Vascular Zip Codes in Nanoparticle Nanoparticle Targeting Targeting - - PowerPoint PPT Presentation

▶
vascular zip codes in nanoparticle nanoparticle targeting
SMART_READER_LITE
LIVE PREVIEW

Vascular Zip Codes in Nanoparticle Nanoparticle Targeting Targeting - - PowerPoint PPT Presentation

Sep 25, 2023 194 likes •630 views

Vascular Zip Codes in Nanoparticle Nanoparticle Targeting Targeting Vascular Zip Codes in Erkki Ruoslahti Erkki Ruoslahti Burnham Institute at UCSB Burnham Institute at UCSB and and Department of Molecular Cellular and Developmental

slide-1
SLIDE 1

Vascular Zip Codes in Vascular Zip Codes in Nanoparticle Nanoparticle Targeting Targeting

Erkki Ruoslahti Erkki Ruoslahti Burnham Institute at UCSB Burnham Institute at UCSB and and Department of Molecular Cellular and Developmental Biology, UCSB Department of Molecular Cellular and Developmental Biology, UCSB

slide-2
SLIDE 2

Ruoslahti lab Santa Barbara Hector Biliran Priya Karmali V.Ramana Kotamraju David Peters Barry Rowan La Jolla Austin Derfus Valentina Fogal Tero Jarvinen Jan Pilch Dmitri Simberg Kazuki Sugahara Tambet Teesalu Miriam Wankell Lianglin Zhang Collaborators Doug Hanahan, UCSF Enrico Giraudo Sangeeta Bhatia, MIT, UCSD, Austin Derfus, Todd Harris Geoffrey von Maltzahn Michael Sailor, UCSD Ji HO Park Michael Schwartz Robert Hoffman, UCSD Matthew Tirrell, UCSB Patrick Daugherty, UCSB Support NCI, NHLBI, DoD, Komen Former lab members Maria Akerman Sven Christian Markus Essler Pirjo Laakkonen Kimmo Porkka

slide-3
SLIDE 3

Ruoslahti, Cancer Cell, 2002

Targeting Nanoparticles into Tumors

Modular Approach to Multi- functional Nanoparticles Physically target:

  • vascular cells
  • tumor cells
  • extracellular matrix

Trigger/engineer:

  • tissue penetration
  • RES avoidance
  • subcellular targeting
  • diagnostic signal(s)
  • release of therapeutic

agent

  • amplification of targeting
slide-4
SLIDE 4

Isolation of homing peptides by ex vivo/in vivo phage screening

Ex vivo

Wash Rescue phage and amplify Incubate phage with cells from target tissue Rescue and amplify phage from target tissue Repeat in vivo screening, sequence individual phage from enriched pool

In vivo

Inject (enriched) phage library into a mouse iv Phage bind to blood vessels in target tissue

Homing peptides

slide-5
SLIDE 5
  • L. Zhang et al. Circulation, 2005

Relative phage titer

Heart Kidney Brain Muscle Lung

50 100 150 200 250 300 350

Heart-homing phage

The blood vessels in individual tissues are distinct

Normal tissues targeted with vascular homing peptides:

Brain Kidneys Lungs Prostate Skin Breast Muscle Pancreas Adrenals Uterus Liver Heart

  • Bacterial 2-hybrid screen for receptors
slide-6
SLIDE 6

Angiogenesis (blood vessels) Lymphangiogenesis (lymphatics) Metastasis

slide-7
SLIDE 7

Tumor-homing Peptides

Peptide Sequence Specificity Receptor Reference

Blood Vessels RGD-4C CDCRGDCFC Angiogenic endothelium αvβ3 and αvβ3 integrins Arap et al., Science, 1998 F3 34-amino acid basic peptide Angiogenic endothelium, tumor cells Cell surface nucleolin Porkka et al., 2002; Christian et al., 2003 CGKRK CGKRK Angiogenic endo- thelium, tumor cells Heparan sulfate? Hoffman et al., 2003 Lymphatic Vessels LyP-1 CGNKRTRGC Breast ca lymphatics, tumor cells Not known Laakkonen et al., 2002; 2004 LSD Melanoma lymphatics CXCR4? Zhang et al, submitted REA Prostate ca lymphatics, tumor cells Zhang et al, submitted Vessel/Tumor ECM CREKA CREKA Breast ca extracellular matrix Not known Essler et al., unpublished Plasma Clot CLTI/2 10-amino acid cyclic peptides Blood clots, fibrin deposits in tumors

Fibro nectin

l.,

Pilch et al., PNAS, 2006

CAR Nonapeptide Angiogenic endothelium Jarvinen et al.

slide-8
SLIDE 8

Tumor Blood Vessels

slide-9
SLIDE 9

F3 + anti- nucleolin F3

F3 is a cell-penetrating peptide specific for tumor endothelial cells and tumor cells

FITC F3 PEPTIDE

Inject i.v. Add to tumor cell culture

F3, HL-60 tumor F3 i.v.- green Blood vessels - red Nuclei - blue Control IgG i.v.- red Blood vessels - green Anti-nucleolin i.v.- red Blood vessels - green

slide-10
SLIDE 10

Tumor-homing peptide CAR recognizes tumor vessels and penetrates into tumor parenchyma

Jarvinen et al.

