (involve mitochondria) apoptosiscanbitrgged m ingatingextm receptorsrevgineigands mythfare Extrinsic: TNFreceptor Fasreceptor seeks molecules Fast TfL a common mechanism S Eigenstate toactivatethepathway Limmeralls during inflammatory responses to Example: apoptosis In viral infections such as hepatitis, control immune cells or damaged tissue. Example: Apoptosis of inflammatory cytotoxic T cells recognize infected cells to limit the immune response in hepatocytes and induce apoptosis ayhg.at diseases like rheumatoid arthritis. ated undis via Fas/FasL signaling to limit viral spread. I infatedlivercells  ligand Extrinsic: receptor Death inducing signaling complex trimerization It Conformation change (expose "death domain” Red square) Recruit “adaptors” (TRADD/FADD) “Death effector domain” Red triangle Procaspase 8 in proximity to each other I C-FLIP: Catalytically inactive caspase-8 homolog blnkgcarparesairatio itthiisit.im Bind to “death effector domain” in FADD or Procaspase 8 (but not TRADD) Inhibit the apoptosis process caspase 8 activation fairpeded deathettendmain caspase cascade recitpuraspates  intrinsic DNA damage/cell stress leadtoopening mitochondrial membrane of Open mitochondria apoptosome releasecomponents ftp.meccmbins Faofatw releasedfun Cytochrome C, and dHPt APAF1 from cytoplasm (and dATP), Procaspase 9 co factor activating procaspase-9 More details Y hdpim Fistases included in figure 7.5 in the book Activated procaspase 9 caspase cascade caspases  an Cytochrome C, Procaspase 9 Storage: Interspace between the 2 membranes Released by outermembrane of mitigdhata Mitochondrial outer membrane permeabilization controlled by BCL2 family proteins BAX, BAK: forming pores  pro-apoptotic i anti-apoptotic: BCL2, MCL1 bind and sequester pro-apoptotic proteins Hiiiii ties  P 97 1 Pl keying Other modulators of Apoptosis Inhibitors of apoptosis proteins (IAPs) bind and inhibit caspases induced by NF-kB (inflammation) to inhibit apoptosis. inhibited by smac/Diablo (released from the mitochondria) 1 smanDiabloputein released funwithordia to during putincomplexthatresponds inflammation apoptosis appt increase IAPlevels to inhibit bindto 1 s preventingIAPs inhibitinhibitorofapoptosis funintilatingcaspases pronteapoptrop  Crosstalk between extrinsic & intrinsic pathway extwininpatheryatimtecaspase8 Harp are8antimtesproteincalledBig Bid tBID translocates to the mitochondrial outer BCL2 family protein extrusin intimsin membrane, where it interacts with pro- apoptotic proteins such as Bax and Bak, hay sat canconverge Converting them from inactive to active (confirmational change) moretwandmitochondria andBAK interactw proapoptoticproteins BAX trigger innotochordial membrane bind BoxandBak activatethem mitochondrial outer membrane permeabilization Cytochrome c release keystepinativating  I may P53: theatplasm Cytoplasm: PUMA releases p53 from binding to BCL- p53antivatesexpression Death receptors 1 xL, enabling p53’s binding to BAX P53 actives BAX to release cytochrome c etc. Pro-apoptotic members of BCL2 family Examples: FAS, BAX, BAK PUMA (p53 upregulated modulated of apoptosis), essential for mediating p53 induced apoptosis ofthesefigapt tIEEE.IT Anti-apoptotic factors Apoptosis and cancer Mutations (M) can be acquired to allow escape from apoptosis Epigenetic alterations makeeasierforcanoncellsto survive fishmities if Induction BCL2: anti-apoptotic Translocation/ microRNA reduction APAF1 (activator of caspase 9) mutated/repressed t this hell app is likelyto occur redupantanimportant inintumipathway Apoptosis in normal vs cancer cells already Tumor cells receive many apoptosis-inducing signals, many caspases are already