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Professor Akseli Hemminki public presentation on oncolytic viruses (May 25, 2010)
1. Syövän uuden geenihoidot
‐ valoa tunnelin päässä
[Gene therapy for cancer]
[G th f ]
Akseli Hemminki, MD, PhD
Specialist in Oncology
Specialist in Oncology
K. Albin Johansson Research Professor,
Finnish Cancer Institute
Cancer Gene Therapy Group
Molecular Cancer Biology Program &
Transplantation Laboratory &
Haartman Institute & FIMM
University of Helsinki and Disclaimer: AH is co‐founder and shareholder of
Disclaimer AH is co founder and shareholder of
Oncos Therapeutics Inc., a company founded for
Helsinki Univ. Central Hospital
facilitating clinical trials with oncolytic viruses
3. Cancer is not a beaten
disease
CANCER
> 1/2 of people alive today will get cancer*
• 1/3 of us will die of cancer
• few disseminated solid tumors can be cured with
currently available treatments
Novel treatments are needed!
* Jemal CA Cancer J Clin 2005 A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 3
4. Human molecular biology
in a nutshell
Most normal and pathological events (diseases) are controlled
by proteins
Proteins are coded by genes
P t i d db
The nucleus of all human cells contain all genes (ca. 30 000),
but expression is different in each cell
but expression is different in each cell
Expression cassette: promoter – gene – polyA signal
Expression of genes ‐> expression of proteins ‐> phenotype
p g p p p yp
and function
Virology in a nutshell:
¬ Viruses are not cells, they use human cells for
reproduction
¬ Viruses reproduce by delivering their own genes into
Viruses reproduce by delivering their own genes into
human cells A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 4
5. Gene therapy is often protein therapy
Vehicle for gene delivery = vector
Gene that codes for
‐ Lack of enzyme causes disease:
Lack of enzyme causes disease: missing or desired
missing or desired
hereditary diseases, eg. protein
hemophilias, immune deficiencies
Promoter Gene pA
‐ Disease can be
Promoter and poly‐A signal
treated with local needed for protein production
needed for protein production
protein
Protein
production: production in
cardiovascular
di l or near target
disease, cancer treats disease
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 5
6. How Far is Clinical Gene Therapy ?
Phase I: Safety and toxicity ?
Phase II: Any evidence of efficacy ?
Phase II: Any evidence of efficacy ?
Phase III: Proof of efficacy
(randomization)
N= 1579
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 6
7. Mutation compensation
Randomized ph. III trial: head & neck ca.
‐ Ad p53 + radiation vs radiation alone
Ad‐p53 + radiation vs. radiation alone
‐ 67% vs. 24% CR (N= 82, P<0.01)
‐ Pan J Clin Oncol 2008
‐ Gendicine® for sale in China
‐ More than 10 000 patients treated
Promoter p53 gene
p53 gene pA
Infection of cells
Infection of cells
Normal cells Cancer cells
Press release 23 Jul 2008: Ad‐p53 (Advexin®) with p53 mutation
with healthy p53
phase III SCCHN trial positive in US: not
approved by FDA Cell death, also sensitation to
chemotherapy and radiation
h h d di i
No cell death
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 7
8. Prodrug converting enzymes
Randomized Phase III trial for glioma (ASPECT):
• Ad-TK + standard care vs. standard care: 1.43
HR (p=0.02)
(p )
Ad coding for
Ad di f
• 40d increase in median survival
thymidine TK • More temozolomide use in control group due to
kinase (TK)
non blinded
non-blinded gene therapy-> dilution of results
therapy >
• EMEA did not approve because non-standard
end-point (time to re-intervention or death)
o to c p od ug
Non‐toxic prodrug
Advantage vs.
Ad t
= ganciclovir
mutation
compensation:
bystander
effect via gap
Activated
Activated junctions
CHALLENGE: even with bystander
toxin
Cell death effect, can we get effective penetration
into established tumors ?
