Cell cycle–dependent chromatin loading of the Fanconi anemia
Updated Results of a European Gene Therapy Trial in ... … · Molecular and Clinical Aspects of...
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Updated Results of a European Gene Therapy Trial in Fanconi Anemia Patients, Subtype A Juan A. Bueren 23 rd ASGCT Annual Meeting, May 12 th , 2020
Transcript of Updated Results of a European Gene Therapy Trial in ... … · Molecular and Clinical Aspects of...
Updated Results of a European Gene Therapy
Trial in Fanconi Anemia Patients, Subtype A
Juan A. Bueren
23rd ASGCT Annual Meeting, May 12th, 2020
Molecular and Clinical Aspects of Fanconi Anemia
• Genetic inherited disorder
• Rare disease 1-3 per million
j)
FANCB
FANCL
FANCC
FANCEFANCF
RAD51C(O)
FANCM
UBE2T(T)
REV7(V) REV3
REV1
RPA
RFWD3(W)
Ub
Ub
Ub
USP1 UAF1
a) ICL recognition b)
c)
e) Unhooking
FA core complexrecruitment
FANCD2 and FANCImonoubiquitination
d) FANCD2-I dimerizationand recruitment
f) Translesionsynthesis g) Homologous
recombination activation
h) Strand invasion andD-loop formation
l) Repaired DNA
FANCD2-Ideubiquitinationk) Complex
inactivation i) Unhooked ICL removal
XRCC2(U)
FANCA
FANCM
FANCC
FANCEFANCF
UBE2T(T)
FANCA
FANCM
Ub
Ub
Ub
FANCB
FANCL
FANCC
FANCEFANCF
UBE2T(T)
FANCA
FANCM
FANCB
FANCL
FANCC
FANCEFANCF
UBE2T(T)
FANCA
FANCM
Ub
FANCB
FANCL
FANCC
FANCEFANCF
UBE2T(T)
FANCA
FANCM
Ub
FANCB
FANCL
FANCC
FANCEFANCF
UBE2T(T)
FANCA
FANCM
Ub
FANCB
FANCL
FANCCFANCB
• Mutations in FA/BRCA Pathway• 22 genes described (FANCA - FANCW)
• Resolution of DNA interstrand–crosslinks
• Genetic instability and cytotoxicity
• Congenital abnormalities
• Bone marrow failure
• Cancer predispositiono Myelodysplastic syndromes/AML
o Solid tumors (HNSCC)Kutler et al, Blood 101:1249, 2003
5. Infusion
(No conditioning)
CD34+ cell
purification
1. Mobilization of CD34+ cells: 2-3 x (G-CSF+Plerixafor)
4. Transduction (24h)
Gene Therapy Trial for FA-A Patients
2. ± Cryopreservation
PGK-FANCA.Wpre* LV
(C: 5 Pts)
(F: 3 Pts)
(9/11 Pts)
(F+C: 1 Pt)
02
00
2
02
00
4
02
00
5
02
00
6
01
00
3
02
00
7
02
00
8
02
01
3
02
00
9
0 .0
0 .2
0 .4
0 .6
0 .8
1 .0
P a t ie n t
VC
N/C
ell
r2=0.78
0 2 0 4 0 6 0 8 0 1 0 0
0 . 0
0 . 2
0 . 4
0 . 6
0 . 8
1 . 0
M M C r e s i s t a n c e
VC
Ns
i
n
CF
Cs
02
00
2
02
00
4
02
00
5
02
00
6
01
00
3
02
00
7
02
00
8
02
01
3
02
00
9
0
2 0
4 0
6 0
8 0
1 0 0
P a t ie n t
MM
C R
es
ista
nc
e (
%)
NE
MMC-resistance at 10nM except for patient 02006 which was 3nM
NE
FANCOLEN-I: Manufacturing Products
02
00
2
02
00
4
02
00
5
02
00
6
01
00
3
02
00
7
02
00
8
