Genes & Cells Therapy

Professor Nagy Habib

Department of Surgical Oncology & Technology
Hammersmith Hospital Campus
Nagy.habib@imperial.ac.uk
 Stem cells

Embryonic Foetal Adult stem cell

Placenta
Umbilical
cord
Different Bone
organs marrow

MSC HSC

MAPC CD34+ CD34-

OmniCyte
CULTURE INITIATION 
Proliferation ­ DAY 3
HAEMOPOIETIC CELLS ­ DAY 7
MESENCHYMAL  STEM  CELLS ­ DAY 14
EPITHELIOID CELL ­ DAY 14
DENDRITE­FORMING CELL ­ DAY 14
 10000

Mean number of cells x 10e-3/plate ±

1000

100

10
sem

1
0 2 4 6 8 10 12 14
Day of culture
FRESHLY ISOLATED CELLS (cont)

Gene OmniCytes Other CD34+

AC133 + ­
Angiopoietin­1 + ­
Angiopoietin­2 + +
CXCR4 + ­
ICAM­2 + +
PECAM + ­
TAL­1 + ­
VEGF + ­
VWF + ­
GATA­4 + ­
Nkx 2.5 + ­
MEF­2D + ­
Phospholamban + ­
Pdx­1 + ­
C­met + ­
Omnicytes express neuronal markers after 7 days in culture

IgG control Nestin 49 ± 10%

NeuN 24 ± 7% NSE 3 ± 2%
Glut2
Amylase
exendin4
Nkx6.1
NeuroD
Pax6
Pax4
Islet1
PDX1
Insulin
1kb DNA ladder
NgN3
Somatostatin
Beta cellulin
IGF1
CK19
Day 0 PANCREAS MARKERS

NKX2.2
Somatostatin
IAP
IGF
Clinical protocol
Insulin C-peptide

Co-localised insulin and c-peptide

 Insulincyte, Blood Glucose, mmol/l
Stem cells injected 01-Dec-2005

42.0
37.0
32.0 Stem Cells
Blood glucose mmol/l

27.0
PBS
22.0
17.0
12.0
7.0
y5
Da 9
Da 9
Da 8
Da 3
Da 7
Da 5
Da 0
Da 5
Da 9
Da 4
Da 8
3
3

y1
y2
y3
y3
y4
y5
y5
y5
y6
y6
y7
y
y-

Da
Da
Da
Mouse Treatment Human Human Mouse Serum Blood
No. Insulin C-Peptide Insulin Sugar
mU/l pmol/l Ug/l

2 Stem cells 0,00 nd 0,60 9,6

3 Stem cells 22,47 nd 0,12 9.9

4 Stem cells 21,72 1134 1,49 9,7

5 Saline 0,00 nd 2,80 15.8

6 Saline 0,00 nd 0,26 32,2

OmniCytes
DAPI-nuclei; pkh26-membrane
Neuroblastoma
(dapi-nucleous; pkh67-membrane)
Fusion OmniCytes and neuroblastoma
 Growth of fibrosarcoma(RHT14-2) in single culture and in
coculture with CD34+ SC

600

500
Number of cells (x10 4)

400
Cocultx104
300 Fibro cultx104
Fibro in cocultx104
200

100

0
0 2 4 6 8
Num ber of days in culture
Non transfected Cells transfected with IFNγ-
cells 10% expressing IFNγ
 IFNgamma expression
% cells expressing IFNgamma
12

10

0
no
8 9 10 11
transfection 10 10 10 10

adn-IFNgamma concentraction
Αlpha1 A trypsin

LDLR

C MET

Glucokinase

Albumin

Nestin

Vimentin

HGF

Cyto p450
HNF3 beta
Day 14 LIVER MARKERS

1kb DNA ladder

Transferrin

AFP

HNF4 alpha

CK18
HUMAN CELLS IN MOUSE LIVER SECTION - DAY 8
 MOUSE LIVER SECTIONS STAINED FOR HUMAN
CYTOKERATIN 18 ONE DAY POST TRANSPLANT

Negative control Transplanted animal

                      
Day 0 Day 1 Day 8
Human nuclei staining
Negative control
 Nu301, Human nuclei staining
Mouse sacrificed 2 days after transplantation
ENGRAFTMENT IN LIVER DAMAGE MODEL
Patient 5

20-04-2005 27-06-2005
Conclusions

• No side effects or procedure-related

complications.
• Subjective improvements reported
• Objective improvements in 3/5 patients.
 3 Alcoholic cirrhosis

190
170 37
Bilirubin (umol/L)

150

Albumin (g/L)
130 32 BILIRUBIN
110 ALB
90 27
70
50 22
Pre WK1` WK2
weeks
 Cirrhosis ( Autoimmune Hepatitis)

70 50
60
40
50
Bilirubin (umol/L)

Albumin (g/L)
40 30 Bilirubin
30 20 Albumin
20
10
10
0 0
Pre WK2 WK6 WK10 WK14
Weeks
 Glycogen Storage Disease
Father to six year old daughter

ALT

350
300

250

200
ALT
150

100

50
0

AST

300

250

200

150 AST

100

50

0
Clinical Trials

• Phase 1 Liver study successfully completed

Phase 2 study started (liver)

• Studies planned in 2006 (MHRA approved):

Heart, Stroke, Diabetes, Phase III liver

• Others: inflammatory bowel disease, peripheral

vascular disease, fracture tibia, back pain, osteo-
arthritis of the knee joint, chronic renal failure,
chronic pain, MS