SOLUTIONS: locally amplifying
systems A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 8
10. Short history of oncolytic viruses
1896. 1st report of response to oncolytic virus ( influenza Dock Am J Med
1896. 1st report of response to oncolytic virus (”influenza” Dock Am J Med
Sci 1904)
1940s. 1st systematic trials with oncolytic viruses (”Egypt 101”, Hoster
Cancer Res 1949)
1950s. Adenovirus used for treatment of cervical cancer patients (Smith
Cancer 1956)
¬ Various serotypes, intratumoral, intravenous and intra‐arterial delivery
¬ Poorly characterized preparations: titers unknown
¬ Treatment with and without immune suppression
¬ Good safety, similar side effects to modern trials
¬ Frequent responses
¬ Approach was abandoned: relapses, advent of chemotherapeutics
1960s & 1970s. Rational development of oncolytic viruses in test animals,
further trials (Asada Cancer 1974)
f th t i l (A d C 1974)
1991 & 1996. Utilization of molecular features for making viruses selective
for tumor cells (Martuza Science 1991, Bischoff Science 1996)
2004. First phase III trial with oncolytic virus completed (”H101”, Yu Curr
2004 First phase III trial with oncolytic virus completed (”H101” Yu Curr
Cancer Drug Targets 2007)
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 1 0
11. H101 (Oncorine®) phase III trial in
advanced head and neck cancer
d dh d d k
H101 (≈dl1520=ONYX‐015 descibed earlier in the US)
Randomized phase III trial (N 105)
Randomized phase III trial (N=105)
H101 + cisplatin + 5‐FU vs. cisplatin + 5‐FU
CR+PR = 79% vs. 38%, P<0.0001
CR PR 79% 38% P 0 0001
Mild tox: flu‐like symptoms, injection site pain
More than 800 patients now enrolled
H101 approved in China
H101 approved in China
Yu Curr Cancer Drug Targets 2007
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 1 1
12. Personalized oncolytic adenovirus treatments
in the Advanced Therapy Access Program
in the Advanced Therapy Access Program
• 190 pts since Nov 2007. 9 different viruses (updated 14 May 2010)
• All had metastatic solid tumors progressing after routine
All had metastatic solid tumors progressing after routine
treatments (chemo, radiation, etc)
• Written informed consent. Full GCP implemented.
• Side effects: gr. 1‐2 flu‐like symptoms, fever, fatigue, pain in all pt
Side effects: gr 1 2 flu like symptoms fever fatigue pain in all pt
• SAE in < 5% (eg. pain, embolus, thrombosis, cholecystitis)
• No treatment related deaths so far (compare to chemo, surgery)
• Clinical benefit (imaging CR, PR, SD): 61% overall, 76% best virus
• Some patients have benefited for > 2 years (= length of follow‐up)
• Additive/synergistic benefits from 2nd ‐ 14th treatments
Additive/synergistic benefits from 2 treatments
• Long term (>300 d) survival in 50% with best virus, best schedule
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 1 2
13. Inclusion and exclusion criteria,
personalization of oncolytic virus treatment
personalization of oncolytic virus treatment
Inclusion criteria
c us o c te a Exclusion criteria
c us o c te a
Refractory solid tumor confirmed brain met. or glioma
Failed treatments for which there is organ transplant, HIV
strong scientific evidence* severe comorbidity
severe comorbidity
Good performance status: WHO 0‐ Elevated serum bilirubin
2. (WHO 3‐4 safe but less efficacy)
( y) Serum AST or ALT > 3 x normal
Written informed consent Thrombocytes < 75.
Personalization:
Personalization:
Selection of virus (out of 9): existing preclinical data on capsids, promoters,
arming, pretreatment efficacy prediction
Dose: tumor burden, comorbidities
Route: ultrasound guided, CT‐guided, i.v.
* In most cases this means 1st line chemotherapy
Virus sensitizers
Vi iti for metastatic disease, and in some cases several
f t t ti di di
lines of chemotherapy (eg. breast, ovarian and
l
colorectal cancer)
Seroswitching when intravenous efficacy sought In practice, the median number of prior chemo 1 3
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 |
regimens is 4 –> heavily pretreated
14. Findings possible only in pts: Mechanisms of anti‐
tumor efficacy
y inflammation
3. Induction of
cytotoxic T‐cells
1. Killing of differentiated tumor cells
1 Killing of differentiated tumor cells against tumors
against tumors
6 CD8+
5
vitiligo
E+8
4
10E
3
2
0 17 41 48
2. Killing of tumor initiating ”stem” cells
4. Induction
of specific
immunity
against
tumor
epitope
(survivin)
Cerullo Cancer Res in press 2010 A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 1 4
Eriksson Mol Ther 2007, Bauerschmitz Cancer Res 2008
15. Case: Systemic efficacy of Ad5/3‐Cox2L‐D24 in
chemo refractory neuroblastoma
chemo refractory neuroblastoma
• Ad5/3‐Cox2L‐D24 replicates in cells overexpressing Cox2 and
defective in the Rb/p16 pathway
Previous
Previous
treatments: • 6 yr old boy, WHO 1
Vincristine + • Progressive disease in bone marrow, left kidney, lymph nodes.
cis/carboplatin • Single oncolytic adenovirus treatment: i v intratumoral
cis/carboplatin Single oncolytic adenovirus treatment: i.v., intratumoral.