02
01
3
02
00
9
0
5 0 0 ,0 0 0
1 ,0 0 0 ,0 0 0
1 ,5 0 0 ,0 0 0
2 ,0 0 0 ,0 0 0
P a t i e n t
CD
34
+ c
ell
s/K
g
02
00
2
02
00
4
02
00
5
02
00
6
01
00
3
02
00
7
02
00
8
02
01
3
02
00
9
0
5 0 0 ,0 0 0
1 ,0 0 0 ,0 0 0
1 ,5 0 0 ,0 0 0
2 ,0 0 0 ,0 0 0
P a t ie n t
cC
D3
4+
/Kg
NE
Infused Number of CD34+ Cells
Review of Patients with a Follow-up of
12-36 months Post-infusion
02002 (Cryo)
(246,000 cCD34+/Kg)
02004 (Cryo)
(163,000 cCD34+/Kg)
02005 (Fresh)
(225,000 cCD34+/Kg)
02006 (Fresh)
(410,000 cCD34+/Kg)
02007 (Cryo)
(48,600 cCD34+/Kg)
02008 (Fresh)
(157,000 cCD34+/Kg)
02013 (Cryo)
(135,000 cCD34+/Kg)
0 .5 1 1 .5 2 4 6 9 1 2 1 8 2 4 2 7 3 0 3 3 3 6
0
5
1 0
1 5
2 0
2 5
3 0
0 .0 0
0 .0 5
0 .1 0
0 .1 5
0 .2 0
0 .2 5
0 .3 0
M o n t h s p o s t G e n e T h e r a p y
%
Ge
ne
ma
rk
ed
ce
lls C
op
ies
pe
r g
en
om
e
0 .5 1 1 .5 2 4 6 9 1 2 1 5 1 8 2 1 2 4 2 7 3 0 3 3 3 6 3 9
0
2 0
4 0
6 0
8 0
1 0 0
0 .0 0
0 .2 0
0 .4 0
0 .6 0
0 .8 0
1 .0 0
M o n t h s p o s t G e n e T h e r a p y
%
Ge
ne
ma
rk
ed
ce
lls C
op
ies
pe
r g
en
om
e
0 .5 1 1 .5 2 4 6 9 1 2 1 5 1 8 2 1 2 4 2 7 3 0 3 3
0
5
1 0
1 5
2 0
2 5
3 0
0 .0 0
0 .0 5
0 .1 0
0 .1 5
0 .2 0
0 .2 5
0 .3 0
M o n t h s p o s t G e n e T h e r a p y
%
Ge
ne
ma
rk
ed
ce
lls C
op
ies
pe
r g
en
om
e
N A
0 .5 1 1 .5 2 4 6 9 1 2
0
5
1 0
1 5
2 0
0 .0 0
0 .0 5
0 .1 0
0 .1 5
0 .2 0
M o n t h s p o s t G e n e T h e r a p y
%
Ge
ne
ma
rk
ed
ce
lls C
op
ies
pe
r g
en
om
e
0 .5 1 1 .5 2 4 6 9 1 2 1 5
0
2
4
6
8
1 0
0 .0 0
0 .0 2
0 .0 4
0 .0 6
0 .0 8
0 .1 0
M o n t h s p o s t G e n e T h e r a p y
%
Ge
ne
ma
rk
ed
ce
lls C
op
ies
pe
r g
en
om
e
* This point requires additional validation as the long-term follow-up study is activated# This data point has been partially validated secondary to Covid-19 related limitations
Progressive Engraftment of Gene-marked Cells in PB
*
#
Correlation Between the Proportion of Marked Cells in PB and BM
Patient 01003 not shown as did not meet the release criteria
Patient 02009 not shown because follow-up < 12 months
02006
02008
02007
02002
02005
02013
02004
Dose-dependent Engraftment of Gene Marked Cells in PB
at 1 year Post Gene Therapy
02002 (Cryo)
(246,000 cCD34+/Kg)
02004 (Cryo)
(163,000 cCD34+/Kg)
02006 (Fresh)
(410,000 cCD34+/Kg)
02005 (Fresh)
(225,000 cCD34+/Kg)
C D 1 4 C D 1 5 C D 3 C D 1 9
0
5
1 0
1 5
2 0
2 5
3 0
0 .0 0
0 .0 5
0 .1 0
0 .1 5
0 .2 0
0 .2 5
0 .3 0
M o n t h s p o s t G e n e T h e r a p y
% G
en
e m
ar
ke
d c
ell
s Co
pie
s p
er
ge
no
me
12 39Months
12 36Months
12 33Months
12 33Months
C D 1 4 C D 1 5 C D 3 C D 1 9
0
2 0
4 0
6 0
8 0
1 0 0