activated, but inhibited by upregulated IAPs. TRAIL receptor ( a subfamily of TNF receptor) + TRAIL (ligand), can induce apoptosis in many cancer cells but not most normal cells tyhoptn Treneptn IRANligand TYntth.tw Hutruralcells spare  Optional reading: https://www.haematologica.org/article/view/4605 treatmentitselfseedsto therapy related tenderer conditionwhere putdamagearmulation leads lenkerva  to averagecell summarythendugsaimcells Apoptotic drugs: deathin car bytargeting prteristhattypnallyprevent apoptosis TRAIL : recombinant trail ligand “dulanermin” Phase III helpsinducecelldeathin samertells BCL2 family protein : redueexpressionofthecells BCL2 overexpression  RNAi redueBlurexpressionincells small molecule to block protein-protein interactions. Phase I, II. Caspase activation mininsSweartocounteract IAB IAP overexpression: SMAC mimetics to relieve IAP inhibition. (Phase I, II) applied inclinicaltrial  BIOL380 : The Biology of Cancer Lecture #10  Cancer stem cells A subpopulation of cells within a tumor that have the ability to: 1) self-renew 2) (differentiate) give rise to phenotypically diverse cancer cells with limited proliferative potential that make up the rest of the tumor 3) functionally: initiate new tumors when transplanted to host animals. aitnentialiiiiiiaiigeem.ae hercellshavelimitedguith ptetistimmae iihediai.lt it inmates  Classic model Neutral competition model Division is intrinsic and asymmetrical Determined by the space within the niche antihistamines are localemonetsanding literature MEEEEE differentiate Entire widentiates Fitment prediction Fighting maintain s Evidences supporting both models have been identified in different studies (different tissues) Fundamental mechanism involved in stem cell regulation are relevant to cancer Cancer stem cells 11 a homogenouspopulation Functionally defined by having the ability to initiate new tumors after transplantation 1/1000000 of AML can develop into new leukemias transplanted  nspamuetmkingmall.to replaneold ones IEisgn.netpiftiatgng p lead to ramer Stem cell functions are linked to cancer cellturnover 1 may sina.tn itiffmiedis  by dg my d normaltissue gene sequencingof mutationsconnected there aremany Self- renewal accumulating Mutations Itocancer mm 7883 covermanymutations YEEYE.IE s and men againIsha iii it.it E ae iaiatome https://www.science.org/doi/10.1126/science.aaa6806 Self-renewal regulatory Mechanisms can be hijacked by Cancer Wnt signaling pathway Hedgehog signaling pathway  The essential parts of a signal transduction pathway ligander Yet Just like a Rube Goldberg machine! ligand budge receptor 1 Specific target genes Signalpathway Iii responseinside  Wnt signal Ligand=Wnt bidatheentt.tt cell pmeanstunsonspenfngen.es Beta-catenin (transcription activator) TCF/LEF Family TF Receptor= Specific target genes Frizzled LRP (Low-Density Lipoprotein Receptor-Related Protein, Co-receptor) MYC Cyclin D1 uppartsnsteperf.co wept to Friated helptellgunt divide Hr toturnoff EETIEAH Co-repressor  Intaghalens instantiated Degradation complex Co-repressor ubiquitinaddedafter mysignal miifi.EEgt inabsence of cateningetsbriendown degraded β AXIN & APC: scaffold for the kinases sinceβcateningive holdkineses inplane Grondo wrapresterstays bundto TLFLEF keeping targetgenesoff Wnt binding  LRP phosphorylation --> disrupt the degradation complex Additional co-activators hE ELkdmtmtsf degredotimaprxfd.ie may Spartan thirtaintiana LGR5, AXIN2 – stem cell specific targets  allthetime 90% of colon cancer --- mutations result in constitutive activation of WNT pathway Inactivation of APC APC (Adenomatous Polyposis Coli) activation of beta-catenin