+ etoposide + • Gr. 1 stomach pain, diarrhea, flu‐like symptoms, liver enzymes
cyclophospham • 4 wk later: complete response in bone marrow, partial
ide response in primary
i i
Doxorubicin +
etoposide +
iphosphamide;
iphosphamide;
Intensive chemo
and autologous
stem cell
stem cell
transplant;
Oral 13‐cis‐retinoic
acid
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 1 5
16. Ad5/3‐
Cox2L‐D24
in neuro‐
blastoma
→ CD56 staining (brown) for
tumor cells in bone
marrow
→ Imaging of primary before
and after treatment
Oncolytic replication alone is usually
Oncolytic replication alone is usually
not enough to cure advanced tumors
→ Increase in cytotoxic T‐cells
→→ Increase in virus
neutralizing antibodies
→→→ Extended presence of
virus in blood
0
→→→→ Cox2 expression in
6540
tumor (reason for
( f
500
selectivity and efficacy)
Pesonen Submitted 2008
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 1 6
Pesonen Acta Oncol 2010
17. Higher efficacy with a second round of
treatment: role of immune response ?
treatment: role of immune response ?
• Metastatic pancreatic ca. WHO 2
• Prior gemcitabine and gemcitabine chemoradiation
• Second round of treatment with Ad5‐24‐RGD (Bauerschmitz
Cancer Res 2002) produced response
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 1 7
Pesonen in preparation
18. Improving antitumor immunity: oncolytic
adenoviruses coding for GM‐CSF
adenoviruses coding for GM CSF
Cerullo Cancer Res 2010
• GM‐CSF is the most potent inducer of anti‐ GM-CSF
GM CSF anti G CS
tumor immunity (Dranoff Immunol Rev 2002)
GM-CSF
• GM‐CSF in E3: expression starts at 8h
⇒ GM‐CSF expressed only in cells that allow
replication of the virus
• Hi h expression at tumor, l systemic
High i low i
GM CSF
GM-CSF
GM-CSF
Cerullo Cancer Res 2010 A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 1 8
19. First immunotherapy product
approved 29 Apr 2010: Provenge
approved 29 Apr 2010: Provenge
(Sipuleucel‐T)
Hormone
Hormone
refactory prostate
cancer
Collection of
Collection of
white blood cells
Transduction w/
PAP & GMCSF to
activate antigen
presenting cells
Return cells into
patient
ti t
First
immunotherapy
product !
product !
PAP = prostatic acid
phosphatase
GMCSF = granulocyte
macrophage colony
stimulating factor
i l i f
A k s e l i H e m mwww.provenge.com | 1 9
i n k i | 2 7 M a y 2 0 1 0
20. GM‐CSF can enhance antigen presentation
and induce NK and cytotoxic T cells
and induce NK and cytotoxic T‐cells
Tumor cells killed with 3 mechanisms:
- Oncolytic effect of virus replication
- NK cell mediated direct cell killing
- DCs mediated tumor specific immunity
NK NK CD8+ CD8+ CD8+
NK NK CD8+ CD8+
CD8+
NK CD8+ CD8+
NK
= personalized
NK Ca Ca cancer vaccine
GM-CSF Ca
Ca Ca Ca
C DC
Ca Ca Ca
GM-CSF A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 2 0
22. Syrian hamsters cured of HapT1 tumors
with Ad5D24 GMCSF: protection from
with Ad5D24‐GMCSF: protection from
HapT1 challenge
N=5
**
***
***
N=5
*
N=5
Cerullo Cancer Res in press 2010 A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 2 2
23. Syrian hamsters cured of HapT1 tumors
with Ad5D24‐GMCSF: no protection
with Ad5D24 GMCSF: no protection
from HaK challenge
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 2 3
Cerullo Cancer Res in press 2010
24. Systemic efficacy of Ad5‐D24‐GMCSF in
injected and non injected tumors: virus
injected and non‐injected tumors: virus
circulation, immune response
• 60 yr mesothelioma patient, asbestos exposure
60 yr mesothelioma patient asbestos exposure
• Prior treatment with cisplatin+pemetrexed
• WHO 1
• Single intrapleural and i.v. injection
Single intrapleural and i v injection
• More prominent reduction of non‐injected tumor than injected tumor
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 2 4
Cerullo Cancer Res in press 2010
25. Complete response in OvCa pt
with small disease burden
ith ll di b d
Operation, adjuvant CEF x6, taxol+carbo x6, docetaxel,
O ti dj t CEF 6 t l b 6 d t l
bevacizumab, topotecan, erlotinib, aromatase inhibitor
Progressive disease, WHO 1
Single intraperitoneal treatment
Complete response (CT, markers) for 9 mo
Cerullo Cancer
Cerullo Cancer
Res in press
2010
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 2 5
26. Rapid response upon re‐treatment with
GM CSF coding oncolytic
GM‐CSF coding oncolytic adenovirus
• Peritoneally metastatic ovarian cancer since 2005.
