Generation of Transgene-Free Lung Disease-Specific Human Induced Pluripotent Stem Cells Using a Single Excisable Lentiviral Stem Cell Cassette

Stem Cells - Tập 28 Số 10 - Trang 1728-1740 - 2010
Aba Somers1,2, Jyh‐Chang Jean1,2, Cesar Sommer3, Amel Omari1,2, Christopher Ford1,2, Jason A. Mills4, Lei Ying4, Andreia Gianotti Sommer3, Brenden W. Smith5, Robert Lafyatis6, George Kroumpouzos7, Daniel C. Chambers8, Deborah L. French4, Paul Gadue4, George J. Murphy9,5, Gustavo Mostoslavsky9,3, Darrell N. Kotton1,9,2
1Boston University Pulmonary Center Boston University School of Medicine, Boston, Massachusetts, USA
2Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, USA.
3Section of Gastroenterology, Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, USA
4Center for Cellular and Molecular Therapeutics, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA
5Section of Hematology and Oncology, Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, USA
6Section of Rheumatology, Department of Medicine, Boston University School of Medicine, Boston, Massachusets, USA
7Department of Dermatology, Boston University School of Medicine, Boston, Massachusets, USA
8Department of Medicine, University of Vermont College of Medicine, Burlington, Vermont, USA.
9Center for Regenerative Medicine (CReM), Boston University School of Medicine, Boston, Massachusets, USA

Tóm tắt

Abstract The development of methods to achieve efficient reprogramming of human cells while avoiding the permanent presence of reprogramming transgenes represents a critical step toward the use of induced pluripotent stem cells (iPSC) for clinical purposes, such as disease modeling or reconstituting therapies. Although several methods exist for generating iPSC free of reprogramming transgenes from mouse cells or neonatal normal human tissues, a sufficiently efficient reprogramming system is still needed to achieve the widespread derivation of disease-specific iPSC from humans with inherited or degenerative diseases. Here, we report the use of a humanized version of a single lentiviral “stem cell cassette” vector to accomplish efficient reprogramming of normal or diseased skin fibroblasts obtained from humans of virtually any age. Simultaneous transfer of either three or four reprogramming factors into human target cells using this single vector allows derivation of human iPSC containing a single excisable viral integration that on removal generates human iPSC free of integrated transgenes. As a proof of principle, here we apply this strategy to generate >100 lung disease-specific iPSC lines from individuals with a variety of diseases affecting the epithelial, endothelial, or interstitial compartments of the lung, including cystic fibrosis, α-1 antitrypsin deficiency-related emphysema, scleroderma, and sickle-cell disease. Moreover, we demonstrate that human iPSC generated with this approach have the ability to robustly differentiate into definitive endoderm in vitro, the developmental precursor tissue of lung epithelia.

Từ khóa


Tài liệu tham khảo

Maherali, 2007, Directly reprogrammed fibroblasts show global epigenetic remodeling and widespread tissue contribution, Cell Stem Cell, 1, 55, 10.1016/j.stem.2007.05.014

Okita, 2007, Generation of germline-competent induced pluripotent stem cells, Nature, 448, 313, 10.1038/nature05934

Takahashi, 2006, Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors, Cell, 126, 663, 10.1016/j.cell.2006.07.024

Wernig, 2007, In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state, Nature, 448, 318, 10.1038/nature05944

Staerk, 2010, Reprogramming of human peripheral blood cells to induced pluripotent stem cells, Cell Stem Cell, 7, 20, 10.1016/j.stem.2010.06.002

Aasen, 2008, Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes, Nat Biotechnol, 26, 1276, 10.1038/nbt.1503

Park, 2008, Reprogramming of human somatic cells to pluripotency with defined factors, Nature, 451, 141, 10.1038/nature06534

Takahashi, 2007, Induction of pluripotent stem cells from adult human fibroblasts by defined factors, Cell, 131, 861, 10.1016/j.cell.2007.11.019

Yu, 2007, Induced pluripotent stem cell lines derived from human somatic cells, Science, 318, 1917, 10.1126/science.1151526

Huangfu, 2008, Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2, Nat Biotechnol, 26, 1269, 10.1038/nbt.1502

Han, 2010, Tbx3 improves the germ-line competency of induced pluripotent stem cells, Nature, 463, 1096, 10.1038/nature08735

Zhou, 2009, Generation of induced pluripotent stem cells using recombinant proteins, Cell Stem Cell, 4, 381, 10.1016/j.stem.2009.04.005