Blood vessels FITC-CAR peptide Nuclei Merge

slide-11
SLIDE 11

Joyce et al., Cancer Cell, 2003; Hoffman et al., Cancer Cell, 2003

Fluorescein-labeled vascular homing peptides from in vivo phage display reveal blood vessel changes that parallel progression in tumor development

1 2 3 Intravenously injected peptide Normal islet Angiogenic islet Tumor

slide-12
SLIDE 12

Tumor Lymphatics

slide-13
SLIDE 13

LyP LyP-

  • 1 peptide co

1 peptide co-

  • localizes with tumor lymphatics

localizes with tumor lymphatics

Laakkonen et al.,Nature Med., 2002

FITC-LyP-1 Lyve-1 FITC-LyP-1 Podoplanin

slide-14
SLIDE 14

LyP-1 and LSD phage have mutually exclusive tumor type specificities

Zhang et al., Cancer Res., 2006

Anti-podoplanin Anti-LYVE-1 Anti-CD31

10 20 30 40 50 C 8 1 6 1 M D A

  • M

B

  • 4

3 5 P y M T t u m

  • r

T r a m p t u m

  • r

P P C 1 H P V s k i n c a n c e r K R I B Fold over non- recombinant phage Fold over non- recombinant phage M D A

  • M

B

  • 4

3 5 20 40 60 80 100 C8161

LyP-1 LSD Breast cancer Prostate cancer Melanoma Lymphatic homing peptides for

slide-15
SLIDE 15

Selective tissue damage by homing peptide-targeted pro-apoptotic peptide D(KLAKLAK)2

Ellerby et al. Nat. Med., 1999 Arap et al. PNAS, 2002 Gerlag et al. Arthr. Res., 2001

RGD-4C = cCDCRGDCFC Receptor: integrins αvβ3 and αvβ5 Target: angiogenesis

slide-16
SLIDE 16

Inhibition of C8161 tumor growth by D(KLAKLAK)2-LSD conjugate

  • L. Zhang et al.

200 400 600 800 1000 1200 1400 1600 16 20 24 28 32 36

days post implantatio

D(KLAKLAK)2-LSD D(KLAKLAK)2 plus LSD

PBS

**

5 10 15 20 25 30 35 40 Blood vessels Lymphatic vesse

Vessels per field

D(KLAKLAK)2-LSD D(KLAKLAK)2 plus LSD

PBS

* **

slide-17
SLIDE 17

Laakkonen et al., PNAS, 2004 T B H Lu Li S K

20 40 60 80 100 120 140

Fluorescence

Control peptide LyP-1 LyP-1b

Tumor imaging with a homing peptide for tumor lymphatics

slide-18
SLIDE 18

Extracellular Matrix

slide-19
SLIDE 19

C L K I H E

tumor kidneybrain liver spleen heart lung

F J G D A

tumor lung brain liver spleen heart

B

A Homing Peptide Binds to Clotted Plasma Proteins in a Tumor

Pilch et al., PNAS, 2006

CLT1 Control peptide CLT1/ Normal tissues

slide-20
SLIDE 20

Nanomedicine

slide-21
SLIDE 21

Specific in vivo homing of quantum dots

LyP-qdots Tumor blood vessels Lung-homing qdots F3 (34 aa peptide) cCGNKRTRGC cCGFECVRQCPERC F3-qdots Tumor blood vessels T L