• 5 lines of chemo (paclitaxel‐carbo, liposomal doxorubicine,
gemcitabine+carbo, gemcitabine, topotecan)
• Progressive disease, WHO 1
• 52.5% tumor size reduction in 17 days after 2nd treatment
y
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 2 6
Cerullo Cancer Res in press 2010
27. Long term survival in 1/3 of patients
treated with Ad5 D24 GMCSF
treated with Ad5‐D24‐GMCSF
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 2 7
Cerullo Cancer Res in press 2010
28. Improving transduction to
improve oncolysis
i l i
Coxsackie‐ LOW CAR ‐
LOW CAR ‐
adenovirus
receptor (CAR):
key to Ad entry
key to Ad entry
LOW GENE
DELIVERY !
CAR IS AN
CAR IS AN
ADHESION
MOLECULE ‐
LOW
LOW
EXPRESSION
IN TUMORS
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 2 8
29. Increasing infectivity of target cells:
transductional targeting
Non-targeted Targeted
T t d
adenovirus adenovirus
Adenovirus
receptor CAR
High Low
transduction Benign cell transduction
Tumor associated
receptor
p
Low High
transduction Cancer cell transduction
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 2 9
30. Serotype chimerism for tumor targeting
120
Ad5 CAR 100 3x 1x108 VP i.p.
Ad3 receptor
80
% Survival
Negative
M1
60
40 Kanerva Mol Ther 2003
20
0
15 25 35 45 55 65 75 85 95 105 115 125 135
Day
Kanerva Clin Cancer Res 2002
1,E+06 Biodistribution
Ad3 receptor CAR 1,E+05
RLU / mg protein
1,E+04
1,E+03
Ad5/3 1,E+02
,
with knob domain
1,E+01 *
from Ad3
1,E+00
Kanerva Mol Ther 2002s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 3 0
Ak
31. Cancer stem cell (CSC) hypothesis
CSC Committed progenitors cells:
Rapid replication
PCa Limited lifespan
Self-renewal: fibro
CSC Slow replication other
Unlimited lifespan inflam vasc
Ca Ca
Most ca. treatments select target Ca
cells based on higher replication Ca Ca Ca Ca
Ca stem cells may not actively
y y
replicate: not killed Ca
C Ca Ca
Differentiated Ca
Ion transporters remove drugs ca. cells
from cells: not killed CSC Ca
Tumors
T mors are mixed
mi ed
Clinical research may have missed populations of cells
CSC specific agents A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 3 1
33. Ad5/3‐D24‐GMCSF = CGTG‐102
Fiber chimerism for enhanced
transduction of cancer cells
CGTG‐102: 76% clinical NK
Replication in cells mutant in Rb‐p16
pathy benefit in advanced ca. pts
benefit in advanced ca pts NK
NK
CD8+
CD8+
CD8+
CD8+
NK CD8+
Includes most human cancers
CD8+ CD8+
CD8+
NK NK
GM‐CSF can enhance antigen
NK Ca Ca
presentation and induce NK and
GM-CSF Ca
cytotoxic CD8+ T‐cells Ca Ca DC
Ca
Expressed under the control of E3
Ca Ca
Starts at 8h Ca
= personalized
Expression coupled to virus GM-CSF cancer vaccine
replication
p
Koski Submitted 2010 A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 3 3
34. Survival of Overall survival
Serial treatment
patients treated
patients treated 50% survival 320 days
50% survival = 320 days
Survival at 300 days = 50%
with CGTG‐102 N= 23
(Ad5/3 D24
(Ad5/3‐D24‐
GMCSF)
Overall survival
All treatments
50% survival = 157 days
Survival at 300 days = 34% All patients were chemo refractory and
N= 144 progressing at treatment
progressing at treatment
Overall clinical benefit in imaging = 76%
Criteria: death due to any cause
Censoring: alive at last follow up
Censoring: alive at last follow‐up
Median overall survival of chemotherapy
resistant patients 30‐115 days (eg.