APLusuallyhelpscurtailBcateninmarking fordatuition keepingwotpathwayinchuk it ftp.gg itarepathay familial adenomatous polyposis (FAP) colon cancer --- familial form (germline mutation in APC) handita high number of polyps in early adulthood - increased risk of colon cancer PMPsgentigay intocancer them sporadic form Giant mandate 1Bang Gleaner  T I momingman ayyygpyyygpygg.gg observedwithohm portersoften immfkf.info cancerprogression Colonoscopy  Inhibitors of the WNT pathway --- several in clinical trials wktsttghltaffd.IT uivte pathway PORCN (porcupine) --- Lipid modification of WNT, Essential for binding to receptor emymethat addslipid II InFEF.TW superty Activate/amplify wnt Disrupt interaction bkkieeatikt.at impurities Co-activator, ntht CREB binding factor Binds to WNT, Block it from binding to the receptor printEither  aug 1 thway 11 top helps guy The hedgehog signaling pathway Ligand=hedgehog Kutrentersinders TF: GLI Receptor= Specific target genes Patched smoothened patched swo work togethertorevive Cyclin D, BCL2, VEGF and SNAIL hedgehogsignal helpcellgun preventcelldeathcreate bloodvessels whichareimportant but cancausecancer unregulated if  allussmatibone Hate great Release Gli mind an released Painden colorgertethered t.atplarmtraliglang.hn mttdpieap.ie I GLI, cyclin D, mᵗt9gaed BCL2, VEGF, snail. peas socaitaticatetargty.mx genes glY Patched localize to cilia, preventing smoothened from moving to cilia genes of production dustriangents Optional reading Primary Cilia and Mammalian Hedgehog Signaling https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5411695/  Tumor suppressor Inactivating mutations inactivecannolonger if blockSnotermit withsignal losinga brake airtatiears activating mutations smhasanticating maritimate Hh always 11 activation 71 apply at 1 oflag g Hedgehog signaling and cancer Basal cell carcinomas (Gli expression) Most common skin cancer rhabdomyosarcomas tissuetours muscle medulloblastoma 4h ofteninchildren appetite Most common childhood malignant brain tumor unintrolled cellguth  Inhibitors of the HH pathway Oral drugs targeting smoothened (clinical approved) inhibitingSmo prevent 4hpathwayoutivation Whittlingthatdepend Antibodies blocking Hh-patched interaction tatra HI thatiteration GANGinhibitor targetsaninvuleus block till finantivating crudens  long g Regulation of differentiation --- PcG proteins leaveprofoundsideettets associated proteins withmanytissues Inhibitors are being tested  Role of lineage-specific TF in differentiation and cancer AML: acute myeloid leukemia Blockage of differentiation towards the granulocyte or monocyte lineage normalhave maw mor versityof Ilmorphology nt at.EE mage impetuating stage attffffins blukage differentiation  1991 Role of lineage-specific TF in differentiation and cancer AML: acute myeloid leukemia Common Lymphoid progenitors Pluripotent Multipotent Hematopoietic progenitors Common Stem cells Myeloid progenitors Differentiation therapy for leukemia i Oncogene – fusion protein after slidefuncontier chromosomal translocation ABL-nuclear Y kinase involved in DNA damage BCR-ABL fusion Philadelphia chromosome Dimerization, autoP  activation in t(9:22) cytoplasm  Novel substrates, interfering with 95% of Chronic normal signal transduction myelogenous leukemia  Abnormal proliferation development linkeries of  Acute promyelocytic leukemia (APL) Philistines tahini mm Chromosomal translocation t(15;17). p - PML-RAR fusion protein normal Higher affinity to HDAC cancer Block differentiation homodimer Degrades PML-RAR High concentration E.EE EEEEEiim Complete remission, but most patients relapsed after a few month RA + arsenic trioxide --- combinational therapy