0 .0
0 .2
0 .4
0 .6
0 .8
1 .0
M o n t h s p o s t G e n e T h e r a p y
% G
en
e m
ar
ke
d c
ell
s Co
pie
s p
er
ge
no
me
C D 1 4 C D 1 5 C D 3 C D 1 9
0
5
1 0
1 5
2 0
2 5
3 0
0 .0 0
0 .0 5
0 .1 0
0 .1 5
0 .2 0
0 .2 5
0 .3 0
M o n t h s p o s t G e n e T h e r a p y
% G
en
e m
ar
ke
d c
ell
s Co
pie
s p
er
ge
no
me
Progressive Increase of Gene-marked Subpopulations in PB
02002 (Cryo)
(246,000 cCD34+/Kg)
02004 (Cryo)
(163,000 cCD34+/Kg)
02006 (Fresh)
(410,000 cCD34+/Kg)
02005 (Fresh)
(225,000 cCD34+/Kg)
T o t a l B M C D 3 4 C D 4 1 C D 4 2 G ly A C D 7 1
0
2 0
4 0
6 0
8 0
1 0 0
0 .0
0 .2
0 .4
0 .6
0 .8
1 .0
% G
en
e m
ar
ke
d c
ell
s Co
pie
s p
er
ge
no
me
M o n t h s p o s t G e n e T h e r a p y
T o t a l B M C D 3 4 C D 4 1 C D 4 2 G ly A C D 7 1
0
5
1 0
1 5
2 0
2 5
3 0
0 .0 0
0 .0 5
0 .1 0
0 .1 5
0 .2 0
0 .2 5
0 .3 0
M o n t h s p o s t G e n e T h e r a p y
% G
en
e m
ar
ke
d c
ell
s Co
pie
s p
er
ge
no
me
T o t a l B M C D 3 4 C D 4 1 C D 4 2 G ly A C D 7 1
0
5
1 0
1 5
2 0
2 5
3 0
0 .0 0
0 .0 5
0 .1 0
0 .1 5
0 .2 0
0 .2 5
0 .3 0
M o n t h s p o s t G e n e T h e r a p y
% G
en
e m
ar
ke
d c
ell
s Co
pie
s p
er
ge
no
me
T o t a l B M C D 3 4 C D 4 1 C D 4 2 G ly A C D 7 1
0
5
1 0
1 5
2 0
2 5
3 0
0 .0 0
0 .0 5
0 .1 0
0 .1 5
0 .2 0
0 .2 5
0 .3 0
M o n t h s p o s t G e n e T h e r a p y%
Ge
ne
ma
rk
ed
ce
lls C
op
ies
pe
r g
en
om
e
6 24Months
6 36Months
6 36Months
6 24Months
Progressive increase of gene-marked subpopulations in BM
FA 02002 FA 02006
Clonal Repopulation Analysis in Patients FA-02002 and FA-02006: Evidence of repopulation by Multipotent HSCs
Wei Wang
Manfred Schmidt
Ning Wu
1
1 0
1 0 0
MM
C S
ur
viv
al
(%
)
0 6 1 2 0 6 1 2 2 4 0 6
F A -0 2 0 0 2 F A -0 2 0 0 6 F A -0 2 0 0 5 F A - 0 2 0 0 4
m o n t h s P o s t - G T 0 6 1 2 1 22 4 2 43 6 3 6 2 4
Progressive Phenotypic correction of BM CFCs (MMC-Resistance)
0 3 6 9 1 2 1 5 1 8 2 1 2 4 2 7 3 0 3 3 3 6
0
2 0
4 0
6 0
8 0
1 0 0
F A -0 2 0 0 5
M o n t h s p o s t G e n e T h e r a p y
%
Ab
err
an
t c
ell
s M e d ia n F A
0 3 6 9 1 2 1 5 1 8 2 1 2 4 2 7 3 0 3 3 3 6
0
2 0
4 0
6 0
8 0
1 0 0
F A -0 2 0 0 4
M o n t h s p o s t G e n e T h e r a p y
%
Ab
err
an
t c
ell
s M e d ia n F A
0 3 6 9 1 2 1 5 1 8 2 1 2 4 2 7 3 0 3 3 3 6
0
2 0
4 0
6 0
8 0
1 0 0
F A - 0 2 0 0 7
M o n t h s p o s t G e n e T h e r a p y
%
Ab
err
an
t c
ell
s M e d ia n F A
0 3 6 9 1 2 1 5 1 8 2 1 2 4 2 7 3 0 3 3 3 6
0
2 0
4 0
6 0
8 0
1 0 0
F A - 0 2 0 0 8
M o n t h s p o s t G e n e T h e r a p y
%
Ab
err
an
t c
ell
s M e d ia n F A
0 3 6 9 1 2 1 5 1 8 2 1 2 4 2 7 3 0 3 3 3 6
0
2 0