Okita, 2008, Generation of mouse induced pluripotent stem cells without viral vectors, Science, 322, 949, 10.1126/science.1164270

Stadtfeld, 2008, Induced pluripotent stem cells generated without viral integration, Science, 322, 945, 10.1126/science.1162494

Heng, 2010, The nuclear receptor Nr5a2 can replace Oct4 in the reprogramming of murine somatic cells to pluripotent cells, Cell Stem Cell, 6, 167, 10.1016/j.stem.2009.12.009

Kaji, 2009, Virus-free induction of pluripotency and subsequent excision of reprogramming factors, Nature, 458, 771, 10.1038/nature07864

Soldner, 2009, Parkinson's disease patient-derived induced pluripotent stem cells free of viral reprogramming factors, Cell, 136, 964, 10.1016/j.cell.2009.02.013

Woltjen, 2009, piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells, Nature, 458, 766, 10.1038/nature07863

Yamanaka, 2007, Strategies and new developments in the generation of patient-specific pluripotent stem cells, Cell Stem Cell, 1, 39, 10.1016/j.stem.2007.05.012

Sommer, 2009, Induced pluripotent stem cell generation using a single lentiviral stem cell cassette, Stem Cells, 27, 543, 10.1634/stemcells.2008-1075

Sommer, 2010, Excision of reprogramming transgenes improves the differentiation potential of iPS cells generated with a single excisable vector, Stem Cells, 28, 64, 10.1002/stem.255

Nostro, 2008, Wnt, activin, and BMP signaling regulate distinct stages in the developmental pathway from embryonic stem cells to blood, Cell Stem Cell, 2, 60, 10.1016/j.stem.2007.10.011

Gadue, 2006, Wnt and TGF-beta signaling are required for the induction of an in vitro model of primitive streak formation using embryonic stem cells, Proc Natl Acad Sci USA, 103, 16806, 10.1073/pnas.0603916103

Yusa, 2009, Generation of transgene-free induced pluripotent mouse stem cells by the piggyBac transposon, Nat Methods, 6, 363, 10.1038/nmeth.1323

Nakagawa, 2008, Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts, Nat Biotechnol, 26, 101, 10.1038/nbt1374

Silva, 2008, Promotion of reprogramming to ground state pluripotency by signal inhibition, Plos Biol, 6, e253, 10.1371/journal.pbio.0060253

Weiss, 2008, Stem cells and cell therapies in lung biology and lung diseases, Proc Am Thorac Soc, 5, 637, 10.1513/pats.200804-037DW

Davies, 2005, Airway gene therapy, Adv Genet, 54, 291, 10.1016/S0065-2660(05)54012-4

Klings, 2006, Abnormal pulmonary function in adults with sickle cell anemia, Am J Respir Crit Care Med, 173, 1264, 10.1164/rccm.200601-125OC

Lynch, 2009, Lung disease related to collagen vascular disease, J Thorac Imaging, 24, 299, 10.1097/RTI.0b013e3181c1acec

Cardoso, 2006, Regulation of early lung morphogenesis: Questions, facts and controversies, Development, 133, 1611, 10.1242/dev.02310

Gouon-Evans, 2006, BMP-4 is required for hepatic specification of mouse embryonic stem cell-derived definitive endoderm, Nat Biotechnol, 24, 1402, 10.1038/nbt1258

Kubo, 2004, Development of definitive endoderm from embryonic stem cells in culture, Development, 131, 1651, 10.1242/dev.01044

D'Amour, 2005, Efficient differentiation of human embryonic stem cells to definitive endoderm, Nat Biotechnol, 23, 1534, 10.1038/nbt1163

Yamamoto, 2004, Nodal antagonists regulate formation of the anteroposterior axis of the mouse embryo, Nature, 428, 387, 10.1038/nature02418

Chan, 2009, Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells, Nat Biotechnol, 27, 1033, 10.1038/nbt.1580

Sridharan, 2009, Role of the murine reprogramming factors in the induction of pluripotency, Cell, 136, 364, 10.1016/j.cell.2009.01.001

Stadtfeld, 2010, A reprogrammable mouse strain from gene-targeted embryonic stem cells, Nat Methods, 7, 53, 10.1038/nmeth.1409

Thông tin
Thông tin xuất bản

Stem Cells

Tập 28 Số 10

1728-1740

Thông tin tác giả