Embed Size (px)
Citation preview
Terapéutica de la enfermedad genética
Angel Raya Profesor d’Investigació ICREA
Institut de Bioenginyeria de Catalunya CIBER-BBN
5 febrero 2010
Induced reprogramming of mouse cells
Generation of iPS cells
Treating human disease with iPS cells
The FA/BRCA2 pathway
Kennedy & D'Andrea, Genes & Dev. 2005
for FANCA
for FANCD2
FA5 Skin High
FA90 Skin High
FA153 Skin High
FA404 Skin Fresh
FA430 Skin Mid
FA431 Skin Low
RV
LV
FANCA
FANCA –EGFP
FANCA-Neo
FANCA
FANCA- Wpre
FANCA – EGFP
RV
LV
FANCD2
FANCD2 –EGFP
FANCD2-Neo
FANCD2- Wpre
viral LTR
internal promoters:
SFFV, CMV, PGK, VAV
internal promoters:
VAV
viral LTR
ID Source Passages Promoters Transgene
Juan Bueren CIEMAT
Raya et al.,Nature 2009
Generation of FA patient-specific iPS cells
Raya et al.,Nature 2009
Generation of FA patient-specific iPS cells
Self-renewal > 20 passages Karyotypic stability Pluripotency Expression of pluripotency-associated markers In vitro differentiation Teratoma formation Molecular DNA fingerprinting Integration of reprogramming transgenes Silencing of reprogramming transgenes Reprogramming of gene expression profile Reprogramming of DNA methylation profile
Criteria for defining bona fide iPS cells
Raya et al.,Nature 2009
Generation of FA patient-specific iPS cells
Raya et al.,Nature 2009
Generation of FA patient-specific iPS cells
Raya et al.,Nature 2009
Generation of FA patient-specific iPS cells
Raya et al.,Nature 2009
Generation of FA patient-specific iPS cells
HDmed: StemSpan +
hBMP-4 (10 ng/ml)
hVEGF (10 ng/ml)
hbFGF (10 ng/ml)
hFL (10 ng/ml)
hTPO (20 ng/ml)
hSCF (25 ng/ml)
d2: medium change
Medium change FACS, CFC & LC
OP9 stroma
d0
d5
d8
d12
d15 Day 5 OP9
FA-specific iPS cells
Raya et al.,Nature 2009
Hematopoietic differentiation of FA iPS cells
Raya et al.,Nature 2009
FA iPS-derived hematopoietic progenitor cells
Raya et al.,Nature 2009
FA iPS-derived hematopoietic progenitor cells
Jordi Surrallés UAB
Raya et al.,Nature 2009
FA iPS-derived cells are disease free
Raya et al.,Nature 2009
Disease-free hematopoietic progenitors
Raya et al.,Nature 2009
Generation of FA patient-specific iPS cells
FA pathway is critical for iPS cell proliferation
Jordi Surrallés UAB
Raya et al.,Nature 2009
FA pathway is critical for iPS cell proliferation
Raya et al.,Nature 2009
Proof-of-concept for iPS-based cell therapy
Derivation of human iPS cells Retroviral integrations Clinical-grade lines Silencing of correcting transgenes
Control of cell proliferation Selective killing Understanding self-renewal
Specific differentiation protocols Directed differentiation protocols Clinical-grade protocols
Shortcomings of iPS cell therapy
Angel Raya Ignasi Rodríguez-Pizà Rita Vassena María José Barrero Antonella Consiglio Eduard Sleep Federico González Gustavo Tiscornia Elena Garreta Trond Aasen Anna Veiga Juan Carlos Izpisúa Belmonte CMRB
Maria Castellà Jordi Surrallés UAB
Guillermo Güenechea Susana Navarro Paula Rio Juan A. Bueren CIEMAT
FA-specific iPS cells
MEC/MICINN EC ʻMarie-Curie Reintegration Grantʼ CIBER-BBN
MEC/MICINN EC FIS (TERCEL) Marató de TV3 Fundación Cellex
Acknowledgements