4 0
6 0
8 0
1 0 0
F A - 0 2 0 1 3
M o n t h s p o s t G e n e T h e r a p y
%
Ab
err
an
t c
ell
s M e d ia n F A
0 3 6 9 1 2 1 5 1 8 2 1 2 4 2 7 3 0 3 3 3 6 3 9
0
2 0
4 0
6 0
8 0
1 0 0
F A -0 2 0 0 2
M o n t h s p o s t G e n e T h e r a p y
%
Ab
err
an
t c
ell
s M e d ia n F A
0 3 6 9 1 2 1 5 1 8 2 1 2 4 2 7 3 0 3 3 3 6
0
2 0
4 0
6 0
8 0
1 0 0
F A -0 2 0 0 6
M o n t h s p o s t G e n e T h e r a p y
%
Ab
err
an
t c
ell
s M e d ia n F A
Progressive Phenotypic correction of PB T Cells (DEB-Induced Chr. Instability)
Months after gene therapy
-6 0 -4 0 -2 0 0
1
1 0
1 0 0
1 0 0 0
1 0 0 0 0
0 6 1 2 1 8 2 4 3 0 3 6
Le
uk
oc
yt
es
/
l02002
02004
02006 02005
Months after gene therapy Months after gene therapy
-9 0 -6 0 -3 0 0
1
1 0
1 0 0
1 0 0 0
1 0 0 0 0
0 6 1 2 1 8 2 4 3 0 3 6
Le
uk
oc
yt
es
/
l
-9 0 -6 0 -3 0 0
1
1 0
1 0 0
1 0 0 0
1 0 0 0 0
0 6 1 2 1 8 2 4 3 0 3 6
Le
uk
oc
yt
es
/
l
0 6 1 2 1 8 2 4 3 0 3 6
0 .1
1
1 0
1 0 0
1 0 0 0
1 0 0 0 0
Le
uk
oc
yt
es
/
l
02007 02008
0 6 1 2 1 8 2 4 3 0 3 6
0 .0 1
0 .1
1
1 0
1 0 0
1 0 0 0
1 0 0 0 0
Le
uk
oc
yt
es
/
l
02013
0 6 1 2 1 8 2 4 3 0 3 6
1
1 0
1 0 0
1 0 0 0
1 0 0 0 0
Le
uk
oc
yt
es
/
l
Months after gene therapyMonths after gene therapy
Months after gene therapy Months after gene therapy Months after gene therapy
-4 0 -2 0 0
1
1 0
1 0 0
1 0 0 0
1 0 0 0 0
0 6 1 2 1 8 2 4 3 0 3 6
Le
uk
oc
yt
es
/
l
Kinetics of corrected and uncorrected PB leukocytes prior to and after Gene Therapy
Months after gene therapyMonths after gene therapyMonths after gene therapy
-9 0 -6 0 -3 0 0
4
6
8
1 0
1 2
1 4
0 6 1 2 1 8 2 4 3 0 3 6
Hb
(g
/dL
)
02005 (Fresh)
(225,000 cCD34+/Kg) Hb
(g
/dL
)
-4 0 -3 0 -2 0 -1 0 0
4
6
8
1 0
1 2
1 4
0 6 1 2 1 8 2 4 3 0 3 6
Ne
utr
op
hil
s (
x1
00
0/
l)
-4 0 -3 0 -2 0 -1 0 0
0
1
2
3
4
0 6 1 2 1 8 2 4 3 0 3 6-4 0 -3 0 -2 0 -1 0 0
0
2 0
4 0
6 0
8 0
1 0 0
0 6 1 2 1 8 2 4 3 0 3 6
1 0 0
2 0 0
3 0 0
Pla
tele
ts (
x1
00
0/
l)
02004 (Cryo)
(163,000 cCD34+/Kg)
-9 0 -6 0 -3 0 0
0
1
2
3
4
0 6 1 2 1 8 2 4 3 0 3 6
Ne
utr
op
hil
s (
x1
00
0/
l)
02006 (Fresh)
(410,000 cCD34+/Kg)
Hb
(g
/dL
)
-8 0 -6 0 -4 0 -2 0 0
6
8
1 0
1 2
1 4
0 6 1 2 1 8 2 4 3 0 3 6
Ne
utr
op
hil
s (
x1
00
0/
l)
-8 0 -6 0 -4 0 -2 0 0
0
1
2
3
4
0 6 1 2 1 8 2 4 3 0 3 6-8 0 -6 0 -4 0 -2 0 0
0
2 0
4 0
6 0
8 0
1 0 0
0 6 1 2 1 8 2 4 3 0 3 6
1 0 0
2 0 0
3 0 0
Pla
te
lets
(x
10
00
/l)
Fe Supplement**
02002 (Cryo)
(246,000 cCD34+/Kg)
Hb
(g
/dL
)
-6 0 -4 0 -2 0 0
6
8
1 0
1 2
1 4
1 6
0 6 1 2 1 8 2 4 3 0 3 6
Ne
utr
op
hil
s (
x1
00
0/
l)
-6 0 -4 0 -2 0 0
0
1
2
3
4
0 6 1 2 1 8 2 4 3 0 3 6-6 0 -4 0 -2 0 0
0
2 0
4 0
6 0
