Discovery and validation of human genomic safe harbor sites for gene and cell therapies

被引:44
作者
Aznauryan, Erik [1 ,2 ,3 ,4 ]
Yermanos, Alexander [1 ,5 ,6 ]
Kinzina, Elvira [7 ]
Devaux, Anna [8 ]
Kapetanovic, Edo [1 ]
Milanova, Denitsa [3 ,4 ]
Church, George M. [3 ,4 ]
Reddy, Sai T. [1 ]
机构
[1] Swiss Fed Inst Technol, Dept Biosyst Sci & Engn, Basel, Switzerland
[2] Life Sci Zurich Grad Sch, Syst Biol Program, Zurich, Switzerland
[3] Harvard Univ, Wyss Inst Biol Inspired Engn, Boston, MA 02115 USA
[4] Harvard Med Sch, Dept Genet, Boston, MA 02115 USA
[5] Swiss Fed Inst Technol, Inst Microbiol, Zurich, Switzerland
[6] Univ Geneva, Dept Pathol & Immunol, Geneva, Switzerland
[7] MIT, McGovern Inst Brain Res, Cambridge, MA 02139 USA
[8] Univ Basel, Dept Biomed, Basel, Switzerland
来源
CELL REPORTS METHODS | 2022年 / 2卷 / 01期
关键词
AAVS1; LOCUS; T-CELLS; EXPRESSION; DESIGN; SPECIFICITY; INTEGRATION; PLATFORM; REVEALS; RNAS;
D O I
10.1016/j.crmeth.2021.100154
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Existing approaches to therapeutic gene transfer are marred by the transient nature of gene expression following non-integrative gene delivery and by safety concerns due to the random mechanism of viral-mediated genomic insertions. The disadvantages of these methods encourage future research in identifying human genomic sites that allow for durable and safe expression of genes of interest. We conducted a bioinformatic search followed by the experimental characterization of human genomic sites, identifying two that demonstrated the stable expression of integrated reporter and therapeutic genes without malignant changes to the cellular transcriptome. The cell-type agnostic criteria used in our bioinformatic search suggest widescale applicability of identified sites for engineering of a diverse range of tissues for clinical and research purposes, including modified T cells for cancer therapy and engineered skin to ameliorate inherited diseases and aging. In addition, the stable and robust levels of gene expression from identified sites allow for the industry-scale biomanufacturing of proteins in human cells.
引用
收藏
页数:19
相关论文
共 70 条
[1]   Timeline - Jurkat T cells and development of the T-cell receptor signalling paradigm [J].
Abraham, RT ;
Weiss, A .
NATURE REVIEWS IMMUNOLOGY, 2004, 4 (04) :301-308
[2]   RTCGD: retroviral tagged cancer gene database [J].
Akagi, K ;
Suzuki, T ;
Stephens, RM ;
Jenkins, NA ;
Copeland, NG .
NUCLEIC ACIDS RESEARCH, 2004, 32 :D523-D527
[3]   Synthetic TRuC receptors engaging the complete T cell receptor for potent anti-tumor response [J].
Baeuerle, Patrick A. ;
Ding, Jian ;
Patel, Ekta ;
Thorausch, Niko ;
Horton, Holly ;
Gierut, Jessica ;
Scarfo, Irene ;
Choudhary, Rashmi ;
Kiner, Olga ;
Krishnamurthy, Janani ;
Le, Bonnie ;
Morath, Anna ;
Baldeyiano, G. Christian ;
Quinn, Justin ;
Tayares, Patrick ;
Wei, Qi ;
Weiler, Solly ;
Maus, Marcela, V ;
Getts, Daniel ;
Schamel, Wolfgang W. ;
Hofmeister, Robert .
NATURE COMMUNICATIONS, 2019, 10 (1)
[4]   Epidermolysis bullosa [J].
Bardhan, Ajoy ;
Bruckner-Tuderman, Leena ;
Chapple, Iain L. C. ;
Fine, Jo-David ;
Harper, Natasha ;
Has, Cristina ;
Magin, Thomas M. ;
Marinkovich, M. Peter ;
Marshall, John F. ;
McGrath, John A. ;
Mellerio, Jemima E. ;
Polson, Rex ;
Heagerty, Adrian H. .
NATURE REVIEWS DISEASE PRIMERS, 2020, 6 (01)
[5]   Promoter less gene targeting without nucleases ameliorates haemophilia B in mice [J].
Barzel, A. ;
Paulk, N. K. ;
Shi, Y. ;
Huang, Y. ;
Chu, K. ;
Zhang, F. ;
Valdmanis, P. N. ;
Spector, L. P. ;
Porteus, M. H. ;
Gaensler, K. M. ;
Kay, M. A. .
NATURE, 2015, 517 (7534) :360-U476
[6]   Gene silencing as a threat to the success of gene therapy [J].
Bestor, TH .
JOURNAL OF CLINICAL INVESTIGATION, 2000, 105 (04) :409-411
[7]   Aberrant expression of c-Jun in glioblastoma by internal ribosome entry site (IRES)-mediated translational activation [J].
Blau, Lior ;
Knirsh, Revital ;
Ben-Dror, Iris ;
Oren, Sivan ;
Kuphal, Silke ;
Hau, Peter ;
Proescholdt, Martin ;
Bosserhoff, Anja-Katrin ;
Vardimon, Lily .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2012, 109 (42) :E2875-E2884
[8]   Selective antagonism of cJun for cancer therapy [J].
Brennan, Andrew ;
Leech, James T. ;
Kad, Neil M. ;
Mason, Jody M. .
JOURNAL OF EXPERIMENTAL & CLINICAL CANCER RESEARCH, 2020, 39 (01)
[9]   Retroviral Insertional Mutagenesis in Humans: Evidence for Four Genetic Mechanisms Promoting Expansion of Cell Clones [J].
Bushman, Frederic D. .
MOLECULAR THERAPY, 2020, 28 (02) :352-356
[10]   Xist recruits the X chromosome to the nuclear lamina to enable chromosome-wide silencing [J].
Chen, Chun-Kan ;
Blanco, Mario ;
Jackson, Constanza ;
Aznauryan, Erik ;
Ollikainen, Noah ;
Surka, Christine ;
Chow, Amy ;
Cerase, Andrea ;
McDonel, Patrick ;
Guttman, Mitchell .
SCIENCE, 2016, 354 (6311) :468-472