Assessing Immune Responses of Stem Cell Transplants with ...€¦ · Imaging Stem Cell Transplants: Assessment of Immune Responses Repeatedly image the location(s) and numbers of

Assessing Immune Responses of Stem Cell Transplants

with clinically applicable Imaging Techniques

Stem Cell Tracking Detection of

stem cell apoptosis Immune Cell Tracking

Imaging Approaches

Imaging Stem Cell Transplants:

Assessment of Immune Responses



Repeatedly image the location(s) and numbers of one or

more specific population(s) of cells in vivo

Allow flexibility to scale across different spatial

resolution levels

Do not significantly perturb the cells while repeatedly

imaging them

Repeatedly image the “status” of stem cells and their

progenies in vivo, including potential interactions with

host cells, such as immune cells

Cellular Imaging Targets / Probes

mRNA

DNA

mRNA

Receptor Mapping

Enzyme Activity:

Inhibition, Conc.,

Synthesis

Accumulation via

AA Transport or

Protein Synthesis Oligonucleotides

mRNA Binding

Reporter Probe Reporter Gene DNA

AAT .

Accumulation via

Phosphorylation

[18F]FDG

mAb,

Fragments Hormones

Drugs and Ligands

Peptides

Accumulation via

DNA-Synthesis

Internalization

Hexokinase

glut 4



1. In vivo tracking of transplanted stem cells

2. In vivo tracking of immune cells

3. In vivo imaging of tissue regeneration

Clinically applicable labels

for in vivo tracking of stem cells

SPECT and PET Imaging

[18F]-FDG (half life 2 h)

Tc99m HMPAO (half life 6 h)

In-111 Oxine (half-life 2.8 days)

[18F]-Fluoro-3-(hydroxymethyl)butyl]guanine

In-111 Oxine-labeled EPC

acute myocardial infarction

Schaechinger et al, Circulation 2008; Hofmann et al, Circulation 2005; Zhu et al, N Engl J Med 2006

FDG-labeled CD34

enriched BMCs

into LAD


for in vivo tracking of stem cells

MR Imaging

Gd-Chelates

Iron Oxide

Nanoparticles






Schaechinger et al, Circulation 2008; Hofmann et al, Circulation 2005; Zhu et al, N Engl J Med 2006

Brain trauma

pre

post

Feridex-

labeled

neural

stem

cells

7 days 14 days 21 days

– FDA approved iron supplement

– Mean hydrodynamic diameter: 30nm

– Non-ionic carboxhydrate coating of polyglucose

sorbital carboxymethylether

– r1: 15 mM-1 sec-1

– r2: 89 mM-1 sec-1 at 1.5 Tesla and 37°C2

Khurana et al (Daldrup-Link lab) Nanomedicine under revision

Ferumoxytol (Feraheme™)

Stem Cell Hypertrophy Stem Cell Loss Labeled Stem Cells

Rudelius M, Daldrup-Link HE et al. Eur J Nuc Med & Mol Imag 30: 1038-44

Normal

differentiation

of CM-labeled

stem cells

Normal

differentiation

of unlabeled

controls

Nestin+ stem cell

Nestin+ stem cell

GFAP+ glia cell

GFAP+ glia cell

MAP2+ (neur.cytoskeletton-prot.) neuronal cell

MAP2+ neuronal cell

Differentiation of hESCs into cardiomyocytes

hESCs

Differentiation into cardiomyocytes

Embryoid Bodies

Castaneda et al (Daldrup-Link lab) Pediatr Radiology 41(11): 1384-1392, 2011

Cardiomyocyte protein expression

maintained post ICG-labeling.

Positive staining was observed for:

Connexin43 (a), Wnt8a (b),

Troponin T (c), GATA4 (d), Lin28

(e), TBX5 (f), Actin (g), Islet1 (h).

Scale bar: 100 µm.