8 0
1 0 0
0 6 1 2 1 8 2 4 3 0 3 6
1 0 0
2 0 0
3 0 0
Pla
te
lets
(x
10
00
/l)
* These particular data points require additional validation as the long-term follow-up study is activated ** Iron supplement is not likely to enable Hb increase in the absence of viable and productive HSPCs
*
-9 0 -6 0 -3 0 0
0
2 0
4 0
6 0
8 0
1 0 0
0 6 1 2 1 8 2 4 3 0 3 6
1 0 0
2 0 0
3 0 0
Pla
te
lets
(x
10
00
/l)
PB Cell Counts in FA Patients prior to and after Gene Therapy
*
*
Conclusions
Open questions
❑ No SAEs associated with the gene correction of FA HSPCs
❑ Infusion of corrected CD34+ cells in non-conditioned FA patients:
✓ Progressive repopulation advantage of gene-corrected HSPCs at 1-3 years post-infusion
(6/6 patients infused with >100,000 cCD34+ cells/kg)
✓ Oligoclonal reconstitution and gene correction of multipotent HSCs
✓ Progressive phenotypic correction of BM progenitors and PB T cells
✓ At 1-3 years post-infusion, stabilization in PBC counts or overall increase in the erythroid
lineage (two patients infused with the highest doses of corrected CD34+ cells)
▪ Additional product metrics that may be predictive of engraftment/phenotypic reversal,
including cCFCs/kg, CD34+CD38-/kg, other.
▪ Fate of uncorrected BM cells?
▪ Management of transfusion-dependent patients with progressive engraftment of cHSCs?
✓ CD34+ cell collection from patients with higher HSC numbers
✓ Modified Immunoselection (Higher yields of CD34+ cells)
✓ No cryopreservation
✓ Improved transduction of HSCs (transduction enhancers)
AIM:
❑ Treatment of patients in earlier stages of the BMF
❑ Infusion of higher numbers of corrected HSCs:
Ongoing Global Phase 2 Registrational Program Trial in the USA and Europe
FANCOLEN II
Acknowledgments
Julián SevillaEva MerinoJosune ZubicarayEva GálvezElena Sebastián
Cristina Díaz de Heredia
Raquel Hladun
Paula Río Susana Navarro
Anne Galy
Fulvio Mavilio
Alan Lamproye
Jordi Surrallés
María Roser Pujol
José A. Casado
Lara Álvarez
G.Güenechea
Rebeca Sánchez
Omaira Alberquilla
José C. Segovia
Yari Giménez
Francisco J. Román
Rosa M. Yáñez Paula Río
Susana Navarro
M. Fernández/ B. Díez
Lara Álvarez
Mluz Lozano
TechnicalResponsible
ProductionDepartment
Quality Control Department
Quality GarantyDepartment
Rosa M. Yáñez
Miriam Hernando
Jean Soulier
Rocío Salgado
Wei Wang
Manfred Schmidt
Ning Wu
Gaurav Shah; Jonathan Schwartz
Kinnari Patel; Brian Beard
Eileen Nicoletti; Miriam Zeini
Adrian Thrasher
Claire Booth
Diego León
Marina Cavazzana
Sol RuízJordi Barquinero
Patients and Families
Aurora de la Cal
Thank you!!!
Acknowledgments
