Metabolic engineering generates a transgene-free safety switch for cell therapy

Wiebking, Volker; Patterson, James O.; Martin, Renata; Chanda, Monica; Lee, Ciaran M.; Srifa, Waracharee; Bao, Gang; Porteus, Matthew H.

doi:10.1038/s41587-020-0580-6

Cited by 44 publications

(30 citation statements)

References 67 publications

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“…Targeted integration can also provide cells with novel functions, such as a “safety-switch” for cell therapy applications. Transgene integration can be directed to inactivate an essential metabolic enzyme, the uridine monophosphate synthetase, which makes T cells dependent on supplemented uridine for their growth and survival (Wiebking et al, 2018 ). This approach could help therapies based on chimeric antigen receptor T cells by introducing a metabolic control of their proliferation and persistence.…”

Section: Disease-modifier Genesmentioning

confidence: 99%

Targeted Gene Delivery: Where to Land

Pavani

Amendola

2021

Front. Genome Ed.

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Genome-editing technologies have the potential to correct most genetic defects involved in blood disorders. In contrast to mutation-specific editing, targeted gene insertion can correct most of the mutations affecting the same gene with a single therapeutic strategy (gene replacement) or provide novel functions to edited cells (gene addition). Targeting a selected genomic harbor can reduce insertional mutagenesis risk, while enabling the exploitation of endogenous promoters, or selected chromatin contexts, to achieve specific transgene expression levels/patterns and the modulation of disease-modifier genes. In this review, we will discuss targeted gene insertion and the advantages and limitations of different genomic harbors currently under investigation for various gene therapy applications.

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Section: Disease-modifier Genesmentioning

confidence: 99%

Targeted Gene Delivery: Where to Land

Pavani

Amendola

2021

Front. Genome Ed.

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“…Selection by 5-FOA has recently been applied to human cell lines ( Wiebking et al, 2020 ). While this selection system has not yet been widely investigated in human cells, one can envision how our screening methodology could be applied to investigate factors which influence protein aggregation and phase separation in human cells.…”

Section: Discussionmentioning

confidence: 99%

Development of a Positive Selection High Throughput Genetic Screen in Dictyostelium discoideum

Williams

Scaglione

2021

Front. Cell Dev. Biol.

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The cellular slime mold Dictyostelium discoideum is a powerful model organism that can be utilized to investigate human health and disease. One particular strength of Dictyostelium is that it can be utilized for high throughput genetic screens. For many phenotypes, one limitation of utilizing Dictyostelium is that screening can be an arduous and time-consuming process, limiting the genomic depth one can cover. Previously, we utilized a restriction enzyme-mediated integration screen to identify suppressors of polyglutamine aggregation in Dictyostelium. However, due to the time required to perform the screen, we only obtained ∼4% genome coverage. Here we have developed an efficient screening pipeline that couples chemical mutagenesis with the 5-fluoroorotic acid counterselection system to enrich for mutations in genes of interest. Here we describe this new screening methodology and highlight how it can be utilized for other biological systems.

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“…One strategy employed by Ferrari S. et al ( 2020 ) was to transiently downregulate p53 with GSE56 in addition to including the E4orf6/7 protein of adenovirus, a known interactor with cellular components involved in survival and cell cycle (Ferrari S. et al, 2020 ) to successfully enhance the efficiency of editing. From another perspective toward advancing safety, Wiebking et al ( 2020 ) disrupted the uridine monophosphate synthetase (UMPS) involved in the pyrimidine de novo synthesis pathway rendering proliferation dependent on external uridine and providing thus the possibility to control cell growth by modulating the uridine supply. However, it should be noted that disruption of UMPS would be an additional genome editing process on top of any other correction, suggesting that to manufacture cell products that have been genetically engineered and present advanced safety features, one would have to edit at least two genomic loci.…”

Section: The Era Of Genome Editing: Challenges and Prospectsmentioning

confidence: 99%

“…However, it should be noted that disruption of UMPS would be an additional genome editing process on top of any other correction, suggesting that to manufacture cell products that have been genetically engineered and present advanced safety features, one would have to edit at least two genomic loci. Although both strategies (Ferrari S. et al, 2020 ; Wiebking et al, 2020 ) certainly assume great potential, they involve genetic manipulation beyond the current state of the art, and the transition to the clinic will probably be challenging from a regulatory standpoint.…”

Section: The Era Of Genome Editing: Challenges and Prospectsmentioning

confidence: 99%

The Promise and the Hope of Gene Therapy

Papanikolaou

Bosio

2021

Front. Genome Ed.

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It has been over 30 years since visionary scientists came up with the term “Gene Therapy,” suggesting that for certain indications, mostly monogenic diseases, substitution of the missing or mutated gene with the normal allele via gene addition could provide long-lasting therapeutic effect to the affected patients and consequently improve their quality of life. This notion has recently become a reality for certain diseases such as hemoglobinopathies and immunodeficiencies and other monogenic diseases. However, the therapeutic wave of gene therapies was not only applied in this context but was more broadly employed to treat cancer with the advent of CAR-T cell therapies. This review will summarize the gradual advent of gene therapies from bench to bedside with a main focus on hemopoietic stem cell gene therapy and genome editing and will provide some useful insights into the future of genetic therapies and their gradual integration in the everyday clinical practice.

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Metabolic engineering generates a transgene-free safety switch for cell therapy

Cited by 44 publications

References 67 publications

Targeted Gene Delivery: Where to Land

Targeted Gene Delivery: Where to Land

Development of a Positive Selection High Throughput Genetic Screen in Dictyostelium discoideum

The Promise and the Hope of Gene Therapy

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