Boddington S et al (Daldrup-Link lab) Cell Transplantation 19: 55–65, 2010

Scaffold +

unlabeled cells

Scaffold +

Fe-labeled cells

Scaffold +

Gd-labeled cells

Chondrogenic differentiation of Fe-labeled hMSC

Henning T et al (Daldrup-Link lab) Contr Media Mol Imaging 4(4): 165-173

Henning T, Gawande R (Daldrup-Link lab), Molecular Imaging 11(3):197-209, 2012

In vivo diagnosis of stem cell proliferation vs apoptosis

Day 0 Week 2 Week 4 Week 8

Prussian Blue Anti-CD44

VIA

BL

E

Prussian Blue Anti-CD44

AP

OP

TO

TIC

Matrix associated stem cell implants

in osteochondral defects






MR Imaging

Gd-Chelates

Iron Oxide

Nanoparticles





[18F]-FHBG

Tracking EpCAM-targeted NK cells

-> EpCAM-positive cancer

CD 57 stain

Fe-stain

CD 57

pre

24 h

Cancer Immunotherapy

Meier R (Daldrup-Link lab), Magn Res Med 65(3):756–763, 2011


for in vivo tracking of leukocytes

MR Imaging of Macrophage Migration into MASI

Khurana A (Daldrup-Link lab), Radiology 264(3):803-11, 2012

Monocyte

Macrophage Imaging

Reticuloendothelial System Bone Marrow

Progenitor cell

Monocyte

Liver Spleen Lymph Nodes

MØ MØ MØ

MØ

i.v. injection

of iron oxide

nanoparticles

Monocyte

Macrophage Imaging


Progenitor cell

Monocyte

Spleen Lymph Nodes

MØ MØ MØ

15-60 min

post USPIO

MØ

Liver

Monocyte

Macrophage Imaging


Progenitor cell

Monocyte


MØ MØ MØ

60 min post

USPIO

MØ

Monocyte

Macrophage Imaging


Progenitor cell

Monocyte


MØ MØ MØ

Stem Cell

Transplant

MØ

Monocyte

Macrophage Imaging


Progenitor cell

Monocyte


MØ MØ MØ

Liver

MØ

Stem Cell

Transplant

No Fe IV ferumoxytol labeled bone marrow macrophages

2a. 2b. 2c.

Week 2 Week 4 Stem Cell Implant

APOPTOTIC

VIABLE MASI

In vivo tracking of macrophages

Stem Cell Implant


wk 0 wk 0 wk 2 wk 2 wk 4 wk 4 0

5

10

15

20

25

VIABLE APOPTOTIC

SN

R

In vivo tracking of macrophages


MR Imaging

Gd-Chelates

Iron Oxide

Nanoparticles






18F-FDG-labeled leukocytes,

Normal scan

Dumarey et al, J Nucl Med 2006

18F-FDG-labeled leukocytes

-> infected aortic valve


for in vivo tracking of leukocytes

PET Reporter Gene Imaging for Tracking CTL

[18F]FHBG

9-[4-[18F]-Fluoro-3-

(hydroxymethyl)butyl]guanine

PET Reporter Gene Imaging for Tracking CTL

Tumor accumulation of CTL

in recurrent Glioma

pre

3d after CTL, 2 h post [18F]FHBG

9-[4-[18F]-Fluoro-3-

(hydroxymethyl)butyl]guanine

courtesy of S. Gambhir, Stanford University






Imaging Cartilage Regeneration

4 years after matrix associated chondrocyte transplant:

Successful cartilage repair

PDW 3200/30 T1 map

dGEMRIC 490/13

Trattnig et al, Radiology 2011, 260: 332

T2 map,

SE 2700/10-20

Anatomy

GAG content proton content

Imaging Tissue Regeneration

reduced

FOV

anatomic

T2 scan

ADCiso Color FA Fibers

MR Tractography

Traumatic Cord Injury

normal

Injury site

courtesy of Greg Zaharchuk, Stanford University

Normal Volunteer

What comes next along the path?

PET

MR Imaging

Radiography

Optical Imaging

SPECT

Fusion

Scans

Clinically applicable techniques

for imaging stem cell transplants

& related host immune responses

are readily available

TAKE HOME MESSAGE

Translational Tumor and Stem Cell Imaging Lab,

Lorry Lokey Stem Cell Research Building @ Stanford

Grant Support:

NIH/NIAMS 2R01AR054458-05, R21AR059861

Stanford Cancer Institute, (CIRM 2007-09)

Aman Khurana

Hossein Nejadnik

Fanny Chapelin

Olga Lenkov

Su Hyun Hong

Jessica Donig

Rakhee Gawande

Qiaoyun Shi

Celina Anasari

Christopher Klenk

Beverly Newman

Richard Barth

Amy Morris

Jennifer Vancil

Jianghong Rao

Deju Ye

Laura Pisani

Brian Rutt

R. Paulmurugan

Nikita Derugin

Greg Zaharchuk

Deqiang Qiu

Michael Moseley

Andrew Quon

John Ronald

Sanjiv Sam Gambhir

Stanford Radiology Institute for Stem Cell

& Regenerative Medicine

Charles Chan

Jeong Hyung

Michael Longaker

Electrical Engineering

Sungmin Lee

Dwight Nishimura

Northwestern University

Thomas Meade

Thank you!

Daldrup-Link Lab, Molecular Imaging Program @ Stanford

Translational Tumor and Stem Cell Imaging Lab,

Lorry Lokey Stem Cell Research Building @ Stanford

Documents

Assessing Immune Responses of Stem Cell Transplants with ...€¦ · Imaging Stem Cell Transplants: Assessment of Immune Responses Repeatedly image the location(s) and numbers of