Vigano Palliat Med 2000, Llobera Eur J of
Cancer 2000)
C 2000)
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 3 4
35. All CGTG‐102 treatments
Summary Overall survival
50% survival=157d
50% survival=157d
Survival @ 300d=34%
N= 144
Clinical proof‐of‐principle available
for many ca. gene therapy approaches
y g py pp
Safety has generally been excellent
Effective gene delivery continues to be key to efficacy
Oncolytic viruses amplify and help in tumor penetration
Anti‐viral and anti‐tumoral immunity key in efficacy
Clinical benefit 76% (radiology) with CGTG‐102 (N=110)
Clinical benefit 76% (radiology) with CGTG 102 (N=110)
Survival up to 500d (ongoing)
50% overall survival at 300d with CGTG‐102 (serial treatments)
( )
Earlier treatment and smaller tumor load increase benefits
CGTG‐102 now being tested in clinical trial (Oncos Therapeutics)
Clinical trials very expensive (3.5 mil€ for phase 1‐2 with 21 pts)
A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 3 5
36. Acknowledgements
Akseli Hemminki Marko Ahonen Suvi Parviainen Institut Catala Univ. Helsinki & HUCH:
d’Oncologica: Petteri Arstila
Sari Pesonen Karoliina Autio Maria Rajecki Petri Bono
Laura Ahtiainen
La ra Ahtiainen Iulia Diaconu
I lia Diacon Tuuli Ranki
T li Ranki Ramon Alemany
a o e a y
Pekka Häyry
P kk Hä
Sophie Escutenaire João Dias Noora Rouvinen U. Washington Krister Höckerstedt
Vincenzo Cerullo Kilian Guse Andre Lieber Helena Isoniemi
Kalevi Kairemo
Anna Kanerva Theresia Gutmann Päivi Hannuksela Tuula Kiviluoto
Minna Oksanen
Minna Oksanen Otto Hemminki
Otto Hemminki Kikka Holm
Kikka Holm Transgene
Monika Lusky Jorma Paavonen
Jorma Paavonen
Anniina Koski Eerika Karli Risto Renkonen
Ilkka Liikanen U. Ottawa Ari Ristimäki
Petri Nokisalmi John Bell Mirja Ruutu
Jarmo Salo
Kalle Saksela
Kalle Saksela
Ulf‐Håkan Stenman
The Patients Mikko Tenhunen
Pekka Virkkunen
Timo Joensuu
Tuomo Alanko
Tuomo Alanko Pekka Simula Grant support:
Grant support:
Saila Eksymä‐Sillman Timo Ahopelto ERC
Anu Koskela Charlotta Backman Academy of Finland
Mauri Kouri Elina Haavisto ASCO
Jenni Kylä‐Kause Lotta Kangasniemi Biocentrum Helsinki
Leena Laasonen
Leena Laasonen Biocenter Fi l d
Bi t Finland
Aila Karioja‐Kallio
Satu Kauppinen Sigrid Juselius Foundation
Kaarina Partanen Maija Salo
University of Helsinki
Marina Rosliakova Mikko Salo
HUCH Research Funds (EVO)
Antti Vuolanto
37. Bert Vogelstein:
Cancer
Cancer
therapeutics
after the cancer
ft th
genome project
(ASCO 2009)
( )
Sequencing of tumor genomes revealed hundreds of mutations in each (Wood
S i ft l dh d d f t ti i h (W d
Science 2007, Parsons Science 2008)
Combination different in each tumor ‐> Each tumor is an individual
‐> Each tumor would require a different combination of inhibitors
‐> For long term efficacy, each pt would have to be treated with hundreds of inhibitors
‐ > Impossible because of side effects
> Impossible because of side effects
All of these mutations seem to fall in 12 pathways (Jones Science 2008).
‐ > Use pathway selective drugs (Vogelstein ASCO 2009)
For example, p16/Rb pathway selective oncolytic virus
http://cgap.nci.nih.gov/ A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 3 7
38. Deletion mutant oncolytic
adenoviruses: ∆24
d i ∆24
Fueyo Oncogene 2000
Heise Nature Med 2000
E2F • S-phase
E2F Rb • Virus replication
• normal cell
• wt Ad Rb E1A & cell lysis
E1A
24 bp deletion in Rb
binding site of E1A
• normal cell E2F Rb E2F Rb • No S phase entry
S-phase
• ∆24 • No virus replication
• Replication in cells ∆24-E1A ∆24-E1A
mutant in Rb-p16
Rb p16
pathway E2F E2F
• cancer cell E2FE2F E2FE2F • S-phase
• ∆24 • Virus replication
• Includes all human ∆24 E1A
∆24-E1A ∆24-E1A
∆24 E1A & cell lysis
cancers
(Sherr Science 1996) A k s e l i H e m m i n k i | 2 7 M a y 2 0 1 0 | 3 8