PEG CdSe ZnS P E G PEG PEG P e p t i d e Peptide Peptide P e p t i d e Akerman et al. PNAS, 2002

slide-22
SLIDE 22

Dextran

Iron oxide

FITC PEPTIDE F I T C P E P T I D E F I T C P E P T I D E FITC PEPTIDE FITC PEPTIDE F I T C P E P T I D E

Peptide-coated magnetic particle

slide-23
SLIDE 23

Proteolytically actuated self-assembly

Harris, von Maltzahn et al. (Bhatia laboratory) Angew. Chem., 2006

slide-24
SLIDE 24

Role of PEG length and characterization of assembly

Harris, von Maltzahn et al. Angew. Chem., 2006

slide-25
SLIDE 25

Triggered self-assembly of nanoparticles by tumor cells

Harris, von Maltzahn et al. Angew. Chem., 2006

HT-1080 HT-1080 + inhibitor HT-1080 HT-1080 + Inhibitor

slide-26
SLIDE 26

Amplification of in vivo homing

Tissue distribution CREKA-coated iron oxide particles vs CREKA peptide

CREKA-IO CREKA peptide tumor bladder tumor liver lymph nodes bladder

  • D. Simberg, T. Duza et al. (Ruoslahti laboratory)
slide-27
SLIDE 27

Treatment of mice with decoy Ni-liposomes enhances tumor homing by CREKA peptide-targeted IO nanoparticles

  • D. Simberg, T. Duza et al.
slide-28
SLIDE 28

Ni-liposomes prolong blood half life of CREKA-IO nanoparticles

  • D. Simberg, T. Duza et al.
slide-29
SLIDE 29

CREKA-IO accumulation in tumor vessels after decoy particle treatment

A x200

  • D. Simberg, T. Duza et al.
slide-30
SLIDE 30

CREKA-IO nanoparticles accumulate in a fibrin meshwork in vivo and in vitro

Plasma clot in vitro Tumor Tumor

Fibrin CREKA-IO DAPI CREKA-IO DAPI CREKA-IO

  • D. Simberg, T. Duza et al.
  • Inhibited by blood clotting inhibitor (heparin, hirudin)
slide-31
SLIDE 31

Intravital imaging confirms clot formation in tumor vasculature in situ

20x

  • D. Simberg, T. Duza et al.

Vessel wall

slide-32
SLIDE 32

Tumor imaging with decoy liposome-enhanced targeting

  • f CREKA-IO nanoparticles

D.Simberg et al.

slide-33
SLIDE 33

Amplification of tumor targeting: CREKA-IO particles induce homing of non-targeted Cy7-IO particles

A B C

D.Simberg et al.

slide-34
SLIDE 34

CREKA-IO particles may induce necrosis in tumor tissue

  • D. Simberg, et al.
slide-35
SLIDE 35
  • Tumor blood vessels differ from vessels in normal tissues
  • angiogenesis-related
  • tumor type-related
  • tumor stage-related
  • Tumor lymphatics differ from lymphatics in normal tissues
  • tumor type-related
  • tumor stage-related
  • Vascular and tumor extracellular matrix differs from normal

ECM

  • Clotted plasma proteins
  • ECM proteins
  • The markers of tumor vessels enable new therapies
  • guided drugs
  • druggable targets
  • Vascular zip code molecules are useful as guidance elements

in nanoparticle targeting

  • Blood and Lymphatic Vessel Zip codes
slide-36
SLIDE 36
slide-37
SLIDE 37

CREKA-IO nanparticles may extravasate from some tumor vessels

Simberg et al.

slide-38
SLIDE 38

Green= FITC conjugates; red=blood vessels; blue=nuclei

Tumor/FITC-CREKA

Homing peptide for Extracellular Matrix in Tumor Vessels and Tumor Stroma

Tumor /FITC-CSG

  • M. Essler, M., H. Biliran et al.,
slide-39
SLIDE 39

Multistage tumorigenesis in RIP-Tag transgenic mice

Normal stage (onc+) <5 wks <5 wks 100% 100% Hyperplastic/ Dysplastic stage 5 5-

  • 7 wks

7 wks ~50% ~50% 7 7-

  • 12 wks

12 wks ~10% ~10% Angiogenic stage Tumor stage 12 12-

  • 14 wks

14 wks 2 2-

  • 4%

4%

slide-40
SLIDE 40

Modular Approach to Multifunctional Nanoparticles Physically target:

  • vascular cells
  • tumor cells
  • extracellular matrix

Trigger/engineer:

  • tissue penetration
  • RES avoidance
  • subcellular targeting
  • diagnostic signal(s)
  • release of therapeutic agent
  • amplification of targeting
slide-41
SLIDE 41

Ni-liposomes reduce RES uptake of CREKA-IO nanoparticles

  • D. Simberg, T. Duza et al.
slide-42
SLIDE 42

Treatment of mice with decoy Ni-liposomes produces only minimal background in control organs

  • D. Simberg, T. Duza et al.
slide-43
SLIDE 43

Heparin pretreatment, but not platelet depletion, inhibits CREKA-IO-induced clotting in tumor vessels

  • D. Simberg, et al.