cell-penetrating apoptotic peptide research presentation

8/19/2019 Cell-Penetrating Apoptotic Peptide Research Presentation

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Cell -penetrating apoptotic peptide/p53 DNA nanocomplex asadjuvant therapy for drug-resistant breast cancer

Presented by Spence

BMS 494 | Missouri Stat

"#an$ %, #an$ %, &ian$ ', et a() Mol


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PurposeTo in!esti$ate the in !itro and in vivo e icacy o chimerica(

modi ied Smac peptides +3 P5 7 , co de(i!ered 8ith p/. p :3on apoptosis and tumor suppression in metastatic and metastaticdru$ resistant breast cancer (ines, as ad;u!ant therapy orantineop(astic +do:S- A!"# +3(a a( Pro 5(e Smac tetrapeptide * $ % +octa ar$inine aa moiety * C"" +ce(( penpeptide * &D$ +mu(ti dru$ resistance * "gp ' 3B? transporter P $(ycoprotein * &( +mitochondria( * aa +am


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>!er!ie85mmorta(i@ation o cancer ce((s by supprese!adin$ ce(( pro$rammed death +apoptosisha((marA o cancer

Mu(ti dru$ resistant +M phenotypes areo .0 0C o irst (ine chemotherapy ai(

esistant to >D

Proapoptotic proteins, inc(udin$ Smac +tetrapeptide mimetics, demonstrate abi(ity too!ercome intrinsic apoptotic si$na((in$

suppressionPre!ious(y, c(inica( e icacy o 3 P5 8as (imitits (o8 permeabi(ity

To o!ercome this, #an$ et a() chimerica((ymodi ied 3 P5 tetrapeptide +3 P5 7 8

urther a((o8in$ its use as !ehic(e or $ene

de(i!ery

'&D$-breast cancer)


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Apoptosis is* 3 ti$ht(y re$u(ated process that maintains thehomeostatic ce((u(ar ba(ance, protectin$ a$ainst-

Eenomic instabi(ity + :3 dama$e, rep(icati!esenescence=n!ironmenta( +patho$en, P3MPs 3MPs5ntrace((u(ar stress + >S, nutrient de iciency, $enoto


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Chemotherapy and radiotherapy exert cytotoxicityprimarily through intrinsic apoptosis signalling

Chemotherapy 'i e . Doxorubicin)

5nduce MT permeabi(i@ation throu$hindirect mechanisms

:3 dama$e +p/. , 5nhibition o apoptosis+53P proteins, >S mediated

$adiotherapy 'gamma. -ray)

Eenera((y be(ie!ed to direct depo(arimembrane H permeabi(i@ation

e(ease o ?ytochrome c, Smac, and opro apoptotic actors

Ihu D, #an$ J, Ihan$ J, et a() Int J Mol Sci. 201.


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A!"#, 0mac derivatives or mimetics 1/ conserved N-terminus 2-aa se uence

:o!e( approach to combat M and apoptotic

e!asion is to reacti!ate apoptosis path8ays53Ps upre$u(ated, ?asp suppression, Smac inhibition

Smac protein is a 2.9 aa po(ypeptide// aa N-terminus encodin$ MT tar$etin$ seKuence isc(ea!ed upon maturation

irect(y binds to 53Ps, thus Smac is pro apoptotic

Smac cut do8n to 4 aa tetra peptides +3 P5 ha!e‐sho8n in vitro and in vivo apoptotic acti!ity, as 8e((as are current(y in L 20 phase 1 2 c(inica( tria(s

3 P5 e ecti!e at inducin$ chemo and radiotherapy

Ma;or dra8bacA is 3 P5 peptides are poor(ypermeabi(ity

ean =', anson M, B(acAha(( F, et a() ?anc


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Chimeric A!"#$ % /p53 nanocomplex

?himeric 3 P5 deri!ati!es 8ere de!e(oped uti(i@in$ a ? terminus octa ar$inine +seKuence + 7

3 P5 7 or 3 P5 7Po(yar$inines + are considered Nce(( penetratin$ seKuencesO

3 P5 7 e


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MethodsSynthesi@in$ 3 P5 7 p :3 :anocomp(e


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"hysicochemical Characteri8ation of A!"#$ % / p53 DNA nanocomplexes via mobility-shift assay'7a). 9eta potential/"article si8e '7b). and (+& '7c)


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(ransfection efficiency in human &C6-: and &C6-:/AD$ '&D$) cells

;ptimal transfection efficiency assayed usingluciferase reporter gene demonstrates 3b)


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Casp-3 'effector caspase) activity in response to D;

6lo1 Cytometry Analysis, Annexin/"#

Buantified 6lo1 Cytometry


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;ff-site toxicity and effect of D; A!"#$ % /p53on body 1eight 'Ea) and organ-specific damage'Eb)


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?onc(usion5n order to o!ercome M breast cancer, code(i!ery o ce(( penetratin$ peptide p/. :3nanocomp(e< 8as de!e(oped as ad;u!ant therapy by re sensiti@in$ M cancer ce((s to apoptosis

Smac inhibits 53Ps, thus acti!atin$ ?asp . eD in non M and M breast cancer mode(s

?oadministration o 3 P5 7 p/. demonstrated u(( arrest o D & /0 3nima( studies usin$ combination o antineop(astic +do


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#orAs ?ited1) Ihu D, #an$ J, Ihan$ J, et a() Iiyu$(ycoside 55 inhibits the $ro8th o human breast carcinoma M 3 MB 4./ ce((s !ia ce(( cyc(e arrest and induction

o apoptosis throu$h the mitochondria dependent path8ay) 5nt ' Mo( Sci) 201.*14+9 -17041 //)

2) oma$o; , #ayne ') Fairbrother Clin Cancer Res. 200 *1.-/99/ 6000

.) in S, Qan$ ?, &i S, Du ?, Ihao Q, en %) Smac- 5ts ro(e in apoptosis induction and use in (un$ cancer dia$nosis and treatment) Can2012*.17+1 -9 1.)

4) 'ames , Parone P3, Terradi((os >, &ucAen ard;omande S, Montessuit S, Martinou '?) Mechanisms o mitochondria( outer membranepermeabi(i@ation) Novartis Found Sym ) 200 *27 -1 0 6)

/) :achmias B, 3shhab Q, Ben yehuda ) The inhibitor o apoptosis protein ami(y +53Ps - an emer$in$ therapeutic tar$et in cancer) Sem2004*14+4 -2.1 4.)

6) ean =', anson M, B(acAha(( F, %o(t S , i!e ?) :o!e( therapeutic tar$ets in (un$ cancer- 5nhibitor o apoptosis proteins rom (aboratory to c(inic)Cancer "reat Rev ) 200 *..+2 -20. 12)

) Shio@aAi =:, Shi Q) ?aspases, 53Ps and Smac 53B&>- mechanisms rom structura( bio(o$y) "rends !iochem Sci. 2004*29+9 -476 9

7) Fu(da S, ebatin JM) =


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ean =', anson M, B(acAha(( F, et a


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cell-penetrating apoptotic peptide research presentation

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